bluuebunny/crossref_metadata_2025_split huggingface dataset:
SUMO (small‐ubiquitin‐like modifier) in Tetrahymena thermophila
Authors: Matejka Stephanie Lynn , Knobbe Amy , Johnson Jacob , Weeks Donald P. , Bailey Cheryl 
Published: June 2021

Cilia and flagella organelles serve important roles in a wide variety of organisms. Outside of Chlamydomonas reinhardtii flagella, there is limited understanding of ciliary protein composition and regulation. We investigate SUMO (Small ubiquitin‐like modifier) protein modification in the cilia of the single celled protozoan, Tetrahymena thermophila. SUMO is a member of the family of ubiquitin‐like polypeptides that become covalently attached to intracellular target proteins in eukaryotic organisms. Bioinformatics have shown a single SUMO gene in the genome of this organism. Real‐time RT‐PCR of T. thermophila samples has confirmed the expression of this SUMO gene. Additionally, western‐blot analysis of ciliary extracts has shown three SUMOylated proteins of approximately 35kD, 50kD and 135kD. The 135kD SUMOylated protein is preferentially found in ciliary extracts and not in cell body extracts. This implies there are specific SUMOylated target proteins in the cilia. Fractionations of ciliary extracts have been submitted for mass‐spectrometry analysis and reveal proteins not previously found in the T. thermophila ciliary proteome.Supported by NSF and the Nebraska Research Council.
Small molecule inhibitors of PHLPP determined by chemical and virtual screening
Authors: Sierecki Emma , Sinko William , Mc Cammon J. Andrew , Newton Alexandra C 
Published: June 2021

PHLPP (PH domain Leucine repeats Protein Phosphatase) dephosphorylates Akt and protein kinase C, poising this phosphatase on two major survival pathways in cells. The catalytic domain of PHLPP belongs to the PP2C family of serine/threonine phosphatases for which there are no specific inhibitors. Using a medium throughput chemical screening combined with a virtual approach, we tested compounds from the NCI compound library and identified small molecules that inhibit PHLPP in vitro at low micromolar range. Several clusters of structures could be identified and structure‐activity relationship studies were initiated. Competitive and uncompetitive inhibitors, specific for PHLPP compared to other serine/threonine phosphatases, were used on cells. Treatment with these molecules resulted in an increase in both basal and agonist‐evoked Akt phosphorylation and activity. These first inhibitors of PHLPP could find their therapeutic relevance in the treatment of diabetes or cardiovascular diseases, two pathologies in which Akt and/or PKC pathways are repressed.This work was supported by the JDFR 3‐2008‐478, NIH 067946, NIH GM08326, NIH GM31749, NSF MCB‐0506593, and The Howard Hughes Medical Institute.
Lutein and Zeaxanthin in Red Blood Cells as a Measure of Their Status in Humans
Authors: Evans Julie A. , Johnson Elizabeth J. 
Published: June 2021

BackgroundIntake of lutein and zeaxanthin (L+Z) may reduce age‐related macular degeneration (AMD) risk. Plasma L+Z, which reflects short‐term intake, is often used to assess status. Red blood cell (RBC) level may reflect long‐term intake. Use of RBC to assess status has not been explored. Macular pigment (L+Z in retinal tissue) is considered reflective of long‐term L+Z status. Evaluation of relationships of L+Z in plasma and RBC with macular pigment density (MPD) may be useful.ObjectiveTo determine the relationship between L+Z in plasma and RBC with MPD.MethodsBlood and MPD were obtained from 15 healthy adults. Extraction and measure of L+Z in plasma and RBC were performed with standard lipid extraction methods and reverse‐phase, gradient HPLC. MPD was measured with a psychophysical method at a 1‐deg target and a 7‐deg parafoveal reference. Pearson correlation coefficients were used to measure relationships with MPD.ResultsPlasma and RBC levels of L+Z were significantly related to MPD (p = 0.050 and 0.018, respectively). The correlation was greater for RBC than plasma (r = 0.622 and 0.532, respectively).ConclusionsA greater correlation of RBC L+Z with MPD may be due to RBC and MPD reflecting long‐term intake and plasma reflecting short‐term intake. Measures of L+Z in RBC in studies examining the relationship between L+Z status and AMD may be useful. Support: USDA grant# 58‐1950‐7‐707
Adiponectin and body weight regulate colon cancer tumorigenesis in obese mice
Authors: Boddicker Rebecca Luchtel , Whitley Elizabeth , Nilsen‐Hamilton Marit , Birt Diane F. , Spurlock Michael E. 
Published: June 2021

Obesity is associated with downregulation of the anti‐inflammatory adipokine, adiponectin (Adn), and concomitant upregulation of proinflammatory cytokine production in adipose tissue. Recently, the chronic inflammatory state characteristic of obesity was causally linked to development of colon cancer. It has been suggested that Adn plays a regulatory role in this association, although results remain controversial. Consequently, we used an Adn knockout (KO) mouse model and wild type C57BL/6J mice (Wt), coupled with a diet high in saturated fat, to investigate the role of Adn in colon tumorigenesis in obesity. Mice were first administered an injection of azoxymethane (10 mg/kg body weight) and provided 1% dextran sodium sulfate in drinking water for 4 wks to induce tumorigenesis, and then provided a high fat lard (HFL) or control (Cont) diet ad libitum for 7 wks. The HFL mice gained more weight than mice fed the Cont diet, regardless of genotype. However, the KO mice had lower blood glucose concentrations, perhaps indicative of early stage hyperinsulinemia. Interestingly, total lesions per mm colon length were higher in Wt mice than KO, although lesions correlated only with body weight. Taken together, these results suggest a role for adiponectin status and body weight in colorectal tumorigenesis. Supported by grant 2008‐34115‐19372 to the Nutrition and Wellness Research Center from the USDA.Grant Funding Source: USDA
Association of Bioscavengers to Human Red Blood Cells
Authors: Calderon Frances Suiyin , Gauchan Sangeeta , Garcia Gregory Edward 
Published: June 2021

We are developing technologies to increase the in vivo retention of bioscavengers by attaching them to red blood cells (RBCs). We have examined cell‐penetrating peptides (CPPs) of 5‐40 amino acids that potentially could either anchor the bioscavengers to the cell surface or internalize them by associated transport. In addition, we tested the binding ability of RBC surface antigens (anti‐type A and B blood group) antibody‐bioscavenger conjugates. RBCs obtained from human blood were incubated with CPPs such as the protein transduction domain of Tat‐HIV (Tat‐PTD), calcitonin, and inverse antennapedia (Inverse‐Antp) biotin congeners in complex with Neutravidin‐fluospheres. No significant differences were observed between control RBCs and CPPs‐incubated RBCs. RBC binding of blood group antibodies was initially monitored with secondary antibodies conjugated to fluorescent marker CY2. Bioscavenger BChE was bound to RBCs by a bridge formed of primary antibody, biotinylated secondary antibody, Neutravidin, and finally biotinylated bioscavenger butyrylcholinesterase (BChE). The preparations were washed and the bound BChE was detected by fluorescence microscopy following incubation with CY2 labeled anti‐butyrylcholinesterase antibodies and BChE enzymatic activity. Our results show that there was significant binding of BChE to the RBCs demonstrating that antibodies can be used as an effective means to bind BChE to RBCs surfaces.This work was supported by DTRA#1.D0011_08_WR_C.
Perinatal high fat diet dysregulates GABAergic signaling to vagal efferent motoneurons regulating gastric motility
Authors: Reagan Zachary K , Browning Kirsteen N 
Published: June 2021

Parasympathetic innervation to the stomach is supplied by dorsal motor nucleus of the vagus (DMV) neurons. Studies have demonstrated that ongoing GABAergic inputs to DMV neurons play a major role in regulating vagal efferent output. The aim of this study was to investigate whether perinatal HFD affects the GABAergic regulation of vagal efferent output to the stomach. Male Sprague Dawley rats were fed a HFD (60%kcal from fat) from E13 until experimentation at 12 weeks of age. Miniaturized strain gauges sutured to the ventral surface of the gastric corpus were used to record gastric motility. The GABAA receptor antagonist, bicuculline methiodide (BMI; 60nl) was microinjected into the dorsal vagal complex and the resulting increase in gastric motility was measured at 3 time points after injection (1.5, 4 and 6.5min) and averaged. Responses were expressed relative to the maximal increase in motility in response to i.v. injection of the non‐selective muscarinic agonist, bethanecol (353pmol; 1ml), which itself was unaffected by diet (9.08±1.20g in control vs 9.27±1.03g in HFD; P>;0.05).In control rats, BMI induced a dose‐dependent increase in gastric motility (3.1±0.6%, 4.8±1.1% and 5.7±1.7% at 25, 50 and 100pmol, n=8,8,4, respectively, P<0.05 for each). In contrast, following HFD, the BMI‐induced increase in gastric motility was significantly attenuated (0.004±0.0001%, 1.2±0.3% and 1.25±0.3% at 25, 50 and 100pmol, n= 4,5,4, respectively, P<0.05 vs control). These data suggest that exposure to a HFD during the perinatal period alters GABAergic activity within brainstem neural circuits controlling gastric functions, which may contribute to the dysregulation of homeostatic reflexes, including appetite and metabolic regulation.Supported by NSF IOS 1148978
Chronology of Early Heat Stress Mediated Changes in Oxidative Skeletal Muscle
Authors: Volodina Olga , Ganesan Shanthi , Pearce Sarah , Gabler Nicholas , Baumgard Lance , Rhoads Robert , Selsby Joshua 
Published: June 2021

We have previously reported that 24 hours of heat stress (HS) induced free radical injury in oxidative skeletal muscle. In addition, we found widespread aberrant intracellular signaling following 12h of HS. The aim of this investigation was to extend our knowledge regarding the early chronology of HS‐mediated intracellular signalling in oxidative skeletal muscle. We hypothesized that 2, 4, and 6 hours of HS would result in increased autophagy and decreased mitochondrial content in oxidative skeletal muscle compared to muscle maintained under thermoneutral (TN) conditions. To address this hypothesis, pigs were assigned to four groups (n=8/group) and housed under TN conditions (21°C) or exposed to HS (37°C) for 2, 4, or 6 hours. Next, animals were euthanized and the semitendinosus red (STR) was collected and analyzed. Compared to TN, relative protein abundance of autophagy initiators phosphoinositide 3‐kinase Class III and Beclin‐1 were increased by 57% and 56%, respectively (p<0.05), following 6 hours of HS. At the same time point, relative abundance of the pre‐autophagosomal structure stabilizer, autophagy‐related protein 16‐1 (ATG16), was increased 59% (p<0.05). Microtubule‐associated protein A/B light chain 3 (LC3)‐I was similar between all four groups but the autophagosome formation marker LC3‐II was increased by 94% (p<0.05) following 6 hours of HS compared to TN. This resulted in a 2‐fold increase in the LC3II/I ratio (p<0.05), which is indicative of increased autophagic flux, though sequstome‐1 (p62) protein abundance was similar between groups. In addition, the mitophagy marker, BCL2/adenovirus E1B protein‐interacting protein 3‐like (BNIP3), was increased by 41% (p<0.05) following 6 hours of HS compared to TN. Relative protein abundance of TCA enzymes, electron transport chain components, and mitochondrial markers was similar between groups. These data demonstrate that short‐term HS induced autophagy/mitophagy. Together with our previous work we also suggest thatbetween 6 and 12 hours of HS the intracellular environment of oxidative skeletal muscle switches from one that promotes autophagy to one that inhibits it.Support or Funding InformationThis work supported by USDA grants 2014‐67015‐21627 and 2011‐67003‐30007.
Take Your Medicine; It's Good For You. Teaching Quantitative Principles To Biologists
Authors: Springer Michael 
Published: June 2021

Recent advances in automation and technology have changed the nature of life science research. Experiments now often generate large amounts of quantitative data – far too much for a person to sort through or to analyze by hand. Both academic institutions and funding agencies have recognized the need for programming and quantitative skills, but most life sciences educational programs have not yet integrated these skills into their curriculum. Compounding this problem, the life sciences, and in particular biology has historically attracted students who were often the least quantitatively inclined science majors. I will discuss our efforts to integrate quantitative methods into our curriculum, with an emphasis on using active learning approaches to engage and motivate students.Support or Funding InformationThis work was supported by the Bertarelli Program in Translational Neuroscience and Neuroengineering and by NSF grant 1349248
A Chemical Glycoproteomics Platform To study the Human Prostate Secretome
Authors: Spiciarich David R. , Purcell Sean , Maund Sophia L. , Peehl Donna M. , Bertozzi Carolyn R. 
Published: June 2021

Accurate diagnosis and treatment of prostate cancer demands a highly specific biomarker of both the disease, progression and prognosis. As glycoproteins have proven to be altered in most cancers, we have focused on studying glycoproteins in human prostate tissue samples. We have developed an efficient method for labeling and enriching glycoproteins, followed by MS‐based analysis of normal and carcinoma tissue. We sought to selectively enrich secreted glycoproteins because of the potential of secreted glycoprotein biomarkers in non‐invasive prostate cancer detection. This is accomplished by culturing human prostate tissue derived from radical prostatectomies, at varying states of prostatic disease, and collecting the cultured media which samples secretome proteins. During culturing, tissue is chemically tagged with an azide functionalized monosaccharide, N‐azidoacetylmannosamine‐tetraacylated (ManNAz). Labeled proteins are subsequently reacted with a biotin‐functionalized probe for the capture of azido‐sialylated glycoproteins. After analysis of confirmed azide‐dependent labeling, glycoproteins were digested and analyzed on a linear ion trap mass spectrometer with collision‐induced dissociation (CID). Greater than 80% of the proteins identified by LC‐MS/MS were secreted proteins, indicating that this strategy achieves enrichment of low abundance proteins. Additionally, a comparative proteomics analysis between diseased and healthy tissue identified proteins unique to each tissue type, which are undergoing validation as potential biomarkers. Virtually all clinically approved cancer biomarkers are cell‐surface or secreted glycoproteins and this work extends the application of existing metabolic labeling techniques, while developing new methodologies for cell surface labeling to discover biomarkers associated with prostate cancer.Support or Funding InformationThis work was funded by grants from the NIH to C.R.B. (GM066047 and GM058867) and to S.L.M. (2T32DK007217‐37). D.R.S. is supported by NSF predoctoral fellowships.
Systematic Review of Child Obesity Prevention and Treatment Trials Addressing Parenting
Authors: Hubbs‐Tait Laura , Kimble Ashley , Hingle Melanie , Novotny Rachel , Fiese Barbara 
Published: June 2021

A systematic review was conducted to determine whether child obesity/overweight treatment and prevention trials that address parenting result in significant changes or differences in child BMIz, BMI percentile, weight status, or related outcomes. On May 14, 2015 Pubmed, PsycINFO, Web of Science, and Scopus were searched with the following search strategy “parenting OR parent OR mother OR father AND intervention OR RCT OR trial AND parenting style OR feeding style OR responsive feeding OR restrictive feeding AND child obesity OR child overweight OR child weight status OR child waist circumference OR child BMI OR child BMI percentile OR child BMI% or child BMI z OR child BMI centile OR child BMIZ or child BMI‐z,” yielding 244 unique articles. Abstract and title screening (α = .90) excluded all protocols as well as completed studies restricted to adolescents. Full text review resulted in 16 studies that included some aspect of parenting in the trial and also provided weight outcomes for children. Study sample was obese and/or overweight children in 12 studies; the remaining four included children from the full range of weight statuses. Ten of 16 studies reported a significant effect of intervention on child BMIz, BMI percentile, or other weight outcome. All 10 studies with significant effects were focused on intervention and treatment of obese/overweight children rather than on obesity prevention. Only two of the 10 were focused solely on children ages 5 and under. Results suggest obesity intervention trials that address parenting styles and similar constructs have been successful, particularly for older children. Additional efforts to increase the success of interventions for the toddler and preschool periods are needed.Support or Funding InformationUSDA/NIFA Hatch/Multistate and Oklahoma Agricultural Experiment Station OKLO2849; Oklahoma Agricultural Experiment Station OKLO2868
Alterations of Gut Microbiota Prior to Changes of Paneth Cell‐antimicrobial Peptides in High Fat‐induced Obese Mice
Authors: Guo Xiulan , Li Jinchao , Tang Renyong , Liu Wei , Liu Zhenhua 
Published: June 2021

It is under intensive argument about the reciprocal relationship between gut microbiota and the altered intestinal metabolic and inflammatory status in the obese state. Gut microbiota composition and Paneth cells is closely associated, especially in obesity. This study aimed to investigate the mutual interaction between the Paneth cell‐produced antimicrobial peptides expression and shift of the intestinal microbiota in obesity. Eight male C57BL/6 mice were fed a low fat diet (LF, 10%) as the control and 24 animals were fed a high fat diet (HF, 60%) and sacrificed sequentially with 8, 12, 16 weeks of feeding. The expression of antimicrobial peptides and the composition of gut microbiota were determined at each stage. The mice with 8‐wk HF feeding had higher fat percentage and significant changes of the gut microbiota comparing to the LF control group (p < 0.05). The abundances of Firmicutes and Bacteroidetes from 8‐ to 16‐wk HF feeding were steady, with a higher abundance of Firmicutes and a lower of Bacteroidetes. However, 8‐wk HF diet feeding did not changed the Paneth antimicrobial peptides: lysozyme (LYZ), α‐defensin 5, and RegIIIγ (p > 0.05), but reduced LYZ and RegIIIγ mRNA expression was observed in animals with 12‐ and 16‐wk HF feeding (p < 0.05), and the results for LYZ protein level, measured by immunohistochemistry, also indicated a decrease after 16‐wk HF feeding. These results indicated that the high fat diet initially alters the profile of gut microbiota in obese mice, and changes in the expression of antimicrobial peptides occurs secondary to alterations in the composition of the intestinal microbiota.Support or Funding InformationThis project is supported by National Natural Science Foundation of China (Grant No.31401488), USDA grants (2014‐67017‐21762 and MAS00454), and Deans Incentive Grant.
Dietary supplementation with tart cherries for prevention of inflammation‐associated colorectal cancer in mice
Authors: Hunter Ashli , Larson Deanna , Phatak Sumira , Hintze Korry , Benninghoff Abby 
Published: June 2021

The cherry fruit is a nutrient‐dense food with comparatively low caloric content and significant amounts of key nutrients and bioactive food chemicals. Much of the health benefit of cherries is attributed to their high amounts of anthocyanins, which have anti‐oxidant and anti‐cancer properties that contribute to changes in cell signaling pathways involved in inflammation, carcinogenesis and angiogenesis. In this project, we aimed to determine whether dietary supplementation with tart cherries prevents colon tumor development in mice consuming a Western diet compared to a prudent diet. Previously, our research team developed the Total Western Diet (TWD) for mice to emulate typical U.S. nutrition with respect to macro‐ (sugars, fats, and protein) and micronutrient (vitamin and mineral) contents (Hintze, et al. 2012 J Ag Food Chem 60(27): 6736–6742). Incorporating the TWD in pre‐clinical studies allows for the analysis of the impact of dietary bioactives or functional foods on tumor development in a physiological environment that reflects nutritional patterns of the average American. We hypothesized that dietary supplementation with freeze‐dried whole tart cherries would suppress development of colon tumors in a model of inflammation‐associated colorectal cancer. A 2×2 factorial design was employed, whereby mice were fed either AIN93G (optimized for rodent health) or the TWD, each with and without Montmorency tart cherry powder added to the diet for a total anthocyanin content of 188 mg/kg diet. Mice were initiated with 10 mg/kg azoxymethane and provided 1% dextran sodium sulfate (DSS) in their drinking water for 4 weeks to promote colonic inflammation and tumorigenesis. Necropsy and tumor assessment were performed after 15 weeks of treatment. TWD consumption markedly enhanced colitis (40‐fold increase in disease activity) and tumor multiplicity (near 6‐fold increase) compared to consumption of the AIN93G diet. Supplementation with tart cherry powder caused a significant 40% reduction (P<0.05) in tumor incidence in mice fed AIN93G, whereas tart cherries had no effect on tumor incidence in mice fed TWD. However, tart cherry powder supplementation did not significantly affect tumor multiplicity or tumor size in mice fed either basal diet. Moreover, addition of the tart cherry supplement did not significantly affect colitis disease activity. These data contrast with a prior observation by our group that green tea supplementation was effective at reducing development of aberrant crypts, but only in mice fed TWD. Collectively, these observations point to important interactions between basal diets and dietary bioactive supplements and underscore the need for careful consideration of the role of basal diet in dietary chemoprevention studies in rodents.Support or Funding InformationSupport was provided by the Utah Agricultural Experiment Station (project UTA‐01178) and USDA NIFA Grant No. 2014‐67017‐21755 to A.B.