bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Novel Gene Crispld2 may Contribute to Facial Dysmophology in a Chicken Model of Crouzon's Syndrome
Authors: Li Xin , Tropp Stephen , Young Nathan , Hallgrimsson Benedikt , Marcucio Ralph 
Published: June 2021

Craniosynostosis causes malformations of the skull that are characterized by premature suture closure, as well as malformations of the brain and face. Recent work in mice, and our data, suggest that the malformations in the brain and face are a primary consequence of mutations that produce craniosynostosis. Mutations in FgfR1–3 account for ~25% of craniosynostosis syndromes. To investigate mechanisms underlying these facial malformations, we infected chick embryos with a retrovirus encoding FgfR2C278F. Compared to controls, treated embryos have widened‐faces and midfacial dysplasia resembling defects seen in craniosynostosis. Further, microarray analysis detected changes in gene expression. In particular, Crispld2 (cysteine rich secretory LCCL domain protein 2), a nonsyndromic cleft lip/palate related gene, is downregulated. The function of Crispld2, and how it may contribute to cleft lip, is unknown. We detected Crispld2 expression in facial mesenchyme in chicken embryos at HH 22. To assess function of this gene, we created an RCAS vector encoding Crispld2. Infected chicken fibroblasts appeared to migrate faster than controls cells, but embryos infected with the virus appeared normal. Therefore, we are currently investigating the effect of knocking‐down Crispld2. In conclusion, we have identified a novel gene Crispld2 and are determining the role this gene plays in craniofacial development.Grant Funding Source: NIH
Anatomy of the Incomplete Ureteral Duplication
Authors: Mehta Mudresh , Bui Trang , Rengasamy Padmanabhan 
Published: June 2021

Ureteral duplications constitute the most common congenital anomalies of the urinary tract. Incomplete duplications are associated with uretero‐uremic reflex or uretero‐pelvic junction obstruction. Here we report an incomplete duplication of the left ureter observed in a 75 year old male cadaver. The medial ureter emerged from the upper end of the hilum, passed posterior to renal, gonadal and left colic vessels and fused with the lateral ureter halfway on the lateral pelvic wall. The lateral ureter emerged from the lower hilum, crossed the psoas, approached the medial one at the left common iliac artery level, both crossed the internal iliac artery, united with each other, crossed the vas, turned medially and dumped into the bladder. Throughout their course, the ureters were retroperitoneal. The common ureter was almost twice as thick as the duplicated segments. Duplicated segments were accompanied by a rich vascular network. A Y‐shaped vein with one arm coming from the renal vein and the other from the lower pole of the kidney connected to the left gonadal vein. The left colic vein dumped into the left gonadal vein. There were two testicular arteries ‐ one from the renal and the other from the aorta. Renal artery divided precociously adjacent to the aorta and behind the renal vein. The right kidney and ureter were normal. Right gonadal vein dumped into right renal close to its union with the inferior vena cava. The cause of anomalous vascular connection of the left gonadal vein associated with ureteral duplication is not known.Grant Funding Source : Mercer University
Hepatocyte maturation is arrested in mouse liver lacking the splicing factor SRSF3 (SRp20)
Authors: Sen Supriya , Webster Nicholas JG 
Published: June 2021

SRSF3 is the smallest member of the highly conserved SR protein family that has numerous roles in mRNA metabolism, from transcription to translation. To investigate the role of SRSF3 in postnatal hepatic function, we used Cre‐loxP‐mediated gene technology to delete SRSF3 specifically in hepatocytes in mice. Deletion of SRSF3 inhibits hepatocyte maturation as livers from mice lacking SRSF3 exhibit a fetal phenotype characterized by increased expression of the fetal isozyme of pyruvate kinase (M‐PK), persistent expression of alpha‐fetoprotein and H19 mRNA. The livers also show increased expression of the fetal isoform of the insulin receptor (IR‐A). The expression of several genes that regulate glucose and lipid metabolism in the mature liver is significantly reduced in SRSF3−/− liver, and glycogen levels are decreased. Global gene expression and exon microarray analyses identified multiple splicing and expression changes in the SRSF3−/− liver. Interestingly, some of these changes alter expression of major hepatic transcription factors that play crucial role in development and metabolism. Thus, SRSF3 directs a genetic program required for morphological and functional differentiation of hepatocytes and the hepatocyte SRSF3 knock‐out mice provide a new model to study metabolic dysregulation and liver disease.This work has been supported by VA Merit Grant Award.
Influx of acetyl‐CoA into the ER lumen regulates the induction of autophagy during the UPR
Authors: Puglielli Luigi , Pehar Mariana 
Published: June 2021

One of the main functions of the UPR is to ensure disposal of large protein aggregates that accumulate in the lumen of the ER while avoiding, under non‐lethal levels of ER stress, cell death. When tightly controlled, ERAD(II) allows the cell to recover from the transient accumulation of protein aggregates; however, when unchecked it can be detrimental and cause autophagic/type 2 cell death. Here we report the results of a proteomic study showing that a large number of ER‐resident and ‐transiting proteins undergoes Nε‐lysine acetylation in the lumen of the organelle. The list of ER‐resident proteins includes chaperones and enzymes involved with post‐translational modification and folding. We also show that during ER stress IRE1/XBP1 controls the influx of acetyl‐CoA into the lumen of the ER by inducing the ER membrane acetyl‐CoA transporter SLC33A1/AT‐1. Failure to induce AT‐1 and maintain the acetylation status of the ER leads to widespread autophagic cell death. Mechanistically, the regulation of the autophagic process involves Nε‐lysine acetylation of Atg9A. In fact, a gain‐of‐acetylation mutant form of Atg9A protected from autophagic cell death while a loss‐of‐acetylation mutant achieved the opposite effect. Finally, we report that a mutant form of AT‐1 that is associated with Autosomal Dominant Spastic Paraplegia is unable to form homodimers and transport acetyl‐CoA across the membrane.Supported by NIH
Using bakersˈ yeast to identify genetic modifiers of alpha‐1 antitrypsin deficiency
Authors: Gelling Cristy L , Brodsky Jeffrey L 
Published: June 2021

Around 1 in 2500 individuals in the US have inherited alpha‐1 antitrypsin deficiency (ATD), which manifests as lung and liver disease. The most common cause of ATD is homozygosity for the Z‐variant of AT (ATZ), which accumulates in the endoplasmic reticulum of liver cells as a native polymer. ATZ is inefficiently secreted to the bloodstream, resulting in AT deficiency in the lungs and polymer accumulation in the liver. However, only a subset of ATD patients suffer from liver disease: 10% of patients show liver dysfunction at infancy, and 50% show signs of cirrhosis in adulthood. The factors that predispose this subset of individuals to liver disease are unknown.We have developed a yeast antitrypsin expression system to find candidate genetic factors that affect the risk for ATD‐associated liver disease. This approach takes advantage of the genetic tools available for yeast, as well as the conservation of pathways implicated in ATD‐associated liver disease. Important disease‐associated characteristics of antitrypsin were recapitulated in yeast: the wildtype form was secreted, while mutant ATZ was not. We then screened mutants representing all non‐essential genes in the yeast genome for increased ATZ secretion. Current work is focused on confirming the effect of the candidate genes in a hepatic cell model.Funding from the Alpha‐1 Foundation, Australian Research Council, and the American Australian Association.
Screening and kinetic studies of new cathepsin D inhibitors
Authors: Sheppard Donald R , Wen Lisa , Huang Jenq-Kuen , McConnell Rose M 
Published: June 2021

Cathepsin D is a ubiquitous aspartyl lysosomal protease. Because it is associated with the mobility and invasiveness of various cancers, inhibitors of this enzyme have the potential for use as chemotherapeutic drugs. Previously prepared synthetic compounds are being evaluated for their ability to inhibit cathepsin D by determining their IC50 values. The studies were performed using human liver cathepsin D and a fluorogenic substrate (Mca‐Gly‐Lys‐Pro‐Ile‐Leu‐Phe‐Phe‐Arg‐Leu‐Lys(Dnp)‐D‐Arg‐NH2).Several of these compounds were found to be strong inhibitors of cathepsin D. The kinetic properties of four potential inhibitors were characterized. These data will be used in the development of new and more effective inhibitors. Supported by National Cancer Institute at NIH (3R15CA08933‐04 and 3R15CA86933‐04A2S1) and Western Illinois University.
Regulation of PI4KIIα distribution between the Golgi and endosomal compartments
Authors: Jovic Marko , Bojjireddy Naveen , Kean Michelle , Gingras Anne-Claude , Brill Julie , Balla Tamas 
Published: June 2021

Phosphatidylinositol 4‐kinases (PI4Ks) are essential for trafficking along the secretory and degradative routes. The importance of PI4K localization is underscored by the role of PI4KIIα, an enzyme localized to trans‐Golgi and endosomes, in EGF receptor degradation and Wnt signaling, and its suspected role in the late‐onset neurodegeneration. Although membrane association of PI4KIIα through a palmitoyl moiety is suggestive of its cycling between endosomes and Golgi, little is known about the regulation of PI4KIIα subcellular localization. Here we performed a proteomic screen of potential binding partners of PI4KIIα and identified a vesicular fusion protein VAMP‐3, that was confirmed by co‐immunoprecipitation. Expression of Tetanus toxin (TeNT), which specifically cleaves VAMP‐3, resulted in accumulation of PI4KIIα on endosomes. Conversely, overexpression of TeNT with the TeNT‐resistant VAMP‐3 mutant recovered the Golgi localization of PI4KIIα. We also found that PI4KIIα rapidly trafficked back to the Golgi within 2–3 min following photobleaching of the Golgi subpopulation, whereas the rate of its reappearance was significantly decreased in the presence of TeNT. Collectively, these data suggest that VAMP‐3 regulates PI4KIIα retrograde transport, uncovering new aspects of the role of PI4KIIα in human health and disease.Supported by the NICHD Intramural research program
The role of an ER‐localized metalloprotease in yeast cell wall integrity
Authors: Hecht Karen Alice , Wytiaz Victoria A. , Brodsky Jeffrey L. 
Published: June 2021

The fungal cell wall protects against environmental stresses, and regulates morphogenesis. It is made up of a polysaccharide layer and a dense network of proteins necessary for cell wall function. Cell wall proteins are extensively glycan and lipid modified, and subject to endoplasmic reticulum (ER) quality control before secretion. When the cell wall is perturbed, stress signaling is essential to up‐regulate secretion of proteins required for cell wall repair. Understanding how yeast synthesize and maintain cell wall integrity is essential to combat fungal pathogens in human disease.We have identified a gene, YBR074, displaying a distinct set of genetic interactions associated with lipid metabolism in the yeast Saccharomyces cerevisiae. Ybr074 is a putative ER‐localized metalloprotease of unknown function. In order to understand the role of Ybr074 in lipid metabolism, we have tested chemical‐genetic interactions of this gene under conditions of cell wall and ER stress. Interactions of YBR074 upon treatment with the cell wall perturbing agent calcofluor white have been observed with CDC48. Cdc48 belongs to the protein family of ATPases Associated with diverse cellular Activities (AAA). It is known to function both in ER quality control and cell wall maintenance. This genetic interaction suggests a role for Ybr074 in regulating components of cell wall maintenance under conditions of stress.NIH grant GM75061
In vitro evidence that peptides derived from the candidate tumor suppressor gene Esophageal Cancer‐Related Gene 4 (Ecrg4) internalize into cells through the innate immunity receptor complex
Authors: Podvin Sonia , Dang Xitong , Kurabi Arwa , Coimbra Raul , Eliceiri Brian , Baird Andrew 
Published: June 2021

In light of data implicating the secreted precursor protein and candidate tumor suppressor Ecrg4 in the inflammatory response, we explored the possibility that predicted ligand domains in its primary sequence can target the innate immunity receptor complex. Predicted fragments of Ecrg4 were displayed on M13 phage and tested for internalization into cultured human embryonic kidney (HEK) cells that normally express low levels of the innate immunity receptor complex and HEK cells that stably overexpress. The receptor‐mediated internalized phage was recovered and phage DNA quantified by PCR. Internalized Ecrg4 targeted phage was compared to untargeted phage or epidermal growth factor (EGF) targeted phage that internalizes through the EGF receptor. Three peptide domains that derive from the Ecrg4 precursor internalized into HEK cells. Two of these, one at the amino terminus and one at the carboxyl terminus internalized 13 to 16 fold higher in HEK cells that overexpress TLR4/MD2/CD14 compared to wild type HEK cells. Untargeted phage internalization was low and EGF‐targeted phage internalization was high, but neither was different between the two cell lines. These data suggest that predicted Ecrg4 peptides derived from the secreted precursor protein can internalize into target cells through the innate immunity receptor complex.Supported by a Mentored Young Investigator award from the Hydrocephalus Association
The Os Peroneum and Accessory Navicular: An integrated radiographic and cadaveric study
Authors: Delamarter Taylor , Benninger Brion 
Published: June 2021

INTRODUCTIONSesamoids and accessory bones commonly exist throughout the human body, especially in the foot. Pain from the foot and ankle is a common symptom, of which the Os Peroneum (OP) and Accessory Navicular (AN) are associated with pathologies that present to clinicians. The objective of this study was to investigate the use of CT as a paradigm in undiagnosed foot and ankle pain and to advocate the teaching the OP and AN in anatomy courses.METHODSLiterature search was conducted on anatomical texts, atlases, websites, and journals regarding the OP and AN. Roentogram and CT scans were conducted on 15 embalmed cadaveric lower limbs, followed by palpation and deep dissection of the distal tibialis posterior and fibularis longus tendons.RESULTSLiterature search revealed a 14 – 26% prevalence of the OP and 5–21% of the AN. Roentograms revealed OP in 2/22 and AN in 5/23 cadavers. CT scans revealed OP in 9/28 and AN in 13/28. Dissection revealed an OP in 9/30 and an AN in 10/28.DISCUSSIONDespite the prevalence of the OP and AN in the literature, they are not addressed in anatomy courses. They are a known cause of pain and are associated with false negatives radiologically.CONCLUSIONThis study suggests that anatomy courses include the OP and AN as clinically relevant structures. CT scans should follow roentograms as part of a diagnostic paradigm when treating unknown foot and ankle pain.
Improvement in Skin Carotenoid Levels Assessed by Reflectance Spectroscopy
Authors: Talbott Shawn , Quan Danyi , Xiong Wade 
Published: June 2021

BACKGROUNDCarotenoids have been directly associated with health benefits, including lutein (eye health), lycopene (cardiovascular and prostate health), and beta‐carotene (ultraviolet skin protection). Various biophotonic spectroscopy techniques are accurate and noninvasive methods of detecting skin carotenoid levels and assessing change in antioxidant levels over time.OBJECTIVEWe used Reflectance Spectroscopy to measure skin carotenoid levels following two supplementation regimens.METHODSSixty‐six healthy volunteers were supplemented with: Group A = 15mg/day of mixed carotenoids (9mg beta‐carotene, 4mg lycopene, and 2mg lutein in a proprietary nano‐emulsion); Group B = 18mg/day of natural beta‐carotene; or Group C = placebo containing no carotenoids. Skin carotenoid levels were measured at Baseline and Weeks 8 & 16.RESULTSSkin carotenoid levels were increased 59% and 57% in Group A, and 37% and 36% in Group B at weeks 8 and 16, respectively, compared to Control.CONCLUSIONThese results show, via Reflectance Spectroscopy, that skin carotenoid levels are significantly increased by supplementation with natural beta‐carotene (18mg/d) or a proprietary nano‐emulsion (15mg/d). The magnitude of skin carotenoid increase was greater with the nano‐emulsion, suggesting a possible enhancement of absorption, delivery, deposition, or retention in the body.
Effect of High‐Fat Meal and High‐Carbohydrate Meal on Daily Total Energy Metabolism in Adults Japanese Men
Authors: Takuya Taira , Hisao Fujii 
Published: June 2021

IntroductionDiet‐induced Thermogenesis (DIT) is the increase in energy expenditure above resting associated with the consumption of a meal, accounts for approximately 10% of an individual fs total daily energy expenditure. The thermic effect of separate nutrients is highest for protein (20–30%), followed by CHO (5–10%), and fat (0–3%).ObjectiveThe objective of this study was to clarify the effect of high‐fat meal and high‐carbohydrate on daily energy metabolism in adult Japanese men using indirect human calorimeter.MethodsThe subjects were fourteen adults Japanese men (21.5 ± 0.3 years). We measured EE (energy expenditure), RQ (respiratory quotient), GOR (glucose oxide rate), LOR (lipid oxide rate), LF/HF (Low Frequency/High Frequency), HF (High Frequency). EE was measured using the IHC for 36 hours. The activity of sympathetic nervous system was assessed by real‐time analysis system of heart rate fluctuation.Results& ConclusionTwenty‐four‐hours total energy expenditure was no different among three trial. On the other, RQ and GOR at supper time were significantly lower in HF trial than HCHO trial during the 120 minutes after ingestion (p < 0.05). LOR at supper time was significantly higher in HF trial than HCHO trial during the 120 minutes after ingestion (p < 0.05). Those results showed that high‐fat meal and high‐carbohydrate meal changed energy substrate.
Trends in dietary intake among U.S. 2–6 year olds, 1989 to 2008
Authors: Ford Christopher Nicholaus , Slining Meghan M. , Popkin Barry M. 
Published: June 2021

ObjectiveDescribe changes in diet among 2–6 year olds from 1989 to 2008 related to sharp rises in obesity during this period.MethodsThis analysis included 10,647 children ages 2–6 from five nationally representative surveys of dietary intake in the U.S.: CSFII 1989–1991; CSFII 1994–1998; and NHANES surveys 2003–2004, 2005–2006 and 2007–2008. Diet data were categorized into groupings using the UNC‐CH approach. Analyses were carried out using a single 24‐h dietary recall with appropriate survey weighting. T‐tests were used to compare means across survey years with P<.05 considered significant.ResultsOver the 20‐year period, there were increases in per capita intake of savory snacks (+51 kcal, P<.01), pizza/calzones (+32 kcal, P<.01), sweet snacks and candy (+25 kcal, P<.01), corn‐based mixed Mexican dishes (+22 kcal, P<.01) and fruit juice (+18 kcal, P<.01), whereas total daily energy intake increased by 212 kcal (from 1833 to 2045) (P<.05). Fruit intake increased marginally (+24 kcal, P<.01).ConclusionsFoods that tend to negatively impact diet quality, and contribute excess calories, such as savory snacks, pizza/calzones, mixed Mexican dishes, sweet snacks and candy, and fruit juice, predominated the top changes in per capita consumption between 1989 and 2008.
The role of cAMP/PKA signaling enhancer Protein Phosphatase Inhibitor 1 (I‐1) in the control of Ca2+ cycling and signaling in VSMCs
Authors: Hadri Lahouaria , Regis Bobe , Lopez Jose J , Turnbull Irene C , Kovacic Jason C , Karakikes Ioannis , Kranias Evangelia , Hajjar Roger J , Lipskaia Larissa 
Published: June 2021

The Ca2+ cycling in “contractile” vascular smooth muscle cells (VSMC) requires the expression of SERCA2a, whereas in “proliferating” ones is associated with the expression of SERCA2b only. Inhibitor‐1 (I‐1), a highly specific inhibitor of protein phosphatase 1 (PP1), enhances both PKA‐dependent Phospholamban (PLB) phosphorylation and SERCA activity.The goal of this study was to elucidate the role of I‐1 in the control of VSMC Ca2+ cycling. In humans and rodents, the expression of I‐1 was found to be specific for contractile VSMC, whereas PP1 was highly expressed in synthetic VSMC. Genetic deletion of I‐1 in mice model resulted in lack of PLB phosphorylation in VSMC. Despite the fact that SERCA2a was expressed, VSMCs from I‐1 deficient mice were locked in the proliferating state, with the development of a vascular proliferative disorder and excessive neointimal proliferation after vascular injury. Gene transfer of constitutively active I‐1 (I‐1c) significantly increased PLB phosphorylation and Ca2+ uptake in both synthetic and contractile VSMCs. Therefore, I‐1c prevented proliferation and remodeling of contractile VSMC, but had no effect on synthetic VSMC, which predominantly express SERCA2b.In conclusion, Gene transfer of I‐1c may be considered as a promising therapeutic strategy for preventing vascular proliferative diseases.
Desflurane‐Induced Coughing in Conscious Guinea Pigs is Mediated through TRPA1
Authors: Daller Justin , McKee Jeffrey S. , Wong Joseph , Brooks Ben 
Published: June 2021

Desflurane is widely used for induction and maintenance of anesthesia, and is associated with a high incidence of perioperative coughing. Desflurane has been shown to activate TRPA1 in vitro and increase total lung resistance in anesthetized guinea pigs that could be mitigated with a selective TRPA1 antagonist.PurposeTo evaluate the mechanism of desflurane‐induced coughing in conscious guinea pigs during anesthetic induction.MethodsGuinea pigs were placed into a transparent chamber where flow rate, pressure, audio and video were recorded. Animals were induced with 1 MAC (6.4%) desflurane or challenged with a selective agonist for TRPV1 (citric acid, 1 M) or TRPA1 (AITC, 80 mM) with or without pre‐treatment with a selective antagonist for TRPV1 (BCTC, 3 mg/kg IP) or TRPA1 (HC‐030031, 300 mg/kg IP). The number of coughs over a fixed, 5‐min exposure period was recorded.ResultsDesflurane caused 4.9 coughs vs. 0 for the carrier air. AITC caused 10.7 coughs vs. 5.8 when pretreated with HC‐030031 (p<0.05). Citric acid caused 12.4 coughs vs. 3.2 when pre‐treated with BCTC (p<0.002). HC‐030031 markedly attenuated desflurane‐induced coughing (4.3 vs. 1.2, p<0.08) whereas BCTC had no effect (5.4 vs. 4.0, p=0.13).ConclusionDesflurane‐induced coughing in conscious guinea pigs appears to be mediated through activation of TRPA1. Supported by Baxter Healthcare Corporation.