Physiological Significance Research Articles

An activity-based sensing fluorescent probe for monitoring endogenous peroxynitrite in Alzheimer's disease models

Alzheimer's disease (AD) is a common neurodegenerative disease that brings great suffering and heavy economic burden to patients. Increasing evidence suggests that peroxynitrite (ONOO−) plays a crucial role in the pathogenesis of AD by mediating neurotoxicity and neuroinflammation. Developing effective peroxynitrite probes to monitor ONOO− is of great significance for the investigation and diagnosis of AD. Herein, two molecular fluorescent probes (NBD-Y and NBD-I) bearing a 7-nitro-2,1,3-benzoxadiazole (NBD) moiety were designed and synthesized for specific ONOO− detection. NBD-Y and NBD-I exhibited a rapid response (within 3 s), low detection limits (11 and 45 nM, respectively), high aqueous solubility (DMSO/PBS = 1: 199, v/v), high selectivity, excellent blood-brain barrier (BBB) penetrability (log P = 3.397), and a ∼78-fold decrease in fluorescence intensity in response to ONOO−. High-resolution mass spectrometry (HRMS), High-performance liquid chromatography (HPLC), and density functional theory (DFT) calculation results indicated that ONOO− may oxidatively break the C-N bond of the probe, resulting in a blue shift of the absorption spectrum due to the principle of intramolecular charge transfer (ICT). Furthermore, NBD-Y has been successfully applied for the detection of endogenous ONOO− in SH-SY5Y cells and is further employed for in vivo imaging in AD mouse models. These results denoted that NBD-Y may serve as a potential biological tool for further understanding the distinct physiological and pathological significance of ONOO−.

The role of the cartilage guanylyl cyclase-B receptor in craniofacial skeletal development

ObjectiveC-type natriuretic peptide (CNP), a member of the natriuretic peptide family, is a potent endochondral bone growth factor that exerts its biological effects via the guanylate cyclase B (GC-B) receptor. We previously demonstrated that CNP knockout (KO) mice exhibited midfacial hypoplasia along the sagittal plane; however, the effects of GC-B (the receptor for CNP) on endochondral ossification in the maxillofacial region remain unclear, and the mechanism of the CNP/GC-B system has not been elucidated. MethodsWe investigated the physiological significance of GC-B in the cartilage of the craniofacial region through analysis of cartilage-specific GC-B KO mice. Morphological assessments were performed at 12 weeks old, with histological analyses performed at 2 weeks old. ResultsGC-B-KO mice exhibited sagittal midfacial hypoplasia, foramen magnum stenosis, and spinal canal stenosis. Histological examination revealed reduced thickness in the spheno-occipital synchondrosis (SOS), a critical growth center in cranio-maxillofacial skeletal development. The hypertrophic zone of the SOS exhibited reduced thickness, accompanied by a reduction in cell count in this area. ConclusionsThis study highlights the essential role of GC-B receptors in craniofacial morphology contributing to our understanding of the mechanisms underlying facial morphological abnormalities, foramen magnum stenosis, and spinal canal stenosis.

Burning Question: How Does Our Brain Process Positive and Negative Cues Associated with Thermosensation?

Whether it is the dramatic suffocating sensation from a heat wave in the summer or the positive reinforcement arising from a hot drink on a cold day; we can certainly agree that our thermal environment underlies our daily rhythms of sensation. Extensive research has focused on deciphering the central circuits responsible for conveying the impact of thermogenesis on mammalian behavior. Here, we revise the recent literature responsible for defining the behavioral correlates that arise from thermogenic fluctuations in mammals. We transition from the physiological significance of thermosensation to the circuitry responsible for the autonomic or behavioral responses associated with it. Subsequently, we delve into the positive and negative valence encoded by thermoregulatory processes. Importantly, we emphasize the crucial junctures where reward, pain, and thermoregulation intersect, unveiling a complex interplay within these neural circuits. Finally, we briefly outline fundamental questions that are pending to be addressed in the field. Fully deciphering the thermoregulatory circuitry in mammals will have far-reaching medical implications. For instance, it may lead to the identification of novel targets to overcome thermal pain or allow the maintenance of our core temperature in prolonged surgeries.

RIPK3 cleavage is dispensable for necroptosis inhibition but restricts NLRP3 inflammasome activation.

Caspase-8 activity is required to inhibit necroptosis during embryogenesis in mice. In vitro studies have suggested that caspase-8 directly cleaves RIPK1, CYLD and the key necroptotic effector kinase RIPK3 to repress necroptosis. However, recent studies have shown that mice expressing uncleavable RIPK1 die during embryogenesis due to excessive apoptosis, while uncleavable CYLD mice are viable. Therefore, these results raise important questions about the role of RIPK3 cleavage. To evaluate the physiological significance of RIPK3 cleavage, we generated Ripk3D333A/D333A mice harbouring apoint mutation in the conserved caspase-8 cleavage site. These mice are viable, demonstrating that RIPK3 cleavage is not essential for blocking necroptosis during development. Furthermore, unlike RIPK1 cleavage-resistant cells, Ripk3D333A/D333A cells were not significantly more sensitive to necroptotic stimuli. Instead, we found that the cleavage of RIPK3 by caspase-8 restricts NLRP3 inflammasome activation-dependent pyroptosis and IL-1β secretion when Inhibitors of APoptosis (IAP) are limited. These results demonstrate that caspase-8 does not inhibit necroptosis by directly cleaving RIPK3 and further underscore a role for RIPK3 in regulating the NLRP3 inflammasome.

Regulation of Autophagosome-Lysosome Fusion by Human Viral Infections.

Autophagy plays a fundamental role in maintaining cellular homeostasis by eliminating intracellular components via lysosomes. Successful degradation through autophagy relies on the fusion of autophagosomes to lysosomes, which leads to the formation of autolysosomes containing acidic proteases that degrade the sequestered materials. Viral infections can exploit autophagy in infected cells to balance virus-host cell interactions by degrading the invading virus or promoting viral growth. In recent years, cumulative studies have indicated that viral infections may interfere with the fusion of autophagosomes and lysosomes, thus benefiting viral replication and associated pathogenesis. In this review, I provide an overview of the current understanding of the molecular mechanism by which human viral infections deregulate autophagosome-lysosome fusion and summarize the physiological significance in the virus life cycle and host cell damage.

Abstract P478: Short-Term Voluntary Exercise Significantly Influences the Diet-Induced Changes in Cecal Microbial Content in Male Mice

Two leading environmental factors that influence the overall health of humans are diet and physical exercise. However, it is still unclear how these factors influence health and disease susceptibility. Both diet and exercise have been shown to modulate the microbial flora that may significantly influence health and disease; however, it is unclear if diet modulates the effects of exercise on gut microflora. Therefore, this study aimed to determine how diet and exercise modulate the microbial content of the mouse cecum. Male, weanling, C57Bl/6 mice were randomly assigned to receive one of three diets ad libitum: standard laboratory chow, AIN93-G, and our novel Americanized diet (AD). Within each diet, mice were then assigned to a voluntary exercise regimen (wheel access, 16-hours per day, 3-days per week) or a control sedentary group (individually housed in cage without wheel for the same duration and frequency). After 2-weeks of exercise, mice were euthanized and cecal microbial content was determined by16S rDNA sequencing and RT-PCR for Firmicutes (FIRM), Bifidobacterium (GBIFD), Lactobacillus (LB) , and Bacteroides (BACT). Data were analyzed using GLM Procedures in SPSS with significance determined at P<0.05. Pilot results found that diet, exercise, and their interaction significantly influenced FIRM expression. Mice fed the standard chow diet had the lowest expression of FIRM , which was significantly lower than mice fed AIN (P<0.001) and AD (P=0.001). Exercise caused a significant decrease (P<0.001) in FIRM presence, especially in mice fed AIN and AD. Diet and exercise also significantly influenced GBIFD , with mice fed AIN having significantly more GBIFD than mice fed chow and AD (P<0.001 for both). Exercise also caused a significant reduction (P=0.016) in GBIFD , and a reduction in LB expression in mice fed chow and AD, but not AIN (P=0.03). The results of this study highlight how two key aspects of lifestyle, diet and exercise, independently and additively influence the gastrointestinal flora in a preclinical mouse model. Additional studies are needed to validate these findings and determine their physiological significance.

Abstract P488: Diet Significantly Alters Metabolic Gene Expression Associated With Probiotic Administration in Mice

The microbial flora in the gastrointestinal tract is known to be mediated by diet and is thus one potential mechanism by which diet influences health. Furthermore, probiotic supplementation also affects various biological markers, however it is unclear if a diet and probiotic interaction exists. Therefore, the goal of our study was to assess how diet and probiotic administration influenced metabolic health in mice. Weanling, male C57Bl/6 mice were randomly assigned to receive one of three diets: chow, American Institute of Nutrition 93 formulation (AIN93), or our novel Americanized diet (AD) formulated to match the 50 th percentile of nutrient intake in Americans. Mice were then administered a commercially-available probiotic for 2-weeks and then euthanized. Insulin sensitivity was determined by performing an insulin-tolerance test (ITT) in a separate cohort. Blood was collected for serum analysis of circulating metabolic markers. Liver and adipose tissue were collected and processed to quantify the mRNA expression of key metabolic genes by real-time RT-PCR. All data were analyzed using General Linear Model Procedures in SPSS with significance determined at P<0.05. Diet (P=0.001), but not probiotic supplementation (P=0.27), significantly affected ITT with the AD group having the lowest responsiveness to insulin ( P ≤0.001). Chow fed mice tended ( P =0.09) to have lower HDL than the AIN-93 and AD fed mice. Probiotic administration alone resulted in an upregulation of liver mRNA expression of SREBF2, particularly the AIN-93 diet group (P=0.024). Probiotic supplementation also modulated liver transcription of NF-κB1 with an increase in chow-fed mice, but decreased in those fed AD (P=0.016). Increased transcription of the key carbohydrate metabolism regulating gene, Mlxipl , was found to occur in liver of the chow and AIN-93 and also tended to differ in mice fed the AD (P=0.06). In the adipose tissue, diet (P<0.001), probiotic (P<0.001), and their interaction (P=0.01) influenced transcription of the androgen receptor ( Ar ) gene. The chow group had the most Ar transcription, while the AD had the lowest transcription in both the diet alone and probiotic treated group. Taken together, our data highlight the significance of diet in mediating the beneficial metabolic effects of probiotic administration. Diet appears to play a greater role in dictating the expression of key metabolic enzymes and transcription factors and future studies are needed to validate these findings and determine their physiological significance.

First- and second-order phase transitions in electronic excitable units and neural dynamics under global inhibitory feedback

The diverse roles of inhibition in neural circuits and other dynamical networks are receiving renewed interest. Here, it is shown that increasing global inhibitory feedback leads to gradual rounding of first-order transition between dynamical phases, turning it into second-order transition. The effect is initially observed in an electronic model consisting of a bi-dimensional array of neon glow lamps, where global inhibition can be simply introduced through a resistor in series with the supply voltage. The experimental findings are confirmed using both an extended numerical model and a mean-field approximation, then replicated across different models of neural dynamics, namely, the Wilson–Cowan model and a network of leaky integrate-and-fire neurons. Across all these systems, a critical point is always found as a function of a pair of parameters controlling local excitability and global inhibition strength, and a general explanation revealing the roles of the shape of the activation function and voltage fluctuations versus the extinction time-scale is provided. It is speculated that the brain could use global inhibition as a versatile means of shifting between first- and second-order dynamics, addressing the conundrum regarding the coexistence in neural dynamics of phenomena stemming from both. Some reflections regarding the comparison with other physical systems and the possible physiological significance are offered, and a hypothetical setup for an optogenetics experiment on cultured neurons is put forward.

Defining Requirements for Heme Binding in PGRMC1 and Identifying Key Elements that Influence Protein Dimerization.

Progesterone receptor membrane component 1 (PGRMC1) binds heme via a surface-exposed site and displays some structural resemblance to cytochrome b5 despite their different functions. In the case of PGRMC1, it is the protein interaction with drug-metabolizing cytochrome P450s and the epidermal growth factor receptor that has garnered the most attention. These interactions are thought to result in a compromised ability to metabolize common chemotherapy agents and to enhance cancer cell proliferation. X-ray crystallography and immunoprecipitation data have suggested that heme-mediated PGRMC1 dimers are important for facilitating these interactions. However, more recent studies have called into question the requirement of heme binding for PGRMC1 dimerization. Our study employs spectroscopic and computational methods to probe and define heme binding and its impact on PGRMC1 dimerization. Fluorescence, electron paramagnetic resonance and circular dichroism spectroscopies confirm heme binding to apo-PGRMC1 and were used to demonstrate the stabilizing effect of heme on the wild-type protein. We also utilized variants (C129S and Y113F) to precisely define the contributions of disulfide bonds and direct heme coordination to PGRMC1 dimerization. Understanding the key factors involved in these processes has important implications for downstream protein-protein interactions that may influence the metabolism of chemotherapeutic agents. This work opens avenues for deeper exploration into the physiological significance of the truncated-PGRMC1 model and developing design principles for potential therapeutics to target PGRMC1 dimerization and downstream interactions.

A novel spatiotemporal graph convolutional network framework for functional connectivity biomarkers identification of Alzheimer’s disease

BackgroundFunctional connectivity (FC) biomarkers play a crucial role in the early diagnosis and mechanistic study of Alzheimer’s disease (AD). However, the identification of effective FC biomarkers remains challenging. In this study, we introduce a novel approach, the spatiotemporal graph convolutional network (ST-GCN) combined with the gradient-based class activation mapping (Grad-CAM) model (STGC-GCAM), to effectively identify FC biomarkers for AD.MethodsThis multi-center cross-racial retrospective study involved 2,272 participants, including 1,105 cognitively normal (CN) subjects, 790 mild cognitive impairment (MCI) individuals, and 377 AD patients. All participants underwent functional magnetic resonance imaging (fMRI) and T1-weighted MRI scans. In this study, firstly, we optimized the STGC-GCAM model to enhance classification accuracy. Secondly, we identified novel AD-associated biomarkers using the optimized model. Thirdly, we validated the imaging biomarkers using Kaplan–Meier analysis. Lastly, we performed correlation analysis and causal mediation analysis to confirm the physiological significance of the identified biomarkers.ResultsThe STGC-GCAM model demonstrated great classification performance (The average area under the curve (AUC) values for different categories were: CN vs MCI = 0.98, CN vs AD = 0.95, MCI vs AD = 0.96, stable MCI vs progressive MCI = 0.79). Notably, the model identified specific brain regions, including the sensorimotor network (SMN), visual network (VN), and default mode network (DMN), as key differentiators between patients and CN individuals. These brain regions exhibited significant associations with the severity of cognitive impairment (p < 0.05). Moreover, the topological features of important brain regions demonstrated excellent predictive capability for the conversion from MCI to AD (Hazard ratio = 3.885, p < 0.001). Additionally, our findings revealed that the topological features of these brain regions mediated the impact of amyloid beta (Aβ) deposition (bootstrapped average causal mediation effect: β = -0.01 [-0.025, 0.00], p < 0.001) and brain glucose metabolism (bootstrapped average causal mediation effect: β = -0.02 [-0.04, -0.001], p < 0.001) on cognitive status.ConclusionsThis study presents the STGC-GCAM framework, which identifies FC biomarkers using a large multi-site fMRI dataset.

Is High-Frequency Activity at Seizure Onset Inhibitory? A Stereoelectroencephalographic Study of Motor Cortex Seizures.

In the era of stereoelectroencephalography (SEEG), many studies have been devoted to understanding the role of interictal high-frequency oscillations. High-frequency activity (HFA) at seizure onset has been identified as a marker of epileptogenic zone. We address the physiological significance of ictal HFAs and their relation to clinical semiology. We retrospectively identified patients with pure focal primary motor epilepsy. We selected only patients in whom SEEG electrodes were optimally placed in the motor cortex as confirmed by electrical stimulation. Based on these narrow inclusion criteria, we extensively studied 5 patients (3 males and 2 females, mean age = 22.4 years) using time-frequency analysis and time correlation with motor signs onset. A total of 157 analyzable seizures were recorded in 5 subjects. The first 2 subjects had tonic or clonic semiology with rare secondary generalization. Subject 3 had atonic onset followed by clonic hand/arm flexion. Subject 4 had clusters of tonic and atonic facial movements. Subject 5 had upper extremity tonic movements. The median frequency of the fast activity extracted from the Epileptogenic Zone Fingerprint pipeline in the first 4 subjects was 76 Hz (interquartile range = 21.9Hz). Positive motor signs did not occur concomitantly with high gamma activity developing in the motor cortex. Motor signs began at the end of HFAs. This study supports the hypothesis of an inhibitory effect of ictal HFAs. The frequency range in the gamma band was associated with the direction of the clinical output effect. Changes from inhibitory to excitatory effect occurred when discharge frequency dropped to low gamma or beta. ANN NEUROL 2024;95:1127-1137.

Association Between Chondrolabral Junction Breakdown and Conversion to Total Hip Arthroplasty After Hip Arthroscopy for Symptomatic Labral Tears: Minimum 8-Year Follow-up

Background: Arthroscopic treatment of femoroacetabular impingement (FAI) and symptomatic labral tears confers short- to midterm benefits, yet further long-term evidence is needed. Moreover, despite the physiological and biomechanical significance of the chondrolabral junction (CLJ), the clinical implications of damage to this transition zone remain understudied. Purpose: To (1) report minimum 8-year survivorship and patient-reported outcome measures after hip arthroscopy for FAI and (2) characterize associations between outcomes and patient characteristics (age, body mass index, sex), pathological parameters (Tönnis angle, alpha angle, type of FAI, CLJ breakdown), and procedures performed (labral management, FAI treatment, microfracture). Study Design: Cohort study; Level of evidence, 3. Methods: This retrospective cohort study included patients who underwent primary hip arthroscopy for symptomatic labral tears secondary to FAI by a single surgeon between 2002 and 2013. All patients were ≥18 years of age with minimum 8-year follow-up and available preoperative radiographs. The primary outcome was conversion to total hip arthroplasty (THA), and secondary outcomes included revision arthroscopy, patient-reported outcome measures, and patient satisfaction. CLJ breakdown was assessed using the Beck classification. Kaplan-Meier estimates and weighted Cox regression were used to estimate 10-year survivorship (no conversion to THA) and identify risk factors associated with THA conversion. Results: In this study of 174 hips (50.6% female; mean age, 37.8 ± 11.2 years) with mean follow-up of 11.1 ± 2.5 years, the 10-year survivorship rate was 81.6% (95% CI, 75.9%-87.7%). Conversion to THA occurred at a mean 4.7 ± 3.8 years postoperatively. Unadjusted analyses revealed several variables significantly associated with THA conversion, including older age; higher body mass index; higher Tönnis grade; labral debridement; and advanced breakdown of the CLJ, labrum, or articular cartilage. Survivorship at 10 years was inferior in patients exhibiting severe (43.6%; 95% CI, 31.9%-59.7%) versus mild (97.9%; 95% CI, 95.1%-100%) breakdown of the CLJ (P < .001). Multivariable analysis identified worsening CLJ breakdown (weighted hazard ratio per 1-unit increase, 6.41; 95% CI, 3.11-13.24), older age (1.09; 95% CI, 1.04-1.14), and higher Tönnis grade (4.59; 95% CI, 2.13-9.90) as independent negative prognosticators (P < .001 for all). Conclusion: Although most patients achieved favorable minimum 8-year outcomes, several pre- and intraoperative factors were associated with THA conversion; of these, worse CLJ breakdown, higher Tönnis grade, and older age were the strongest predictors.

Clerodendrum Glandulosum Lindl.: A Review of Ethnopharmacology, Pharmacological Potentials, and their Mechanism of Action.

Clerodendrum glandulosum Lindl. is popularly known for its traditional herbal remedies and therapeutic properties among the people of Northeast Indian communities, as well as Chinese traditional medicine. For the myriad pharmacological properties, viz., anti-hypertensive, hypolipidemic, hepatoprotective, anti-inflammatory, and neuroprotective, planting this species in kitchen gardens is a common practice to treat various ailments, especially hypertension, diabetes, and other metabolic complications. Different phytochemicals, representing the diverse classes of secondary metabolites comprising physiological and phytopharmaceutical significance, have been reported from C. glandulosum. Compounds with terpenoids, steroids, and phenolics are in demand in the pharmaceutical industry. An overview of the mechanism of action of the prominent compounds has also been collated for future research on C. glandulosum-based therapeutics. Current information focuses on this important medicinal plant's ethnomedicinal use, phytochemistry, pharmacology, associated mechanisms, and toxicology. This review will help explore this potential medicinal plant, which can pave the path for its application in the pharmaceutical industry.

Light Wavelength as a Contributory Factor of Environmental Fitness in the Cyanobacterial Circadian Clock.

A circadian clock is an essential system that drives the 24-h expression rhythms for adaptation to day-night cycles. The molecular mechanism of the circadian clock has been extensively studied in cyanobacteria harboring the KaiC-based timing system. Nevertheless, our understanding of the physiological significance of the cyanobacterial circadian clock is still limited. In this study, we cultured wild-type Synechococcus elongatus PCC 7942 and circadian clock mutants in day-night cycles at different light qualities and found that the growth of the circadian clock mutants was specifically impaired during 12-h blue light/12-h dark (BD) cycles for the first time. The arrhythmic mutant kaiCAA was further analyzed by photosynthetic measurements. Compared with the wild type, the mutant exhibited decreases in the chlorophyll content, the ratio of photosystem I to II, net O2 evolution rate and efficiency of photosystem II photochemistry during BD cycles. These results indicate that the circadian clock is necessary for the growth and the maintenance of the optimum function of the photosynthetic apparatus in cyanobacteria under blue photoperiodic conditions.

Predisposal of Interferon Regulatory Factor 1 Deficiency to Accumulate DNA Damage and Promote Osteoarthritis Development in Cartilage.

Interferon regulatory factor 1 (IRF1) is a transcriptional regulator conventionally associated with immunomodulation. Recent molecular analyses mapping DNA binding sites of IRF1 have suggested its potential function in DNA repair. However, the physiologic significance of this noncanonical function remains unexplored. Here, we investigated the role of IRF1 in osteoarthritis (OA), a condition marked by senescence and chronic joint inflammation. OA progression was examined in wild-type and Irf1-/- mice using histologic assessments and microcomputed tomography analysis of whole-joint OA manifestations and behavioral assessments of joint pain. An integrated analysis of assay for transposase-accessible chromatin with sequencing and whole transcriptome data was conducted for the functional assessment of IRF1 in chondrocytes. The role of IRF1 in DNA repair and senescence was investigated by assaying γ-H2AX foci and senescence-associated beta-galactosidase activity. Our genome-wide investigation of IRF1 footprinting in chondrocytes revealed its primary occupancies in the promoters of DNA repair genes without noticeable footprint patterns in those of interferon-responsive genes. Chondrocytes lacking IRF1 accumulated irreversible DNA damage under oxidative stress, facilitating their entry into cellular senescence. IRF1 was down-regulated in the cartilage of human and mouse OA. Although IRF1 overexpression did not elicit an inflammatory response in joints or affect OA development, genetic deletion of Irf1 caused enhanced chondrocyte senescence and exacerbated post-traumatic OA in mice. IRF1 offers DNA damage surveillance in chondrocytes, protecting them from oxidative stress associated with OA risk factors. Our study provides a crucial and cautionary perspective that compromising IRF1 activity renders chondrocytes vulnerable to cellular senescence and promotes OA development.

α-Methylacyl-CoA Racemase from Mycobacterium tuberculosis-Detailed Kinetic and Structural Characterization of the Active Site.

α-Methylacyl-CoA racemase in M. tuberculosis (MCR) has an essential role in fatty acid metabolism and cholesterol utilization, contributing to the bacterium's survival and persistence. Understanding the enzymatic activity and structural features of MCR provides insights into its physiological and pathological significance and potential as a therapeutic target. Here, we report high-resolution crystal structures for wild-type MCR in a new crystal form (at 1.65 Å resolution) and for three active-site mutants, H126A, D156A and E241A, at 2.45, 1.64 and 1.85 Å resolutions, respectively. Our analysis of the new wild-type structure revealed a similar dimeric arrangement of MCR molecules to that previously reported and details of the catalytic site. The determination of the structures of these H126A, D156A and E241A mutants, along with their detailed kinetic analysis, has now allowed for a rigorous assessment of their catalytic properties. No significant change outside the enzymatic active site was observed in the three mutants, establishing that the diminution of catalytic activity is mainly attributable to disruption of the catalytic apparatus involving key hydrogen bonding and water-mediated interactions. The wild-type structure, together with detailed mutational and biochemical data, provide a basis for understanding the catalytic properties of this enzyme, which is important for the design of future anti-tuberculosis drug molecules.

Issue Information

The pyramidal muscle (PM) is a paired small triangular muscle of the anterior abdominal wall. Its physiological significance is unclear. The present study used high‐resolution MRI to determine when the PM emerged and revealed its features. The PM was first detected at Carnegie stage (CS)20 and in five of the seven samples between CS21 and CS23. 81.4% of samples during the early fetal period had PM on both the right and left sides. The frequency, side differences, sex dimorphism, and spatial position of the PM in the early fetal period were similar to those in adults.FIGURE: Representative images of the PM (CRL: 58.7 mm). (A) T1W MR image. (B) 3D‐reconstructed image showing the PM (red), rectus abdominis muscle (purple). (C) Diffusion tensor image showing the PM and rectus abdominis muscles. Further details can be seen in the article by Iwasa et al. in this issue.image

Mechanoregulation and function of calponin and transgelin.

It is well known that chemical energy can be converted to mechanical force in biological systems by motor proteins such as myosin ATPase. It is also broadly observed that constant/static mechanical signals potently induce cellular responses. However, the mechanisms that cells sense and convert the mechanical force into biochemical signals are not well understood. Calponin and transgelin are a family of homologous proteins that participate in the regulation of actin-activated myosin motor activity. An isoform of calponin, calponin 2, has been shown to regulate cytoskeleton-based cell motility functions under mechanical signaling. The expression of the calponin 2 gene and the turnover of calponin 2 protein are both under mechanoregulation. The regulation and function of calponin 2 has physiological and pathological significance, as shown in platelet adhesion, inflammatory arthritis, arterial atherosclerosis, calcific aortic valve disease, post-surgical fibrotic peritoneal adhesion, chronic proteinuria, ovarian insufficiency, and tumor metastasis. The levels of calponin 2 vary in different cell types, reflecting adaptations to specific tissue environments and functional states. The present review focuses on the mechanoregulation of calponin and transgelin family proteins to explore how cells sense steady tension and convert the force signal to biochemical activities. Our objective is to present a current knowledge basis for further investigations to establish the function and mechanisms of calponin and transgelin in cellular mechanoregulation.

The global regulation of c-di-GMP and cAMP in bacteria.

Nucleotide second messengers are highly versatile signaling molecules that regulate a variety of key biological processes in bacteria. The best-studied examples are cyclic AMP (cAMP) and bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP), which both act as global regulators. Global regulatory frameworks of c-di-GMP and cAMP in bacteria show several parallels but also significant variances. In this review, we illustrate the global regulatory models of the two nucleotide second messengers, compare the different regulatory frameworks between c-di-GMP and cAMP, and discuss the mechanisms and physiological significance of cross-regulation between c-di-GMP and cAMP. c-di-GMP responds to numerous signals dependent on a great number of metabolic enzymes, and it regulates various signal transduction pathways through its huge number of effectors with varying activities. In contrast, due to the limited quantity, the cAMP metabolic enzymes and its major effector are regulated at different levels by diverse signals. cAMP performs its global regulatory function primarily by controlling the transcription of a large number of genes via cAMP receptor protein (CRP)in most bacteria. This review can help us understand how bacteria use the two typical nucleotide second messengers to effectively coordinate and integrate various physiological processes, providing theoretical guidelines for future research.

Immune priming modulates Galleria mellonella and Pseudomonas entomophila interaction. Antimicrobial properties of Kazal peptide Pr13a.

Galleria mellonella larvae repeatedly infected with Pseudomonas entomophila bacteria re-induced their immune response. Its parameters, i.e. the defence activities of cell-free hemolymph, the presence and activity of antimicrobial peptides, and the expression of immune-relevant genes were modulated after the re-challenge in comparison to non-primed infected larvae, resulting in better protection. No enhanced resistance was observed when the larvae were initially infected with other microorganisms, and larvae pre-infected with P. entomophila were not more resistant to further infection with other pathogens. Then, the peptide profiles of hemolymph from primed- and non-primed larvae infected with P. entomophila were compared by quantitative RP-HPLC (Reverse Phase - High Performance Liquid Chromatography). The level of carbonic anhydrase, anionic peptide-1, proline peptide-2, and finally, unknown so far, putative Kazal peptide Pr13a was higher in the primed infected animals than in the larvae infected with P. entomophila for the first time. The expression of the Pr13a gene increased two-fold after the infection, but only in the primed animals. To check whether the enhanced level of Pr13a could have physiological significance, the peptide was purified to homogeneity and checked for its defence properties. In fact, it had antibacterial activity: at the concentration of 15 µM and 7.5 µM it reduced the number of P. entomophila and Bacillus thuringiensis CFU, respectively, to about 40%. The antibacterial activity of Pr13a was correlated with changes observed on the surface of the peptide-treated bacteria, e.g. surface roughness and adhesion force. The presented results bring us closer to finding hemolymph constituents responsible for the effect of priming on the immune response in re-infected insects.