Search

Abstract Enhancing thermogenic brown adipose tissue (BAT) function is a promising therapeutic strategy for metabolic disease. However, predominantly thermoneutral modern human living conditions deactivate BAT. We demonstrate that selective adipocyte deficiency of the oxygen-sensor HIF-prolyl hydroxylase (PHD2) gene overcomes BAT dormancy at thermoneutrality. Adipocyte-PHD2-deficient mice maintain higher energy expenditure having greater BAT thermogenic capacity. In human and murine adipocytes, a PHD inhibitor increases Ucp1 levels. In murine brown adipocytes, antagonising the major PHD2 target, hypoxia-inducible factor-(HIF)−2a abolishes Ucp1 that cannot be rescued by PHD inhibition. Mechanistically, PHD2 deficiency leads to HIF2 stabilisation and binding of HIF2 to the Ucp1 promoter, thus enhancing its expression in brown adipocytes. Serum proteomics analysis of 5457 participants in the deeply phenotyped Age, Gene and Environment Study reveal that serum PHD2 associates with increased risk of metabolic disease. Here we show that adipose-PHD2-inhibition is a therapeutic strategy for metabolic disease and identify serum PHD2 as a disease biomarker.

The activation of brown adipose tissue by cold exposure in paternal mice has been reported to improve the metabolic health of offspring. Notably, dietary components for activating BAT have drawn considerable attention owing to their high feasibility for human trials. Therefore, here the potential metabolic improving effects of dietary capsaicin intervention on diet-induced obese paternal mice and their offspring were measured. The results support that paternal preconceptional capsaicin intervention not only elicited the anticipated enhancement of brown fat activity in obese paternal mice, but also improved the metabolic profiles in male offspring evidenced by their elevated resistance to high-fat diet challenge. Moreover, changes in sperm DNA methylation and small non-coding RNA profile were noted in paternal mice with capsaicin intervention, though additional investigation is needed to further delve into the mechanisms. In summary, paternal dietary intervention activating BAT could represent a promising strategy for enhancing the metabolic health of offspring.

Abstract Autophagy is a complex self‐degrading process, and its imbalance adversely affects local (metabolically active organs) or systemic metabolism, resulting in metabolic dysfunction correlated with obesity, fatty liver, and type 2 diabetes. Polyphenols are plant secondary metabolites and their ingestion may offer beneficial effects on biological processes, including cell proliferation, cell cycle, apoptosis and autophagy. In this review, the effects of plant‐based polyphenols on autophagy and the underlying mechanisms in the regulation of glucolipid metabolic diseases (GLMDs) are examined in the context of potential health benefits. The available evidence, mainly from animal models, strongly supports that polyphenols improve the metabolic functions of different organs by acting on various molecular targets related to autophagic flux. Multiple autophagic pathways, including cAMP, AMPK, MAPK, AKT, SIRT1, PI3K, Nrf2/HO‐1, PINK1/Parkin, PPARδ, and miRNAs have been implicated in the improvement of GLMDs by polyphenols. Moreover, the potential utilization of polyphenol‐meditated autophagy in the food industry was discussed. The current review provides a comprehensive understanding of the prospective role and mechanisms of polyphenol‐regulated autophagy in GLMDs and opens up future insights into its application to the healthcare and food industries.

Abstract Background Dysfunctional adipose tissue (AT) is known to contribute to the pathophysiology of metabolic disease, including type 2 diabetes mellitus (T2DM). This dysfunction may occur, in part, as a consequence of gut-derived endotoxaemia inducing changes in adipocyte mitochondrial function and reducing the proportion of BRITE (brown-in-white) adipocytes. Therefore, the present study investigated whether endotoxin (lipopolysaccharide; LPS) directly contributes to impaired human adipocyte mitochondrial function and browning in human adipocytes, and the relevant impact of obesity status pre and post bariatric surgery. Methods Human differentiated abdominal subcutaneous (AbdSc) adipocytes from participants with obesity and normal-weight participants were treated with endotoxin to assess in vitro changes in mitochondrial function and BRITE phenotype. Ex vivo human AbdSc AT from different groups of participants (normal-weight, obesity, pre- and 6 months post-bariatric surgery) were assessed for similar analyses including circulating endotoxin levels. Results Ex vivo AT analysis (lean & obese, weight loss post-bariatric surgery) identified that systemic endotoxin negatively correlated with BAT gene expression (p

Prior to its implantation into the uterus, the mammalian embryo stores substantial lipids. Here the authors unveil how lipid storage and morphogenesis of pluripotent stem cells are fundamentally connected during peri-implantation development.

Methods of isolating mitochondria commonly utilise mechanical force and shear stress to homogenize tissue followed by purification by multiple rounds of ultracentrifugation. Existing protocols can be time-consuming with some physically impairing integrity of the sensitive mitochondrial double membrane. Here, we describe a method for the recovery of intact, respiring mitochondria from murine skeletal muscle tissue and cell lines using nitrogen cavitation. This protocol results in high-yield, pure and respiring mitochondria without the need for purification gradients or ultracentrifugation. The protocol takes under an hour and requires limited specialised equipment. Our methodology is successful in extracting mitochondria of both cell extracts and skeletal muscle tissue. This represents an improved yield in comparison to many of the existing methods. Western blotting and electron microscopy demonstrate the enrichment of mitochondria with their ultrastructure well-preserved and an absence of contamination from cytoplasmic or nuclear fractions. Using respirometry analysis we show that mitochondria extracted from murine skeletal muscle cell lines (C2C12) and tibialis anterior tissue have an appropriate respiratory control ratio. These measures are indicative of healthy coupled mitochondria. Our method successfully demonstrates the rapid isolation of functional mitochondria and will benefit researchers studying mitochondrial bioenergetics as well as providing greater throughput and application for time-sensitive assays.

Abstract Background Plant glycine-rich proteins are categorized into several classes based on their protein structures. The glycine-rich RNA binding proteins (GRPs) are members of class IV subfamily possessing N-terminus RNA-recognition motifs (RRMs) and proposed to be involved in post-transcriptional regulation of its target transcripts. GRPs are involved in developmental process and cellular stress responses, but the molecular mechanisms underlying these regulations are still elusive. Results Here, we report the functional characterization of rice GLYCINE-RICH PROTEIN 3 (OsGRP3) and its physiological roles in drought stress response. Both drought stress and ABA induce the expression of OsGRP3. Transgenic plants overexpressing OsGRP3 (OsGRP3 OE ) exhibited tolerance while knock-down plants (OsGRP3 KD ) were susceptible to drought compared to the non-transgenic control. In vivo, subcellular localization analysis revealed that OsGRP3-GFP was transported from cytoplasm/nucleus into cytoplasmic foci following exposure to ABA and mannitol treatments. Comparative transcriptomic analysis between OsGRP3 OE and OsGRP3 KD plants suggests that OsGRP3 is involved in the regulation of the ROS related genes. RNA-immunoprecipitation analysis revealed the associations of OsGRP3 with PATHOGENESIS RELATED GENE 5 (PR5), METALLOTHIONEIN 1d (MT1d), 4,5-DOPA-DIOXYGENASE (DOPA), and LIPOXYGENASE (LOX) transcripts. The half-life analysis showed that PR5 transcripts decayed slower in OsGRP3 OE but faster in OsGRP3 KD , while MT1d and LOX transcripts decayed faster in OsGRP3 OE but slower in OsGRP3 KD plants. H2O2 accumulation was reduced in OsGRP3 OE and increased in OsGRP3 KD plants compared to non-transgenic plants (NT) under drought stress. Conclusion OsGRP3 plays a positive regulator in rice drought tolerance and modulates the transcript level and mRNA stability of stress-responsive genes, including ROS-related genes. Moreover, OsGRP3 contributes to the reduction of ROS accumulation during drought stress. Our results suggested that OsGRP3 alleviates ROS accumulation by regulating ROS-related genes’ mRNA stability under drought stress, which confers drought tolerance.

Optogenetics is a powerful tool for controlling biological functions using light. Optical fibers have been extensively utilized in optical stimulation devices for optogenetics. However, the use of optical fibers results in a small photo-stimulation region. In this study, micro-LED array devices were developed to achieve large-area photo-stimulation in the brain of a large animal, such as macaque monkeys. Planar and linear micro-LED array devices were designed and fabricated to photo-stimulate the prefrontal cortex (PFC) and ventral tegmental area (VTA) of the brain and induce a neurochemical response. Device operation, optical intensity, and safety were first characterized using rats. Subsequently, the devices were used to photo-stimulate the brain of macaque monkeys. In addition, microdialysis in the PFC was performed. The devices detected modulated levels of dopamine in the brains. Thus, the photo-stimulation of both the PFC and VTA were successfully achieved, and the effectiveness of the developed micro-LED array devices was demonstrated. The study will help facilitate further studies on micro-LED array stimulation for system-wide optogenetic manipulation in large animals.

In this study, we propose a complementary-metal-oxide-semiconductor (CMOS) image sensor with a self-resetting system demonstrating a high signal-to-noise ratio (SNR) to detect small intrinsic signals such as a hemodynamic reaction or neural activity in a mouse brain. The photodiode structure was modified from N-well/P-sub to P+/N-well/P-sub to increase the photodiode capacitance to reduce the number of self-resets required to decrease the unstable stage. Moreover, our new relay board was used for the first time. As a result, an effective SNR of over 70 dB was achieved within the same pixel size and fill factor. The unstable state was drastically reduced. Thus, we will be able to detect neural activity. With its compact size, this device has significant potential to become an intrinsic signal detector in freely moving animals. We also demonstrated in vivo imaging with image processing by removing additional noise from the self-reset operation.

Obesity is characterized by a state of chronic inflammation in adipose tissue mediated by the secretion of a range of inflammatory cytokines. In comparison to WAT, relatively little is known about the inflammatory status of brown adipose tissue (BAT) in physiology and pathophysiology. Because BAT and brown/beige adipocytes are specialized in energy expenditure they have protective roles against obesity and associated metabolic diseases. BAT appears to be is less susceptible to developing inflammation than WAT. However, there is increasing evidence that inflammation directly alters the thermogenic activity of brown fat by impairing its capacity for energy expenditure and glucose uptake. The inflammatory microenvironment can be affected by cytokines secreted by immune cells as well as by the brown adipocytes themselves. Therefore, pro-inflammatory signals represent an important component of the thermogenic potential of brown and beige adipocytes and may contribute their dysfunction in obesity.

Objective: Imprinted genes are crucial for the growth and development of fetal and juvenile mammals. Altered imprinted gene dosage causes a variety of human disorders, with growth and development during these crucial early stages strongly linked with future metabolic health in adulthood. Neuronatin (Nnat) is a paternally expressed imprinted gene found in neuroendocrine systems and white adipose tissue and is regulated by the diet and leptin. Neuronatin expression is downregulated in obese children and has been associated with stochastic obesity in C57BL/6 mice. However, our recent studies of Nnat null mice on this genetic background failed to display any body weight or feeding phenotypes but revealed a defect in glucose-stimulated insulin secretion due to the ability of neuronatin to potentiate signal peptidase cleavage of preproinsulin. Nnat deficiency in beta cells therefore caused a lack of appropriate storage and secretion of mature insulin. Methods: To further explore the potential role of Nnat in the regulation of body weight and adiposity, we studied classical imprinting-related phenotypes such as placental, fetal, and postnatal growth trajectory patterns that may impact upon subsequent adult metabolic phenotypes. Results: Here we find that, in contrast to the lack of any body weight or feeding phenotypes on the C57BL/6J background, deletion of Nnat in mice on 129S2/Sv background causes a postnatal growth restriction with reduced adipose tissue accumulation, followed by catch up growth after weaning. This was in the absence of any effect on fetal growth or placental development. In adult 129S2/Sv mice, Nnat deletion was associated with hyperphagia, reduced energy expenditure, and partial leptin resistance. Lack of neuronatin also potentiated obesity caused by either aging or high fat diet feeding. Conclusions: The imprinted gene Nnat plays a key role in postnatal growth, adult energy homeostasis, and the pathogenesis of obesity via catch up growth effects, but this role is dependent upon genetic background. Keywords: Obesity, Postnatal growth, Imprinted genes, Neuronatin, Genetic background, Energy homeostasis

Abstract Background To compare femtosecond laser-assisted versus conventional phacoemulsification in terms of visual and refractive outcomes, cumulative dissipated energy, anterior chamber inflammation and endothelial cell loss. Methods In this retrospective cohort study, records of eyes that underwent femtosecond laser-assisted cataract surgery (FLACS) or conventional phacoemulsification (CP) were reviewed. The Victus femtosecond laser (Bausch and Lomb, Germany) was used to carry out corneal incisions, anterior capsulotomy, and lens fragmentation in FLACS procedures. Manifest refraction spherical equivalence (MRSE), uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), cumulative dissipated energy (CDE), postoperative cells and flare and endothelial cell count data were collected. Subgroup analysis of the visual acuity tests was performed based on the type of intraocular lens implanted (monofocal, monofocal toric, multifocal, multifocal toric, accommodating). Results A total of 735 eyes were included in the study (296 eyes for the FLACS group and 439 eyes for the CP group). At one year follow-up, 120 eyes comprised the FLACS group and 265 eyes for the CP group. MRSE in the FLACS group was − 0.16 ± 0.58 D and − 0.20 ± 0.52 D in the CP group (P = 0.50). UDVA in the FLACS group was 20/25 (mean logMAR 0.12 ± 0.13) and 20/25 (mean logMAR 0.11 ± 0.13) in the CP group (P = 0.48). CDVA was 20/20 (mean logMAR 0.03 ± 0.07) in the FLACS group and 20/20 (mean logMAR 0.02 ± 0.06) in the CP group (P = 0.15). No statistically significant trend was seen for FLACS versus CP by intraocular type for visual acuity. CDE for the different cataract grades ranged from 6.97 ± 5.74 to 29.02 ± 16.07 in the FLACS group and 7.59 ± 6.42 to 35.69 ± 18.30 in the CP group. The FLACS group was significantly lower for post-operative central corneal edema (P = 0.05), cells and flare (P = 0.01), and endothelial cell loss (P = 0.04). Conclusions Femtosecond laser-assisted cataract surgery and conventional phacoemulsification had similar refractive and visual outcomes. Phacoemulsification energy, anterior chamber inflammation and corneal endothelial cell loss were less in the femtosecond laser group.

Abstract Brown adipose tissue (BAT) activation stimulates energy expenditure in human adults, which makes it an attractive target to combat obesity and related disorders. Recent studies demonstrated a role for G protein‐coupled receptor 120 (GPR120) in BAT thermogenesis. Here, we investigated the therapeutic potential of GPR120 agonism and addressed GPR120‐mediated signaling in BAT. We found that activation of GPR120 by the selective agonist TUG‐891 acutely increases fat oxidation and reduces body weight and fat mass in C57Bl/6J mice. These effects coincided with decreased brown adipocyte lipid content and increased nutrient uptake by BAT, confirming increased BAT activity. Consistent with these observations, GPR120 deficiency reduced expression of genes involved in nutrient handling in BAT. Stimulation of brown adipocytes in vitro with TUG‐891 acutely induced O2 consumption, through GPR120‐dependent and GPR120‐independent mechanisms. TUG‐891 not only stimulated GPR120 signaling resulting in intracellular calcium release, mitochondrial depolarization, and mitochondrial fission, but also activated UCP1. Collectively, these data suggest that activation of brown adipocytes with the GPR120 agonist TUG‐891 is a promising strategy to increase lipid combustion and reduce obesity.

Brown adipose tissue uptake of glucose and fatty acids is very high during nonshivering thermogenesis. Adrenergic stimulation markedly increases glucose uptake, de novo lipogenesis, and FA oxidation simultaneously. The mechanism that enables this concerted response has hitherto been unknown. Here, we find that in primary brown adipocytes and brown adipocyte-derived cell line (IMBAT-1), acute inhibition and longer-term knockdown of DGAT2 links the increased de novo synthesis of fatty acids from glucose to a pool of TAG that is simultaneously hydrolyzed, providing FA for mitochondrial oxidation. DGAT1 does not contribute to this pathway, but uses exogenous FA and glycerol to synthesize a functionally distinct pool of TAG to which DGAT2 also contributes. The DGAT2-dependent channelling of 14C from glucose into TAG and CO2 was reproduced in β3-agonist-stimulated primary brown adipocytes. Knockdown of DGAT2 in IMBAT-1 affected the mRNA levels of UCP1 and genes important in FA activation and esterification. Therefore, in β3-agonist activated brown adipocytes, DGAT2 specifically enables channelling of de novo synthesized FA into a rapidly mobilized pool of TAG, which is simultaneously hydrolyzed to provide substrates for mitochondrial fatty acid oxidation

Thermal imaging has shown to be a better tool for the quantitative measurement of temperature than single spot infrared thermometers. However, thermographic cameras can encounter errors in acquiring accurate temperature measurements in the presence of other environmental heat sources. Some of these errors arise due to the inability of the thermal camera to detect objects and features in the infrared domain. In this paper, the thermal image is registered as a stereo image from a Kinect system prior to depth-based correction. Experiments demonstrating the error are presented together with the determination of the measurement errors under prior knowledge of the thermographed scene. The proposed correction scheme improves the accuracy of the thermal image through augmentation using the Kinect system.

Background Diets containing fats of different proportions and types have been demonstrated to influence metabolism. These fats differ in long chain fatty acids (LCFAs) or medium chain fatty acids (MCFAs) content. In our laboratory using swimming as the training modality, MCFAs increased endurance attributed to increased activities of oxidative enzymes. How it affects whole-body metabolism remains unexplored. The present study investigated the metabolic, biochemical and genetic adaptations with treadmill running as the training modality. Methods C57BL/6N mice were divided into untrained and trained groups and provided with low-fat (10% kcal from soybean oil), coconut oil (10% kcal from soybean oil, 20% kcal from coconut oil) or soybean oil (30% kcal from soybean oil) diet. Training was performed on a treadmill for 30 days. After recovery, whole-body metabolism at rest and during exercise, endurance, substrate metabolism, mitochondrial enzyme activities, and gene expression of training-adaptive genes in the muscle and liver were measured. Results At rest, medium-fat diets decreased respiratory exchange ratio (RER) (p

Objective: To evaluate the visual and refractive outcomes of LASIK, PRK, phakic IOL, and Supracor as treatment for errors of refraction, including presbyopia, performed at a private eye center. Method: This is a retrospective, single-center, single-surgeon study that reviewed the surgical outcomes of patients who underwent LASIK, PRK, phakic IOL, and Supracor from January 2010 to December 2014. Main outcome measures were postoperative uncorrected and corrected distance, intermediate, and near visual acuity (for Supracor), and mean manifest-refraction spherical equivalent (MRSE) of patients who had at least 1 month follow up. Complications and enhancements were analyzed independently. Results: Data were analyzed from 1,366 eyes of 771 patients. LASIK was the most commonly performed procedure (68%), followed by PRK (18.3%), Supracor (10.2%), and phakic IOL (3.7%). The postoperative mean MRSE at 1 month for LASIK, PRK, and phakic IOL were -0.08 ± 0.36, +0.06 ± 0.52 and -0.11 ± 0.44, respectively. The mean postoperative logMAR uncorrected distance visual acuity (UDVA) at 1 month for LASIK, PRK, and phakic IOL were 0.02 ± 0.10, 0.07 ± 0.12, 0.001 ± 0.09, respectively. In the Supracor group, the mean preoperative and postoperative spherical equivalent were +1.12 ± 0.8 and -0.76 ± 0.62, respectively. The mean postoperative logMAR UDVA and uncorrected near visual acuity (UNVA) for Supracor were 0.24 ± 0.19 and 0.02 ± 0.08, respectively. The most common postoperative complication was symptomatic dry eye (13%). Regression and off-target outcomes occurred in 24 (1.8%) and 13 eyes (1%), respectively. Overall enhancement rate was 2%. Conclusion: LASIK remained to be the most common refractive procedure, with femtosecond laser-created flap becoming the norm. Refractive and visual outcomes showed good efficacy with all the refractive procedures. Complication and enhancement rates were low.

Objective: To describe the demographic and clinical profile of patients who underwent refractive surgery screening. Method: Medical records of patients who sought consult for refractive surgery from January 2010 to December 2014 at a refractive center were reviewed and analyzed. The preoperative clinical conditions, optical characteristics of myopes and hyperopes, refractive screening tests, and causes for disqualification were determined. Results: A total of 1215 patients who sought consult for refractive surgery had a mean age of 36.45 ± 11.60 years. Seven hundred ten (58.44%) were females and 860 (70.78%) were Filipinos. Nine hundred eighty eight (81%) were myopes. The mean manifest refractive spherical equivalent (MRSE) for myopic patients was -4.41D±2.98 with mean uncorrected distance vision (UCDVA) of 20/400 (logMAR 1.26) and mean best-corrected distance vision (BCDVA) of 20/20 (logMAR 0.02). For hyperopic patients, the mean MRSE was +1.33D±3.76 with mean UCDVA of 20/40 (logMAR 0.33) and mean BCDVA of 20/20 (logMAR 0.001). Reasons for disqualification from undergoing a refractive procedure included thin cornea (5.27%), irregular corneal topography (2.39%), steep cornea (0.78%), high refractive errors (0.41%), optic nerve (0.41%), and retina (0.25%) pathologies. Conclusion: Patients who underwent screening for refractive surgery were young, mostly female, with myopic refractive errors. LASIK remained the most popular refractive surgery procedure.

Abstract Although circulating fatty acids are utilized as energy substrates, they also function as ligands to the peroxisome‐proliferator activated receptors (PPARs), a family of fatty acid sensing transcription factors. Exercise training leads to various adaptations in the muscle such as elevation of glycogen content, mitochondrial number as well as upregulation of fatty acid uptake and utilization through downstream transcriptional adaptations. In line with this, CD36 has been shown to be critical in controlling fatty acid uptake and consequently, fatty acid oxidation. We show that exercise training could not ameliorate impaired endurance performance in CD36 KO mice despite intact adaptations in muscle glycogen storage and mitochondrial function. Changes in whole‐body metabolism at rest and during exercise were also suppressed in these animals. Furthermore, there was inefficient upregulation of PPAR and PPAR‐related exercise‐responsive genes with chronic training in CD36 KO mice despite normal upregulation of Pgc1a and mitochondrial genes. Our findings supplement previous observations and emphasize the importance of CD36 in endurance performance, energy production and efficient downstream transcriptional regulation by PPARs.

Arachidonic acid (AA) is a major PUFA that has been implicated in the regulation of adipogenesis. We examined the effect of a short exposure to AA at different stages of 3T3-L1 adipocyte differentiation. AA caused the upregulation of fatty acid binding protein 4 (FABP4/aP2) following 24 h of differentiation. This was mediated by the prostaglandin F2α (PGF2α), as inhibition of cyclooxygenases or PGF2α receptor signaling counteracted the AA-mediated aP2 induction. In addition, calcium, protein kinase C, and ERK are all key elements of the pathway through which AA induces the expression of aP2. We also show that treatment with AA during the first 24 h of differentiation upregulates the expression of the transcription factor Fos-related antigen 1 (Fra-1) via the same pathway. Finally, treatment with AA for 24 h at the beginning of the adipocyte differentiation is sufficient to inhibit the late stages of adipogenesis through a Fra-1-dependent pathway, as Fra-1 knockdown rescued adipogenesis. Our data show that AA is able to program the differentiation potential of preadipocytes by regulating gene expression at the early stages of adipogenesis.

The study aimed to assess the students' extent of knowledge and identify their perspectives towards biodiversity and its protection and conservation. A total of 268 randomly selected students at Aurora State College of Technology were involved in the study. Survey questionnaires were used to obtain data and information which were subjected to statistical tests. The students had a moderate knowledge level on biodiversity with a mean score of 6.65 out of 10 items (SD = 1.50). Their perspective on biodiversity was leaning toward its protection and conservation, with a mean score of 7.2 out of 10 items (SD = 1.29). Factors affecting the students' knowledge were gender (p =0 .003) and academic department (p = 0.003). Females and those associated with the Department of Forestry and Environmental Sciences and Department of Industrial technology were found to have more knowledge than the others. Males, on the other hand, were found to have more positive perspectives towards biodiversity. Knowledge and perspective had a weak correlation with r = 0.39. Students were not well-aware, but were in support of the Philippines' biodiversity-related laws, which could help shape their mindset and actions towards biodiversity conservation and protection. Thus, the college administration must revisit the curricula of all degree programs and ensure that students from each degree program are environmentally educated, emphasizing biodiversity conservation.

The accurate diagnosis and clinical management of the growth restriction disorder Silver Russell Syndrome (SRS) has confounded researchers and clinicians for many years due to the myriad of genetic and epigenetic alterations reported in these patients and the lack of suitable animal models to test the contribution of specific gene alterations. Some genetic alterations suggest a role for increased dosage of the imprinted CYCLIN DEPENDENT KINASE INHIBITOR 1C (CDKN1C) gene, often mutated in IMAGe Syndrome and Beckwith-Wiedemann Syndrome (BWS). Cdkn1c encodes a potent negative regulator of fetal growth that also regulates placental development, consistent with a proposed role for CDKN1C in these complex childhood growth disorders. Here, we report that a mouse modelling the rare microduplications present in some SRS patients exhibited phenotypes including low birth weight with relative head sparing, neonatal hypoglycemia, absence of catch-up growth and significantly reduced adiposity as adults, all defining features of SRS. Further investigation revealed the presence of substantially more brown adipose tissue in very young mice, of both the classical or canonical type exemplified by interscapular-type brown fat depot in mice (iBAT) and a second type of non-classic BAT that develops postnatally within white adipose tissue (WAT), genetically attributable to a double dose of Cdkn1c in vivo and ex-vivo. Conversely, loss-of-function of Cdkn1c resulted in the complete developmental failure of the brown adipocyte lineage with a loss of markers of both brown adipose fate and function. We further show that Cdkn1c is required for post-transcriptional accumulation of the brown fat determinant PR domain containing 16 (PRDM16) and that CDKN1C and PRDM16 co-localise to the nucleus of rare label-retaining cell within iBAT. This study reveals a key requirement for Cdkn1c in the early development of the brown adipose lineages. Importantly, active BAT consumes high amounts of energy to generate body heat, providing a valid explanation for the persistence of thinness in our model and supporting a major role for elevated CDKN1C in SRS.

Maintenance of energy homeostasis depends on the highly regulated storage and release of triacylglycerol primarily in adipose tissue, and excessive storage is a feature of common metabolic disorders. CIDEA is a lipid droplet (LD)-protein enriched in brown adipocytes promoting the enlargement of LDs, which are dynamic, ubiquitous organelles specialized for storing neutral lipids. We demonstrate an essential role in this process for an amphipathic helix in CIDEA, which facilitates embedding in the LD phospholipid monolayer and binds phosphatidic acid (PA). LD pairs are docked by CIDEA trans-complexes through contributions of the N-terminal domain and a C-terminal dimerization region. These complexes, enriched at the LD–LD contact site, interact with the cone-shaped phospholipid PA and likely increase phospholipid barrier permeability, promoting LD fusion by transference of lipids. This physiological process is essential in adipocyte differentiation as well as serving to facilitate the tight coupling of lipolysis and lipogenesis in activated brown fat.

Spray drying is one of the common methods of preserving eggs. A study had shown that addition of 20% tapioca starch to liquid egg prior to drying increases the shelf-life of the powdered product. However, although the said factor has been identified, production in a larger capacity is still a problem. One way to solve this predicament is to identify the optimum drying parameter that would produce a higher yield of dried eggs but retain the quality of the desired product such as the moisture content and water activity. The key issue in optimizing the dryer operation is flow stability wherein the airflow patterns, such as velocity, temperature, and relative humidity, existing inside the drying chamber are considered as the primary factor that influences the histories of the droplets produced. In this article, the author used ANSYS-Fluent version 14.5, a computational fluid dynamics software, to analyze the flow pattern of the continuous phase and to track the particle histories of the disperse phase inside the short-form drying chamber that produces powdered egg with 20% tapioca starch additive. Simulations for the short-form dryer showed that the optimized inlet air temperature and outlet particle temperature were 438 and 385 K, respectively. Based on the results, the calculated moisture content and water activity are 4.55% and 0.02 a w , respectively. Note that the critical moisture content and water activity for powdered eggs are 5% and 0.4 a w , respectively. The values beyond these limits will cause abrupt spoilage of the dried egg.

Here, we describe a robust method for mapping the number and type of neuro-chemically distinct synaptic inputs that a single reconstructed neuron receives. We have used individual hypoglossal motor neurons filled with Neurobiotin by semi-loose seal electroporation in thick brainstem slices. These filled motor neurons were then processed for excitatory and inhibitory synaptic inputs, using immunohistochemical-labeling procedures. For excitatory synapses, we used anti-VGLUT2 to locate glutamatergic pre-synaptic terminals and anti-PSD-95 to locate post-synaptic specializations on and within the surface of these filled motor neurons. For inhibitory synapses, we used anti-VGAT to locate GABAergic pre-synaptic terminals and anti-GABA-A receptor subunit α1 to locate the post-synaptic domain. The Neurobiotin-filled and immuno-labeled motor neuron was then processed for optical sectioning using confocal microscopy. The morphology of the motor neuron including its dendritic tree and the distribution of excitatory and inhibitory synapses were then determined by three-dimensional reconstruction using IMARIS software (Bitplane). Using surface rendering, fluorescence thresholding, and masking of unwanted immuno-labeling, tools found in IMARIS, we were able to obtain an accurate 3D structure of an individual neuron including the number and location of its glutamatergic and GABAergic synaptic inputs. The power of this method allows for a rapid morphological confirmation of the post-synaptic responses recorded by patch-clamp prior to Neurobiotin filling. Finally, we show that this method can be adapted to super-resolution microscopy techniques, which will enhance its applicability to the study of neural circuits at the level of synapses.

Decidualization renders the endometrium transiently receptive to an implanting blastocyst although the underlying mechanisms remain incompletely understood. Here we show that human endometrial stromal cells (HESCs) rapidly release IL-33, a key regulator of innate immune responses, upon decidualization. In parallel, differentiating HESCs upregulate the IL-33 transmembrane receptor ST2L and other pro-inflammatory mediators before mounting a profound anti-inflammatory response that includes downregulation of ST2L and increased expression of the soluble decoy receptor sST2. We demonstrate that HESCs secrete factors permissive of embryo implantation in mice only during the pro-inflammatory phase of the decidual process. IL-33 knockdown in undifferentiated HESCs was sufficient to abrogate this pro-inflammatory decidual response. Further, sequential activation of the IL-33/ST2L/sST2 axis was disordered in decidualizing HESCs from women with recurrent pregnancy loss. Signals from these cultures prolonged the implantation window but also caused subsequent pregnancy failure in mice. Thus, Il-33/ST2 activation in HESCS drives an autoinflammatory response that controls the temporal expression of receptivity genes. Failure to constrain this response predisposes to miscarriage by allowing out-of-phase implantation in an unsupportive uterine environment.

Skeletal muscle constitutes the major site of glucose uptake leading to increased removal of glucose from the circulation in response to insulin. Type 2 diabetes and obesity are often associated with insulin resistance that can be counteracted by exercise or the use of drugs increasing the relative proportion of oxidative fibers. RIP140 is a transcriptional coregulator with a central role in metabolic tissues and we tested the effect of modulating its level of expression on muscle glucose and lipid metabolism in two mice models. Here, we show that although RIP140 protein is expressed at the same level in both oxidative and glycolytic muscles, it inhibits both fatty acid and glucose utilization in a fiber-type dependent manner. In RIP140-null mice, fatty acid utilization increases in the extensor digitorum longus and this is associated with elevated expression of genes implicated in fatty acid binding and transport. In the RIP140-null soleus, depletion of RIP140 leads to increased GLUT4 trafficking and glucose uptake with no change in Akt activity. AMPK phosphorylation/activity is inhibited in the soleus of RIP140 transgenic mice and increased in RIP140-null soleus. This is associated with increased UCP1 expression and mitochondrial uncoupling revealing the existence of a signaling pathway controlling insulin-independent glucose uptake in the soleus of RIP140-null mice. In conclusion, our findings reinforce the participation of RIP140 in the maintenance of energy homeostasis by acting as an inhibitor of energy production and particularly point to RIP140 as a promising therapeutic target in the treatment of insulin resistance.