Rapid flotation of Microcystis wesenbergii mediated by high light exposure: implications for surface scum formation and cyanobacterial species succession.

Increasing occurrences of Microcystis surface scum have been observed in the context of global climate change and the increase in anthropogenic pollution, causing deteriorating water quality in aquatic ecosystems. Previous studies on scum formation mainly focus on the buoyancy-driven floating process of larger Microcystis colonies, neglecting other potential mechanisms. To study the non-buoyancy-driven rapid flotation of Microcystis, we here investigate the floating processes of two strains of single-cell species (Microcystis aeruginosa and Microcystis wesenbergii), which are typically buoyant, under light conditions (150 µmol photons s-1 m-2). Our results showed that M. wesenbergii exhibited fast upward migration and formed surface scum within 4 hours, while M. aeruginosa did not form visible scum throughout the experiments. To further explore the underlying mechanism of these processes, we compared the dissolved oxygen (DO), extracellular polymeric substance (EPS) content, and colony size of Microcystis in different treatments. We found supersaturated DO and the formation of micro-bubbles (50-200 µm in diameter) in M. wesenbergii treatments. M. aeruginosa produces bubbles in small quantities and small sizes. Additionally, M. wesenbergii produced more EPS and tended to aggregate into larger colonies. M. wesenbergii had much more derived-soluble extracellular proteins and polysaccharides compared to M. aeruginosa. At the same time, M. wesenbergii contains abundant functional groups, which was beneficial to the formation of agglomerates. The surface scum observed in M. wesenbergii is likely due to micro-bubbles attaching to the surface of cell aggregates or becoming trapped within the colony. Our study reveals a species-specific mechanism for the rapid floatation of Microcystis, providing novel insights into surface scum formation as well as succession of cyanobacterial species.

Effect of extracellular polymeric substances on the colony size and morphological changes of Microcystis.

Surface blooms of colony-forming Microcystis are increasingly occurring in aquatic ecosystems on a global scale. Recent studies have found that the Microcystis colonial morphology is a crucial factor in the occurrence, persistence, and dominance of Microcystis blooms, yet the mechanism driving its morphological dynamics has remained unknown. This study conducted a laboratory experiment to test the effect of extracellular polymeric substances on the morphological dynamics of Microcystis. Ultrasound was used to disaggregate colonies, isolating the cells and of the Microcystis suspension. The single cells were then re-cultured under three homologous EPS concentrations: group CK, group Low, and group High. The size, morphology, and EPS [including tightly bound EPS (TB-EPS), loosely bound EPS (LB-EPS), bound polysaccharides (B-polysaccharides), and bound proteins (B-proteins)] changes of colonies were closely monitored over a period of 2 months. It was observed that colonies were rapidly formed in group CK, with median colony size (D50) reaching 183 µm on day 12. The proportion of colonies with a size of 150-500 µm increased from 1% to more than 50%. Colony formation was also observed in both groups Low and High, but their D50 increased at a slower rate and remained around 130 µm after day 17. Colonies with a size of 50-150 µm account for more than 50%. Groups CK and Low successively recovered the initial Microcystis morphology, which is a ring structure formed of several small colonies with a D50 of 130 µm. During the recovery of the colony morphology, the EPS per cell increased and then decreased, with TB-EPS and B-polysaccharides constituting the primary components. The results suggest that colony formation transitioned from adhesion driven to being division driven over time. It is suggested that the homologous EPS released into the ambient environment due to the disaggregation of the colony is a chemical cue that can affect the formation of a colony. This plays an important but largely ignored role in the dynamics of Microcystis and surface blooms.

A modified nucleoside O6-methyl-2'-deoxyguanosine-5'-triphosphate exhibits anti-glioblastoma activity in a caspase-independent manner.

Resistance to temozolomide (TMZ), the frontline chemotherapeutic agent for glioblastoma (GBM), has emerged as a formidable obstacle, underscoring the imperative to identify alternative therapeutic strategies to improve patient outcomes. In this study, we comprehensively evaluated a novel agent, O6-methyl-2'-deoxyguanosine-5'-triphosphate (O6-methyl-dGTP) for its anti-GBM activity both in vitro and in vivo. Notably, O6-methyl-dGTP exhibited pronounced cytotoxicity against GBM cells, including those resistant to TMZ and overexpressing O6-methylguanine-DNA methyltransferase (MGMT). Mechanistic investigations revealed that O6-methyl-dGTP could be incorporated into genomic DNA, disrupting nucleotide pools balance, and inducing replication stress, resulting in S-phase arrest and DNA damage. The compound exerted its anti-tumor properties through the activation of AIF-mediated apoptosis and the parthanatos pathway. In vivo studies using U251 and Ln229 cell xenografts supported the robust tumor-inhibitory capacity of O6-methyl-dGTP. In an orthotopic transplantation model with U87MG cells, O6-methyl-dGTP showcased marginally superior tumor-suppressive activity compared to TMZ. In summary, our research, for the first time, underscores the potential of O6-methyl-dGTP as an effective candidate against GBM, laying a robust scientific groundwork for its potential clinical adoption in GBM treatment regimens.

Operando dynamics of trapped carriers in perovskite solar cells observed via infrared optical activation spectroscopy.

Conventional spectroscopies are not sufficiently selective to comprehensively understand the behaviour of trapped carriers in perovskite solar cells, particularly under their working conditions. Here we use infrared optical activation spectroscopy (i.e., pump-push-photocurrent), to observe the properties and real-time dynamics of trapped carriers within operando perovskite solar cells. We compare behaviour differences of trapped holes in pristine and surface-passivated FA0.99Cs0.01PbI3 devices using a combination of quasi-steady-state and nanosecond time-resolved pump-push-photocurrent, as well as kinetic and drift-diffusion models. We find a two-step trap-filling process: the rapid filling (~10 ns) of low-density traps in the bulk of perovskite, followed by the slower filling (~100 ns) of high-density traps at the perovskite/hole transport material interface. Surface passivation by n-octylammonium iodide dramatically reduces the number of trap states (~50 times), improving the device performance substantially. Moreover, the activation energy (~280 meV) of the dominant hole traps remains similar with and without surface passivation.

A simple and low-energy strategy for the separation of water and acetonitrile.

As acetonitrile is a widely used solvent for the chemical industry, the recovery of acetonitrile from acetonitrile wastewater is significant for both industrial cost reduction and environmental protection. In this article, a simple, low-energy, and low-cost strategy is proposed for the effective separation of acetonitrile from high-concentration acetonitrile wastewater. The approach is based on a sequential combination of two steps: salt-induced phase separation and hydrophobic filtration. The acetonitrile wastewater was first induced to split into two phases by salt, that is, the acetonitrile-rich phase and the water-rich phase, then the above two phases were poured into the hydrophobic filter paper funnel for the separation. It was shown that NaCl is a suitable salting-out reagent, and that hydrophobic filter papers-obtained from modification by butyltrichlorosilane and octyltrichlorosilane were the optimal choice for hydrophobic filtration. The salt-induced phase separation process is able to increase the volume fraction of acetonitrile in the acetonitrile-rich phase up to 92%. The acetonitrile-rich phase can pass through the hydrophobic filter paper, whereas the water-rich phase was intercepted. The hydrophobic filter paper retained strong hydrophobicity and high acetonitrile-separating capacity after 3 months storage, or upon immersion in acetonitrile-water mixtures for 12 h, or applied for 25 consecutive separations.

Effect of Repetitive Peripheral Magnetic Stimulation on Patients With Low Back Pain: A Meta-analysis of Randomized Controlled Trials.

OBJECTIVE: The purpose of this meta-analysis was to investigate the effects of repetitive peripheral magnetic stimulation (rPMS) on pain intensity, functional mobility, and kinesiophobia in individuals with low back pain (LBP). DATA SOURCES: The PubMed, Physiotherapy Evidence Database, Embase, Cochrane Library, and Web of Science databases were systematically searched from inception until November 25, 2022. STUDY SELECTION: Eligible randomized controlled trials contained information on the population (LBP), intervention (rPMS), and outcomes (pain intensity, functional mobility, and kinesiophobia). Participants in the rPMS intervention group were compared with those in sham or other control groups. Two independent researchers searched for, screened, and qualified the articles. DATA EXTRACTION: Two independent researchers extracted key information from each eligible study. The authors' names, year of publication, setting, total sample size, rPMS parameters, baseline/mean difference (MD), and 95% confidence interval (CI) were extracted using a standardized form, and the methodological quality was assessed using the Physiotherapy Evidence Database score and GRADE (Grading of Recommendations, Assessment, Development, and Evaluation) system. DATA SYNTHESIS: Of 733 studies identified, 6 randomized controlled trials (n = 139) were included for meta-analysis. Compared with sham rPMS or other therapy, rPMS showed significant efficacy in reducing pain intensity (visual analog scale: MD, -1.89; 95% CI, -3.32 to -0.47; P<.05; very low-quality evidence). Significant efficacy was also found in terms of functional disability (Oswestry Disability Index: MD, -8.39; 95% CI, -13.65 to -3.12; P<.001; low-quality evidence). However, there was no statistically significant between-group difference on the Tampa scale of kinesiophobia (MD, -1.81; 95% CI, -7.60 to 3.98; P>.05; very low-quality evidence). CONCLUSIONS: This meta-analysis found very low- to low-quality evidence that rPMS can be used to reduce pain intensity and improve functional disability in individuals with LBP. However, no significant effect of rPMS on kinesiophobia was found.

Effects of repetitive peripheral magnetic stimulation on spasticity evaluated with modified Ashworth scale/Ashworth scale in patients with spastic paralysis: A systematic review and meta-analysis.

Background: Spasticity is a common motor disorder resulting from upper motor neuron lesions. It has a serious influence on an individual's motor function and daily activity. Repetitive peripheral magnetic stimulation (rPMS) is a non-invasive and painless approach developed for therapeutic intervention in clinical rehabilitation. However, the effectiveness of this intervention on spasticity in patients with spastic paralysis remains uncertain. Objective: This study aimed to investigate the effectiveness of rPMS on spasticity, motor function, and activities of daily living in individuals with spastic paralysis. Methods: PubMed, PEDro, Embase, Cochrane Library, and Web of Science were searched for eligible papers with date up to March 31, 2022. Two independent researchers conducted study screening, data extraction, and methodological quality assessment. RCTs that explored the effects of rPMS on spasticity, motor function, and activities of daily living in patients with spastic paralysis were included for review. The Cochrane collaboration tool was used to assess methodological quality. The cumulative effects of available data were processed for a meta-analysis using Reedman software. Results: Eight studies with 297 participants were included. Most of the studies presented low to moderate risk of bias. Compared with the control group, the results showed that rPMS had a significant effect on spasticity (all spasticity outcomes: standardized mean difference [SMD] = -0.55, 95% confidence interval [CI]: -0.94 to -0.16, I 2 = 40%, and P = 0.006, Modified Ashworth Scale: mean difference [MD] = -0.48, 95% CI: -0.82 to -0.14, I 2 = 0%, and P = 0.006), motor function (Fugl-Meyer Assessment: MD = 4.17, 95% CI: 0.89 to 7.46, I 2 = 28%, and P = 0.01), and activities of daily living (Barthel Index: MD = 5.12, 95% CI: 2.58 to 7.67, I 2 = 0%, and P < 0.0001). No side effect was reported. Conclusion: The meta-analysis demonstrated that the evidence supported rPMS in improving spasticity especially for passive muscle properties evaluated with Modified Ashworth Scale/Ashworth Scale, as well as motor function and daily activity of living in individuals with spastic paralysis. Study registration: The reviewed protocol of this study is registered in the international prospective register of systematic reviews (PROSPERO) (CRD42022322395). Systematic review registration: https://www.crd.york.ac.uk/PROSPERO/#recordDetails, identifier CRD42022322395.

The Effectiveness of High-Frequency Repetitive Transcranial Magnetic Stimulation on Patients with Neuropathic Orofacial Pain: A Systematic Review of Randomized Controlled Trials.

Background: Repetitive transcranial magnetic stimulation (rTMS) has been widely used in the treatment of neuropathic orofacial pain (NOP). The consistency of its therapeutic efficacy with the optimal protocol is highly debatable. Objective: To assess the effectiveness of rTMS on pain intensity, psychological conditions, and quality of life (QOL) in individuals with NOP based on randomized controlled trials (RCTs). Methods: We carefully screened and browsed 5 medical databases from inception to January 1, 2022. The study will be included that use of rTMS as the intervention for patients with NOP. Two researchers independently completed record retrieval, data processing, and evaluation of methodological quality. Quality and evidence were assessed using the PEDro scores and the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) system. Results: Six RCTs with 214 participants were included in this systematic review: 2 studies were considered level 1 evidence, and 4 were considered level 2 evidence. Six studies found that high-frequency rTMS had a pain-relieving effect, while 4 studies found no improvement in psychological conditions and QOL. Quality of evidence (GRADE system) ranged from moderate to high. No significant side effects were found. Conclusions: There is moderate-to-high evidence to prove that high-frequency rTMS is effective in reducing pain in individuals with NOP, but it has no significant positive effect on psychological conditions and QOL. High-frequency rTMS can be used as an alternative treatment for pain in individuals with NOP, but further studies will be conducted to unify treatment parameters, and the sample size will be expanded to explore its influence on psychological conditions and QOL.

The Expression of miR-34a in Gingival Crevicular Fluid of Chronic Periodontitis and Its Connection with the TLR/NF-κB Signaling Pathway.

Objective: This study analyzed the expression of miR-34a in gingival crevicular fluid (GCF) of patients with chronic periodontitis and its connection with the Toll-like receptor (TLR)/nuclear factor kappa-B (NF-κB) signaling pathway. Methods: We collected the GCF of the two groups of subjects, using RT-PCR to detect the expression of miR-34a and NF-κB p65 mRNA and TLR4 mRNA and ELISA to detect the inflammatory factor degree in GCF, and performed periodontal examinations on both groups. Results: The gingival index, bleeding index, probe depth, and attachment loss indexes of periodontal examination in the observation group were remarkably superior to those in the control group (P < 0.05). The levels of IL-1ß, IL-6, and TNF-α in the GCF of the observation group were higher than those of the control group (P < 0.05). The mRNA relative expression levels of miR-34a, NF-κB p65, and TLR4 in the GCF of the observation group were dramatically higher than those of the control group (P < 0.05). Correlation analysis showed that miR-34a was highly expressed in patients with chronic periodontitis. Conclusion: There is an abnormally high expression of miR-34a in GCF of chronic periodontitis. Its expression is associated with the degree of periodontal inflammation, periodontal tissue damage, and the activation of the TLR/NF-κB signaling pathway and could be used as a potential index for auxiliary diagnosis and severity of the disease.

Computed Tomography Three-Dimensional Reconstruction Algorithm in the Diagnosis of Periodontitis and Its Correlation with Hypertension.

This study was aimed at exploring the value of iterative reconstruction (IR) algorithm to treat the periodontitis using computed tomography (CT) image and analyze the relationship between periodontitis and hypertension. 95 patients with periodontitis were selected, including 43 patients with periodontitis, 41 patients with advanced periodontitis and hypertension, and 11 patients with periodontitis and nonhigh blood pressure (NBP). The IR algorithm was introduced to the CT image scanning of them to reduce the noise. In addition, the CT value was statistically analyzed. High-sensitivity C-reactive protein (hs-CRP) and interleukin 6 (IL-6) were dramatically increased compared with periodontitis patients with NBP and with hypertension (P < 0.05). After the IR algorithm of the image, the quality, information, and mean square error (MSE) of the image were all effectively improved. Image with a 50% dose showed the lowest noise, but the reconstruction algorithm improved the low-contrast resolution. Moderate and severe periodontitis was independently related to hypertension. Inflammatory cytokines were independently related to hypertension of periodontitis patients (P < 0.05). Therefore, it was concluded that the IR algorithm could effectively improve the spatial resolution of the CT image when it was adopted to treat periodontitis and showed a high accuracy rate; the incidence of hypertension in patients with periodontitis was relatively high, and it had a certain relationship with periodontitis; and inflammatory cytokines were related to periodontitis and hypertension of patients.

Incubating green turtle (Chelonia mydas) eggs at constant temperatures: Hatching success, hatchling morphology and post-hatch growth.

Past studies applying constant-temperature incubation of eggs have involved all species of sea turtles, but rarely can we find a single one incubating eggs at three or more temperatures. Here, we incubated green turtle (Chelonia mydas) eggs from Ganquan Island, South China Sea, at five constant temperatures (26, 28, 30, 32 and 34 °C) to determine hatching success, incubation length and hatchling phenotype at each test temperature and temperatures optimal for embryonic development. Temperature affected hatching success, incubation length and all seven examined hatchlings traits, and clutch origin affected three (head length, fore-flipper length and hind-flipper length) of the seven. Hatching success was lowest at 34 °C and none of hatchlings hatched at this temperature was normal and survived over one week. The rate of embryonic development and the rate of post-hatch growth both were lowest at 26 °C. Given that low survival and growth rates can translate into reduced individual fitness, we conclude that both 26 °C and 34 °C are unsuitable for incubation of C. mydas eggs. Post-hatch growth was fastest in hatchlings incubated at 30 °C, and eggs of C. mydas incubated at temperatures around 30 °C are more likely to produce mixed sexes. Accordingly, we conclude that temperatures within the range from 28 °C to 32 °C are generally optimal for embryonic development of C. mydas.

Inhibition of circular RNA circ_0138959 alleviates pyroptosis of human gingival fibroblasts via the microRNA-527/caspase-5 axis.

Circular RNA (circRNA) plays a regulatory role in periodontitis. This study explored whether circ_0138959 affected lipopolysaccharide (LPS)-induced pyroptosis in human gingival fibroblasts (HGFs). The periodontal ligament (PDL) tissues and HGFs were derived from patients with periodontitis and healthy volunteers. HGFs treated with LPS were considered to mimic periodontitis in vitro. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to evaluate the mRNA expression levels of circRNAs, miR-527, and caspase-5 (CASP5), and Western blotting assay was used to measure protein expression levels of caspase-1, caspase-4, and cleaved N-terminal gasdermin D (GSDMD-N). Cell viability was evaluated by cell counting kit-8 (CCK-8) assay. The concentration of lactate dehydrogenase (LDH), interleukin (IL)-1ß, and IL-18 and the pyroptosis rate were determined to evaluate pyroptosis. The interaction between miR-527 and circ_0138959 or CASP5 was verified by dual-luciferase reporter and RNA pull-down assays. Circ_0138959 expression was higher in the PDL tissues of patients with periodontitis than in the healthy group; likewise, circ_0138959 was also upregulated in LPS-treated HGFs. Suppressed circ_0138959 increased cell viability and decreased pyroptosis of HGFs induced by LPS. miR-527 was a target of circ_0138959, and inhibition of miR-527 contributed to the dysfunction of LPS-treated HGFs and reversed the protective effects of downregulated circ_0138959. Additionally, miR-527 targeted CASP5. Increased CASP5 abrogated the effects of overexpressed miR-527 on cell viability and pyroptosis of LPS-treated HGFs. Inhibition of circ_0138959 promoted cell viability and suppressed pyroptosis of HGFs via the miR-527/CASP5 axis. Therefore, knockdown of circ_0138959 may be a promising therapy for periodontitis.

Structure-Based Optimization of Multifunctional Agonists for Opioid and Neuropeptide FF Receptors with Potent Nontolerance Forming Analgesic Activities.

The opioid and neuropeptide FF pharmacophore-containing chimeric peptide 0 (BN-9) was recently developed and produced potent nontolerance forming analgesia. In this study, 11 analogues of 0 were designed and synthesized. An in vitro cAMP assay demonstrated that these analogues behaved as multifunctional agonists at both opioid and NPFF receptors. In mouse tail-flick test, most of the analogues produced potent nontolerance forming antinociception. Notably, 11 (DN-9) was 33-fold more potent than 0 at analgesic effects, which was mediated by µ- and κ-opioid receptors. In addition, 11 also produced powerful analgesic effects in the formalin pain and CFA-induced chronic inflammatory pain models. Strikingly, following its repeated administration for 6 days, 11 did not produce antinociceptive tolerance in the tail-flick test and CFA-induced pain model. The present work indicates that it is reasonable to design multifunctional peptide ligands for opioid and NPFF receptors in a single molecule producing effective nontolerance forming antinociception.

Effects of neuropeptide FF and related peptides on the antinociceptive activities of VD-hemopressin(α) in naive and cannabinoid-tolerant mice.

Neuropeptide FF (NPFF) system has recently been reported to modulate cannabinoid-induced antinociception. The aim of the present study was to further investigate the roles of NPFF system in the antinociceptive effects induced by intracerebroventricular (i.c.v.) administration of mouse VD-hemopressin(α), a novel endogenous agonist of cannabinoid CB1 receptor, in naive and VD-hemopressin(α)-tolerant mice. The effects of NPFF system on the antinociception induced by VD-hemopressin(α) were investigated in the radiant heat tail-flick test in naive mice and VD-hemopressin(α)-tolerant mice. The cannabinoid-tolerant mice were produced by given daily injections of VD-hemopressin(α) (20 nmol, i.c.v.) for 5 days and the antinociception was measured on day 6. In naive mice, intracerebroventricular injection of NPFF dose-dependently attenuated central analgesia of VD-hemopressin(α). In contrast, neuropeptide VF (NPVF) and D.NP(N-Me)AFLFQPQRF-NH2 (dNPA), two highly selective agonists for Neuropeptide FF1 and Neuropeptide FF2 receptors, enhanced VD-hemopressin(α)-induced antinociception in a dose-dependent manner. In addition, the VD-hemopressin(α)-modulating activities of NPFF and related peptides were antagonized by the Neuropeptide FF receptors selective antagonist 1-adamantanecarbonyl-RF-NH2 (RF9). In VD-hemopressin(α)-tolerant mice, NPFF failed to modify VD-hemopressin(α)-induced antinociception. However, both neuropeptide VF and dNPA dose-dependently potentiated the antinociception of VD-hemopressin(α) and these cannabinoid-potentiating effects were reduced by RF9. The present works support the cannabinoid-modulating character of NPFF system in naive and cannabinoid-tolerant mice. In addition, the data suggest that a chronic cannabinoid treatment modifies the pharmacological profiles of NPFF, but not the cannabinoid-potentiating effects of neuropeptide VF and dNPA.

Neuropeptide VF Enhances Cannabinoid Agonist WIN55,212-2-Induced Antinociception in Mice.

BACKGROUND: Cannabinoids produce analgesia in several pain models, but the undesirable side effects from high doses of cannabinoid drugs limit their clinic use. Our recent results indicate that cannabinoid-induced antinociception was enhanced by neuropeptide VF (NPVF). Here, we investigate whether low-dose cannabinoid agonists combined with NPVF can produce effective antinociception with limited side effects. METHODS: The in vivo properties of (R)-(-1)-[2,3-dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-1-napthalenylmethanone (WIN55,212-2) given alone and its combination with NPVF were evaluated in nociceptive modulation, locomotor activity, gastrointestinal transit, and tolerance development assays after intracerebroventricular administration in mice. RESULTS: In the radiant tail-flick test, the antinociception of combination of WIN55,212-2 and NPVF was more potent than that of cannabinoid agonist given alone, with an ED50 shift from 3.51 to 0.69 nmol; 9 nmol WIN55,212-2 alone and 3 nmol WIN55,212-2 combined with NPVF induced equivalent antinociception after supraspinal administration. The cannabinoid-potentiating effects of NPVF were reduced by both the cannabinoid receptor type 1 and the neuropeptide FF receptor antagonists. In the formalin assay, WIN55,212-2 combined with NPVF also significantly reduced pain-related behaviors. However, the combination of WIN55,212-2 with NPVF exerted significant hypoactivity in a manner similar to high doses of WIN55,212-2. It was important to note that the combination of WIN55,212-2 with NPVF produced nontolerance-forming antinociception and weaker inhibition of gastrointestinal transit compared with high dose of WIN55,212-2. CONCLUSIONS: These data suggest that the cannabinoid agonist combined with NPVF produces effective antinociception-lacking tolerance via both cannabinoid receptor type 1 and neuropeptide FF receptors in the brain.

Opposite effects of neuropeptide FF on central antinociception induced by endomorphin-1 and endomorphin-2 in mice.

Neuropeptide FF (NPFF) is known to be an endogenous opioid-modulating peptide. Nevertheless, very few researches focused on the interaction between NPFF and endogenous opioid peptides. In the present study, we have investigated the effects of NPFF system on the supraspinal antinociceptive effects induced by the endogenous µ-opioid receptor agonists, endomorphin-1 (EM-1) and endomorphin-2 (EM-2). In the mouse tail-flick assay, intracerebroventricular injection of EM-1 induced antinociception via µ-opioid receptor while the antinociception of intracerebroventricular injected EM-2 was mediated by both µ- and κ-opioid receptors. In addition, central administration of NPFF significantly reduced EM-1-induced central antinociception, but enhanced EM-2-induced central antinociception. The results using the selective NPFF1 and NPFF2 receptor agonists indicated that the EM-1-modulating action of NPFF was mainly mediated by NPFF2 receptor, while NPFF potentiated EM-2-induecd antinociception via both NPFF1 and NPFF2 receptors. To further investigate the roles of µ- and κ-opioid systems in the opposite effects of NPFF on central antinociception of endomprphins, the µ- and κ-opioid receptors selective agonists DAMGO and U69593, respectively, were used. Our results showed that NPFF could reduce the central antinociception of DAMGO via NPFF2 receptor and enhance the central antinociception of U69593 via both NPFF1 and NPFF2 receptors. Taken together, our data demonstrate that NPFF exerts opposite effects on central antinociception of endomorphins and provide the first evidence that NPFF potentiate antinociception of EM-2, which might result from the interaction between NPFF and κ-opioid systems.

Analgesic tolerance and cross-tolerance to the cannabinoid receptors ligands hemopressin, VD-hemopressin(α) and WIN55,212-2 at the supraspinal level in mice.

The nonapeptide hemopressin and its N-terminal extension VD-hemopressin(α) were reported as an antagonist/inverse agonist and an agonist of CB1 receptor, respectively. These novel cannabinoid peptides have been demonstrated to modulate the acute pain. In the present study, hemopressin (11, 22 and 45 nmol, i.c.v.) dose-dependently produced antinociception after supraspinal administration in the radiant heat tail-flick test. Furthermore, the development of antinociceptive tolerance to hemopressin, VD-hemopressin(α) and WIN55,212-2, and cross-tolerance among these cannabinoids were investigated in mice. The tolerance developed on day 4 after supraspinal injection of hemopressin (45 nmol), VD-hemopressin(α) (20 nmol) and WIN55,212-2 (7.5 nmol). Our results indicated symmetrical cross-tolerance between hemopressin, VD-hemopressin(α) and WIN55,212-2 at the supraspinal level in mice. These results demonstrate that both hemopressin and VD-hemopressin(α) have a time-course and extent of tolerance similar to the synthetic cannabinoid WIN55,212-2. In addition, our data imply that a common mechanism is involved in the antinociception of the three cannabinoid ligands.

The hypotensive effect of intrathecally injected (m)VD-hemopressin(α) in urethane-anesthetized rats.

Previous studies suggest that cannabinoids system plays an important role in cardiovascular regulation. (m)VD-hemopressin(α) (VD-Hpα), an 11-residue peptide originating from the α1 chain of hemoglobin, was recently reported as a selective agonist of cannabinoid CB1 receptor. The present study was undertaken to investigate the intrathecal (i.t.) action of (m)VD-Hpα on blood pressure in urethane-anesthetized rats. Our results demonstrated that injections of (m)VD-Hpα (5-30 nmol, i.t.) produced a dose-dependent decrease in mean arterial pressure (MAP), similar to that of the non-peptidic cannabinoid receptor agonist WIN55212-2 (1.25-10 nmol, i.t.). The hypotensive effect of (m)VD-Hpα was not influenced by the CB1 receptor antagonist AM251 (20 nmol, i.t.) or the CB2 receptor antagonist AM630 (20 nmol, i.t.). However, WIN55212-2-induced hypotension was almost completely prevented by i.t. administration of AM251, not by AM630. The spinal hypotension of (m)VD-Hpα and WIN55212-2 was significantly reduced by pretreatment with the α-adrenoceptor antagonist phentolamine (1 mg/kg, i.v.), but not by the ß-adrenoceptor antagonist propranolol (2 mg/kg, i.v.) or the muscarinic receptor antagonist atropine (2 mg/kg, i.v.). In addition, L-NAME (50 mg/kg, i.v.), the inhibitor of nitric oxide (NO) synthase, significantly reduced WIN55212-2-induced hypotension, but had no effect on the hypotensive response to (m)VD-Hpα. Collectively, the results show that i.t. administration of (m)VD-Hpα induces a decrease in MAP via a non-CB1 and non-CB2 mechanism.

Antinociceptive effects of central administration of the endogenous cannabinoid receptor type 1 agonist VDPVNFKLLSH-OH [(m)VD-hemopressin(α)], an N-terminally extended hemopressin peptide.

The cannabinoid system has been demonstrated to modulate the acute and chronic pain of multiple origins. Mouse VD-hemopressin(α) [(m)VD-Hpα], an 11-residue α-hemoglobin-derived peptide, was recently reported to function as a selective agonist of the cannabinoid receptor type 1 (CB1) in vitro. To characterize its behavioral and physiological properties, we investigated the in vivo effects of (m)VD-Hpα in mice. In the mouse tail-flick test, (m)VD-Hpα dose-dependently induced antinociception after supraspinal (EC50 = 6.69 nmol) and spinal (EC50 = 2.88 nmol) administration. The antinociceptive effects of (m)VD-Hpα (intracerebroventricularly and intrathecally) were completely blocked by N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3- carboxamide (AM251; CB1 antagonist), but not by 6-iodo-2-methyl-1-[2-(4-morpholinyl)ethyl]-1H-indol-3-yl(4-methoxyphenyl)-methanone (AM630; CB2 antagonist) or naloxone (opioid antagonist), showing its selectivity to the CB1 receptor. Furthermore, the central nervous system (CNS) effects of (m)VD-Hpα were evaluated in body temperature, locomotor activity, tolerance development, reward, and food intake assays. At the highly antinociceptive dose (3 × EC50), (m)VD-Hpα markedly exerted hypothermia and hypoactivity after supraspinal administration. Repeated intracerebroventricular injection of (m)VD-Hpα resulted in both development of tolerance to antinociception and conditioned place aversion. In addition, central injection of (m)VD-Hpα dose-dependently stimulated food consumption. These findings demonstrate that this novel cannabinoid peptide agonist induces CB1-mediated central antinociception with some CNS effects, which further supports a CB1 agonist character of (m)VD-Hpα. Moreover, the current study will be helpful to understand the in vivo properties of the endogenous peptide agonist of the cannabinoid CB1 receptor.