bluuebunny/crossref_metadata_2025 huggingface dataset:
Origin, development, function, and evolution of cypriniform trophic novelties
Authors: Hernandez L Patricia
Published: June 2021

Cypriniform fishes are characterized by a number of evolutionary trophic novelties. These feeding novelties have allowed cypriniforms to take full advantage of a number of unique trophic niches. One such feeding innovation, the palatal organ, is a complex muscular structure located on the pharyngeal roof of cypriniform fishes. The palatal organ is a dorsal mass of complexly arranged muscle fibers within the buccopharyngeal cavity innervated by the vagal lobe and covered by a taste bud coated epithelium. Here we use developmental data from zebrafish, functional data from carp and goldfish, and histological data and modeling approaches from assorted cypriniform species to address how the palatal organ developed and explain some of its functional evolution. Using zebrafish we first describe the origin and ontogenetic development of the palatal organ. Comparative data on histological structure and myosin composition of the palatal organ across a wide phylogenetic sampling of cypriniforms is then used to determine the variation in structure and potential function of the palatal organ across Cypriniformes. The general assumption has been that the function of the palatal organ is conserved across cypriniforms, and requires the careful sensory control made possible by a hypertrophied vagal lobe to function properly. Few have considered the possibility that the palatal organ may have become adapted for different functions during the course of cypriniform evolution. Its possible role in other feeding modes has been largely overlooked as it has been presumed that the palatal organ's only function is in sorting during benthic feeding events. Using electromyography and high‐speed video to examine palatal organ activity in the common carp and goldfish, muscle activation patterns demonstrate that the palatal organ is likely employed just prior to suction generation, during suction feeding behaviors, during prey processing events, and may even play a role in respiration during stressful conditions. Furthermore, the palatal organ shows behavioral modulation when different food types and positions are used. These data suggest that the palatal organ is a multi‐functional structure with a much wider range of functional repertoires than previously shown. While sorting during benthic feeding may have been the primitive function of the palatal organ, it has likely been secondarily adapted for increasing dietary breadth during the course of cypriniform evolution. This muscular pad has become secondarily modified for different forms of feeding but may even play a more generalized role in suction feeding and prey processing within cypriniforms.
PMJs and TMJs: convergence in the craniomandibular joints of crocodilians and mammals
Authors: Holliday Casey , Sellers Kaleb , Porro Laura , Ross Callum , Witmer Lawrence , Vickaryous Matthew
Published: June 2021

Crocodyliforms evolved a series of key features responsible for their Mesozoic adaptive radiation. Among these is the pterygoid buttress, the characteristic hypertrophied pterygoid flange in the skull. Although historically approached simply as a palatal element, the buttress forms a prominent articulation with the mandible here referred to as the pterygomandibular joint (PMJ). The joint bears the necessary anatomical features, such as a capsule and fibrocartilage, to be considered an enthesis organ. Biomechanically, the joint acts as a fulcrum between the dentary and jaw muscle attachments and stabilizes the mandible from medial bending and long axis rotation during orthal and twist feeding. Although all jaw muscles act about the jaw joint (i.e., the quadratoarticular joint), they also create strong mediolaterally and vertically‐oriented moments about the pterygomandibular joint such that this secondarily‐acquired craniomandibular joint is heavily loaded during most feeding behaviors. The joint originates early in pseudosuchian evolution prior to the evolution of an akinetic skull, laterally‐wrapping pterygoideus muscles, sutured mandibular symphysis, secondary palate, and skull flattening suggesting it was the key factor that stabilized the mandible and released the lineage to evolve high bite forces and robust skulls. Finally, the crocodyliform pterygomandibular joint bears resemblance to the eutherian temporomandibular joint as both joints possess sesamoid‐like fibrocartilages, such as the TMJ articular disc, within evolutionarily conserved muscle attachments. Thus, crocodyliforms and mammals convergently evolved dual craniomandibular joint systems with similar joint morphologies albeit at different locations. These new findings enable significant new insights into cranial biomechanical modeling, skeletal development and vertebrate evolution.
New Insights into Biosynthesis of Cuticular Wax
Authors: Kunst Ljerka
Published: June 2021

The outer epidermal cell walls of plant shoots are covered with a cuticle, a continuous lipid structure that protects plants from desiccation, UV‐light, pathogens, and insects. The cuticle is composed of cutin polymer matrix embedded and coated with waxes. Major wax components are derived from very‐long‐chain fatty acid (VLCFA) precursors with chain lengths between C24 and C34. We are taking advantage of wax‐deficient eceriferum (cer) mutants in Arabidopsis to identify and functionally characterize gene products involved in wax biosynthesis, wax export to the cuticle, and regulation of these processes. I will describe our work on the cer2 mutant, which resulted in the identification of a novel component of fatty acid elongation machinery required for the extension of acyl chains beyond C28. This process is unique to plants where C30‐C34 VLCFAs are used for the production of cuticular wax and the pollen coat, critical for pollen hydration. The core VLCFA elongation complex involves four proteins. Condensing enzymes catalyze the first elongation reaction and determine the chain length of fatty acid products. While necessary for the production of all VLCFAs, known condensing enzymes cannot produce VLCFAs longer than C28. Heterologous expression in yeast revealed that CER2 protein could modify the chain‐length specificity of a condensing enzyme, CER6, from C28 to C30. We identified two CER2 homologs in Arabidopsis. Results of our investigation of the specificities and physiological roles of this protein family will be discussed.
Reprogramming Factor OCT4 is Crucial for Transdifferentiation of Hepatocytes to Biliary Epithelial Cells.
Authors: Bhave Vishakha , Doffou Mboya , Adams George , Bowen William , Donthamsetty Shashikiran , Michalopoulos George
Published: June 2021

Transdifferentiation of liver epithelial cells (hepatocytes and biliary cells) into each other provides a rescue mechanism in liver disease under situations where either cell compartment fails to regenerate by itself. The mechanisms underlying such transdifferentiation of terminally differentiated epithelial cells are not fully elucidated. Recently we reported that adult rat liver expresses reprogramming factors OCT4 and its expression is important for hepatocyte proliferation and liver regeneration. To test if reprogramming factor OCT4 plays a role in transdifferentiation of hepatocytes to BECs, we used the hepatocyte organoid culture. In this model, primary hepatocytes when plated on collagen coated roller bottles and cultured in presence of HGF, EGF, and dexamethasone, transdifferentiate to form BECs between 6‐8 days in culture. We found that OCT4 is upregulated at day 6 in this model as assessed by mRNA and protein levels suggesting its possible involvement in transdifferentiation. When expression of OCT4 was blocked in hepatocytes using adenovirus containing OCT4 shRNA, there was a significant decrease in the number of biliary cells as compared to controls as assessed by biliary marker HNF1‐beta, strongly supporting the role of OCT4 in transdifferentiation of hepatocyte to biliary epithelial cells. Moreover, OCT4 inhibited group showed significantly less SOX9 expression as compared to controls. Experiments to investigate mechanisms through which OCT4 might regulate transdifferentiation and mechanisms upstream of OCT4 are under way.
MiR‐376c Inhibits Metastasis by Controlling RUNX2‐INHBA Axis in Oral Squamous Cell Carcinoma
Authors: Chang Wei‐min , Shiah Shine‐Gwo , Hsiao Michael
Published: June 2021

Aberrant microRNA expression is a common phenotype in cancer cell progression. MicroRNA is the negative regulator of hundred to thousand gene expressions. Here we report chr14q32.2 miRNA cluster significantly down‐regulated in clinical OSCC samples. 19 miRNAs were found with coordinated silencing in OSCC patients. However, their specific target and regulation mechanism have not clearly identified. Here we showed miR‐376c is the most significant microRNA down‐regulated by RT‐Q‐PCR validation in the miRNA cluster. Ectopic expression of miR‐376c not only inhibited oral cancer cells proliferation, but also migration and invasion. MiR‐376c also inhibited the in vivo tumorigenesis and metastasis. Furthermore, miR‐376c was found directly binding to the 3′‐UTR of oncogenic transcription factor, RUNX2, which was found significantly up‐regulated in OSCC patients. Silencing of RUNX2 inhibited OSCC proliferation and invasion through INHBA expression. Taken together, our study demonstrates the tumor suppressor role of miR‐376c, which regulates RUNX2 transcriptomes in OSCC. Moreover, our results indicate RUNX2‐INHBA axis may play important roles in OSCC progression.
On Modulating the Regulatory Role of Intrinsically Disordered Proteins Using Small Molecules
Authors: Camacho Carlos
Published: June 2021

Intrinsically disordered regions (IDRs) in proteins play a variety of critical roles in complex regulatory networks of higher organisms. Most importantly, perhaps, is that they are necessary to provide specificity in promiscuous interactions. I will show that both the specificity of IDRs for their targets and their entropy loss upon binding are essential to provide discrimination among similar physical interactions. Attempts to temper with this class of interacting proteins using small molecules will necessarily comprise densely functionalized compounds. Implications for the challenges involve in the rational design of small molecules targeting IDRs will be discussed.
Sirtuins and Novel Protein Post Translational Modifications
Authors: Lin Hening
Published: June 2021

Sirtuins are a class of enzymes known as nicotinamide adenine dinucleotide (NAD)‐dependent deacetylases. Sirtuins regulate aging, transcription, and metabolism, and are important targets for treating several human diseases. There are seven sirtuins in humans, SIRT1‐7. Four of them (SIRT4‐7) have very weak deacetylase activity, which have caused many confusions and debates in the biological community. My laboratory has recently discovered several novel enzymatic activities, such as desuccinylation and defatty‐acylation, for several sirtuins with no robust deacetylase activity. This have led to the identification of previously unknown protein posttranslational modifications and revealed new biological functions of sirtuins, such as the regulation of protein secretion.
Using Facebook to Engage Biology Undergraduates and Keep Alumni Connected
Authors: Halpin Patricia
Published: June 2021

The UNH Manchester Biology Program launched its Facebook (Fb) page, “UNHM Biology Majors,” in 2012. Many of our students were using Fb, and we immediately saw the value of Fb as a virtual bulletin board where all relevant communications could be posted in one place. Like many professional Fb pages, this page is closed to non‐members, and only Biology majors – students and alumni – are allowed to join. For our non‐residential campus, the page has become an important way to keep our undergraduates engaged in our science community on campus, and our alumni connected with our current students. Examples of posts include notices of advising information sessions for students interested in graduate programs in health professions and veterinary medicine. The Biology Club posts their meeting announcements and events. The page also connects our commuter campus to events on the main campus. Another use is linking undergraduates with alumni who can provide information on careers and graduate programs. Current students and alumni have also provided valuable advice to first‐year students about making the transition to college and studying for their first college science course. Opportunities for volunteer work, research, independent study, internships, jobs, study abroad, and scholarships are posted. To keep the community connected, good news items regarding awards, scholarships, entrance to graduate school, and jobs of undergraduates and alumni are posted. In the summer, students use Fb to stay in contact with each other, and those working on campus are shown in posts describing their projects. To date over 250 members have joined and continue to be involved. This positive response indicates that Fb will continue to be a valuable resource for engaging our undergraduates and keeping alumni connected.
The use of Independent‐Study Modules to Review Areas of Misconception in Physiology
Authors: Laudadio Rachel
Published: June 2021

Several authors have noted areas in which students continuously hold misconceptions in physiology. This may be due to personal experience, an inability to transfer a general principle to more specific scenarios, or imprecise (or frankly deficient) explanations by texts, cartoons, or professors. Known topics of concern include: 1. Homeostasis, 2. Cardiovascular and pulmonary changes occurring during exercise and ascent to altitude, 3. The difference between oxygen saturation and oxygen partial pressure, 4. How lung CO2 content (and pressure) change with different percentages of inspired O2, and 5. The relationship between metabolic activity and ventilation. Focusing on subjects of difficulty and being cognizant of potential complications, short modules which students could review independently were provided along with quiz questions, both general and applied, to test their understanding of the material.
An Approach to Teaching Histology and Embryology Efficiently in English in China
Authors: Sha Ou , Guan Youfei , Xie Miao , Liu Zhigang , Fan Xinmin
Published: June 2021

Aims

The aim of this article is to encourage further discussion on different approaches to equip students in a non‐English environment to study medical topics in English by reporting on the initial experience and valuable lessons learned teaching Histology and Embryology in English at the medical school of Shenzhen University of China.

Methods

The course contents of traditional histology and embryology were rearranged into integrated general and system courses, and were taught in English. Various initiatives have been introduced to increase the teaching and learning efficiency. The histology and English teaching staff collaborated to support the teaching of histology and embryology in English. The final students mark was analyzed. Three questionnaires were designed to evaluate from an academic perspective and the student's attitude and perception of the efficacy of methods employed.

Results

The majority of the students (85%) reported they understood the histology lecture in English and (79%) hoped to continue to learn histology in English. Communicating in a social setting with English speaking people ranked as the primary aim to learn English, achieving a high GPA (grade point average) score second. 98% agreed that it is important to learn histology in English for their professional careers, 67% prefer to be exposed to the topic and terminology in English lessons prior to or at the same time as the histology lesson. Discussing in English a related topic, labeling diagrams, and analyzing the anatomy of medical terms ranked as the top three preferred tasks.

Conclusion

The experience gained thus far at Shenzhen Medical School has been very valuable and provides a sound foundation to further explore this important topic (GDJG20141144, 0000233202).
Undergraduate Research ‐ a Key to Advancing Interest in STEM
Authors: Carroll Margaret , Catapane Edward , Skeete Dereck
Published: June 2021

In 2006 Medgar Evers College received an NSF grant (0622197 of the DUE Program) designed to increase the number of students earning BS degrees in Biology and Environmental Science. The goals of our STEP into Science program were to: recruit new students and non‐STEM students into Biology or Environmental Sciences; improve retention by providing academic, financial and mentoring support; foster integration of research and technology to better equip majors to be successful applicants to graduate/professional programs; and increase the number of students graduating with BS degrees. We use peer recruiters to attract more high school, transfer, and non‐science college students into STEM majors and place emphasis on undergraduate research experiences to increase the quality and retention of science majors through their BS degree. Since the inception of the program, STEM enrollment more than doubled. Our program evaluations consistently showed that a key factor in our successful recruitment and retention was student involvement in undergraduate research activities. The number of majors actively engaged in research has risen more than 90% with a concurrent increase in student research presentations at scientific conference, and an 87% increase in the number of students receiving external research internships and travel awards to attend national conferences. STEM graduates have also increased and many of them are continuing on to Masters and Doctoral programs in STEM and hopefully will ultimately enter rewarding careers in the science enterprise.
A Computational Study of the Contributions to the Relative Stability of the α and β Conformers of D‐Glucopyranose
Authors: Balilli Rabih , Abdulsalam Maryam , Baello Rofe , Booth Robert , Suresh Sandeep , Kasner Marc
Published: June 2021

In order to better evaluate the contributors to the conformational energy of D‐Glucopyranose, the geometry‐energy relationships for a sequence of model compounds was determined. The conformational energies of 1‐Hydroxy (equatorial) 3‐Hydroxymethylcyclohexane and 2‐Hydroxy 6‐ Hydroxymethyltetrahydro‐2H‐pyran were used to model D‐Glucopyranose. The conformational energies were determined at the B3LYP/6‐311G++(d,p) and MP2/6‐311++(d,p) levels in the gas phase and solution. The relative B3LYP energy as a function of the orientation of the substituents were determined by relaxed scans as the –CH2‐OH at C5 was rotated about the OCCO and HOCC dihedrals and the –OH at C1 was rotated about the HOCO dihedral. The lowest energy rotamers for the axial (α) and equatorial (β) conformers were used to calculate the conformational energy. Because solute‐solvent interactions have significant effects on the relative stability of the axial and equatorial conformers, the energy versus dihedral scans were repeated using a continuum solvent model with water (ε = ~80), acetone (ε = ~20) and cyclohexane (ε = ~2) selected as the solvent.
Analysis of the Helicase Activity of Escherichia Coli RNase R
Authors: Sk Tofajjen Hossain , Deutscher Murray P.
Published: June 2021

RNase R, which belongs to the RNB family of enzymes, is a 3′ to 5′ hydrolytic exoribonuclease able to digest highly structured RNA without the help of an added RNA helicase. It was previously reported that RNase R possesses an intrinsic helicase activity that is independent of its ribonuclease activity. However, the basis of this helicase activity and whether it has any relation to the ribonuclease activity is not yet known. Here, we identify a P‐loop domain consisting of Walker A and Walker B motifs present in the E. coli RNase R sequence, and show that it is responsible for ATP binding and the helicase activity. Interestingly, the Walker A motif is located in the C‐terminal region of RNase R while the Walker B motif is in the N‐terminal region indicating that the two parts of the protein must interact to generate a functional ATP binding site. Detailed analysis of the helicase activity of RNase R revealed that it is dependent on binding of ATP, but that ATP hydrolysis is not required. Thus, both ATP‐γ‐S and AMP‐PNP can function for helicase activity. While the nuclease activity of RNase R is not needed for the helicase activity, the helicase activity is important for the nuclease activity against a dsRNA substrate, particularly at lower temperatures. Although there is little effect of the helicase activity at 37⁰C, its stimulation of the nuclease activity becomes much more pronounced as the temperature is lowered. Mutation of either the Walker A or Walker B motifs or the absence of ATP eliminates stimulation of the nuclease activity confirming the role of each motif in the helicase activity. These findings indicate that the helicase activity plays an important role in the catalytic function of RNase R.

This work was supported by Grant GM16317 from the National Institute of Health.
A Quick, Affordable Method for the Quantification of Gene Isoforms
Authors: Bistulfi Gaia , Sutherland Catherine
Published: June 2021

Most genes are transcribed as several variants known as isoforms. Isoforms are transcripts from the same locus with either a different transcription start site or post‐transcriptional modifications, for example alternative splicing. As a result, isoforms might be characterized by different functions and/or stability. The differences between gene isoforms span from a few bases to several kilobases. In some cases, no region of the transcript might be specific to only one isoform, making quantification problematic. We devised a quick, reliable, one‐step method for the accurate quantification of gene isoforms, even in the latter case. We compared the isoforms' sequences using bioinformatic tools freely available online. We analyzed RNA samples with a one‐step real‐time RT‐PCR protocol, using primer sets specific to one or more isoforms and RNA standard curves as a reference. Differential analysis of the data provided reliable information on the copy number of each isoform in the original sample. This method might provide a first step in understanding the importance of differential expression of gene isoforms

This work was supported by a D'Youville College research grant.
Investigating EAAT3 trafficking through the secretory pathway
Authors: Saha Kusumika , Steinkellner Thomas , Hofmaier Tina , Kudlacek Oliver , Sucic Sonja , Freissmuth Michael , Sitte Harald
Published: June 2021

The eukaryotic secretory pathway allows synthesized proteins to leave the endoplasmic reticulum and makes its way to the cell surface where it carries out its individual functions. The intracellular pathway through the various cellular compartments comprises different coat protein complexes, namely coat protein complex II (COPII) and coat protein complex I (COPI).

The excitatory amino acid transporter 3 (EAAT3) mediates the regulation of the synaptic transmission at the glutamatergic synapse by transporting the neurotransmitter glutamate. Our goal is to understand the key mediators of the secretory pathway who play a pivotal role in transporting the EAAT3 to the surface. The work focuses on looking at the EAAT3 as a cargo within the cells and following it through the anterograde pathway mediated by COPII. The knowledge gained from this work will be able to shed some light in the pathway of EAAT3 through the cell and can be further extrapolated to the other glutamate transporter family members. Furthermore, the aim of the presented work is to understand key factors that regulate the surface expression of EAAT3 and the role they may play in health and disease.
Bile Acids and Bile Salts of the Arabian Gulf Catfish Arius Bilineatus, Val
Authors: Al‐Hassan Jassim , Oommen Sosamma , Paul Bincy , Afzal Mohammad
Published: June 2021

Catfish gall bladders were collected from freshly caught catfish. Bile solution was collected by rupturing the gall bladder. The bile solution was freeze dried and extracted with chloroform:methanol (1:2). The solvent mixture was evaporated and the residue was extracted with methanol. The extracted material was subjected to hydrolysis to remove the conjugated parts from the bile acids and together with standards were methylated. MS analysis was performed on the methylated products. The following bile acids were obtained: Cholic acid, chenodeoxycholic acid, deoxycholic acid, and lithocholic acid. Trimethylsilyl (TMS) derivatization was done on the extracted material and analyzed by GCMS to give allocholic acid. The extracted bile salts and bile acids were also separated in LC/MS QTOF using acetonitrile: water as solvent. The following conjugated bile acids were obtained: Taurochenodeoxycholic acid, glycolithocholic acid, glycodeoxycholic acid, glycocholic acid, and cholic acid conjugated with Cysteinolic acid.

(This project was funded by The Research Sector of Kuwait University Grant # SL04/09. Spectroscopic analysis was performed by Science analytical Facilities, KU).
Specific Transient Interactions Between a Bacillus Virulence Factor and Phosphatidylcholine in Membranes
Authors: Gershenson Anne , Yang Boqian , He Tao , Khan Hanif , Grauffel Cédric , Reuter Nathalie , Roberts Mary
Published: June 2021

How do phospholipase Cs associated with bacterial virulence bind to membranes and search for substrates? The virulence factor Bacillus thuringiensis phosphatidylinositol‐specific phospholipase C (BtPI‐PLC) cleaves GPI‐anchored proteins off the outer leaflet of eukaryotic cells, helping the bacteria evade host defenses. Single molecule fluorescence microscopy studies of fluorescently labeled BtPI‐PLC interacting with model membranes reveal a koff of 2.64 ± 0.34 s‐1 at 22°C. This relatively fast dissociation is correlated with shallow anchorage of the protein in the membrane revealed by molecular dynamic simulations. Experimental and computational results suggest that cation‐π interactions between phosphatidylcholine (PC) headgroups and BtPI‐PLC Tyr residues play an important role in membrane binding. To further test this hypothesis, we substituted 3,5‐difluoroTyr for individual Tyr residues, thus perturbing cation‐π electrostatic interactions while increasing Tyr hydrophobicity. We also performed 31P NMR experiments with spin‐labeled BtPI‐PLC, to determine the location of PC binding sites on the protein surface. Both sets of experiments identify the same regions as important for PC binding. These biophysical and computational studies coupled with changes in enzyme activity provide a detailed, quantitative picture of how PC interactions with BtPI‐PLC influence its transient binding, PI cleavage and the search for substrates on the membrane.
Exploring the Serine Protease Involved in Cell‐mediated Immunity in Fish
Authors: Matsuura Yuta , Yabu Takeshi , Shiba Hajime , Moritomo Tadaaki , Nakanishi Teruyuki
Published: June 2021

Viral and intracellular bacterial diseases are big problem in aquaculture. Defense against these diseases is largely dependent on cell‐mediated immunity in which cytotoxic T lymphocytes and NK cells play major roles. These cells kill cellular targets via either of two pathways, i.e. secretary (e.g. perforin/granzyme) and non‐secretary (e.g. Fas/FasL) one. In fish, biochemical mechanisms of the killing remain unknown. Here we tried to identify the protease involved in the killing adopting ginbuna crucian carp which has been used as a nice model for cell‐mediated immunity in fish. In order to identify the protease involved in the killing of allogeneic target cells, we performed cytotoxicity assays in the presence of serine or cysteine protease inhibitors using lymphocytes of ginbuna. Cytotoxicity was completely inhibited by the serine protease inhibitor “DCI”. By contrast cytotoxicity was partially inhibited with cysteine protease inhibitor, E‐64d. These findings suggest that serine proteases play a major role in the killing of target cells. We further analyzed the substrate specificity of the protease involved in cytotoxicity. Protease activity of lymphocyte lysate was enhanced by allo‐sensitization when Ac‐IETD‐MCA, Bz‐R‐MCA, Z‐GPR‐MCA, K‐MCA, and Suc‐AAPF‐MCA were used as substrates. To classify the group of protease involved in the cytotoxicity, protease inhibitor was added in enzyme assay. Serine protease inhibitor “AEBSF” completely inhibited enzyme activity only when Z‐GPR‐MCA was used. Furthermore, there was no inhibition with cysteine protease inhibitor “E‐64”. These results suggest that the serine protease which cleaves Z‐GPR‐MCA is involved in cytotoxicity.
Isolation and Characterization of Mycobacteriophage ZoeJ, a K2 Cluster Phage
Authors: Jancevski Alicia , Dubois Heloise , Scola Shelby , Lebel Eric , Pinches Seth , Brown Melissa , Cavedon William , Fernando Michelle , Perri Christina , Rogers Stephen , Austriaco Nicanor , Cornely Kathleen
Published: June 2021

Mycobacteriophages are pathogenic viruses that infect and kill mycobacteria, many of which cause diseases including tuberculosis and leprosy. Mycobacteriophage ZoeJ was isolated from a soil sample at Providence College via an enrichment procedure. Plaque morphology and electron microscopy photos suggest that ZoeJ is a Siphoviridae lysogenic phage. DNA sequencing and annotation of the genome indicates ZoeJ contains 57,315 bp and 92 probable genes. Based on homology, ZoeJ has been assigned to the K2 subcluster, a new subcluster consisting of ZoeJ, Mufasa, and TM4. Genomic comparison of the K2 subcluster has revealed a gene sequence containing an integrase which is present in both ZoeJ and Mufasa, yet absent from TM4, a phage commonly used in the study of mycobacteria. Immunity assay results suggest that the integrase of ZoeJ is functional, allowing it to form lysogens. The putative attP/attB sites of phage integration were identified for both ZoeJ and Mufasa. A PCR assay designed to amplify the regions of phage integration confirmed that the integrase in both ZoeJ and Mufasa is functional, allowing these K2 cluster phages to form lysogens.
The Total Amino Acid Profile of Human Milk is Stable Over the First Year of Lactation and Consistent Across Geographical Populations: a GEHM Study
Authors: Labhart Beau , Gray Michael , Maria Sarah , Phillips Shay , McMahon Robert , Davidson Barbara , Morrow Ardythe
Published: June 2021

The amino acid composition of human milk is thought to be critical in the development of breastfed infants. Part of the Global Exploration of Human Milk (GEHM) study examines variability in amino acid profiles over the course of lactation, both between individual mothers and across geographical populations. Total and free amino acid profiles, determined using specialized ion exchange chromatography followed by post column derivitization, are reported for longitudinal samples collected from three global cohorts (Mexico, China, and USA) at 4, 13, 26, and 52 weeks lactation. This study focused on utilization of rigorously controlled sample collection techniques as well as statistically monitored process control during analysis to ensure the accuracy of generated results. Although the amino acid profiles of human milk have been reported previously, these extra control measures as well as the integration of numerous samples from different geographical populations provides additional insights into amino acids in human milk. While the free amino acid profiles exhibit higher variability amongst individuals, these findings suggest that the total amino acid profiles remain remarkably consistent even when investigated in distinct geographical locations, amongst multiple donors, and across lactation. This characteristic pattern, observed in all amino acids, can be illustrated by the essential amino acid Threonine (Thr), which has a variability of 55% for free Thr and 4% for total Thr. Support for this research was provided by Mead Johnson Pediatric Nutrition Institute.
Vitamin B6 Status is Associated with Depression Among Women with Inflammation
Authors: Faraj Joycelyn , Troy Lisa , Bertone‐Johnson Elizabeth , Ronnenberg Alayne
Published: June 2021

Depression is the leading cause of disease burden in U.S. women of reproductive age. Inflammation is associated with depression, yet the mechanism for this association remains unclear. Additionally, vitamin B6 status has been linked to depression and found to be compromised during inflammation. In secondary data analyses, we evaluated the association between inflammation, vitamin B6, and depression in non‐pregnant women (19‐45 yrs) from NHANES 2005‐2006. Depression was evaluated with the Patient Health Questionnaire 9 (PHQ‐9); a score of 10 or above denotes clinically significant depression. Inflammation was defined as a C‐reactive protein level above the median (1.8 mg/L). Vitamin B6, as plasma pyridoxine‐5′phosphate, was categorized as deficient (<20 nmol/L), insufficient (20‐30 nmol/L) or normal (>30 nmol/L). Overall, 7.7% of women had depression, but this proportion varied by inflammation status: 4.8% of women with no inflammation, compared to 10.4% of women with inflammation had depression (p=0.008). Vitamin B6 levels were significantly higher (82.1±5.7 vs 54.1±4.6 nmol/L, p=0.005) and PHQ‐9 scores were lower (2.6±0.2 vs 3.7±0.4; p=0.01) in women without inflammation compared to those with inflammation. The PHQ‐9 score for women with no inflammation did not vary by vitamin B6 status. However, among those with inflammation, women with vitamin B6 deficiency had significantly higher PHQ‐9 score (6.1±6.8), followed by those with vitamin B6 insufficiency (3.9±4.9), and those with normal vitamin B6 status (3.4±4.5) (p<0.01). Understanding the mechanism linking inflammation and vitamin B6 to depression could lead to new approaches to prevent and treat depression.
Self‐Weighing Increases Weight Loss in Free‐Living Adults: A Double‐Blind Randomized Field Trial among 200,000 Health App Users
Authors: Novello Sarah , Cash Sean , Roberts Susan , Masters William
Published: June 2021

Self‐weighing is closely associated with sustained weight loss, but randomized trials to test causality have been limited to small samples in controlled settings subject to Hawthorne effects. This study employs a simple behavioral nudge to induce more frequent self‐weighing among new users of the Lose It! app, randomly assigned among all individuals who enrolled over a four week period in 2014. The study population is similar in age, sex and socio‐economic status to other groups seeking weight‐loss assistance in the United States and Canada. There were no differences between treatment (T) and control (C) other than the nudge, and subjects were not told about differences between T and C. Among the 184,955 users who recorded at least one plausible data point, we find mean self‐weighing frequency (days) was T=4.27±0.07 and C=3.85±0.07, and mean weight loss (lbs) was T=1.95±0.04 and C=1.90±0.04, for a difference in weight loss of 0.054±0.052 over the first three months after enrollment. Using two‐stage least squares to estimate the causal effect of self‐monitoring, we find that each additional weigh‐in led to an average increase in weight loss of 0.131±0.119 lbs. These results suggest that encouraging frequent self‐weighing can improve the efficacy of weight loss programs at a national level, at very low additional cost.
Obesity Significantly Intensifies the Depressive Symptom among Adults with Neck or Back Pain
Authors: Lawrence Wayne , Badana Adrian , Barnett Gloria , Sexton Jessica , Oliver Kirsten , Hogges Jamesa , Zhang Jian
Published: June 2021

Objective

Obesity and back or neck pain are the leading chronic conditions among adults in the United States, yet the interaction between these two conditions and the role in the development of depressive symptom have not yet been investigated.

Methods: National Health and Nutrition Examination Survey data from 2009‐2010 of 1,952 men and 1,971 women aged 20 years or older was used to calculate odds ratios (OR) of depression among normal, overweight, and obese men and women that reported a presence or absence of neck or back pain.

Results: The prevalence of moderate/severe depression were 1.9 (SE: 0.18) and 3.46 (SE: 0.27) percent among men and women, respectively. Using neck or back pain‐free adults with similar body mass index (BMI) level as the reference, the odds ratios (OR) of depression were 2.60 (1.04‐6.51), 3.03 (1.10‐8.37), and 11.6 (6.01‐22.5) among normal, overweight, and obese men that reported neck or back pain. The ORs of depression were 2.28 (1.08‐4.81), 2.97 (1.14‐7.80), and 3.05 (2.03‐4.58) among normal, overweight, and obese women that reported neck or back pain. These estimates were obtained after adjustments for relevant health history and socioeconomic factors.

Conclusions

Excess body weight significantly intensifies the depressive symptoms and increases the prevalence of depression among adults diagnosed with neck or back pain. Obesity prevention may improve the both mental and physical well‐being among adults.
Quantifying the Bioefficacy of β‐Carotene‐biofortified Sorghum Using a Mongolian Gerbil Model
Authors: You Hong , Zhang Yinghui , Zhao Zuo‐Yu , Che Ping , Albertsen Marc , Glassman Kimberly , White Wendy
Published: June 2021

β‐Carotene‐biofortified transgenic sorghum is being developed to alleviate vitamin A deficiency in semiarid regions of Africa. Our objective was to quantify the bioefficacy of the biofortified sorghum using a Mongolian gerbil model. Gerbils were fed a vitamin A (VA)‐free control diet (45% by wt non‐transgenic sorghum flour) for 4 wk; 8 animals were then killed at baseline. The remaining animals were randomly assigned to treatments for 6 wk (n = 11 per group): 1) control diet dosed daily with cottonseed oil vehicle; 2) biofortified diet (45% by wt transgenic sorghum flour) dosed daily with cottonseed oil vehicle (95.8 nmol retinol equivalents/d); 3) control diet dosed daily with β‐carotene (84.5 nmol retinol equivalents/d); 4) control diet dosed daily with VA (84.5 nmol retinol equivalents/d). Liver VA stores in the baseline (1.27 ± 0.29 μmol), β‐carotene‐dosed (1.30 ± 0.26 μmol), and biofortified (1.31 ± 0.23 μmol) groups were not different; liver stores in the vehicle‐dosed control group were lower (0.92 ± 0.22 μmol) (P < 0.03). Liver stores were highest in the VA‐dosed group (2.48 ± 0.23 μmol) (P < 0.0001). The calculated bioconversion efficiency for the β‐carotene in the biofortified sorghum (4.3 μg β‐carotene to 1 μg retinol) was similar to that of the β‐carotene dose (3.8 μg to 1 μg retinol). The β‐carotene in the biofortified sorghum effectively restored liver vitamin A stores in an animal model.
Examining the causal effects of changes in iron status on brain and attentional functioning using structural equation modeling
Authors: Wenger M , Rhoten S , Scott S , Murray‐Kolb L , Haas J
Published: June 2021

Iron deficiency (ID) in otherwise healthy adults can impair perceptual and cognitive functioning, along with brain dynamics (as measured by EEG). We used structural equation modeling (SEM) to test the hypothesis that changes in iron status lead to changes in brain functioning and then to changes in behavioral measures. The data used here were taken from 54 ID (sFt <15 µg/L) participants in a randomized double‐blind efficacy study. Blood, behavioral, and EEG measures were taken at baseline, endline, and at one of eight random time‐points in between. Five measures of iron status, behavioral data from three tasks measuring attentional capture and control, and peak amplitudes of a set of event related potentials (ERP) measures taken during the tasks at baseline were used as indicators of separate latent constructs in a set of contrasting SEMs. Thus far, a model with three latent factors provides the best description of the data, with the three latent factors being (a) systemic iron (sFt and sTfR) directly influencing (b) brain dynamics (ERP amplitudes), with brain dynamics then influencing (c) attentional function (reaction times from the three tasks). This model provides a better description of the data than does a model representing no causal link from blood to brain to behavior, or a model in which oxygen delivery (as indicated by hemoglobin, red blood cell distribution width, and mean cell volume) influences brain dynamics and then attentional functioning. These results suggest the utility of SEMs to relate changes in blood, brain, and behavior.
Aqueous cinnamon extract protects against oxygen‐glucose deprivation‐induced damage in bEnd.3 cells
Authors: Qin Bolin , Zhao Aiping , Liu Chao , Anderson Richard
Published: June 2021

Oxidative stress may contribute to vascular endothelial injury leading to impaired cerebral vascular function following stroke related to diabetes. A cinnamon extract (Cinnulin PF) has been shown to reverse the negative effects of oxidative stress. The aim of this study was to investigate the effects of Cinnulin in an oxygen glucose deprivation (OGD)‐induced injury model, of mouse brain microvascular endothelial (bEnd.3) cells. Cell viability and the expression of the Bcl‐xl; a structural integrity protein, HSP90; and TNFα and phospho‐Nf‐kB p65 were evaluated. A 3h OGD/24h reperfusion insult decreased cell viability and induced Bcl‐xl, HSP90, TNFα, and phospho‐p65 proteins, while the addition of Cinnulin significantly attenuated these effects. A 3h OGD treatment decreased glucose uptake measured by fluorescence microscopy which was also reversed by Cinnulin. PGE2 is an important mediator of many biological functions, including vasodilation, antiinflammatory actions. OGD/24h reperfusion down regulated PGE2 secretion into the media and cellular PGE2 expression. Cinnulin attenuated these effects and also induced SIRT1 protein expression, which is required for the inhibition of apoptosis and inflammatory effects. These effects were inhibited by SIRT1 inhibitor IV. In summary, Cinnulin protects against ischemic injury in bEnd.3 cells and thus cinnamon extract may be beneficial in oxidative stress related chronic diseases.
Myricetin Ameliorates Liver Steatosis in Diabetic Obese Mice
Authors: Kim Jung‐In , Jin Ga‐Yeong , Kang Soo‐Jung , Lee Ah‐Yeon
Published: June 2021

Non‐alcoholic fatty liver disease (NAFLD), which includes simple fatty liver (hepatic steatosis), non‐alcoholic steatohepatitis, and advanced fibrosis and cirrhosis in the absence of alcohol abuse, is the most common cause of chronic liver disease. The purpose of this study was to investigate the protective effect of myricetin against fatty liver in diabetic obese mice. Four‐week‐old male C57BL/KsJ‐db/db mice were fed AIN‐93G diet or a diet containing myricetin at 0.08% of the diet for 7 weeks after one week of adaptation. The animals were sacrificed after an overnight‐fast. Liver samples were homogenized in saline, and total lipids were extracted. Total lipids and triglycerides in the liver and serum alanine transaminase (ALT) activities were measured. Hepatic contents of total lipids and triglycerides were significantly lowered in the myricetin group compared with the control group (p<0.05). Consumption of myricetin also significantly reduced ALT activities compared with the control group, suggesting that myricetin can improve liver function. These results indicate that myricetin could exert protective effects against development of NAFLD in obese db/db mice. This work was supported by the National Research Foundation of Korea (NRF) Grant funded by the Korean Government (MSIP) (No. 2012015763).
Pneumococcal protein PspA facilitates Streptococcus Pneumoniae‐induced pyrotosis
Authors: Jung Yun Joo , Pyo Suhkneung
Published: June 2021

Pyroptosis is a process in which a programmed cell death occurs via caspase‐1‐dependent mechanism which is triggered by various pathological stimuli. Streptococcus pneumoniae is gram‐negative bacteria that bring about a variety of serious diseases. D39 is serotype 2 strain of Streptococcus pneumoniae and virulent. D39 is widely used to study pathogenesis of Streptococcus pneumoniae. Nevertheless, proteins which are involved in Streptococcus Pneumoniae‐induced pyroptosis are not revealed yet. The cleavage of procaspase‐1 into the active form of caspase‐1 was reduced in PspA mutant‐infected BV2 cells, compared with D39‐infected BV2 cells. Western blot analysis demonstrated that the expression of IL‐18 was decreased in PspA mutant‐infected BV2 cells, compared with D39‐infected BV2 cells. When BV2 cells were treated with various doses of PspA recombinant protein alone, no significant change in cell proliferation was observed, comparing with uninfected cells. On the other hand, the cell proliferation of BV2 cells treated with both PspA recombinant protein and 1:10 of MOI of D39 was gradually decreased with time and this change is not statistically important when comparing with D39 only‐infected cells. This suggests that PspA is unable to act as a pyroptosis‐inducing factor. Immunoblotting analysis indicated that the expression of NLRP3 was increased in the presence of D39 or PspAmutants, compared with uninfected BV2 cells. Western blot analysis also demonstrated that the expression of NLRP3 was increased in D39‐infected after 30 min of infection, whereas an increased expression of NLRP3 in PspA mutant‐infected BV2 cells was exhibited but later than 30 min of infection. Taken together, these results suggest that pneumococcal PspA facilitates pyroptosis via NLRP3 expression in BV2 cells.
in vivo Tracking and Effects of SPIO‐labeled Mesenchymal Stem Cells in a Porcine Model of Abdominal Aortic Aneurysm
Authors: Li Jennifer , Lee Dong , Han Daniel , Rawal Bhakti , Briley‐Saeboe Karen , Mani Venkatesh , Gillespie Virginia , Hajjar Roger , Fayad Zahi , Tadros Rami , Faries Peter
Published: June 2021

Abdominal aortic aneurysm (AAA) pathogenesis involves extracellular matrix remodeling, degeneration, and inflammation, making it a promising target for cell‐based therapy. This study demonstrates, in a large animal model, the feasibility of tracking implanted mesenchymal stem cells (MSC) on in vivo MRI with Superparamagnetic Iron Oxide (SPIO) labeling and then characterizing potential reparative effects.

Dosimetry and cell viability experiments with SPIO were performed on porcine BM‐MSCs. Tracking of SPIO‐labeled MSCs was performed with 3T MRI. Histological changes in iron, elastin, collagen, and smooth muscle actin were assessed, and levels of inflammatory markers were quantified using RT‐PCR.

Optimal imaging and cell viability were achieved with SPIO conc of 500ug/mL. SPIO‐labeled MSCs demonstrated positive CD90 and negative CD45 markers, consistent with the MSC phenotype. MRI tracking demonstrated circumferential migration of the implanted MSCs. Histological analysis demonstrated an increase in collagen, elastin, and smooth muscle cell synthesis in treatment groups compared to controls, as well as mild decrease in inflammatory markers such as MMPs ‐2, ‐3, ‐9, IL‐1B, and TNF‐alpha.

MSCs can be successfully labeled, implanted, and tracked by in vivo MRI in a large animal model of AAA. The MSCs may stabilize AAA degeneration through the production of new collagen, elastin, and smooth muscle cells, as well as decreasing inflammation.