bluuebunny/crossref_metadata_2025 huggingface dataset:
Activation of Adenylyl Cyclase in Pulmonary Microvascular Endothelial Cells Increases Microparticle cAMP Content
Authors: Sayner Sarah L , Hargett Leslie , Scruggs April , Ramila KC , Yarbrough Thomas , Bauer Natalie
Published: November 2023

Rationale

Microparticles (MPs) are submicron intact vesicles released from many cell types constitutively or following activation. MPs play a role in cellular homeostasis, injury and act as biomarkers. These submicron vesicles can encapsulate a variety of functional proteins and RNAs/miRNAs; however, little is known about whether they encapsulate signaling molecules, such as cyclic adenosine monophosphate (cAMP). Thus, we investigated the hypothesis that MPs from endothelial cells contain cAMP and that pulmonary microvascular endothelial cells (PMVEC) can be stimulated to increase MP‐cAMP content.

Methods

MPs were collected from the media of unstimulated pulmonary and aortic endothelial cells as well as pulmonary artery smooth muscle cells (PASMCs) by ultracentrifugation. MPs were also collected from (PMVECs) following treatment with agonists to stimulate endogenous cAMP production from the transmembrane adenylyl cyclase (AC) or bicarbonate‐stimulation of the soluble AC (AC10) in the presence or absence of the phosphodiesterase (PDE) 4 inhibitor, rolipram. Cyclic AMP in the MPs as well as cellular lysates was measured by immunoassay or mass spectrometry and normalized to protein concentration.

Results

While cAMP was detected in MPs derived from various endothelial cell types as well as PASMCs, the cAMP content was highest in PMVECs compared to the other cell types. Further, stimulation of PMVECs with agents that increase the near membrane cAMP pool led to an increase in cAMP content in MPs.

Conclusion

Microparticles released from the vasculature contain cAMP while stimulation of transmembrane ACs in PMVECs increases MP‐cAMP content.

Support or Funding Information

This work was supported by the Parker B. Francis Fellowship Foundation Award (SLS) and HL121513 (SLS), AHA 11SDG7390037 (NB) and T32 HL076125 (LH, AS, TY)
Lipopolysaccharide and Isoproterenol Alter Sodium Bicarbonate Cotransporter Phosphorylation in Pulmonary Microvascular Endothelial Cells
Authors: Sayner Sarah L.
Published: November 2023

The hallmark of acute respiratory distress syndrome (ARDS) is damage to alveolar capillaries leading to increased endothelial permeability. Bacterial induced sepsis is a primary cause of pulmonary endothelial barrier dysfunction. Our recent studies reveal that the lipopolysaccharide (LPS)‐induced increase in pulmonary endothelial permeability is dependent upon bicarbonate stimulation of soluble adenylyl cyclase isoform 10 (AC10 a.k.a. sAC). Further, bicarbonate stimulation of AC10 is dependent upon sodium, suggesting a role for Na+‐bicarbonate cotransporters (NBCs). Further, it has been reported that forskolin stimulation of transmembrane ACs and generation of subplasma membrane cAMP inactivates NBCn2 via PKA, potentially reducing the availability of bicarbonate for stimulation of AC10. Thus, we investigated the expression of NBCs in the lung and pulmonary endothelium and the phosphorylation status of NBCn2 in response to barrier disruptive (LPS) or barrier protective (isoproterenol) agents. To examine the phosphorylation status of NBCn2, endothelial cells were treated with either LPS or isoproterenol and rolipram, the phosphodiesterase 4 inhibitor. Immunoprecipitation was performed on whole cell lysate with the NBCn2 or control IgG antibodies and western blot analysis with PKA Phospho‐Ser/Thr antibody. Our results revealed the differential expression of NBCs in the pulmonary circulation. Further, while LPS led to a decrease in the phosphorylation of NBCn2 at a PKA specific site, isoproterenol and rolipram increased PKA‐phosphorylation of NBCn2. Thus, while transmembrane AC activity increases NBCn2 phosphorylation, LPS decreases NBCn2 phosphorylation and could act as a mechanism to regulate the bicarbonate source necessary to stimulate AC10.

Support or Funding Information

This work is supported by the Parker B. Francis Fellowship Foundation Award (SLS) and HL121513 (SLS)
Characterization of Ectonucleotidase Expression in the Rat Carotid Body: Potential Regulation by Hypoxia?
Authors: Salman Shaima , Vollmer Cathy , Nurse Colin A
Published: November 2023

The carotid body (CB) maintains homeostasis in the respiratory and cardiovascular systems of mammals by initiating reflex responses during exposures to blood‐borne stimuli such as low O2 (hypoxia). Thus, stimulation of CB chemoreceptors during hypoxia results in compensatory hyperventilation and activation of the sympathoadrenal system. CB hyperactivity has been linked to several clinical morbidities including sleep apnea, chronic obstructive pulmonary disease, hypertension, obesity, and heart failure. The purine ATP and its derivative, adenosine, contribute significantly to the excitatory CB afferent discharge during hypoxia (Conde et al., 2012, J Appl Physiol 112: 2002–10; Nurse and Piskuric, 2013, Semin Cell Dev Biol 24: 22–30). Given that extracellular nucleotides and products of their hydrolysis by ectonucleotidases are known to play a key role in purinergic signaling in the brain and tissues innervated by the autonomic nervous system, we investigated ectonucleotidase expression in the rat CB. Ectonucleoside triphosphate diphosphohydrolase (E‐NTPDase) is the dominant family of ectonucleotidases and subtypes of this enzyme family catalyze the hydrolysis of purines and regulate their levels in the synaptic cleft. In extracts of whole rat CBs, RT‐PCR and sequence analyses revealed the presence of E‐NTPDase‐1 and ‐2, as well as ecto‐5′‐ectonucleotidase (Nt5e) mRNA. Interestingly, expression of ectonucleotidases, E‐NTPDase‐1 (CD39) and Nt5e (CD73), has been proposed to have a protective role in mouse intestinal epithelia during hypoxia (Synnestvedt et al., 2002, J Clin Invest 110: 993–02). This raised the possibility that ectonucleotidases might be regulated by hypoxia in O2‐sensitive organs such as the CB and its neural crest‐derived counterpart, the adrenal medulla. Indeed, microarray analysis of a rat O2‐sensitive immortalized adrenomedullary chromaffin cell line (MAH cells) suggests that chronic hypoxia exerts a selective hypoxia inducible factor‐2 alpha (HIF2a)‐dependent upregulation of E‐NTPDase‐2 transcript. Future studies will determine the cellular localization of ectonucleotidases in the rat CB and the potential role of HIF2a in the regulation of E‐NTPDase‐2 expression during chronic hypoxia.

Support or Funding Information

Supported by grants from Canadian Institutes of Health Research and Natural Sciences and Engineering Research Council of Canada
Early Postnatal Exposure to Intermittent Hypoxia Results in Significant Alterations in White Matter Integrity in a Rat Pup Model of Apnea of Prematurity
Authors: Darnall Robert , Chen Xi , Sirieix Chrystelle , Xia Luxi , Gimi Barjor
Published: November 2023

Exposure to severe hypoxemia in the human newborn infant results in white matter (WM) injury. However, premature infants are exposed to multiple episodes of less severe intermittent hypoxia (IH) associated with apnea of prematurity (AOP) and immature respiratory control. Often these events are not detected clinically and it is unknown whether IH during this period is associated with WM injury. To answer this question we exposed newborn rat pups to IH from postnatal day (P) 2 to P12 (6 decreases to 5–8% oxygen per hour alternating with an hour of normoxia, 24 hours a day, 7 days a week). A control group was treated identically, except that the periods of hypoxia were replaced with room air. This period is roughly equivalent to a preterm infant from 28 to 40 weeks postmenstrual age. At P20 we performed Diffusion Tensor Imaging (DTI) under isofluorane anesthesia in 11 IH exposed and 9 control pups using a 9.4 T small animal magnetic resonance imaging system, using 6 gradient directions with b=1000 with a slice thickness of 1 mm and an in‐plane resolution of ~160 × 160 μm. Compared to controls, exposure to IH resulted in a decrease in fractional anisotropy (FA) and radial diffusion (RD) and an increase in axial diffusion (AD) in several major WM regions including the corpus callosum (P=0.004) and cingulate (P<0.001) consistent with the loss of white matter integrity. We conclude that in the rodent repeated exposures to even mild hypoxia in the early postnatal period results in white matter injury. Future studies should be done to determine whether similar MRI changes occur in premature infants with apnea of prematurity and if so, whether IH associated with AOP can be treated more effectively.

Support or Funding Information

NIH PO1 HD36379, American SIDS Institute
Chronic Hypoxia Downregulates Colocalization of PKG with its Target BK Channel Proteins in Ovine Cerebral Arteries
Authors: Thorpe Richard Burdette , Williams James M , Pearce William J
Published: November 2023

BACKGROUND

Previous studies have shown that chronic hypoxia ablates the endothelium‐dependent NO‐mediated vasorelaxation pathway of which protein kinase G (PKG) is a key effector. Direct activation of PKG likewise demonstrates attenuated PKG influence on 5HT‐induced vascular tone in chronic hypoxic cerebral arteries compared with normoxic arteries. Furthermore, these effects have been shown to correlate with reduced BK channel activity (potassium currents). Taken together, this suggests that the direct influence of PKG on BK channel function is diminished in conditions of chronic hypoxia.

OBJECTIVE

Given that PKG phosphorylates multiple known targets, this present study explores the hypothesis that the proximal association of PKG with its target BK‐channel proteins is altered by chronic hypoxia, and that this diminished colocalization accounts for a significant loss of function with regards to cerebral vasorelaxation.

METHODS

Pregnant and Non‐Pregnant ewes were held at sea level (normoxic) or at 3820m (hypoxic) for the final 110 days of gestation. Middle cerebral arteries from non‐pregnant adults and term fetal sheep were prepared for organ culture with ±8‐pCPT‐cGMP. Colocalization of PKG with the alpha and beta isoforms of the BK channel proteins, as well as colocalization of BK‐alpha and ‐beta proteins with each other, was determined by confocal microscopy.

RESULTS

Chronic hypoxia reduced the ability of PKG to localize with its target BK channel proteins. Chronic hypoxia also reduced the colocalization of BK‐alpha with BK‐beta channel proteins. Activation of PKG with 8‐p‐CPT‐cGMP in organ culture did not overcome these effects of chronic hypoxia on PKG colocalization.

CONCLUSIONS

PKG‐mediated activation of BK channels accounts for a significant portion of PKG‐induced vasorelaxation in ovine middle cerebral arteries. Chronic hypoxia attenuates the ability of PKG to associate with BK channel proteins, and thereby attenuates the influence of PKG on BK channel activity. This loss of association explains in part how chronic hypoxia ablates the ability of PKG to relax 5HT‐induced vascular tone.

Support or Funding Information

The work reported here was supported by USPHS Grants HL54120, HD31266, HL64867 and the Loma Linda University School of Medicine
Activation of the hypoxia‐inducible factor pathway induced by PHD2 deficiency enhances the effect of running training in mice
Authors: Nunomiya Aki
Published: November 2023

Background

Recently, altitude training has widely been accepted by elite athletes as an efficient way to improve the sea‐level performance. The adaptation to the deprivation of O2, which is called hypoxic response and mediated by hypoxia‐inducible factors (HIFs), seems to contribute to the benefit of altitude training. However, the veracity of this benefit is still controversial, and the effect of biological processes mediated by the HIF pathway on the improvement in exercise performance after exercise training, if any, needs to be demonstrated. The main regulator of HIFs activity is prolyl hydroxylase domain protein 2 (PHD2). Previous reports showed that Phd2 deficiency induced HIFs activation and subsequent hematopoietic and angiogenic adaptations (Minamishima et al., 2008, Takeda et al., 2007). Overall, we believe that the Phd2‐deficient mouse model enables us to elucidate the influence of the activated HIF pathway on the exercise training effect. We therefore hypothesised that the stabilization of HIFs and the subsequent hypoxic response induced by Phd2 deficiency contribute to the improvement in exercise training. To examine this hypothesis, we investigated the maximal exercise capacity of Phd2‐deficient mice before and after a 4‐week regimen of exercise training.

Methods

Because Phd2 knockout is embryonic lethal, estrogen receptor (ER) agonist‐induced Phd2 cKO mouse using Cre‐loxP system was used. Two weeks after tamoxifen, an estrogen‐agonist, maximal running test using a rodent treadmill was performed. Control mice (n = 16) and Phd2 cKO mice (n = 15) were then divided into two groups: trained and non‐trained. The mice in the training group underwent 4‐week treadmill training on a rodent treadmill. After 4‐week period, all mice of both groups were tested again for their maximal running capacity and were sacrificed for blood and tissue examination.

Results

Because of Phd2 deficiency, HIF‐α proteins accumulated. In line with increases in plasma erythropoietin levels (P < 0.01), the Phd2 cKO mice exhibited marked increases in haematocrit values (P < 0.01) and haemoglobin concentrations (P < 0.01). Increased expression of plasma vascular endothelial growth factor (P < 0.05) induced angiogenesis in the skeletal muscle of the Phd2 cKO mice (P < 0.05). Importantly, trained Phd2 cKO mice showed a significantly higher gain in running time by a factor of approximately 1.53 compared with the trained control mice (P < 0.05, Figure). On the other hand, we did not find any difference between the non‐trained control and non‐trained Phd2 cKO mice (P = 0.21). Collectively, these data suggest that the combination of training and the activation of the HIF pathway is important for maximizing the effect of running training.

Conclusions

We concluded that the activation of the HIF pathway induced by PHD2 deficiency enhanced the effect of running training.

Support or Funding Information

This work was supported by a JSPS Grant‐in‐Aid for JSPS fellows (26‐3209).
Neonatal maternal separation increases microglial activation in brainstem regions controlling the laryngeal chemoreflex
Authors: Baldy Cécile , Boisjoly‐Villeneuve Samuel , Fournier Stéphanie , Tremblay Marie‐Ève , Kinkead Richard
Published: November 2023

Laryngeal chemoreflex (LCR) is immature in preterm infants and results in prolonged apneas, O2 desaturations and bradycardias. Previous work from our laboratory has shown that in newborn rat, neonatal maternal separation (NMS) interferes with LCR development and aggravates cardio‐respiratory complications. This effect was more important in males. Microglia are the immune cells of the central nervous system; during early life, they contribute to brain development by playing an important role in synaptic pruning. These cells are activated during stress and their activation is characterized by morphological changes. Indeed, activated microglia are more round (ameboid shaped) with short and few processes like macrophages, whereas inactivated microglia have several ramified processes scanning for abnormal cells and contacting synapses. Because neonatal stress (in the form of NMS) affects LCR development, we hypothesised that NMS increases microglial activation in regions which are involved in the LCR, particularly the nucleus tractus solitarius (NTS) and dorsal motor nucleus of the vagus nerve (NX). Rat pups were separated from their mother 3 hours each day during 10 days from post‐natal day 3 to 12 (P3 to P12). Control animals were left undisturbed. At post‐natal days 14–15, brains were harvested for Iba‐1 immunostaining. Microglia were counted in the NTS and NX by light microscopy. Cell morphology was assessed in each microglia by measuring morphological index (ratio between cell body and arborisation areas), cell density and scattering (nearest neighbour distance) in NTS and NX. Stress increase significantly morphological index in both region of interest and in males and females. NMS increases microglia density in males in NTS and NX. Stress increases also microglial scattering in the N10 but this effect is significant only on P15 males and females. We conclude that NMS elicits microglial activation in the NTS and NX in a sex‐dependent fashion. Based on these morphological changes we propose that NMS may interfere with microglia's ability to perform synaptic pruning during early life. This, in turn may contribute to abnormal LCR function in pups.

Support or Funding Information

This research was supported by CIHR (RK).
Neuroprotective Properties of the Coffee Component EHT
Authors: Huber Kristen , Voronkov Michael , Shen Miles , Zhang Sherry , Fernandez Jose , Rouzard Karl , Webb Corey , Healy Jason , Perez Eduardo , Stock Maxwell , Stock Jeffry
Published: November 2023

Coffee is a complex mixture of more than eight hundred compounds with a variety of health benefits including reducing the risk of neurodegenerative diseases such as Alzheimer's and Parkinson's disease. A coffee compound, unrelated to caffeine, called eicosanoyl‐5‐hydroxytryptamide (EHT) has been identified as a regulator of the major serine/threonine phosphatase in the brain, protein phosphatase 2A (PP2A). In diseased tissue, PP2A exhibits reduced activity and methylation of its carboxy‐terminal tail. Previous reports indicate EHT is able to protect PP2A's methylation state and activity in vitro in addition to improving cognitive and motor function in animal models. In order to further understand the role of EHT in neuroprotection we sought to evaluate its ability to combat oxidative injury, structural damage and chemical toxicity in neuronal cell cultures. Utilizing lipid peroxidation as a marker for oxidative injury, we directly measured the formation of lipid hydroperoxides in the presence and absence of EHT. This colorimetric assay reports on the redox reaction between iron and hydroperoxides. Our results indicate that EHT is able to significantly reduce the production of the ferric ion suggesting EHT possesses antioxidant activity that may aid in maintaining membrane integrity. Next we studied the potential of EHT to reinforce the structural integrity of neurons, specifically analyzing their neuronal processes. Differentiated SH‐SY5Y neuroblastoma cells were treated with lysophosphatidic acid (LPA) to induce neurite retraction. Upon treatment with EHT, an increase in neurite length was observed suggesting protection from LPA. Lastly, the neuroprotection of SH‐SY5Y neuroblastoma cells against the chemical neurotoxin MPTP was evaluated. Dose‐dependent treatments with EHT resulted in maximal neuroprotection at a concentration of 100 nM. These data further support EHT as a component from coffee with neuroprotective activity.
In the neurodegenerative disease INAD, infantile neuroaxonal dystrophy, glutamate‐ and ATP‐induced Ca2+ signaling is disturbed in neurons and astrocytes, with mitochondrial dysfunction
Authors: Reiser Georg , Strokin Mikhail
Published: November 2023

About 85% of cases of INAD (infantile neuroaxonal dystrophy, OMIM #256600), an autosom al recessive inherited degenerative disease with early onset can be associated with mutations of the PLA2G6 gene . Ca2+‐independent phospholipase A2 (VIA iPLA2) is encoded in PLA2G6 . However, connections of these mutations to disease are unclear. For the mainly phospholipid metabolizing VIA iPLA2 also so‐called non‐canonical functions were discussed and reported. These are regulation of store‐regulated Ca2+ entry in cells and change in mitochondrial Ca2+ homeostasis. In this study, we investigate glutamate‐evoked Ca2+ signals in neurons and astrocytes in co‐culture from three INAD mouse models with Pla2g6 gene mutations. The strain‐1 has a hypomorphic Pla2g6 allele with reduced transcript levels (5% of wild type), the strain‐2 has knocked‐out Pla2g6, and strain‐3 has a Pla2g6 point mutation with the VIA iPLA2 inactive. It is shown that homozygous offspring from all strains develop pathology similar to that observed in INAD patients. We demonstrate in our present study a strongly reduced glutamate‐induced Ca2+ response in mutants in astrocytes compared to control cells. When we elucidate the mechanism we find that reduced Ca2+ responses are due to about 2‐fold‐reduced Glu‐induced capacitative Ca2+ entry in astrocytes in all three mouse strains. This effect is based on the lack in VIA iPLA2 activity since we could mimic the decrease by pharmacologically inhibiting iPLA2 with the compound S‐BEL enantiomer. On the contrary, in neurons the Pla2g6 mutation completely disrupted the influence of mitochondrial Ca2+ uptake on the extracellular Glu‐induced Ca2+ influx . We see it in neurons treated with Ru360, a blocker of mitochondrial Ca2+ uniporter or treated with rotenone. The dependency of Glu‐induced Ca2+ influx on mitochondrial Ca2+ uptake was wiped out in neurons with Pla2g6 mutation, whereas a strong reduction was seen in wild‐type cells. Thus, all INAD models have comparable changes, but they are different for astrocytes and neurons, respectively, in Glu‐induced Ca2+ signaling. These modifications in Ca2+ signaling in cells with mutated Pla2g6 are critical for adequate neuron‐astrocyte communication. Thus our data help to understand molecular mechanisms of INAD pathology.

Support or Funding Information

Deutsche Forschungsgemeinschaft (Re 563‐22‐1)
An LC‐MS/MS‐Based Protocol for the Absolute Quantitation of Histone Post‐Translational Modifications
Authors: Kingsley Philip John , Galligan James J. , Camarillo Jeannie M. , Marnett Lawrence J.
Published: November 2023

Post‐translational modifications (PTMs) of histones are utilized by eukaryotic cells to modulate chromatin and effect DNA‐based cellular processes. Identification and quantification of these PTMs provides essential information on both chromatin structure and gene expression in various pathophysiological conditions. Here we report a method for (1) the isolation of core histones, (2) their digestion to single amino acid residues, and (3) the quantitative analysis of the resultant digest for both modified and unmodified amino acids.

Nuclei are isolated from cultured cells or murine tissue and chromatin is isolated via high‐salt extraction. The four core histones (H2B, H4, H2A, and H3s) are purified by preparative reversed‐phase HPLC with UV detection. The histones are baseline resolved and the elution order is H2B, H4, H2A, H3.2/3, and H3.1.

Isolated histones are spiked with stably labeled internal standards, proteolytically digested to their component amino acids and analyzed via LC‐MS/MS. Chromatographic separation is in the reversed‐phase mode where the ion‐pairing agent heptafluorobutyric acid facilitates retention. The analytes are detected by selected reaction monitoring in positive ion mode and quantification is achieved by stable isotope dilution with internal standards previously added to the samples.

To validate this methodology, HEK293 cells were treated with the histone deacetylase inhibitor, trichostatin A, and histones were collected and subjected to the methodologies described. We observed quantitative differences in histone lysine acetylation, as expected. In addition, we quantized histone–specific alterations in lysine mono‐, di‐, and tri‐methylation, and arginine methylation.

This methodology may be used to probe for novel or unanticipated PTMs with data‐dependent LC‐MS/MS acquisition methods (DDMs). Lysine and many lysine PTMs such as carboxyethyl lysine and trimethyl lysine generate a strong fragment at m/z 84 upon collisionally induced dissociation. The survey scan of a DDM can utilize this common fragment by selecting its precursors for subsequent isolation and fragmentation, potentially revealing new PTMs.

Support or Funding Information

NIH: 2R01CA087819‐16
The YycFG Two‐Component System As A Regulatory Feedback Loop For The Expression Of Autolysin Activity
Authors: Keidel Victoria , Valderrama Andres , Kobir Ahasanul , Szurmant Hendrik
Published: November 2023

The YycFG (aka WalRK) system is present in all aerobic and some aenaerobic Gram‐positive organisms of the phylum Firmicutes. This two‐component signal transduction system is essential for viability and now known to conserve a function in regulating the expression of enzymes involved in cell wall metabolism. Our previous efforts have focused on the molecular pathways that lead to activation or inhibition of the YycG kinase in Bacillus subtilis. We have identified numerous interactions of the kinase with proteins of the cell division apparatus and two dedicated inhibitory proteins YycH and YycI and have revealed some of the molecular determinants of YycG activity regulation. As a consequence of these studies the knowledge has emerged that the YycFG system serves to regulate growth dependent cell wall restructuring by coordinating autolysin expression with cell division. We present novel data that suggests that this essential signaling system also responds to the autolytic activities of enzymes under direct transcription control by the YycFG system thus providing a feedback loop to fine tune autolysin expression according to its current need. In addition, evidence has emerged suggesting that this feedback loop might be the culprit of antimicrobial activities of a class of human chemokines against Bacillus anthracis and perhaps other Gram‐positive bacteria.

Support or Funding Information

Supported by grant GM106085 from the US National Institute of General Medical Sciences, National Institutes of Health
Anatomical substract of the Gluteus medius muscle myofascial trigger points: Preliminary results
Authors: Akamatsu Flávia Emi , Macaya Daniela Medina , Itezerote Ana Maria , Saleh Samir Omar , Hojaij Flávio , Andrade Mauro , Jacomo Alfredo Luiz
Published: November 2023

Myofascial pain syndrome (MPS) is a common cause of chronic musculoskeletal pain and is characterized by myofascial trigger points (MTP), limited range of motion in joints, referred pain and local twitch response during mechanical stimulation of MTP. MTP are clinically identified by palpation of a taut band of muscle or fascia. Physiopathology of MPS remains unclear and it has been suggested that MTP occur when a nociceptor and a muscular motor endplate coincide. The gluteus medius muscle MTPs are prevalent in patellofemoral pain, back pain and can occur spontaneously gluteal pain, weakness in the lower limbs abduction and increased sensitivity. Three different MTP are clinically described for the gluteus medius muscle, but anatomical correlation is still lacking.

Gluteus medius muscle from five cadavers were carefully dissected. We divided the gluteus medius muscle in four topographic áreas (I, II, III and IV) to describe the exact entry points of nerve fibers into the muscle.

As these values vary with muscle size, the relative values of the insertion point was calculated from the average longitudinal (from the highest point of iliac crest to greater trochanter of the femur) and transverse dimension (the medium point of the longitudinal dimension) and categorized into four areas: I, II, III and IV. For a comparison of the number of points between the quadrants we have used generalized estimation equations with correlation matrix exchangeable between the quadrants, with marginal Poisson distribution and identity link function. For comparison of the quadrants 2 to 2 multiple Bonferroni comparisons were carried out. We observed that most of the gluteus medius muscle innervation occurred in quadrants I, II and IV of the muscle. These locations correspond to the clinically described MTP. Anatomical correlation of the MTP may be useful for a better understanding of the physiopathology of these disorders and provide basis for their treatment.
Arachidonic and Oleic Acid Exerts Distinct Effects on DNA Methylome
Authors: Silva‐Martinez Guillermo Antonio , Rodriguez‐Rios Dalia , Alvarado‐Caudillo Yolanda , Barbosa‐Sabanero Gloria , Vaquero Alejandro , Esteller Manel , Carmona F. Javier , Moran Sebastian , Nielsen Finn C. , Wickström‐Lindholm Marie , Wrobel Katarzyna , Wrobel Kazimierz , Zaina Silvio , Lund Gertrud
Published: November 2023

Abnormal fatty acid metabolism and availability are landmarks of metabolic diseases, which in turn are associated with aberrant DNA methylation profiles. To understand the role of fatty acids in disease epigenetics, we sought DNA methylation profiles specifically induced by arachidonic or oleic acid (AA and OA) in cultured cells and compared those with published profiles of normal and diseased tissues. THP‐1 monocytes were stimulated with AA or OA and analyzed using Infinium HumanMethylation450 BeadChip (Illumina) and Human Exon 1.0 ST array (Affymetrix). Data were corroborated in mouse embryonic fibroblasts. Comparisons with publicly available data were conducted by standard bioinformatics. AA and OA elicited a complex response marked by a general DNA hyper‐ and hypomethylation in the 1–200 μM range, respectively, with a maximal differential response at the 100 μM dose. The divergent response to AA and OA was prominent within the gene body of target genes, where it correlated positively with transcription. AA‐induced DNA methylation profiles were similar to the corresponding profiles described for palmitic acid, atherosclerosis, diabetes, obesity and autism, but relatively dissimilar from OA‐induced profiles. A set of genes was identified, that display gene body hypermethylation in response to AA and in human atherosclerosis. Biochemical evidence pointed to beta‐oxidation, PPAR‐alpha and sirtuin 1 as important mediators of AA‐induced DNA methylation changes. In conclusion, AA and OA exert distinct effects on the DNA methylome. The observation that AA may contribute to shape the epigenome of important metabolic diseases supports and expands current diet‐based therapeutic and preventive efforts.

Support or Funding Information

CONCyTEG grant no. 08‐03‐K662‐020‐A01, Mexican Council for Science and Technology (CONACyT) Basic Science (Ciencia Básica) grant no. 83401 to GL; CONACyT Sabbatical Fellowships no. 166209 to G.L. and no. 166058 to S.Z.; CONACyT Ph.D. Fellowship no. 334370 to G.A.S.‐M.; the Spanish Ministry of Ministry of Economy and Competitiveness (MINECO) grant SAF2011‐25619 to A.V.
Seaweed Supplementation Attenuates Long‐term High Fat Diet‐induced Chronic Inflammation and Insulin Resistance Regardless of Obesity in C57BL/6N Mice
Authors: Lee Yunkyoung , Oh Ji‐Hyun , Han A‐reum , Hwang Yeonji , Choi Ji‐Hey , Yang Hyoseon , Kang Sae‐im
Published: November 2023

Antidiabetic and anti‐inflammatory effects of brown seaweeds have been reported in various experimental models, however information is scarce on their effects combined with a long‐term high fat diet. In this present study, we investigated that effects and mechanisms of seaweed supplementation (Undaria pinnatifida (UP), Laminaria japonica (LJ), Sargassum fulvellum (SF), Hizikia fusiforme (HF)) along with high‐fat (HF) diet in C57BL/6N male mice for 16 weeks. Body weight, food intake, and fasting blood glucose concentrations were measured. Insulin tolerance test was performed at week 14. Animals were sacrificed at week 16 and tissues were dissected, weighted, and snap frozen for analysis. Bone‐marrow (BM) cells were isolated from femoral and tibial BM of a representative mouse from each group. As a result, 5% seaweed supplementation did not prevent HF‐induced obesity. Although there was a trend to have less weight gain in HF+LJ and HF+HF groups compared to HF group at week 8, this trend disappeared as HF continued. In spite of being obese, mice in HF+LJ group had significantly improved insulin resistance, lowered cholesterol in plasma and liver, and increased TG in feces compared to mice in HF group that were consistent with histological observations. In addition, BM‐derived macrophages(BMDM) from mice fed HF with seaweeds showed to have less inflammatory cytokines compared to BMDM from mice fed HF only. Increased phosphorylation of insulin signaling proteins was observed in liver of the seaweed supplemented HF groups compared to the HF group. In addition, levels of TG and cholesterol excretion were altered by seaweed supplementation. Conclusively, seaweed supplementation attenuated insulin resistance and chronic inflammation induced by a long‐term HF feeding partially due to altered insulin signaling and lipogenesis in liver and muscle as well as reduced inflammation in adipose tissue regardless of obesity in C57BL/6N mice.

Support or Funding Information

This work was supported by NRF‐2013R1A1A10575730.
Weight Loss and Program Satisfaction in Response to the Weight Watchers SmartPoints Plan
Authors: Rogers Renee J , Kovacs Sara J , Byard Thomas D , Collins Katherine A , Creasy Seth A , Gomberg Monica A , McCoy Stephanie M , Peluso Anna , Raybuck Shawn D , Rupp Kristie , Jakicic John M
Published: November 2023

Background

As commercial weight loss programs adapt their approaches, it is important to evaluate the protocols for satisfaction and efficacy. This study evaluates changes in body weight and body mass index (BMI) along with program satisfaction in participants enrolled in a Weight Watchers program with the updated SmartPoints plan.

Methods

Participants (N=150; mean age: 47.3 ± 12.7 years; mean BMI: 33.9 ± 4.3 kg/m2) were enrolled in a 6‐month Weight Watchers program. The intervention was delivered by trained Weight Watchers leaders and evaluated by independent research staff that was not engaged in intervention delivery. The Weight Watchers intervention included weekly group sessions, instructions on calculating points for foods and beverages, and use of an app to log points. Assessment of body weight and BMI were completed at baseline and 6 months, attendance at group sessions was tracked weekly, and program satisfaction was reported using a survey at 6 months.

Results

Of the 150 enrolled participants, 143 (95.3%) completed the 6‐month assessment. There was a significant reduction in weight of 2.8 ± 5.0 kg (3.1 ± 5.5%) from 0 at 6 months (p<0.001). This corresponded to a reduction in BMI from 33.8 ± 4.3 kg/m2 to 32.8 ± 4.7 kg/m2 (p<0.001). A clinically meaningful threshold of 5% weight loss was achieved in 32.9% (N=47) of the sample. There was a significant correlation between attendance and weight loss (r=0.59, p<0.001) with 11.9 ± 6.4 group sessions attended. Participants who attended >75% of sessions (N=21) lost more weight (−9.4 ± 6.6%) than those who attended >50–<75% (N=52, −4.2 ± 4.5%), 25–50% (N=39, −1.5±3.3%), and <25% (N=31, −0.9 ± 3.7%) of group sessions (p<0.001); those attending >50–<75% lost significantly more weight than those attending either 25–50% or <25% of group sessions (p<0.001). There was no significant difference in weight loss between those attending 25–50% or <25% of group sessions. Weight change was significantly correlated with overall program satisfaction (r=0.52, p<0.001), satisfaction with the amount of weight lost (r=0.54, p<0.001), and with the likelihood of recommending the program to friends and colleagues (r=0.36, p<0.001).

Conclusions

The Weight Watchers SmartPoints plan appears to be effective at eliciting significant 6‐month reductions in body weight and BMI. It appears that attendance at the weekly sessions is an important component to success when participating in this commercial weight loss program.

Support or Funding Information

Weight Watchers International, Inc.
Pancreatic protein tyrosine phosphatase 1B deficiency exacerbates acute pancreatitis in mice
Authors: Whitehead Katherine , Chahed Samah , Koike Shinichiro , Bachaalany Santana , Haj Fawaz , Bettaieb Ahmed
Published: November 2023

Acute pancreatitis (AP) is one of the most common gastrointestinal disorders and causes significant morbidity. The disease starts as local inflammation in the pancreas that may progress to systemic inflammation and complications. Protein tyrosine phosphatase 1B (PTP1B) is implicated in inflammatory signaling but its significance in AP remains unclear. To investigate whether PTP1B may have a role in AP we utilized pancreas PTP1B knockout (panc‐PTP1B KO) mice to determine the effects of pancreatic PTP1B deficiency on cerulein‐ and arginine‐induced pancreatitis. PTP1B protein expression was increased in the early phase of AP in mice and rats. In addition, histological analyses of pancreas samples revealed enhanced features of AP in cerulein‐treated panc‐PTP1B KO mice compared with controls. Moreover, cerulein‐ and arginine‐induced serum amylase and lipase were higher in panc‐PTP1B KO mice compared with controls. Similarly, pancreatic mRNA and serum concentrations of the inflammatory cytokines IL‐1β, IL‐6 and TNFα were increased in panc‐PTP1B KO mice. Moreover, panc‐PTP1B KO mice exhibited enhanced cerulein‐ and arginine‐induced NF‐κB inflammatory response accompanied by increased MAPKs activation and elevated ER stress. These findings revealed a novel role for pancreatic PTP1B in cerulein‐ and arginine‐induced acute pancreatitis.
Effect of Rice Cake, Seolgitteok, on Glucose, Insulin, and Satiation Response in Humans
Authors: Pottgen Ellen , Yoon Suk Hoo , Pham Tung , Seo Han‐Seok , Lee Sun‐Ok
Published: November 2023

A research study was conducted to determine the effects of Seolgitteok (Korean rice cake) on postprandial glucose, insulin, and appetite in humans. Using a randomized‐crossover design, 23 participants consumed one of three Seolgitteok (white rice, brown rice, and mixture of equal parts white and brown rice) with a one‐week washout period. Seolgitteok contained 50 g of total starch based on rice flour analysis, and additional ingredients were consistent. Blood samples were collected intravenously at 15 minutes prior and 0, 30, 60, 90, 120, and 180 minutes after consumption and self‐reported feelings of satiety were collected at each interval using a visual analog scale (VAS). The brown rice Seolgitteok contained a significantly greater percentage of insoluble fiber (3.3±0.3%) compared to the mixed (1.6±0.3%) and white (0.8±0.3%) rice treatments (p<0.05). The brown rice cake produced the lowest glucose response at 30–120 minute intervals and the lowest insulin response at 60–180 minute intervals. Glucose incremental area under the curve (iAUC) for the brown rice Seolgitteok (1610±326 mg (~3h) dL−1) was reduced ~38% compared to the mixed rice Seolgitteok (2580±450 mg (~3h) dL−1) and ~44% compared to the white rice Seolgitteok (2871±553 mg (~3h) dL−1). Insulin iAUC for the brown rice Seolgitteok (2968±493 μU (~3h) L−1) was reduced by approximately 13 and 18 percent compared to the mixed rice (3407±607 μU (~3h) L−1) and white rice (3595±633 μU (~3h) L−1) treatments, respectively. Based on the iAUC of VAS, the brown rice cake was reportedly 10–15% more satiating over the 3 hour postprandial time period compared to the white and mixed rice Seolgitteok. These findings suggest that brown rice Seolgitteok has a potential for use as functional food to reduce postprandial glucose and insulin levels and suppress appetite.

Support or Funding Information

Funding provided by Korean Food Research Institute is greatly appreciated.
Moderate and Hard Physical Activity are Associated with Increased Total Energy Expenditure in Overweight and Obese Individuals with Knee Osteoarthritis
Authors: Akhavan Neda S , Ormsbee Lauren T , Elam Marcus L , Johnson Sarah A , Panton Lynn B , Arjmandi Bahram H
Published: November 2023

Osteoarthritis (OA) is the leading cause of chronic disability in the elderly and the most common degenerative joint disorder in the United States. OA is characterized by cartilage degeneration leading to joint pain and tenderness, loss of mobility in joints, and inflammation. A major modifiable risk factor for the development of OA is obesity, where those with a body mass index (BMI) greater than 30 kg/m2 are 6.8 times more likely to develop knee OA compared to men and women of normal weight. OA is a progressive disease, where those who have mild‐to‐moderate OA may respond better to lifestyle modifications, such as physical activity or dietary interventions, than those who have severe OA. The objective of this study was to investigate the relationship between different levels of physical activity (light, moderate, hard, and very hard) and total energy expenditure in 94 overweight and obese men and women (average BMI = 32.77±6.34 kg/m2) aged 40–90 years with clinically diagnosed mild‐to‐moderate OA of the knee. A Seven‐Day Physical Activity Recall was used to assess overall pattern and level of physical activity and total energy expenditure (TEE) of participants. Out of all the levels of physical activity, moderate (r=0.580) and hard (r=.539) physical activity were shown to have a positive correlation with TEE, whereas light (r=−0.581) physical activity had a negative correlation with TEE. These findings implicate the potential use of hard and moderate physical activity as beneficial exercise intensities for increasing TEE for overweight and obese men and women with mild‐to‐moderate knee OA. Further studies are needed to fully understand the relationship between obesity and knee OA with regards to physical activity and the microenvironment of knee cartilage.
Dietary Supplement Label Database (DSLD) Captures Dietary Supplements Used by Cancer Survivors: The Case of Fiber Supplements
Authors: Emenaker Nancy J , Sorkin Barbara C , Dwyer Johanna T , Rodriguez LM
Published: November 2023

Background

More than 50% of all U.S. adults consume dietary supplements (DS) contributing to an estimated $35 billion in sales in 2015. Among these are DS often recommended by oncology practitioners for the reduction of disease risks, including risk of colorectal cancer. The Dietary Supplement Label Database (DSLD) contains over 50,000 DS labels representing a large, and ever‐increasing, fraction of those sold in the U.S. Supplements touted as possessing anti‐cancer properties include multivitamins, botanical extracts, and isolated dietary components such as fiber.

Objective

Describe, for the specific case of dietary fiber supplements, how the oncology community (e.g., oncology health care professionals, researchers and patients) can use DSLD in assessing DS consumption hypothesized to affect cancer outcomes.

Description

The DSLD provides ready access to label data for dietary supplements sold in the U.S. using a web‐based interface.

Method

We searched DSLD for all products including the terms “fiber”, “colon cleanse” and “colon health” as either a product name, dietary ingredient, brand name, or label element.

Results

DSLD yielded a total of 4,873 products containing the search term “fiber” anywhere on the product label. A total of 181 products contained ingredients classified as “fiber” and 196 products contained “fiber” in the product name. 109 products contained “colon cleanse” anywhere on the product label; 58 contained “colon cleanse” on in the product name; and one contained “colon cleanse” in the dietary ingredient name. Finally, 235 products contained “colon health” anywhere on the label; and 5 contained “colon health” in the product name. DSLD can also be used in some cases to determine the type(s) of fiber claimed as present in each product, providing the potential for assessment of intake of soluble vs insoluble fiber.

Conclusion

DSLD serves as an online resource supplying a broad range of users, including the oncology community, with relevant DS information for research and clinical use

Support or Funding Information

Office of Dietary Supplements and the National Library of Medicine, National Institutes of Health.
The Relationship between Waist‐to‐Height Ratio and Femoral Neck Bone Mineral Density in Men and Women
Authors: Clark Elizabeth , Johnson Sara A , Pourafshar Shirin , Navaei Negin , Akhavan Neda , Elam Marcus L , Foley Elizabeth , Arjmandi Bahram H
Published: November 2023

In the United States, adults over the age of 45 years old represent about 40% of the population. Over half of this population is afflicted with one or more chronic health conditions, among which cardiometabolic dysregulation and decreased bone mineral density (BMD) are two of the highest in prevalence. Trabecular bone, as found in the femoral neck, is regulated via metabolic activity in the human body. Therefore, age‐related bone loss, at sites such as femoral neck may lead to complications such as fracture. However, there is a conflict of existing data regarding the nature of how cardiometabolic risk factors are related to BMD. High body mass index is one of the cardiometabolic risk factors that has been shown to have a positive correlation with total BMD; however, other metabolic factors have been identified to be inversely correlated with BMD. For example, high waist‐to‐height ratio (WHtR), which is a strong indication of cardiometabolic risk, has been implicated as having an inverse relationship with BMD. On the contrary, some studies suggest that as the number of cardiometabolic risk factors increases, so does BMD. Thus, the purpose of this cross‐sectional study was to investigate the extent to which WHtR may have a relationship with femoral neck BMD in an aging population. For this purpose, forty‐eight men and women (age 45 to 65 years old) were assessed for their baseline right and left femoral neck BMD measurements. Duel‐energy x‐ray absorptiometry was used to determine participants’ femoral neck BMD. For the statistical analysis, SPSS bivariate two‐tailed correlation and linear regression was used to compare the data. Results from this study showed a significant (P <0.05) positive correlation between WHtR and femoral neck BMD. Therefore, the findings of this study suggest that WHtR is positively correlated with femoral neck BMD; however, these findings are limited and more research is needed to determine how visceral fat is causally associated with BMD.