Search Results (154)

Polymicrobial biofilms involving fungal and bacterial species are increasingly recognized as contributors to persistent infections, particularly in the oral cavity. Candida albicans and Aggregatibacter actinomycetemcomitans are two commensals that can turn into opportunistic pathogens and are able to form robust biofilms. Objectives: This study aimed to assess the interaction dynamics between these two microorganisms and to evaluate their susceptibility to fluconazole and azithromycin in single- and mixed-species forms. Methods: Biofilm biomass was quantified using crystal violet assays, while biofilm cell viability was assessed through CFU enumeration (biofilm viability assay). To assess the resistance properties of single versus mixed-species coincubations, we applied the antimicrobial susceptibility test (AST) to each drug, and analysed spatial organization with confocal laser scanning microscopy, using PNA-FISH. Results: The results indicated that both species can coexist without significant mutual inhibition. However, a non-reciprocal synergism was also observed, whereby mixed-species biofilm conditions promoted the growth of A. actinomycetemcomitans, while C. albicans growth remained stable. As expected, antimicrobial tolerance was elevated in mixed cultures, likely due to enhanced extracellular matrix production and potential quorum-sensing interactions, contributing to increased resistance against azithromycin and fluconazole. Conclusions: This study provides novel insights into previously rarely explored interactions between C. albicans and A. actinomycetemcomitans. These findings underscore the importance of investigating interspecies interactions within polymicrobial biofilms, as understanding their mechanisms, such as quorum-sensing molecules and metabolic cooperation, can contribute to improved diagnostics and more effective targeted therapeutic strategies against polymicrobial infections. Full article

Guvermectin, a Streptomyces-derived purine nucleoside compound, exhibits dual bioactivities as a plant growth regulator and an antibacterial agent. While its biosynthetic gene cluster (BGC) is regulated by the cluster-situated activator GvmR and the adjacent repressor GvmR2, the role of distal transcriptional regulators (TRs) in guvermectin biosynthesis remains unexplored. Here, we identified ScnR1, a highly conserved LacI-family TR located far from the guvermectin BGC, which is directly activated by GvmR. Overexpression of scnR1 significantly suppressed guvermectin biosynthesis. Further investigations revealed that ScnR1 competitively binds to the gvmR, gvmA, and O1 promoters (overlapping with the GvmR-binding sites), thereby inhibiting the guvermectin BGC transcription. Moreover, ScnR1 formed a reciprocal feedback loop with the adjacent repressor GvmR2, where each repressor inhibits the other’s expression. These findings reveal a multi-layered regulatory mechanism wherein LacI-family TRs fine-tune guvermectin biosynthesis through competitive DNA binding and reciprocal feedback control. This study offers new perspectives on the hierarchical control of secondary metabolism in Streptomyces and provides valuable theoretical guidance for the engineering of strains with enhanced natural product production. Full article

Magnetic shape memory alloys have attracted considerable attention due to their multifunctional properties. Among these materials, Ni-Mn-Ga alloys are distinguished by their ability to achieve up to 10% strain when exposed to a magnetic field, a characteristic predominantly observed in single-crystal samples. Consequently, it is essential to develop nanomaterials with a crystal structure closely resembling that of a single crystal. In this study, an epitaxial Ni-Mn-Ga thin film was fabricated using Pulsed Laser Deposition on an Al₂O₃ $\left(11\overline{2}0\right)$ single-crystal substrate. The crystal structure was characterised through X-ray diffraction methodologies, such as symmetrical $2\theta -\omega$ scans, pole figures, and reciprocal space maps. The results indicated that the sample was mainly in a slightly distorted cubic austenite phase, and some incipient martensite phase also appeared. A detailed microstructural analysis, performed by transmission electron microscopy, confirmed that certain regions of the sample exhibited an incipient transformation to the martensite phase. Regions closer to the substrate retained the austenite phase, suggesting that the constraint imposed by the substrate inhibits the phase transition. These results indicate that it is possible to grow high crystalline quality thin films of Ni-Mn-Ga by Pulsed Laser Deposition. Full article

The bone marrow microenvironment is a dynamic and complex niche that plays a central role in the development, progression, and therapeutic resistance of leukemia. Among the various stromal and immune cells that compose this microenvironment, adipocytes are increasingly recognized as active participants rather than passive bystanders. These cells contribute to leukemia pathophysiology by supplying leukemic cells with vital metabolic fuels such as free fatty acids and glutamine, which support cellular bioenergetics and biosynthesis. Furthermore, adipocytes secrete adipokines—including leptin, adiponectin, and others—that influence leukemic cell proliferation, apoptosis, and chemoresistance. Leukemic cells, in turn, are not merely recipients of these signals, but actively remodel the marrow niche to their advantage. They can suppress adipogenesis, inhibit the differentiation of mesenchymal stem cells into adipocytes, or reprogram existing adipocytes to adopt a tumor-supportive phenotype. These transformed adipocytes may enhance leukemic cell survival, dampen immune responses, and create a metabolic sanctuary that enables resistance to standard chemotherapies. This reciprocal and dynamic interaction between leukemic cells and adipocytes contributes significantly to minimal residual disease and relapse, posing a major challenge for durable remission. Recent advances in tissue engineering—such as organ-on-chip and 3D co-culture systems—offer promising platforms to recapitulate and study these leukemia–adipocyte interactions with high fidelity. These models facilitate mechanistic insights and provide a foundation for developing novel therapeutic strategies aimed at disrupting the metabolic and paracrine crosstalk within the leukemic niche. Targeting the adipocyte–leukemia axis represents a compelling and underexplored avenue for improving leukemia treatment by sensitizing malignant cells to existing therapies and overcoming the protective influence of the bone marrow microenvironment. Full article

Background: In addition to its known antioxidant function, the reduced form of vitamin C, ascorbate, also acts as a neuromodulator in the nervous system. Previous work showed a reciprocal interaction of ascorbate with glutamate in chicken embryo retinal cultures. Ascorbate modulates extracellular glutamate levels by inhibiting excitatory amino acid transporter 3 and promoting the activation of NMDA receptors and the consequent activation of intracellular signaling pathways involved in transcription and survival. Objective: In the present work, we investigated the regulation of AKT phosphorylation by ascorbate in chicken embryo retina cultures. Methodology: Cultures of chicken embryo retina cells were tested using Western blot, immunocytochemistry, fluorescent probe transfection, and cellular imaging techniques. Results: Our results show that ascorbate induces a concentration and time-dependent increase in AKT phosphorylation via the accumulation of extracellular glutamate, the activation of glutamate receptors, and the activation of the PI3K pathway. Ascorbate produces an increase in intracellular calcium accumulation and, accordingly, AKT phosphorylation by ascorbate is blocked by the calcium chelator BAPTA-AM. Moreover, AKT phosphorylation is also blocked by the nitric oxide synthase inhibitor 7-nitroindazole, indicating that it is mediated by calcium and nitric oxide-dependent mechanisms. Conclusions: We demonstrate that ascorbate modulates the PI3K/AKT pathway in retinal cultures through the activation of glutamate receptors and NO production in a calcium-dependent manner. Given that previous research has shown that glutamate induces ascorbate release in retinal cultures, our findings emphasize the significance of the reciprocal interactions between ascorbate and glutamate in retinal development. These findings provide further evidence supporting the role of ascorbate as a neuromodulator in retinal development. Full article

Background/Objectives: Sepsis has been shown to depress arterial baroreceptor function, and this effect is counterbalanced by the cholinergic anti-inflammatory pathway. Considering the importance of central adenosine receptors in baroreceptor function, this study tested whether central adenosine A3 receptors (A3ARs) modulate the cholinergic-baroreflex interaction in sepsis and whether this interaction is modulated by mitogen-activated protein kinases (MAPKs) and related proinflammatory cytokines. Methods: Sepsis was induced by cecal ligation and puncture (CLP) and rats were instrumented with femoral and intracisternal (i.c.) catheters. Baroreflex sensitivity (BRS) was measured 24 h later in conscious animals using the vasoactive method, which correlates changes in blood pressure caused by i.v. phenylephrine (PE) and sodium nitroprusside (SNP) to concomitant reciprocal changes in heart rate. Results: The reduction in reflex bradycardic (BRS-PE), but not tachycardic (BRS-SNP), responses elicited by CLP was reversed by i.v. nicotine in a dose-related manner. The BRS-PE effect of nicotine was blunted following intracisternal administration of IB-MECA (A3AR agonist, 4 µg/rat). The depressant action of IB-MECA on the BRS facilitatory action of nicotine was abrogated following central inhibition of MAPK-JNK (SP 600125), PI3K (wortmannin), and TNFα (infliximab), but not MAPK-ERK (PD 98059). Additionally, the nicotine suppression of sepsis-induced upregulation of NFκB and NOX2 expression in the nucleus tractus solitarius (NTS) was negated by A3AR activation. The molecular effect of IB-MECA on NFκB expression disappeared in the presence of SP 600125, wortmannin, or infliximab. Conclusions: The central PI3K/MAPK-JNK/TNFα pathway contributes to the restraining action of A3ARs on cholinergic amelioration of sepsis-induced central neuroinflammatory responses and impairment of the baroreceptor-mediated negative chronotropism. Full article

Reciprocating liquid hydrogen pumps are essential equipment for hydrogen refueling stations with liquid hydrogen stored. The valves play a crucial role in facilitating unidirectional flow and the pressurization of liquid hydrogen within the pump. This paper establishes a comprehensive numerical model to simulate the whole working cycle of a reciprocating liquid hydrogen pump. The influence of valve parameters and pump operating conditions on the motion characteristics of valves, including lift, closing lag angle, and impact velocity, is investigated. The results indicate that with the maximum lift of the suction valve at 10 mm and the discharge valve at 5 mm, the closing lag angle is minimal, and the impact velocity of the valve falls within an acceptable range. The optimal rotation speed range is between 200 and 300 rpm, within which both the closing lag angle and impact velocity of valves are minimized. Excessive maximum lift and low rotational speed lead to significant oscillations and high impact velocity in valve movement with the effects being more pronounced in the suction valve. The effects of the subcooling degree of inflow liquid hydrogen on the valve motion are further analyzed. The findings suggest that the subcooling degree of inflow liquid hydrogen helps inhibit the vaporization in the pump operation and ensures the valves work correctly. This work would contribute to pump optimization and valve collision failure analysis in reciprocating liquid hydrogen pumps. Full article

Background: Reciprocal inhibition (RI) is a spinal reflex that controls posture and movement. The modulation of spinal RI represented by the H-reflex has been studied, before and after voluntary contraction and electrical nerve stimulation but not during voluntary, electrically induced muscle contraction or a combination of voluntary and electrically induced muscle contractions. This study investigates the effects of the ongoing voluntary isometric contraction, the electrically induced isometric contraction, and the combination of voluntary with electrically induced isometric contraction of the Tibialis Anterior (TA) muscle on spinal RI represented by Soleus H-reflex. Methods: Eighteen healthy adults participated. Soleus H-reflex and M-response were measured during four different conditions as follows: (1) at rest, (2) electrically induced isometric contraction of the TA, (3) voluntary isometric contraction of the TA with a 1 kg force, and (4) combined voluntary and electrically induced isometric contraction of the TA with a 1 kg force. Results: The ANOVA clearly demonstrated significant differences in Soleus H-reflex amplitude across the four recording conditions (F3,16, 17.28, p < 0.001). The amplitude at rest was significantly higher than during electrically induced isometric contraction, voluntary isometric contraction, and the combined contraction conditions (p < 0.05). Furthermore, the amplitude recorded during the electrically induced isometric contraction condition significantly surpassed that of voluntary isometric contraction and the combined contraction conditions (p < 0.05). Moreover, there was no significant difference between Soleus H-reflex amplitude recorded during voluntary isometric contraction and the combined voluntary isometric contraction and electrically induced isometric contraction (p < 0.87). The combined voluntary isometric contraction and electrically induced isometric contraction condition had a higher inhibitory effect on the Soleus H-reflex with no significant differences from voluntary isometric contraction. Moreover, both were significantly better than electrically induced isometric contraction (p = 0.05). In terms of Soleus H-reflex latency, there was no significant difference among all four conditions (p > 0.05), meaning Soleus H-reflex latency was not influenced by the conditions. Conclusions: RI can be best modulated by combining voluntary with electrically induced isometric muscle contractions. Full article

Pancreatic cancer is an aggressive tumor with dismal prognosis. Neural invasion is one of the pathological hallmarks of pancreatic cancer. Peripheral nerves can modulate the phenotype and behavior of the malignant cells, as well as of different components of the tumor microenvironment, and thus affect tumor growth and metastasis. From a clinical point of view, neural invasion is translated into intractable pain and represents a predictor of tumor recurrence and poor prognosis. Several molecules are implicated in neural invasion and pain onset in PDAC, including neutrophins (e.g., NGF), chemokines, adhesion factors, axon-guidance molecules, different proteins, and neurotransmitters. In this review, we discuss the role of nerves within the pancreatic cancer microenvironment, highlighting how infiltrating nerve fibers promote tumor progression and metastasis, while tumor cells, in turn, drive nerve outgrowth in a reciprocal interaction that fuels tumor advancement. We outline key molecules involved in neural invasion in pancreatic cancer and, finally, explore potential therapeutic strategies to target neural invasion, aiming to both inhibit cancer progression and alleviate cancer-associated pain. Full article

Soluble epoxide hydrolase (sEH) is a bifunctional enzyme with epoxide hydrolase activity in the C-terminal domain (C-EH) and lipid phosphate phosphatase activity in the N-terminal domain (N-phos). The C-EH hydrolyzes bioactive epoxy fatty acids such as epoxyeicosatrienoic acid (EET). The N-phos hydrolyzes lipid phosphomonesters, including the signaling molecules of lysophosphatidic acid (LPA). Here, we report that the C-EH and N-phos are reciprocally regulated by their respective substrates. Full-length sEH (sEH-FL) showed positive cooperativity toward the substrate for each domain. Similar cooperativity was found when truncated enzymes having only C- and N-terminal domains, sEH-C and sEH-N, respectively, were used, suggesting an intra-domain nature of the cooperativity. In addition, the N-phos substrate LPA inhibited C-EH activity in sEH-FL and sEH-C equally. Similarly, the C-EH substrate EET inhibited N-phos activity. Structural and kinetic data suggest the presence of allosteric sites in each domain of the sEH enzyme, which share the binding of LPA and EET. Thus, each of the two sEH activities is regulated by a substrate of its own and by that of the other domain. This mechanism may explain why sEH has evolved to have two different enzyme activities, which possibly allows sEH to balance the metabolism of bioactive lipids. Full article

Reciprocal inhibition is often diminished in elderly individuals and those with upper motor neuron disorders. This reduction in reciprocal inhibition can hinder smooth joint movement. For subjects who have increased muscle tone and a limited range of motion in the joints, we focused on visual kinesthetic illusions as an intervention to increase reciprocal inhibition. We aimed to investigate the effects of visual kinesthetic illusions on reciprocal inhibition and motor function in the ankle joint. Participants participated in two experiments measuring reciprocal inhibition, namely reciprocal Ia inhibition and D1 inhibition, as well as motor functions related to ankle dorsiflexion and plantar flexion. Visual kinesthetic illusion was induced by displaying an image of each subject’s foot on a monitor. Our results showed that the visual kinesthetic illusion enhanced D1 inhibition and improved motor function in the ankle joint by prioritizing agonist muscle activity. We also observed a correlation between reciprocal inhibition and the muscle activity ratio. These findings suggest that visual kinesthetic illusions may improve motor function by increasing reciprocal inhibition. This study is the first to demonstrate the effects of visual kinesthetic illusion on reciprocal inhibition, and we believe that these findings can be applied in rehabilitation. Full article

Extensive and profound landscape alterations significantly contribute to ecological vulnerability in environmentally delicate regions. Existing research primarily emphasizes ecological risks caused by landscape alterations, while overlooking vulnerable characteristics of landscape functions; particularly lacking are studies on the driving mechanism of landscape ecological risk through the reciprocal relationship between landscape pattern risk and function risk. Based on these issues, this paper constructed a landscape pattern risk index (LPRI), a landscape function risk index (LFRI), and a landscape ecological risk index (LERI) in the counties of the dry–hot valley of the Jinsha River in southwest China. By employing a coupling degree and a coordination model, we analyzed temporal and spatial variations in the interaction between two types of ecological risk, thereby revealing the driving mechanisms of landscape ecological risk. The results indicated that the average LPRI values of the study area were 0.373, 0.327, and 0.427, respectively, while the average LFRI values were 0.451, 0.356, and 0.442 in 2000, 2010, and 2020, respectively. More than 90% of the study area exhibited a medium coupling relationship between the two types of ecological risks. The area proportion of the coupling coordination regions has increased from 25.58% to 31.07% from 2010 to 2020. The two types of risk exhibited a low level of constraint inhibition. Extremely evident expansion of high pattern–function risk areas and the area increase of coupling coordination region resulted in the acceleration of regional landscape ecological risk level. Increasing competition between market-driven land-use activities and ecological regulations from the government has rendered the diversification of landscape ecological risk sources and its underlying mechanisms intricate. This study serves as a model reference for assessing landscape ecological risk and a theoretical basis for sustainable landscape management and ecological regulation in the Yangtze River basin. Full article

Background/Objectives: Carcinoma-associated fibroblasts (CAFs), a prominent cell type in the tumor microenvironment (TME), significantly contributes to cancer progression through interactions with cancer cells and other TME components. Consequently, targeting signaling pathways driven by CAFs has potential to yield new therapeutic approaches to inhibit cancer progression. However, the mechanisms underlying their long-term interactions with cancer cells in vivo remains poorly understood. Methods: To address this, we developed a three-dimensional (3D) parallel coculture model of human triple-negative breast cancer (TNBC) cells and CAFs using our innovative TAME devices. This model allowed for the analysis of TNBC paracrine interactions via their secretome over extended culture periods (at least 70 days). Results: Using TNBC cell lines (MDA-MB-231, MCF10.DCIS, and HCC70), we found that TNBC spheroids in 3D parallel cocultures with CAFs exhibited more pronounced invasive finger-like outgrowths than those in cocultures of TNBC cells and normal fibroblasts (NFs) over a period of 50–70 days. We also established that the CAF-derived secretome affects TNBC migration towards the CAF secretome region. Additionally, we observed a preferential migration of CAFs, but not NFs, toward TNBC spheroids. Conclusions: Overall, our results suggest that paracrine interactions between TNBC cells and CAFs enhance TNBC invasive phenotypes and promote reciprocal migration. Full article

Host restriction factor GBP2 suppresses the replication of the ecotropic Moloney murine leukemia virus (E-MLV) by inhibiting furin protease, which cleaves the viral envelope glycoprotein (Env) into surface (SU) and transmembrane (TM) subunits. We analyzed the impacts of GBP2 on the infection efficiency mediated by MLV Envs of different strains of ecotropic Moloney, polytropic Friend, amphotropic, and xenotropic MLV-related (XMRV) viruses. Interestingly, the Envs of ecotropic Moloney and polytropic Friend MLV were sensitive to the antiviral activity of GBP2, while XMRV and amphotropic Envs showed resistance. Consistent with the sensitivity to GBP2, the amino acid sequences of the sensitive Envs at the SU-TM cleavage site were similar, as were the sequences of the resistant Envs. SU-TM cleavage of the GBP2-sensitive Env protein was inhibited by furin silencing, whereas that of GBP2-resistant Env was not. The substitution of the ecotropic Moloney cleavage site sequence with that of XMRV conferred resistance to both GBP2 and furin silencing. Reciprocally, the substitution of the XMRV cleavage site sequence with that of the ecotropic sequence conferred sensitivity to GBP2 and furin silencing. According to the SU-TM cleavage site sequence, there were sensitive and resistant variants among ecotropic, polytropic, and xenotropic MLVs. This study found that the dependence of MLV Env proteins on furin cleavage and GBP2-mediated restriction is determined by the amino acid sequences at the SU-TM cleavage site. Full article

Fibromyalgia (FM) is a disorder characterized by widespread chronic pain, significant depression, and various neural abnormalities. Recent research suggests a reciprocal exacerbation mechanism between chronic pain and depression. In patients with FM, dysregulation of tryptophan (Trp) metabolism has been identified. Trp, an essential amino acid, serves as a precursor to serotonin (5-HT), a neuromodulator that influences mood, appetite, sleep, and pain perception through the receptors 5-HT1, 5-HT2, and 5-HT3. Additionally, Trp is involved in the kynurenine pathway, a critical route in the immune response, inflammation, and production of neuroactive substances and nicotinamide adenine dinucleotide (NAD+). The activation of this pathway by pro-inflammatory cytokines, such as tumor necrosis factor α (TNF-α) and interferon gamma (IFN-γ), leads to the production of kynurenic acid (KYNA), which has neuroprotective properties, and quinolinic acid (QA), which is neurotoxic. These findings underscore the crucial balance between Trp metabolism, 5-HT, and kynurenine, where an imbalance can contribute to the dual burden of pain and depression in patients with FM. This review proposes a novel therapeutic approach for FM pain management, focusing on inhibiting QA synthesis while co-administering selective serotonin reuptake inhibitors to potentially increase KYNA levels, thus dampening pain perception and improving patient outcomes. Full article