bluuebunny/crossref_metadata_2025_split huggingface dataset:
The effects of angiotensin receptor blockade on functional recovery and inflammatory gene expression following spinal cord injury
Authors: Robbins Emily A , Shlifer I. G. , Manning D. , Jones C. B. , Jones T. B. 
Published: June 2021

The role of the renin‐angiotensin system (RAS) in inflammation has been documented. However, its effect on neuroinflammation following spinal cord injury (SCI) has not yet been studied. We sought to evaluate the effect of inhibiting RAS on recovery of function and markers of inflammation following SCI. Female, Sprague‐Dawley rats were treated with losartan (angiotensin II type 1 receptor (AT1R) blocker), captopril (angiotensin converting enzyme (ACE) inhibitor) or vehicle (i.p., sid), beginning one day after lateral compression SCI. Functional recovery was assessed using the Basso, Beattie, Bresnahan (BBB) Locomotor Rating Scale. Rats were sacrificed at 7, 14 or 28 days post injury (dpi) and the tissues processed for PCR or immunohistochemical analyses. We show that rats treated with losartan or captopril have improved functional recovery compared to vehicle‐treated rats. Quantitative PCR array revealed the expression of AT1R and AT2R in the spinal cords of injured and uninjured rats and the induction of pro‐inflammatory genes over 28 dpi. AT1R appears to be localized to infiltrating immune cells following SCI. These data indicate that manipulation of RAS by blocking AT1R or ACE improves functional recovery and causes changes in gene expression in rats following SCI. Thus, further exploration of therapeutic intervention of RAS following SCI appears warranted.Grant Funding Source: Office of Research and Sponsored Programs ‐ Midwestern University
Distribution of Nicastrin in the central nervous system of APP/PS1 double transgenic mice
Authors: Luo Shifang , He Guiqiong 
Published: June 2021

Nicastrin (NCT) is an integral component of gamma‐secretase complex that also contains presenilin (PS), Aph‐1 and Pen‐2. gamma‐secretase is involved in the processing of over 60 substrates (eg. amyloid precursor protein, APP). Studies show that NCT is required for substrate recognition and selection of gamma‐secretase. However, little is known about its distribution in central nervous system (CNS) of APP/PS1 transgenic AD mice model and its wildtype controls. We report here, using immunoblotting and immunohistochemical methods, that NCT is widely expressed in all major areas of both AD and control mice, including olfactory bulb, cortex, hppocampus, basal ganglion, thalamus, cerebellum, brainstem and spinal cord. The NCT expression level is elevated in AD mice, and it is distributed much more extensively than circumscribed amyloid deposits. At the early stages of postnatal development, the NCT expression level is relatively high in CNS, but declined gradually in adult mice. At cellular level, NCT is localized predominantly in cell membrane and cytoplasma at early postnatal stages of AD mice, while only distributed at cell membrane in controls. Our results provide an anatomical basis to suggest that NCT may play a multifunctional role for gamma‐secretase in the developing and adult AD mice brain.This work was supported by National Natural Science Foundation of China (30700885)
Hypoplastic Left Heart Syndrome: Molecular Consequences of Transcription Factor Mutations
Authors: Shay Ashley Elizabeth , Kirwin Susan , Prospero Carol , Pizarro Christian , Funanage Vicky 
Published: June 2021

Hypoplastic left heart syndrome (HLHS; OMIM #241550) is characterized by abnormal hypoplasia of the left ventricle and aorta, stenosis or atresia of the aortic arch and mitral valve, an atrial septal defect, and a patent ductus arteriosus. The incidence of this rare disorder is about 1 in every 5,000 live births. These abnormalities lead to a decrease in proper blood flow from the left ventricle to the systemic circulatory system. Failure of the systemic circulation leads to profound cyanosis in patients. Without intervention, this condition is fatal within the first days of life. Studies have indicated that HLHS has a strong genetic basis when compared to other congenital heart defects. We are examining the role of several genes associated with cardiac development by screening exons and partial intronic regions for mutations by direct screening of genomic DNA from HLHS patients. Candidate genes for this study include TBX5, NKX2.5, GATA4, and GJA1 due to the correlation between mutations in these genes and cardiac defects. These results will be compared within a cohort of HLHS patients to determine if there is a genetic basis for their abnormal cardiovascular development. We have identified three mutations in exon 8 of TBX5: Val263Met, Ser276Asn, and Arg279Gln, as well as three variations within intronic regions: c. 148‐19 A/T (Intron 2), c.755‐29 C/T (Intron 7), and c.983‐8 C/T (Intron 8).Funding: Nemours Research Programs and NIH‐INBRE (Delaware)
Identification of Protein‐Interacting Partners of USF2 in Erythroid Cells
Authors: Masannat Jude , Barrow Joeva , Bungert Jorg 
Published: June 2021

Adult hemoglobin, comprised of α and β‐globin protein chains, is the major constituent of red blood cells that functions to transport oxygen around the body. Mutations of these genes lead to hemoglobinopathies such as sickle cell anemia. It has been previously demonstrated that the Upstream Stimulatory Factor (USF) activates globin gene expression. USF, composed of USF1 and USF2, is a ubiquitously expressed protein and likely interacts with erythroid‐specific activities to regulate globin gene expression. The goal of this project is to identify protein‐interacting partners of USF in order to derive a molecular pathway of adult β‐globin activation.Using the murine erythroleukemia (MEL) and the human erythroleukemia (K562) cell lines as models for erythroid globin expression, we have successfully expressed a biotin‐flag‐tagged USF protein in these cells in an inducible manner. USF interacting proteins are identified by tandem affinity purification followed by mass spectrometry analyses. Here, we present data on the novel identification and characterization of the multiple protein‐interacting partners of USF. This knowledge provides insight into developing novel therapies for hemoglobinopathies.
The promise of CD44v6, a therapeutic target for colon cancer
Authors: Misra Suniti , Hascall Vincent C. , Markwald Roger R. , Ghatak Shibnath 
Published: June 2021

ObjectiveThe proposed study links CD44v6 and its cross‐talk with growth factor receptor signaling pathways in colon tumorigenesis, and the use of colon tumor specific delivery of CD44v6shRNA through nanoparticles will suppress tumorigenesis.MethodsMechanism of sustained production of CD44v6 in response to stimulation of hepatocyte growth factor (HGF) were studied in non metastatic primary intestinal epithelial cell line Apc10.1 isolated from Apc Min/+ mice. Involvement of CD44v6 in clonogenic growth, sphere formation, motility and in vitro angiogenesis were studied in human colon rectal cancer (CRC) cell lines. CD44v6shRNA through nanoparticles were delivered intraperitonially to immunodeficient mice carrying HT29 xenograft and the effect of CD44v6shRNA on tumorigenesis and CD44v6 signaling were studied.ResultsMechanistically, CD44v6 acts as co‐receptor for c‐Met and VEGFR2 were observed. HGF effect on CD44v6 is inducible whereas CCD44v6 physically binds with VEGFR2 for its activation. CD44v6 forms a positive feed‐back loop with PI3K for sustained production of CD44v6. CD44v6shRNA reduces xenograft growth by 80–90% within ten treatments.ConclusionThe data provide new insights into growth factor induced CD44v6 splicing mechanism and its role in CRC and suggest that CD44v6shRNA/nanoparticles delivery is a promising strategy for targeting CRC tumor growth and progression.
Ligand induced activation of VEGFR during angiogenesis
Authors: Li Xiaobo , Xu Ping , Toombs Jason E. , Parker Matthew W. , Brekken Rolf A. , Vander Kooi Craig W. 
Published: June 2021

Vascular endothelial growth factor (VEGF) is one of the most potent pro‐angiogenic growth factors and a key regulator of both physiological and pathological angiogenesis. VEGF signals through VEGF receptor (VEGFR) tyrosine kinase family members. VEGFR‐2 is a main regulator of angiogenic effects of VEGF under both normal and abnormal conditions. VEGF binding to VEGFR‐2 induces specific dimerization and subsequent activation of angiogenesis. A model of VEGFR‐2 activation shows that ligand binding induces specific dimerization of the juxtamembrane D7 domain of VEGFR‐2. We report here the high‐resolution structure of the VEGFR‐2 Ig d7 dimer with a molecular interface centered around R724 that is primarily composed of salt‐bridges and hydrogen bonds. We further demonstrate a monoclonal antibody that binds to the D7 dimerization interface in a manner that is mutually exclusive with dimerization. We further show that antibody binding reduces VEGFR‐2 signaling. Taken together, these results indicate that ligand binding is necessary but not sufficient for VEGFR‐2 activation and define a new potential mechanism for therapeutic intervention.Supported by NIH Grant GM094155.
In vivo microdialysis sampling of adipokines from rat mammary fat pad. Comparison of dialysates to whole tissue concentrations
Authors: Bajpai Geetika , Stenken Julie A 
Published: June 2021

ObjectiveMicrodialysis sampling was used to collect adipokines, CCL2 (MCP‐1) and IL‐6, from mammary fat pads of female rats (Figure 1). A critical issue with microdialysis sampling is the calibration and determining actual tissue concentrations. Tissue concentrations of the adipokines were obtained and compared with dialysates.MethodsCMA 20 polyethersulfone, 100 kDa MWCO, probes were implanted in the right abdominal mammary fat pad of female Sprague‐Dawley rats (220–250g). Tissue around the probe was harvested for protein extraction. CCL2 and IL‐6 were quantified using Luminex and ELISA kits.Results and discussionConcentrations of IL‐6 and CCL2 increased after probe implantation (Table 1). Interestingly, IL‐6 levels were not different, but CCL2 levels were between the implanted vs. control tissue. This may suggest that IL‐6 levels are not influenced by the probe insertion trauma.ConclusionsAdipokines can be collected from mammary fat pad using microdialysis sampling. Up to ~50 fold differences exist between tissue and dialysate levels.Funding: COBRE 1P30RR031154‐01 and Arkansas Biosciences Institute.
Structural properties of functional HDL and variants of apoA‐I
Authors: Petrlova Jitka , Zhu Lin , Mörgelin Matthias , Hebert Hans , Jegerschöld Caroline , Voss John C. , Lagerstedt Jens O. 
Published: June 2021

Apolipoprotein A‐I (apoA‐I) is the main protein involved in the formation of high density lipoprotein (HDL). HDL is the principal mediator of the reverse cholesterol transfer (RCT) pathway and provides cardio‐protection. The RCT process involves several sub‐species of HDL where structural plasticity of apoA‐I is critical. This property, and the general high hydrophobicity of apoA‐I, has made structural analyses of lipid‐free and lipid‐bound apoA‐I challenging. We have previously used electron paramagnetic resonance spectroscopy to analyze the structure of apoA‐I [J Biol Chem 282:9143; 286:2966]. We here extend those studies by use of Cryo‐Electron Microscopy (Cryo‐EM). Cryo‐EM is a stain‐free 3D‐structure determination technique and thus highly suitable for structural analysis of the different species of HDL protein‐lipid complexes. In addition to functional apoA‐I, several variants of apoA‐I have been shown to form tissue‐specific amyloid deposits in vivo. We previously characterized G26Rvariant [Biochemistry 46:9693] and L178H variant, which possesses decreased stability. Currently, we are analyzing the structural and functional properties of the C‐terminal variant R173C that is associated with low level of HDL and in contrast, with low prevalence of developing atherosclerosis, and longevity.This work was funding by grants from the Swedish Research Council (522‐2008‐3724,7480).
Can fluorescently labeled cytokinins become a new approach for receptor domain mapping?
Authors: Szucova Lucie , Plihal Ondrej , Zahajska Lenka , Mik Vaclav , Dolezal Karel , Spichal Lukas , Strnad Miroslav 
Published: June 2021

Cytokinins belong to a class of plant hormones structurally based on adenine. They play important roles in many aspects of plant growth and development. Isoprenoid cytokinin N6‐ isopentenyladenine was accompanied with various spacers in C2, C8 and N9 position of adenine and fluorescently labeled with NBD, FITC, rhodamine B and many other common fluorescent labels. The activation of signaling pathway was assessed with Arabidopsis cytokinin receptors AHK3 and AHK4 and maize cytokinin receptors ZmHK1 and ZmHK3a in a bacterial receptor test where specific activation of the cytokinin signaling pathway (YojN> RcsB > cps::lacZ) was quantified. Although the structural changes led to the loss of the ability to activate signaling pathway through the receptors, some derivatives were still able to interact with the receptor binding site as was evident from competitive ligand receptor testing. One of the N9 substituted iP derivatives was promising in staining procedures: it was shown that the derivative was transported to the cell cytosol and the signal was associated with subcellular structures, especially with the endoplasmic reticulum, where these receptors are probably localized.The work was supported by GACR P501/10/1450.
Natural ethanol availability directed the evolution of short‐chain alcohol dehydrogenase in animals
Authors: Julián-Sánchez Adriana , Hernández-Tobías Aída , Piña Enrique , Riveros-Rosas Héctor 
Published: June 2021

The presence of alcohol dehydrogenase (ADH) in animals has been assumed to be a consequence of chronic exposure to ethanol. Therefore, in this work we investigate if natural ethanol availability directed the evolution of type II or short‐chain ADHs evolution. In animals, 15‐hydroxyProstaglandin dehydrogenase (PGDH) is the closest homologous to type II ADHs. In contrast, bona fide type II ADHs and ADH‐like proteins appear to be restricted to Diptera (flies). Thus, it can be assumed that the origin of PGDH predates the origin of type II ADHs in Diptera. Adh locus in Drosophila has undergone at least five duplication events, and all them occurred within the Diptera. Because point mutations in Adh can abolish ADH activity in this fly, it can be considered the unique fly enzyme involved in ethanol metabolism.The natural breeding substrates of Drosophila are fermenting fruits. The Drosophila genus is among the most diverse of the Diptera, encompassing >2,000 species. The efficient exploitation of such niches is the reasons for their successful worldwide spreading. Thus, induction of type II Adh by ethanol exposure, a positive correlation between ADH activity and ethanol resistance, ethanol utilization as a nutrient, and the fact that flies and type II Adh diversification occurred in concert with angiosperm diversification, strongly suggest that type II ADHs were recruited to allow larval flies to exploit new restricted niches with high ethanol content. Supported by DGAPA‐UNAM grant IN208510.
Construction of synthetic protein‐based library by combinatorial approach
Authors: Su Yue , Parameswaran Hariharan , Guan Lan 
Published: June 2021

Protein‐capture reagents have been shown to be a useful tool for scientific studies and clinical applications. We have constructed a repeat protein‐based combinatorial library for further structural and functional study of membrane proteins. This library was constructed by an ordered assembly of repeat DNA fragments containing random amino acid codons at certain positions. Theoretically, it should contain a diversity of ~2 × 1022. We obtained 45 % error‐free full‐length proteins, reducing the theoretic diversity to ~1 × 1022. Although feasible diversity size is limited by a screening method, a fraction of such library is still sufficient for many applications. By DNA sequencing analysis, 48 out of 49 correct clones are unique in protein sequence; among them, 90 % individuals have >10 positions that are different with one and another. Biophysical analyses of purified proteins from four randomly selected clones suggested correct Mr values and protein folding. Thermostability study using circular dichroism spectroscopy revealed that the melting temperatures for all four proteins are > 95 °C. Taken together, the diversity and stability of this library provide a pool of candidates for further screening of capture reagents that specifically bind to a protein of interest with high affinity.This work was supported by National Institutes of Health Grants R01 GM095538 to LG.
Use of the Medifast meal replacement program for weight loss in overweight and obese clients: A retrospective chart review of three Medifast Weight Control Centers (MWCC)
Authors: Coleman Christopher David , Kiel Jessica R. , Davis Lisa M. , Mitola Andrea , Sonzone Carmine M. 
Published: June 2021

ObjectiveA chart review was performed to evaluate the effectiveness of the Medifast meal replacement plan (MD) on body weight and body composition at 4, 12, 24 weeks (wk), and final weight loss visit (19.6 ± 13.5 wk) (FV) at 3 MWCCs for the years 2007–10.MethodsCharts included adults aged 18–70 following MD with a BMI ≥ 25 that signed a patient health information consent form. 453 charts (males n=60 (13.5%); females n= 386 (86.5%)) were included. Data were collected electronically and included weight (Wt), systolic and diastolic blood pressure (SBP, DBP), lean muscle mass (LMM), body fat mass (BFM), % body fat (%BF), and abdominal circumference (AC). Additional data collected included: demographics, side effects, and compliance.ResultsSignificant improvements (p<0.0001) were seen at all time‐points: wk 4, 12, 24, and FV. Wt loss: −12.7, −26.3, −38.2, −25.7 lbs; % Wt loss: −6.0, −12.1, −17.0, −12.3%; %BF: −2.2, −5.2, −8.5, −6.6; BFM: −10.1, −20.1, −31.6, −23.5 lbs; AC: −2.2, −4.0, −6.0, −4.7 in; SBP: −8.0, −8.6, −15.6, −8.8 mmHG; DBP: −5.3, −6.4, −9.2, −6.0 mmHG. Percent total body weight that was LMM improved by 3.5, 9.8, 16.0, and 13.9% (P<0.0001) respectively.ConclusionMedifast's meal replacement plan, along with the support available from MWCC, is an efficacious model that promotes significant weight loss and improvements in body composition. This research was funded by Medifast, Inc., Owings Mills, MD.
Insulin Resistance and Food Cravings
Authors: Fieselmann Krista N. , Sharma Rashmi S. , Maroney Owen H. , Ebner Rachel E. , McCrory Megan A. 
Published: June 2021

BACKGROUNDIndividuals with insulin resistance (IR) have been suggested to have more food cravings than those without IR, particularly sweet cravings. However, little research has been done in this area. We tested the hypothesis that individuals with IR would have more frequent and stronger cravings, perceive themselves as less able to ignore cravings, and ‘give in’ to cravings more often than those without IR.METHODSWe analyzed data from healthy, non‐smoking, weight stable adults (aged 18–58 y; BMI 19–35 kg/m2) who had participated in recent studies in our lab and reported energy intakes (EI) plausibly (within ±25% of energy requirement (Huang et al 2005)). Cravings were assessed by questionnaire (Hill et al 1991). IR was defined as homeostasis model assessment (HOMA) >1.775 (Esteghamati et al 2010).RESULTSIndividuals with IR (n=28) had higher BMI (p=0.014) and were slightly more active (p=0.08) and younger (p=0.11) than those without IR (n=34). After controlling for confounders, those with IR had 45% less frequent cravings than those without IR (p=0.04), but no other craving variables differed significantly by IR status.CONCLUSIONSOn average, cravings were not experienced differently in those with IR versus without IR. Nevertheless, perceived cravings may be important if they trigger intake of foods that may exacerbate IR. [Funding: DK075862]
The Relationship between Changes in Serum Leptin Levels and Changes in Bone Mineral Density (BMD) in Diabetes En Balance Program
Authors: Chukwueke Ihuoma Okezie , Fashola Yomi , Beeson Larry , Schulz Eloy , Firek Anthony , DeLeon Marino , Cordero-MacIntyre Zaida 
Published: June 2021

ObjectiveTo access the relationship between changes in serum leptin levels and changes in bone mineral density (BMD) in diabetic HispanicsMethodsThirty‐four Hispanic diabetic subjects participated in the three‐month study. The fan beam dual energy X‐ray absorptiometry (DXA) hologic 4500 version software was used to measure body composition. Paired t‐test was used to evaluate the mean difference between baseline and 3‐months. Non parametric test was used to evaluate the mean difference at body composition and serum leptin at baseline and three months. Spearman correlation coefficient was used to assess the change in Serum Leptin and Bone Mineral Density.ResultsSignificant negative correlations (Spearman's) were observed at three months between transformed values of leptin (adjusted for trunk fat (r = −0.369, p=0.035) and pelvis BMD; leptin (adjusted for total fat), r = −0.372, p=0.033) versus pelvis BMD. The values obtained after three months when compared to baseline were: Total fat (30.41± 10.68 vs 29.50 ± 10.52, p< 0.001), trunk fat (16.54± 5.73 vs 15.93 ± 5.60., p<0.001), serum leptin (19.55±14.99 vs 16.34 ± 12.67 p<0.002).ConclusionThere is a significant inverse relationship serum leptin and bone mineral density in diabetic Hispanics.Sponsor: CMS 03‐00335 Health Services Research and NIH award 5P20MD001632
Parity and Pre‐pregnancy Obesity are Independently Associated with Delayed Lactogenesis in Women with History of Gestational Diabetes: Preliminary Results from the SWIFT Study
Authors: Matias Susana Lilian , Chiang Vicky , Dewey Kathryn G , Gunderson Erica P 
Published: June 2021

We used baseline data (n=1014) from SWIFT, a prospective study of women with gestational diabetes mellitus (GDM) within Kaiser Permanente (KP) in California, to determine the incidence of delayed lactogenesis and its association with selected maternal prenatal and delivery characteristics. Onset of lactogenesis II was assessed at baseline (6–9 weeks postpartum) by maternal report as well as pre‐pregnancy and pre‐delivery weights, and height was measured. Delayed lactogenesis was defined as onset of lactogenesis II occurring after 72 hours postpartum. Results from a prenatal 3‐hour 100 g oral glucose tolerance test (OGTT) were obtained from KP electronic records. Severity of GDM was measured by number of abnormal OGTT plasma values. Delayed lactogenesis occurred in 33% of postpartum nondiabetic women. After adjusting for maternal, labor and delivery characteristics, primiparity and pre‐pregnancy obesity were independently associated with delayed lactogenesis in this population. High priority for early postpartum lactation support should be given to GDM women who are obese before pregnancy and primiparous.This study was funded by the National Institute of Child Health and Human Development (NICHD), R01 HD050625, R01 HD050625‐03S1, and R01 HD050625‐05S. This project was also partially supported by the National Institutes of Health National Center for Research Resources UCSF‐CTSI UL1 RR024131.