Glutamatergic melanocortin-4 receptor neurons regulate body weight.

The locus coeruleus (LC), enriched in vesicular glutamate transporter 2 (VGlut2) neurons, is a potential homeostasis-regulating hub. However, the identity of melanocortin-4 receptor (MC4R) neurons in the paraventricular nucleus (PVN) of the hypothalamus, PVNVGlut2::MC4R and LCVGlut2::MC4R regulation of body weight, and axonal projections of LCVGlut2 neurons remain unclear. Conditional knockout of MC4R in chimeric mice was used to confirm the effects of VGlut2. Interscapular brown adipose tissue was injected with pseudorabies virus to study the central nervous system projections. We mapped the LCVGlut2 circuitry. Based on the Cre-LoxP recombination system, specific knockdown of MC4R in VGlut2 neurons resulted in weight gain in chimeric mice. Adeno-associated virus-mediated knockdown of MC4R expression in the PVN and LC had potential superimposed effects on weight gain, demonstrating the importance of VGlut2 neurons. Unlike these wide-ranging efferent projections, the PVN, hypothalamic arcuate nucleus, supraoptic nucleus of the lateral olfactory tegmental nuclei, and nucleus tractus solitarius send excitatory projections to LCVGlut2 neurons. The PVN → LC glutamatergic MC4R long-term neural circuit positively affected weight management and could help treat obesity.

Effects of amylin on food intake and body weight via sympathetic innervation of the interscapular brown adipose tissue.

Objective: Amylin acts on the lateral dorsal tegmental nucleus (LDT), resulting in anorexic and weight-loss effects and activates thermogenesis in the interscapular brown adipose tissue (IBAT). In addition, it induces neuronal nitric oxide synthase (nNOS) and choline acetyltransferase (ChAT)-mediated feeding. However, the influence of the intact sympathetic nervous system (SNS) in mediating amylin's effects has not been fully characterised. We investigated whether extracellular signal-regulated kinase (ERK), nNOS, and ChAT activities in the LDT are responsible for amylin's anorexigenic effects and whether this requires an intact SNS.Methods: C57BL/6J mice [wild-type (WT), sham, and sympathetic denervation of IBAT] were used. Food consumption, body weight, and distribution of pERK, nNOS, and ChAT positive neurons in the brain were examined following acute and chronic amylin administration.Results: Food intake was significantly decreased in WT and sham animals following acute amylin injection, but not in the denervated mice. Chronic amylin reduced body weight and serum glucose levels after 6 weeks, but increased insulin levels; no changes were observed in the denervated mice. Acute amylin increased the expression of nNOS, ChAT, and uncoupling protein-1 in the IBAT of WT and sham mice, while no changes were observed in the denervated mice and pERK from the above effect.Conclusions: Intact SNS of IBAT influences amylin-induced suppression of food intake and body weight, thus affecting nNOS and ChAT signalling in the LDT and locus coeruleus.

Dynamic Digital Twin: Diagnosis, Treatment, Prediction, and Prevention of Disease During the Life Course.

A digital twin (DT), originally defined as a virtual representation of a physical asset, system, or process, is a new concept in health care. A DT in health care is not a single technology but a domain-adapted multimodal modeling approach incorporating the acquisition, management, analysis, prediction, and interpretation of data, aiming to improve medical decision-making. However, there are many challenges and barriers that must be overcome before a DT can be used in health care. In this viewpoint paper, we build on the current literature, address these challenges, and describe a dynamic DT in health care for optimizing individual patient health care journeys, specifically for women at risk for cardiovascular complications in the preconception and pregnancy periods and across the life course. We describe how we can commit multiple domains to developing this DT. With our cross-domain definition of the DT, we aim to define future goals, trade-offs, and methods that will guide the development of the dynamic DT and implementation strategies in health care.

Response of the expression of oxytocin neurons to ghrelin in female mice.

Ghrelin is an orexigenic agonist that acts directly on neurons in the hypothalamus, controlling appetite and energy balance. Although its role in appetite-associated neurons has been described, the relationship between peripheral ghrelin stimulation and oxytocin expression in the paraventricular nucleus is not fully understood. We evaluated the suppressive function of ghrelin in oxytocin-positive paraventricular nucleus neurons in ovariectomized C57BL/6 mice 2 h after ghrelin injection. The results showed that, in intact mice, peripheral ghrelin stimulation activated estrogen receptor alpha-expressing neurons during the estrous cycle and that agouti-related peptide mRNA expression was remarkably increased. Agouti-related peptide neuron axons co-localized with oxytocin neurons in the paraventricular nucleus. Moreover, the response of oxytocin-positive paraventricular nucleus neurons to ghrelin was suppressed in the proestrus period, while ghrelin decreased the serum concentration of estradiol in the proestrus phase. These data suggest that ghrelin may suppress oxytocin-positive neuron expression via the arcuate nucleus agouti-related peptide circuit, with the possible influence of estradiol in the murine estrous cycle. Unraveling the mechanism of ghrelin-induced oxytocin expression in the hypothalamus paraventricular nucleus broadens the horizon for ghrelin-related appetite research.

p66Shc is associated with hydrogen peroxide-induced oxidative stress in preimplantation sheep embryos.

The low efficiency of in vitro embryo production is associated with oxidative stress induced by suboptimal culture conditions. p66Shc is a 66-kDa protein of the ShcA (Src homologous-collagen homolog) adaptor protein family, which is involved in signaling pathways involved in oxidative stress regulation, apoptosis induction, and aging. However, the functional role of p66Shc during the preimplantation development of sheep embryos is not understood. Our results showed that early-cleavage (≤28 hr) embryos had a higher developmental potential than late-cleavage (>28 hr) embryos. The poor quality of these late-cleavage embryos was associated with increased the transcripts and protein of p66Shc and decreased mitochondrial activity. In addition, exogenous hydrogen peroxide-induced oxidative stress significantly increased p66Shc protein abundance and suppressed embryonic development, which was ameliorated by antioxidant treatment. Notably, oxidative stress induced the nuclear localization of p66Shc and phosphorylated (Ser-36) p66Shc. Collectively, these observations suggest that p66Shc may be playing an important role in the regulation of oxidative stress during the preimplantation development of sheep embryos.

Melanocortin 4 receptor-mediated effects of amylin on thermogenesis and regulation of food intake.

AIMS: Amylin, a pancreatic hormone cosecreted with insulin, exerts important anorexic and weight-loss effects. Melanocortin 4 receptor (MC4R) signalling plays a critical role in energy homeostasis; however, its role on amylin-dependent regulation of food intake and adaptive thermogenesis of interscapular brown adipose tissue (IBAT) are unclear. In this study, we examined the effects of amylin on food intake and thermogenesis on IBAT via the MC4R pathway in mice. MATERIALS AND METHODS: Acute food consumption and thermogenesis in IBAT were measured in male wild-type (WT) and MC4R-deficient mice following intraperitoneal injection of amylin and SHU9119, an MC3R/4R antagonist, to determine the role of the central melanocortin system on the hypothalamus and IBAT. RESULTS: Amylin (50 µg/kg) suppressed feeding and stimulated thermogenesis on IBAT via activation of the MC4R system in mice. Pharmacological blockade of MC4R using SHU9119 (50 µg/kg) attenuated amylin-induced inhibition of feeding and stimulation of thermogenesis in IBAT. No changes were observed when SHU9119 was injected alone. Moreover, amylin significantly increased MC4R expression and c-Fos neuronal signals in the arcuate nucleus and significantly increased acetyl-CoA carboxylase (ACC) phosphorylation in the hypothalamus and IBAT and uncoupling protein-1 (UCP1) expression in the IBAT of WT mice via the MC4R pathway. CONCLUSION: The melanocortin system was involved in amylin-induced suppression of food intake and activation of thermogenesis in both the hypothalamus and IBAT via modulation of ACC phosphorylation and UCP1 expression.

Erratum: HOXC10 promotes carboplatin resistance of ovarian cancer by regulating ABCC3.

[This corrects the article on p. 4602 in vol. 12, PMID: 36381312.].

The role of circadian clocks in cancer: Mechanisms and clinical implications.

Circadian rhythm refers to the inherent 24-h cycle oscillation of biochemical, physiological and behavioral functions, which is almost universal in eukaryotes. At least 14 core clock genes have been reported to form multiple chain feedback loops that confer intrinsic circadian rhythmicity onto the molecular clock. Accumulating evidence has shown that the circadian gene dysfunction resulted from single nucleotide polymorphisms (SNPs), deletions, epigenetic modification, and deregulation is strongly associated with cancer risk. In the present review, we describe the composition of circadian rhythm system. We highlight the function and mechanism of clock genes in cancer pathogenesis and progression. Moreover, their potential clinical implications as prognostic biomarkers and therapeutic targets have been addressed.

3D Printing a Low-Cost Miniature Accommodating Optical Microscope.

This decade has witnessed the tremendous progress in miniaturizing optical imaging systems. Despite the advancements in 3D printing optical lenses at increasingly smaller dimensions, challenges remain in precisely manufacturing the dimensionally compatible optomechanical components and assembling them into a functional imaging system. To tackle this issue, the use of 3D printing to enable digitalized optomechanical component manufacturing, part-count-reduction design, and the inclusion of passive alignment features is reported here, all for the ease of system assembly. The key optomechanical components of a penny-sized accommodating optical microscope are 3D printed in 50 min at a significantly reduced unit cost near $4. By actuating a built-in voice-coil motor, its accommodating capability is validated to focus on specimens located at different distances, and a focus-stacking function is further utilized to greatly extend depth of field. The microscope can be readily customized and rapidly manufactured to respond to task-specific needs in form factor and optical characteristics.

SBC (Sanhuang Xiexin Tang combined with Baihu Tang plus Cangzhu) alleviates NAFLD by enhancing mitochondrial biogenesis and ameliorating inflammation in obese patients and mice.

In the context of non-alcoholic fatty liver disease (NAFLD), characterized by dysregulated lipid metabolism in hepatocytes, the quest for safe and effective therapeutics targeting lipid metabolism has gained paramount importance. Sanhuang Xiexin Tang (SXT) and Baihu Tang (BHT) have emerged as prominent candidates for treating metabolic disorders. SXT combined with BHT plus Cangzhu (SBC) has been used clinically for Weihuochisheng obese patients. This retrospective analysis focused on assessing the anti-obesity effects of SBC in Weihuochisheng obese patients. We observed significant reductions in body weight and hepatic lipid content among obese patients following SBC treatment. To gain further insights, we investigated the effects and underlying mechanisms of SBC in HFD-fed mice. The results demonstrated that SBC treatment mitigated body weight gain and hepatic lipid accumulation in HFD-fed mice. Pharmacological network analysis suggested that SBC may affect lipid metabolism, mitochondria, inflammation, and apoptosis-a hypothesis supported by the hepatic transcriptomic analysis in HFD-fed mice treated with SBC. Notably, SBC treatment was associated with enhanced hepatic mitochondrial biogenesis and the inhibition of the c-Jun N-terminal kinase (JNK)/nuclear factor-kappa B (NF-κB) and extracellular signal-regulated kinase (ERK)/NF-κB pathways. In conclusion, SBC treatment alleviates NAFLD in both obese patients and mouse models by improving lipid metabolism, potentially through enhancing mitochondrial biogenesis. These effects, in turn, ameliorate inflammation in hepatocytes.

The emerging roles of HDACs and their therapeutic implications in cancer.

Deregulation of protein post-translational modifications is intensively involved in the etiology of diseases, including degenerative diseases, inflammatory injuries, and cancers. Acetylation is one of the most common post-translational modifications of proteins, and the acetylation levels are controlled by two mutually antagonistic enzyme families, histone acetyl transferases (HATs) and histone deacetylases (HDACs). HATs loosen the chromatin structure by neutralizing the positive charge of lysine residues of histones; whereas HDACs deacetylate certain histones, thus inhibiting gene transcription. Compared with HATs, HDACs have been more intensively studied, particularly regarding their clinical significance. HDACs extensively participate in the regulation of proliferation, migration, angiogenesis, immune escape, and therapeutic resistance of cancer cells, thus emerging as critical targets for clinical cancer therapy. Compared to HATs, inhibitors of HDAC have been clinically used for cancer treatment. Here, we enumerate and integratethe mechanisms of HDAC family members in tumorigenesis and cancer progression, and address the new and exciting therapeutic implications of single or combined HDAC inhibitor (HDACi) treatment.

HOXC10 promotes carboplatin resistance of ovarian cancer by regulating ABCC3.

HOXC10 has been reported to be upregulated in ovarian cancer (OC) tissues, attributing to the metastasis of OC. However, the specific functions of HOXC10 in OC, especially its role in chemoresistance, remain to be determined. Therefore, in this study, we explored the function and the underlying mechanisms of HOXC10 in carboplatin resistance of OC. A variety of approaches were utilized to analyze the expression of HOXC10 and its related genes. The effect of HOXC10 in cell growth and chemoresistance was investigated in carboplatin-resistant OC subline TOV21G-R and the parental TOV21G-P cells. ROC curve and survival analysis were conducted to determine the predictive value of HOXC10 and ABCC3 combination in carboplatin resistance and the prognosis of OC. Luciferase reporter assay and Chromatin immunoprecipitation (ChIP) assay were used to explore the direct regulation of ß-catenin by HOXC10. Our results demonstrated that the expression of HOXC10 was upregulated both in the carboplatin-resistant OC tissues and TOV21G-R cells. Furthermore, the upregulation of HOXC10 could promote the expression of ABCC3 by transcriptionally upregulating ß-catenin. Moreover, overexpression of HOXC10 could decrease the sensitivity of cells to carboplatin, while knocking down HOXC10 had the opposite effect both in vitro and in vivo. Therefore, the expression of HOXC10/ABCC3 could be a novel biomarker for predicting the carboplatin resistance and the prognosis of OC patients.

Amylin regulates testosterone levels via steroidogenesis-related enzymes in the central nervous system of male mice.

Amylin is a peripheral satiation signal polypeptide co-secreted with insulin by pancreatic ß-cells in response to nutrient ingestion. Amylin participates in the eating-inhibitory effect and regulates energy metabolism by acting on the central nervous system (CNS). However, the role of amylin in regulating the biosynthesis of steroid hormones, such as testosterone, through the hypothalamic-pituitary-gonadal axis (HPG) remains unexplored. However, only limited evidence is available on the involvement of amylin in steroid synthesis, we hypothesize that amylin regulates testosterone levels via steroidogenesis-related enzymes in the CNS. In this study, we elucidated the effect of intraperitoneal injection of amylin on the protein expression of steroidogenesis-related enzymes, including 3ß-hydroxysteroid dehydrogenase (3ß-HSD), cytochrome P450 17A1 (CYP17A1), and steroidogenic acute regulatory protein (StAR), and phospho-extracellular signal-regulated kinase (pERK). Additionally, the effect of amylin on testosterone levels in male mice was examined. Our results suggested that 3ß-HSD and CYP17A1 neurons were widely expressed in the CNS of male mice, whereas StAR neurons were mainly expressed in the zona incerta (ZI) and locus coeruleus (LC) regions. Intraperitoneal injection of amylin significantly reduced (p < 0.01) the expression of 3ß-HSD, CYP17A1, and StAR in ZI and other areas near the third ventricle (3 V) but increased (p < 0.01) pERK expression, brain testosterone levels, serum FSH, serum LH, and decreased (p < 0.01) serum testosterone levels in mice. In conclusion, amylin regulates testosterone levels via steroidogenesis-related enzymes in the central nervous system of male mice.

Characterization of Histone Deacetylase Mechanisms in Cancer Development.

Over decades of studies, accumulating evidence has suggested that epigenetic dysregulation is a hallmark of tumours. Post-translational modifications of histones are involved in tumour pathogenesis and development mainly by influencing a broad range of physiological processes. Histone deacetylases (HDACs) and histone acetyltransferases (HATs) are pivotal epigenetic modulators that regulate dynamic processes in the acetylation of histones at lysine residues, thereby influencing transcription of oncogenes and tumour suppressor genes. Moreover, HDACs mediate the deacetylation process of many nonhistone proteins and thus orchestrate a host of pathological processes, such as tumour pathogenesis. In this review, we elucidate the functions of HDACs in cancer.

Conformal Geometry and Multimaterial Additive Manufacturing through Freeform Transformation of Building Layers.

3D printing, formally known as additive manufacturing, creates complex geometries via layer-by-layer addition of materials. While 3D printing has been historically perceived as the static addition of build layers, 3D printing is now considered as a dynamic assembly process. In this context, here a new 3D printing process is reported that executes full degree-of-freedom (DOF) transformation (translating, rotating, and scaling) of each individual building layer while utilizing continuous fabrication techniques. Transforming individual building layers within the sequential layered manufacturing process enables dynamic transformation of the 3D printed parts on-the-fly, eliminating the time-consuming redesign steps. Preserving the locality of the transformation to each layer further enables the discrete conformal transformation, allowing objects such as vascular scaffolds to be optimally fabricated to properly fit within specific patient anatomy obtained from the magnetic resonance imaging (MRI) measurements. Finally, exploiting the freedom to control the orientation of each individual building layer, multimaterials, multiaxis 3D printing capability are further established for integrating functional modules made of dissimilar materials in 3D printed devices. This final capability is demonstrated through 3D printing a soft pneumatic gripper via heterogenous integration of rigid base and soft actuating limbs.

Neuroanatomy of melanocortin-4 receptor pathway in the mouse brain.

OBJECTIVE: Melanocortin-4 receptors (MC4Rs) are key regulators of energy homeostasis and adipose deposition in the central nervous system. Considering that MC4R expression regions and function-related research mainly focus on the paraventricular nucleus (PVN), little is known about their distribution throughout the mouse brain, although its messenger RNA distribution has been analyzed in the rat. Therefore, MC4R protein localization in mouse neurons was the focus of this study. METHODS: MC4R protein distribution was assessed in mice through immunofluorescence and Western blotting. RESULTS: MC4R was differentially expressed throughout the arcuate nucleus (ARC), nucleus of the solitary tract (NTS), raphe pallidus (RPa), medial cerebellar nucleus, intermediolateral nucleus, and brainstem. The highest MC4R protein levels were found in the ARC and ventromedial hypothalamic nucleus, while they were significantly lower in the parabrachial nucleus and NTS. The lowest MC4R protein levels were found in the PVN; there was no difference in the protein levels between the area postrema and RPa. CONCLUSIONS: These data provide a basic characterization of MC4R-expressing neurons and protein distribution in the mouse brain and may aid further research on its role in energy homeostasis.

The Relationship between Different Types of Alarm Sounds and Children's Perceived Risk Based on Their Physiological Responses.

Due to differences in cognitive ability and physiological development, the evacuation characteristics of children are different from those of adults. This study proposes a novel method of using wearable sensors to collect data (e.g., electrodermal activity, EDA; heart rate variability, HRV) on children's physiological responses, and to continuously and quantitatively evaluate the effects of different types of alarm sounds during the evacuation of children. In order to determine the optimum alarm for children, an on-site experiment was conducted in a kindergarten to collect physiological data for responses to different types of alarm sounds during the evacuation of 42 children of different ages. The results showed that: (1) The alarm sounds led to changes in physiological indicators of children aged 3-6 years, and the effects of different types of alarm sounds on EDA and HRV activities were significantly different (p < 0.05). Skin conductance (SC), skin conductance tonic (SCT) and skin conductance level (SCL) can be used as the main indicators for analysing EDA of children in this experiment (p < 0.05), and the indicators of ultralow frequency (ULF) and very low frequency (VLF) for HRV were not affected by the type of alarm sounds (p > 0.05). (2) Unlike adults, kindergarten children were more susceptible to the warning siren. The combined voice and warning alarm had optimal effects in stimulating children to perceive risk. (3) For children aged 3-6 years, gender had a significant impact on children's reception to evacuation sound signals (p < 0.05): Girls are more sensitive than boys in receiving evacuation sound signals, similar to findings of studies of risk perception of adult males and females. In addition, the higher the age, the greater the sensitivity to evacuation sound signals, which accords with results of previous studies on the evacuation dynamics of children.

Disposable ultrasound-sensing chronic cranial window by soft nanoimprinting lithography.

Chronic cranial window (CCW) is an essential tool in enabling longitudinal imaging and manipulation of various brain activities in live animals. However, an active CCW capable of sensing the concealed in vivo environment while simultaneously providing longitudinal optical access to the brain is not currently available. Here we report a disposable ultrasound-sensing CCW (usCCW) featuring an integrated transparent nanophotonic ultrasonic detector fabricated using soft nanoimprint lithography process. We optimize the sensor design and the associated fabrication process to significantly improve detection sensitivity and reliability, which are critical for the intend longitudinal in vivo investigations. Surgically implanting the usCCW on the skull creates a self-contained environment, maintaining optical access while eliminating the need for external ultrasound coupling medium for photoacoustic imaging. Using this usCCW, we demonstrate photoacoustic microscopy of cortical vascular network in live mice over 28 days. This work establishes the foundation for integrating photoacoustic imaging with modern brain research.

Effects of maternal undernutrition on the growth, development and antioxidant status of ovine placentome subtypes during late pregnancy.

This study investigated the effects of maternal undernutrition on the growth, development and antioxidant status of ovine placentome subtypes during late pregnancy. Eighteen time-mated Mongolian ewes with singleton fetuses were allocated to three groups at d 90 of pregnancy: Restricted Group 1 (RG1, 0.18 MJ ME · kg BW-0.75 · d-1, n = 6), Restricted Group 2 (RG2, 0.33 MJ ME · kg BW-0.75 · d-1, n = 6) and Control Group (CG, ad libitum, 0.67 MJ ME · kg BW -0.75 · d -1, n = 6). All animals were supplied chopped hay during the restriction period. At 140 days of pregnancy, the gravid uterus was removed; then individual fetuses and placentae were weighed. The numbers and weight of the placentome subtypes were examined. The number of vessels of the placentome subtypes were observed in sections stained with hematoxylin-eosin. The indexes of antioxidant capacity and oxidative stress, including total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity and mal-ondialdehyde (MDA) concentration, were measured by spectrometric kits. The fetal weights, placental weights and the numbers of Type A placentomes in both restricted groups were reduced (P < .05). Decreased fetal/placental weight ratio and increased total weight of the Type A placentome were found in the RG1 group compared to CG (P < .05). The total weights and the numbers of the Type B, Type C and Type D increased with severe maternal undernutrition in RG1 compared to CG (P < .05), as well as the increased numbers of Type B in the RG2 group compared to the CG group (P < .05). The placentome subtypes in RG1, including Type A, Type B and Type C, exhibited decreased concentrations of T-AOC and SOD activities, but higher MDA concentration and GSH-Px activity than those in CG (P < .05). For the RG2 group, the decreased T-AOC and SOD and increased GSH-Px were found in Type A placentome compared to CG (P < .05). For the same subtype placentome, there were no differences in the number of vessels among CG, RG2 and RG1 (P > .05); however, the number of vessels and MDA concentration of Type C and Type D placentome were higher than those of the Type A and Type B (P < .05). These results indicate that the growth and development of ovine Type A placentome was altered during late pregnancy. More and more Type B, Type C and Type D were formed from the Type A associated with maternal undernutrition. Oxidative stress occurred in all types of placentomes for the RG1 group, and the more flattening placentomes that were converted, the worse the antioxidant/pro-oxidant imbalance that was induced.