DCPIB Attenuates Ischemia-Reperfusion Injury by Regulating Microglial M1/M2 Polarization and Oxidative Stress.

The inflammatory response plays an indispensable role in ischemia-reperfusion injury, the most significant of which is the inflammatory response caused by microglial polarization. Anti-inflammatory therapy is also an important remedial measure after failed vascular reconstruction. Maintaining the internal homeostasis of the brain is a crucial measure for suppressing the inflammatory response. The mechanism underlying the relationship between DCPIB, a selective blocker of volume-regulated anion channels (VRAC), and inflammation induced by cerebral ischemia-reperfusion injury is currently unclear. The purpose of this study was to investigate the relationship between DCPIB and microglial M1/M2 polarization-mediated inflammation after cerebral ischemia-reperfusion injury. C57BL/6 mice were subjected to transient middle cerebral artery occlusion (tMCAO). DCPIB was administered by a lateral ventricular injection within 5 min after reperfusion. Behavioral assessments were conducted at 1, 3, and 7 days after tMCAO/R. Pathological injuries were evaluated using TTC assay, HE and Nissl staining, brain water content measurement, and immunofluorescence staining. The levels of inflammatory cytokines were analyzed using qPCR and ELISA. Additionally, the phenotypic variations of microglia were examined using immunofluorescence staining. In mouse tMCAO/R model, DCPIB administration markably reduced mortality, improved behavioral performance, and alleviated pathological injury. DCPIB treatment significantly inhibited the inflammatory response, promoted the conversion of M1 microglia to M2 microglia via the MAPK signaling pathway, and ultimately protected neurons from the microglia-mediated inflammatory response. In addition, DCPIB inhibited oxidative stress induced by cerebral ischemia-reperfusion injury. In conclusion, DCPIB attenuates cerebral ischemia-reperfusion injury by regulating microglial M1/M2 polarization and oxidative stress.

Progress in the application of graphene and its derivatives to osteogenesis.

As bone and joint injuries from various causes become increasingly prominent, how to effectively reconstruct and repair bone defects presents a difficult problem for clinicians and researchers. In recent years, graphene and its derivatives have been the subject of growing body of research and have been found to promote the proliferation and osteogenic differentiation of stem cells. This provides a new idea for solving the clinical problem of bone defects. However, as as numerous articles address various aspects and have not been fully systematized, there is an urgent need to classify and summarize them. In this paper, for the first time, the effects of graphene and its derivatives on stem cells in solution, in 2D and 3D structures and in vivo and their possible mechanisms are reviewed, and the cytotoxic effects of graphene and its derivatives were summarized and analyzed. The toxicity of graphene and its derivatives is further reviewed. In addition, we suggest possible future development directions of graphene and its derivatives in bone tissue engineering applications to provide a reference for further clinical application.

Effect of comfort nursing in the perioperative period of free flap transplantation to repair the defects after surgery for malignant head and neck tumors.

To explore the effect of comfort nursing in the perioperative period of free flap transplantation to repair defects of malignant head and neck tumors. This retrospective study included 242 patients with postoperative defects of malignant head and neck tumors repaired using free flap transplantation from December 2017 to December 2022. Among them, 113 patients received routine nursing and were included in the control group, and 129 patients received comfort nursing and were included in the observation group. The perioperative indexes of skin flap transplantation, skin flap necrosis, hospital stay, and cortisol levels were compared. Albumin and pre-albumin levels were compared 1 hour before and 24 hours after surgery. The nursing quality, Self-Rating Depression Scale and Self-Rating Anxiety Scale scores, and nursing satisfaction were compared between the 2 groups. The success rate of skin flap transplantation in the observation group was significantly higher than that in the control group, whereas the rates of skin flap necrosis, length of stay, and cortisol levels were significantly lower in the observation group than in the control group. The Albumin and pre-albumin levels in the observation group were significantly higher than those in the control group 24 hours after surgery. The complication rate was significantly lower in the observation group than in the control group. The Self-Rating Depression Scale and Self-Rating Anxiety Scale scores in the observation group were significantly lower than those in the control group 24 hours after surgery. The satisfaction with nursing was significantly higher in the observation group than in the control group. Comfort nursing improved the success rate of free flap transplantation, reduced complications, reduced stress, improved nutritional, and psychological status of patients during the perioperative period, and improved nursing quality and patient satisfaction with nursing.

Multimorbidity Patterns and Associations with Gait, Balance and Lower Extremity Muscle Function in the Elderly: A Cross-Sectional Study in Northwest China.

Purpose: Fall is a common geriatric syndrome leading to various adverse outcomes in the elderly. Gait and balance disorders and decreased lower extremity muscle function are the major intrinsic risk factors of falls, and studies suggested that they were closely related to the underlying chronic conditions. This study aimed to explore the patterns of multimorbidity and determine the associations of these multimorbidity patterns with gait, balance and lower extremity muscle function. Patients and Methods: A cross-sectional survey of 4803 participants aged ≥60 years in Shaanxi Province, China was conducted and the self-reported chronic conditions were investigated. The 6-m walk test, timed-up-and-go test (TUG) and 5-sit-to-stand test (5-STS) were conducted to evaluate gait, balance, and lower extremity muscle function respectively. Latent class analysis was used to explore patterns of multimorbidity, and multivariate regression analysis was used to determine the associations of multimorbidity patterns with gait, balance, and lower extremity muscle function. Results: Five multimorbidity patterns were identified: Degenerative Disease Class, Cardio-metabolic Class, Stroke-Respiratory-Depression Class, Gastrointestinal Class, and Very sick Class, and they were differently associated with gait and balance disorders and decreased lower extremity muscle function. In particular, the multimorbidity patterns of Degenerative Disease Class and Stroke-Respiratory-Depression Class were closely associated with all the three risk factors of falls. Conclusion: There are significant differences in the impact of different multimorbidity patterns on the major intrinsic risk factors of falls in the elderly population, and appropriate multimorbidity patterns are closely related to the prediction of falls and can help to develop fall prevention strategies in the elderly.

Red blood cell distribution width is a risk factor for multiple organ dysfunction syndrome in elderly patients with infection: a case control study.

BACKGROUND: Infection is the leading cause of multiple organ dysfunction syndrome (MODS) in the elderly. Red blood cell distribution width (RDW) was considered to be associated with many diseases. We aimed to explore whether RDW was associated with MODS in elderly infected patients. METHODS: We retrospectively collected data from elderly patients (≥ 65 years old) with infection. In this study, we conducted a 1:3 case-control match based on age and gender and utilized binary Logistic regression to investigate the impact of variables such as RDW on MODS. RESULTS: A total of 576 eligible patients were included in this study. RDW in the case group was significantly higher than that in control group (p < 0.001). Multivariate analysis found that RDW was an independent risk factor for MODS in elderly infected patients (OR = 1.397, 95% CI: 1.166-1.674, p < 0.001). CONCLUSIONS: RDW was an independent risk factor for MODS in elderly patients with infection.

Toward the favorable consequences of academic motivation and L2 enjoyment for students' willingness to communicate in the second language (L2WTC).

Students' willingness to communicate in the second language (L2WTC) is perceived to be the ultimate goal of L2 acquisition in that high levels of L2WTC facilitate L2 use. For this reason, factors leading learners to higher levels of L2WTC have been widely scrutinized. Yet, the role of psycho-emotional factors like academic motivation and L2 enjoyment in promoting students' L2WTC has remained elusive. Moreover, as existing literature reveals, no inquiry has conceptually reviewed the impacts of these factors on students' L2WTC. To respond to this gap, this conceptual review strived to elucidate the consequences of academic motivation and L2 enjoyment for students' L2WTC. The favorable effects of academic motivation and L2 enjoyment on students' L2WTC levels were shown in the light of empirical and theoretical evidence. Finally, the potential implications of the findings are highlighted.

Novel Perspective of Cardiovascular Diseases: Volume-Regulatory Anion Channels in the Cell Membrane.

Cardiovascular diseases (CVDs) are the leading cause of morbidity and mortality worldwide. Although there are established mechanisms and preventions for CVDs, they are not totally elucidative and effective. Emerging evidence suggests that the dysregulation of ion channels in the cell membranes underpins the dysfunction of the cardiovascular system. To date, a variety of cation channels have been widely recognized as important targets for the treatment of CVDs. As a critical component of the anion channels, the volume-regulated anion channel (VRAC) is involved in a series of cell functions by the volume regulation and maintenance of membrane homeostasis. It has been confirmed to play crucial roles in cell action potential generation, cell proliferation, differentiation and apoptosis, and the VRAC appears to be a major participant in metabolic processes during CVDs. This review summarizes the current evidence and progress concerning the VRAC, to determine the future directions and challenges for CVDs for both preventive and therapeutic purposes.

A double-J stent misguided by zebra guidewire into ileum: A case report and literature review.

The case is a 70-year-old man who underwent a left nephroureterectomy and cutaneous ureterostomy on the contralateral side for invasive bladder cancer had to be accepted replacement of the double-J stent because of stomal stenosis.When replacing the double-J stent, a severe complication that the double-J stent misguided into the ileum occurred. The patient underwent gastrointestinal motility drugs, and the double-J stent was excreted with the feces after 12 hours. Unfortunately，patient suffered a uretero-ileal fistula and died of septic shock finally.The diagnosis and management of Uretero-ileal fistula as an iatrogenic complication of zebra guidewire use is discussed.

Transurethral Incision of the Bladder Neck at Three Points with a Needle-Type Electrode for Bladder Neck Contracture.

Purpose: This study aims to evaluate the efficacy of transurethral incision of the bladder neck (TUIBN) at three points with a needle-type electrode for treatment of bladder neck contracture (BNC). Materials and Methods: Between January 2016 and April 2021, the bladder necks of 53 patients with BNC after surgery were incised by the needle-type electrode at the 5, 7, and 12 O'clock positions. Patient's preoperational characteristics, peri- and postsurgical outcomes, such as time of operation, postoperative bladder irrigation, and postoperative hospital stay, and data of the international prostate symptom score (IPSS), maximum flow rate (Qmax), and postvoid residual (PVR) were recorded 3 and 6 months after surgery. Results: All 53 cases of BNC were successfully treated in 35.00 (25.00, 45.00) min with 18.00 (14.00, 21.00) h for postoperative bladder irrigation with little intraoperative bleeding (less than 50 mL). The postoperative hospital stay ranged from 2 to 8 days, a mean of 3.50 (3.00, 5.00) days. No major intraoperative or postoperative complications were observed. All cases that underwent follow-up assessment at 3 and 6 months after the surgery showed significantly decreased IPSS and PVR and increased Qmax compared to preoperation ones (p ≤ 0.001). Of these 53 patients, there was no recurrence in severe BNC patients, but 5 of 53 (9.4%) BNC patients developed BNC again within 6 months and required repeated TUIBN. Thirty patients comprised five recurrent cases with a follow-up period of more than 1 year. Conclusions: TUIBN at three points provides a safe, effective, and reliable option in treating patients with BNC.

Coating CoCrMo Alloy with Graphene Oxide and ε-Poly-L-Lysine Enhances Its Antibacterial and Antibiofilm Properties.

INTRODUCTION: With increases in implant infections, the search for antibacterial and biofilm coatings has become a new interest for orthopaedists and dentists. In recent years, graphene oxide (GO) has been extensively studied for its superior antibacterial properties. However, most of these studies have focused on solutions and there are few antibacterial studies on metal surfaces, especially the surfaces of cobalt-chromium-molybdenum (CoCrMo) alloys. ε-Poly-L-lysine (ε-PLL), as a novel food preservative, has a spectrum of antimicrobial activity; however, its antimicrobial activity after coating an implant surface is not clear. METHODS: In this study, for the first time, a two-step electrodeposition method was used to coat GO and ε-PLL on the surface of a CoCrMo alloy. Its antibacterial and antibiofilm properties against S. aureus and E. coli were then studied. RESULTS: The results show that the formation of bacteria and biofilms on the coating surface was significantly inhibited, GO and ε-PLL composite coatings had the best antibacterial and antibiofilm effects, followed by ε-PLL and GO coatings. In terms of classification, the coatings are anti-adhesive and contact-killing/inhibitory surfaces. In addition to oxidative stress, physical damage to GO and electrostatic osmosis of ε-PLL are the main antibacterial and antibiofilm mechanisms. DISCUSSION: This is the first study that GO and ε-PLL coatings were successfully prepared on the surface of CoCrMo alloy by electrodeposition. It provides a promising new approach to the problem of implant infection in orthopedics and stomatology.

Rare complication of cushing's syndrome in the elderly: a case report.

PURPOSE: Cushing's syndrome (CS) is a rare and severe disease caused by sustained hypercortisolism. The clinical manifestations of CS can be atypical in the elderly, and the diagnosis in these patients is often missed. Infectious Purpura Fulminans (PF) is a life-threating, thrombotic form of disseminated intravascular coagulation with high mortality. To our knowledge, PF occurring in a patient with CS has not been reported previously. METHODS: We described an 84-year-old female presented with severe infection, but normal temperature. She suffered from a variety of diseases especially personality change. Physical examination revealed thin skin, general edema, and multiple scattered ecchymosis. Combined with obviously elevated serum cortisol (36.85 ug/dl) and adenoma revealed by adrenal CT scanning, endogenous CS was diagnosed. During hospitalization, the patient developed serious subcutaneous hemorrhage on the right thigh and back. The skin biopsy showed multiple small vessel thrombosis suggesting that the patient developed the rare complication of CS, Purpura Fulminans (PF). RESULTS: Chronic hypercortisolism can cause immune suppression, low-grade inflammation, endothelial damage, and a hypercoagulable state, which together increased susceptibility of PF. Fluid resuscitation, antibiotics, infusion of blood product, and debridement were effective treatment measures when CS complicated with infectious PF. CONCLUSION: Severe subcutaneous hemorrhage due to PF could occur in the patients of CS, especially in the elderly. Clinicians should be alert to the diagnosis of CS in older adults with cognitive decline and personality change.

Antibacterial and antibiofilm properties of graphene and its derivatives.

Infections resulting from bacteria and biofilms have become a huge problem threatening human health. In recent years, the antibacterial and antibiofilm effects of graphene and its derivatives have been extensively studied. However, there continues to be some controversy over whether graphene and its derivatives can resist infection and biofilms. Moreover, the antibacterial mechanism and cytotoxicity of graphene and its derivatives are unclear. In the present review, antibacterial and antibiofilm abilities of graphene and its derivatives in solution, on the surface are reviewed, and their toxicity and possible mechanisms are also reviewed. Furthermore, we propose possible future development directions for graphene and its derivatives in antibacterial and antibiofilm applications.

Involvement of α7nAChR in the Protective Effects of Genistein Against ß-Amyloid-Induced Oxidative Stress in Neurons via a PI3K/Akt/Nrf2 Pathway-Related Mechanism.

Abnormal excessive production and deposition of ß-amyloid (Aß) peptides in selectively susceptible brain regions are thought to be a key pathogenic mechanism underlying Alzheimer's disease (AD), resulting in memory deficits and cognitive impairment. Genistein is a phytoestrogen with great promise for counteracting diverse Aß-induced insults, including oxidative stress and mitochondrial dysfunction. However, the exact molecular mechanism or mechanisms underlying the neuroprotective effects of genistein against Aß-induced insults are largely uncharacterized. To further elucidate the possible mechanism(s) underlying these protective effects, we investigated the neuroprotective effects of genistein against Aß-induced oxidative stress mediated by orchestrating α7 nicotinic acetylcholine receptor (α7nAChR) signaling in rat primary hippocampal neurons. Genistein significantly increased cell viability, reduced the number of apoptotic cells, decreased accumulation of reactive oxygen species (ROS), decreased contents of malondialdehyde (MDA) and lactate dehydrogenase (LDH), upregulated BCL-2 expression, and suppressed Caspase-3 activity occurring after treatment with 25 µM Aß25-35. Simultaneously, genistein markedly inhibited the decreases in α7nAChR mRNA and protein expression in cells treated with Aß25-35. In addition, α7nAChR signaling was intimately involved in the genistein-mediated activation of phosphatidylinositol 3-kinase (PI3K)/Akt and Nrf2/keap1 signaling. Thus, α7nAChR activity together with the PI3K/Akt/Nrf2 signaling cascade likely orchestrates the molecular mechanism underlying the neuroprotective effects of genistein against Aß-induced oxidative injury.

Mating experience modifies locomotor performance and promotes episodic motor activity in Drosophila melanogaster.

Sexual behavior is a routine among animal species. Sexual experience has several behavioral consequences in insects, but its physiological basis is less well-understood. The episodic motor activity with a periodicity around 19 s was unintentionally observed in the wildtype Canton-S flies and was greatly reduced in the white-eyed mutant w1118 flies. Episodic motor activity co-exists with several consistent locomotor performances in Canton-S flies whereas reduced episodic motor activity is accompanied by neural or behavioral abnormalities in w1118 flies. The improvements of both episodic motor activity and locomotor performance are co-inducible by a pulsed light illumination in w1118. Here we show that mating experience of w1118 males promoted fast and consistent locomotor activities and increased the power of episodic motor activities. Compared with virgin males, mated ones showed significant increases of boundary preference, travel distance over 60 s, and increased path increments per 0.2 s. In contrast, mated males of Canton-S showed decreased boundary preference, increased travel distance over 60 s, and increased path increments per 0.2 s. Additionally, mated males of w1118 displayed increased power amplitude of periodic motor activities at 0.03-0.1 Hz. These data indicated that mating experience promoted fast and consistent locomotion and improved episodic motor activities in w1118 male flies.

"The unusual suspects"-Mammographic, sonographic, and histopathologic appearance of atypical breast masses.

Breast malignancy is the second most common cause of cancer death in women. However, less common breast masses can mimic carcinoma and can pose diagnostic challenges. This case-based review describes a spectrum of rare breast neoplastic and non-neoplastic masses ranging from malignant to benign entities. Malignant masses in this review include adenoid cystic carcinoma, spindle cell lipoma, granular cell tumor, angiosarcoma, glomus tumor, adenosquamous carcinoma, and myofibroblastoma. Benign masses include sarcoidosis, diabetic mastopathy, and cat scratch disease. Demographics and, when relevant, clinical presentation are summarized. Breast imaging appearance on mammography and ultrasound are highlighted along with radiology-pathology correlation with the appearance and characteristics of the histopathological specimen of these rare masses.

Transplantation of activated olfactory ensheathing cells by curcumin strengthens regeneration and recovery of function after spinal cord injury in rats.

BACKGROUND AIMS: The pro-regeneration capabilities of olfactory ensheathing cells (OECs) remain controversial. However, little is known regarding whether the transplantation of activated OECs by curcumin (CCM) elicits neural regeneration and functional recovery after spinal cord injury (SCI) in rats, and the possible molecular mechanisms have never been investigated. METHODS: Primary OECs were treated with 1µM CCM for 1-3 days. Concomitantly, activated OECs were transplanted into the traumatic spinal cord of Sprague Dawley rats. One to 9 weeks after surgery, the assessment of behavior recovery was made using the Basso, Beattie and Bresnahan (BBB) locomotor scale; electrophysiology tests, such as somatosensory evoked potential (SEP) and motor evoked potential (MEP); and the cylinder test. Pathological study, including hematoxylin and eosin staining and immunofluorescence staining for neurofilaments (NFs), was conducted at 5 weeks post-surgery. In addition, activation profiles of OECs by CCM stimulus were assessed and levels of transglutaminase-2 (TG2) and phosphatidylserine receptor (PSR) in OECs stimulated by CCM were further determined. RESULTS: CCM remarkably enhanced OEC proliferation, improved cell viability and strengthened secretion of neurotrophins and anti-inflammatory factors. In addition, the levels of TG2 and PSR in CCM-treated OECs were significantly elevated. More importantly, beyond 1 week post-transplantation of CCM-treated OECs into lesioned spinal cord, BBB score and cylinder test score were significantly higher than that seen in the other three groups and a more postponed latent SEP and MEP period was noted. Furthermore, 5 weeks later, numerous, well-arranged NF-positive nerve fibers, lesions with less cavities and reduced levels of pro-inflammatory cytokines were found in activated OEC implantation groups. In addition, the number of NF-positive fibers was significantly improved and the number and area of both cavities and gliotic scars were remarkably decreased compared with the corresponding controls. CONCLUSIONS: Transplantation of OECs activated by CCM promotes neural regeneration and functional recovery following SCI, the underlying mechanisms of which are intimately associated with the elevated production of neurotrophic factors and anti-inflammatory factors in OECs stimulated by CCM as well as reduced pro-inflammatory cytokines from the post-contusion spinal cord. In addition, OECs activated by CCM were mediated through TG2 and PSR.

Total knee arthroplasty for a valgus deformity angle of >90°: A case report.

RATIONALE: Valgus knees are relatively rare in the clinic. Treatments for valgus deviations >90° represent a surgical challenge to achieve a balance between the soft tissue and bone and prevent nerve damage. PATIENT CONCERNS: A 63-year-old woman with valgus deviations >90° in both knees complained that she had been unable to walk for 50 years. DIAGNOSES: Congenital malformation valgus deformity. INTERVENTIONS: Bilateral total knee arthroplasty (TKA) was performed using a rotating hinge knee instrument from Endo-Model for axial correction and stabilization of the joint. OUTCOMES: The patient fully recovered 3 months after surgery. At the follow-up 6 years after the operation, the function of the knee joint clearly improved. The knee society score (KSS) increased from 35 to 90. LESSONS: Constrained implants are commonly used to stabilize the joint and correct the bone axis in patients with severe ligamental instability, gross deformity, bone loss, and extreme deviation of the straight leg axis. Intraoperative exploration of the common peroneal nerve and the postoperative flexed position of the knee joints could help prevent nerve injuries.

Tomato lectin-modified nanoemulsion-encapsulated MAGE1-HSP70/SEA complex protein vaccine: Targeting intestinal M cells following peroral administration.

Vaccines administered orally enable the stimulation of both the mucous membrane and system immune responses. However, tumor vaccines, whose effective elements are antigen protein molecules or gene-encoding antigens, are hardly accustomed to the harsh gastrointestinal environment. Here, we explored an oral nanoecapsulated tumor vaccine complex to evaluate the anti-tumor effect. Tomato lectin (TL) was modified on the surface of a nanoemulsion (NE) composed of MAGE1-HSP70/SEA (MHS). C57BL/6 mice were immunized with NE (-), NE (MHS) and TL-NE (MHS) via po. or sc. administration. Additionally, the cellular immunocompetence was detected by the enzyme-linked immunospot assay and lactate dehydrogenase release assay. Serum antibody titers were analyzed using the enzyme-linked immuno sorbent assay. Next, the therapeutic and tumor challenge assays were performed. The TL-NE (MHS) particles were 20 ± 5 nm in diameter and could resist pepsin and trypsin digestion. The cellular immune responses elicited by TL-NE (MHS) perioral were stronger than those by TL-NE (MHS)-sc. (p < 0.05) when targeted to B16-MAGE1 tumor cells. The levels of MAGE-1 antibody induced by TL-NE (MHS) via the oral route was higher than control group (p < 0.05). The percentage of CD4+ and CD8+ T cells in TL-NE (MHS)-po. group was more than other groups (p < 0.05). Furthermore, oral TL-NE (M)HScould delay tumor growth and defer tumor occurrence and tumor recurrence after resection in mice challenged with B16-MAGE-1 tumor cells. The study suggested that the oral TL-NE (MHS) vaccine delivery system is feasible to improve the vaccine protection effect and may have broad application in cancer therapy.

ApoE4 May be a Promising Target for Treatment of Coronary Heart Disease and Alzheimer's Disease.

Alzheimer disease (AD) is most common neurodegenerative disorder of dementia, as we all know that ApoE4 is the greatest genetic risk factor of late-onset Alzheimer's disease (LOAD). Coronary heart disease (CHD) leads to one-fourth of all deaths in industrialized countries, it is reported that ApoE4 increases the risk of coronary heart disease as well. Furthermore, evidence show that coronary heart disease also increases the incidence of Alzheimer's disease. Whether ApoE4 is a bridge connecting AD with CHD or not? And what are the special mechanism and therapeutic methods? Researchers found that cholesterol metabolic disorder is the common cause and risk factor of AD and CHD. Epidemiological studies demonstrate that carriers of the ApoE4 allele have higher cholesterol plasma concentration. More evidence indicate that hypercholesterolemia accelerates the progression of coronary atherosclerosis, damages the central nervous system blood-brain barrier, promotes Aß protein production and Tau deposition in brain. Therefore, ApoE4 is likely to be the bridge between AD and CHD, and may be a potentially promising therapeutic target.

Effect of temperature on anoxic metabolism of nitrites to nitrous oxide by polyphosphate accumulating organisms.

Temperature is an important physical factor, which strongly influences biomass and metabolic activity. In this study, the effects of temperature on the anoxic metabolism of nitrite (NO2(-)) to nitrous oxide (N2O) by polyphosphate accumulating organisms, and the process of the accumulation of N2O (during nitrite reduction), which acts as an electron acceptor, were investigated using 91% +/- 4% Candidatus Accumulibacter phosphatis sludge. The results showed that N2O is accumulated when Accumulibacter first utilize nitrite instead of oxygen as the sole electron acceptor during the denitrifying phosphorus removal process. Properties such as nitrite reduction rate, phosphorus uptake rate, N2O reduction rate, and polyhydroxyalkanoate degradation rate were all influenced by temperature variation (over the range from 10 to 30 degrees C reaching maximum values at 25 degrees C). The reduction rate of N2O by N2O reductase was more sensitive to temperature when N2O was utilized as the sole electron acceptor instead of N2O, and the N2O reduction rates, ranging from 0.48 to 3.53 N20-N/(hr x g VSS), increased to 1.45 to 8.60 mg N2O-N/(hr x g VSS). The kinetics processes for temperature variation of 10 to 30 degrees C were (theta1 = 1.140-1.216 and theta2 = 1.139-1.167). In the range of 10 degrees C to 30 degrees C, almost all of the anoxic stoichiometry was sensitive to temperature changes. In addition, a rise in N2O reduction activity leading to a decrease in N2O accumulation in long term operations at the optimal temperature (27 degrees C calculated by the Arrhenius model).