bluuebunny/crossref_metadata_2025 huggingface dataset:
Desensitization of cough during exercise in anesthetized rabbits
Authors: Poussel Mathias , Bosser Gilles , Tiotiu Angelica , Chalon Bernard , Chenuel Bruno
Published: June 2021

The ‘cough center’ exhibits plasticity at the sensor and integration levels leading to modulation of the reflex in its strength or in its pattern. Little is known about interactions between cough and human activities, especially during exercise. The present study was designed to determine whether exercise, mimiked by electrically‐induced muscular contractions, is able to change the occurrence and/or the strenght of cough following a mechanical stimulation of the trachea in anesthetized rabbits.

Thirteen anesthetized, tracheotomized rabbits were studied allowing 329 tracheal stimulations : 208 at rest and 121 during exercise.

During exercise, the incidence of the cough reflex (CR) decreased and the expiration reflex (ER) increased (p < 0.0001). Whereas the sensitivity of the CR and the ER are both decreased during exercise compared to the sensitivity of the CR at rest (p < 0.02), the strength of the expulsive response is unchanged.

These results indicate that adjustments occurring during exercise likely down‐regulates tracheal defensive reflexes in anesthetized rabbits.
Persistent activity and interactions in the brainstem respiratory network during hyperventilatory apnea
Authors: Nuding Sarah Catherine , Segers Lauren S. , O'Connor Russell E. , Dean Jay B. , Bolser Donald C. , Morris Kendall F. , Lindsey Bruce G.
Published: June 2021

Brainstem respiratory neuron interactions during hyperventilatory apnea (HA) and the dissolution and reemergence of the motor pattern for breathing remain poorly understood. We used multi‐array recording, cross‐correlation, and gravity clustering methods to evaluate spike train data from the ventral respiratory column (VRC), pontine respiratory group, and medullary raphé nuclei of decerebrate vagotomized neuromuscularly‐blocked cats. Phrenic activity, blood pressure, end‐tidal CO2, and arterial blood gas measures of O2, CO2 and pH were monitored. During HA: (i) tonic activities persisted in subsets of VRC inspiratory (I) and expiratory (E) neurons; (ii) short‐time scale correlations indicated interactions among neurons that expressed E and non‐respiratory modulated discharge patterns during control conditions, including neurons with functional inhibitory actions on I neurons; (iii) selective stimulation of carotid chemoreceptors by close injection of CO2‐saturated saline transiently evoked rhythmic respiratory network activity and suppressed tonic E neuron activity; (iv) respiratory and slower (0.04 – 0.1 Hz) network oscillations were disrupted. We conclude that VRC tonic E neurons contribute to I drive inhibition during HA (cf. Sears et al. 1982) and document the loss of multiple rhythms during HA. Support: NIH grant NS19814
Sustained apnea induced by GABAA receptor blockade at ventrolateral pons/A5 region is reversed by subsequent blockade of glutamatergic receptors
Authors: Song Gang , Ning Yinghui , Poon Chi‐Sang
Published: June 2021

The A5 region in ventrolateral pons (vl‐pons/A5) is thought to be an auxiliary substrate for the putative pneumotaxic center in the dorsolateral pons that modulates inspiratory‐expiratory phase switching (inspiratory off‐switch), post‐inspiratory activity (post‐I) and expiratory duration (TE). In urethane‐anesthetized, vagotomized and ventilated adult SD rats, we observed that blockade of GABAA receptors at vl‐pons/A5 by unilateral microinjection of bicuculline (BIC) caused prolongation of vagal post‐I motor activity and TE, an effect that was similar to that evoked by electrical or chemical stimulations in this region. Bilateral microinjections of BIC caused further protracted prolongation of TE or even sustained apnea with complete cessation of phrenic activity and prolongation of post‐I activity. Remarkably, the apnea so induced was reversed and rhythmic phrenic discharges gradually resumed upon subsequent blockade of excitatory glutamatergic receptors by bilateral microinjections of CNQX and AP‐5 at the same vl‐pons/A5 sites, or by the application of hypoxic or hypercapnic stimuli. These findings indicated that the vl‐pons/A5 plays an important role in promoting post‐I activity and inspiratory off‐switch subject to a delicate balance between GABAA receptor‐mediated inhibitory and glutamatergic receptor‐mediated excitatory modulations. (Supported by HL093225)
Responses of vestibular nucleus neurons to hindlimb movement in decerebrate cats
Authors: Arshian Milad S , Puterbaugh Sonya R , Cotter Lucy A , Yates Bill J , McCall Andrew A
Published: June 2021

Several lines of evidence suggest that both vestibular and proprioceptive inputs play an important role in maintaining balance stability during locomotion. However, little research has been devoted to how these two signals are integrated by the nervous system. In the current study, we tested the hypothesis that limb movement affects the firing rate of vestibular nucleus neurons in decerebrate cats. Recordings were made from vestibular nucleus neurons during rhythmic flexion and extension of the hindlimb as well as static positioning of the limb. Over half of neurons in the caudal aspect of the vestibular nuclei responded to limb movement, with some being sensitive to the velocity of flexion or extension, and others having firing rates proportional to static limb position. The firing rate of other units changed during both flexion and extension, such that the neuron's sensitivity to vestibular inputs was altered throughout the step cycle. There was an inverse relationship between the gain of responses to limb movement and whole‐body tilt (vestibular stimulation). These findings show that proprioceptive inputs from the limbs profoundly affect the sensitivity of vestibular nucleus neurons to inputs from the inner ear, and underscore that the vestibular system integrates multiple sensory inputs regarding body position in space.
Basolateral transport of bicarbonate by avian renal proximal tubule cells in culture
Authors: Ghanem Kahina , Laverty Gary
Published: June 2021

Bicarbonate (HCO3−) is an important component of acid/base regulation. Therefore, it is crucial to understand the mechanism of bicarbonate reabsorption in the kidney proximal tubule (PT). In mammals HCO3−reabsorption is well understood, but evidence suggests that birds may use a different mechanism. We hypothesize that in the avian PT HCO3−crosses the luminal membrane in ionic form, rather than as CO2, and that HCO3− ions then leave the basolateral side via an NBC1‐like transporter, as in the mammalian PT. Experiments used primary cell cultures of chick PT and electrophysiological studies to measure currents (ISC) associated with ion transport. Monolayers were first stimulated with 1μM forskolin, which activated a chloride secretory current in the avian PT. This was followed by basolateral application of 100 μM DIDS, an inhibitor of NBC1. With bicarbonate in the bathing solution DIDS caused an increase in ISC of 6.25 ± 1.55 μAmps/cm2 (n = 6), but only 2.30 ± 0.58 μAmps/cm2 (n = 5) in the absence of bicarbonate. The increased ISC is consistent with inhibition of electrogenic transport of HCO3− ions via the NBC1 transporter, and the decreased effect of DIDS in the nominal absence of bicarbonate further supports this. Additional studies support both mRNA expression and NBC1 protein expression (western blotting) in chick PT cultures and native tissue. Experiments are underway to test for an alternative apical transporter.
Ano6 regulates C2C12 myoblast proliferation and differentiation
Authors: Zhao Piming , Xu Li , Mariano Andrew T. , Han Renzhi
Published: June 2021

Anocatamin 6 (Ano6) is a member of TMEM16, a conserved gene family predicted to code for eight transmembrane proteins with putative Ca2+‐activated chloride channel activity. Ano6 mutations lead to Scott syndrome (SCTS) in patients, a bleeding disorder caused by a defect in Ca2+‐activated Ano6‐mediated phosphatidylserine exposure in platelets and other blood cells. However, the function of ANO6 remains unknown in most other tissues including skeletal muscle. Here we show that Ano6 regulates muscle cell proliferation and differentiation using C2C12 cells as a model system. Ano6 is expressed in adult mouse skeletal muscle and C2C12 myocytes. To investigate the functions of Ano6 in muscle cells, we established Ano6 knockdown (KD) C2C12 cell line using lentiviral shRNA targeting Ano6. Ano6 KD significantly reduced C2C12 myoblast proliferation and promoted myoblast differentiation to form myotubes, suggesting that Ano6 plays an essential role in muscle growth. In the future, we will further define the mechanisms underlying the roles of Ano6 in muscle cells.
RNA degradation is elevated with age‐, but not disuse‐associated skeletal muscle atrophy
Authors: Shah Aman , Confides Amy L , Richards‐White Jena , Dupont‐Versteegden Esther E
Published: June 2021

Aging and inactivity are both associated with decreased muscle size and protein content. The possible role of RNA degradation in the loss of protein has not yet been investigated. Therefore, we hypothesized that RNA degradation was elevated with muscle atrophy in aging and disuse. Brown Norway/Fisher344 male rats at 6 and 32 months were hindlimb suspended (HS) for 14 days to induce muscle atrophy or remained weight bearing (WB). Cytosolic extracts from gastrocnemius muscles were prepared for Western analysis of Dcp‐2 protein (marker of p‐bodies) and RNA degradation assay. In vitro total RNA decay assay was performed using 30μg of total RNA (from tibialis anterior) incubated with 20μg of S15 extracts from gastrocnemius. RNA integrity was determined using the Agilent Technologies algorithm to calculate the RNA Integrity Number (RIN); decay rate and half‐life were calculated for each sample. Results indicated an increase in Dcp‐2 protein at 32 months of age in both HS and WB groups. In addition, an almost 2‐fold increase in decay rate and 48% decrease in half‐life of total RNA was observed in muscle from 32 month old rats. However, no significant difference in decay rate and half‐life was observed with disuse at either 6 or 32 months. We conclude that muscle atrophy associated with aging, but not disuse, may be due to a decrease in total RNA because of increased RNA degradation. Supported by APS UGSRF and AG028925.
The role played by Nav1.8 channels in the somatic pressor chemoreflex
Authors: Stone Audrey J , Kim Joyce S , Kaufman Marc P
Published: June 2021

Thin fiber muscle afferents (Aδ and C) are supplied by NaV 1.8 channels, whereas thick fiber afferents (spindles) are not. We determined the role played by NaV 1.8 channels on the thin fiber afferents evoking the reflex pressor response to femoral arterial injections of lactic acid (24mM; 0.1mL) and capsaicin (0.1ug) in decerebrated rats with patent or ligated femoral arteries. This was done before and after femoral arterial injection of the A803467 (500μg, 1mg, and 2mg), a NaV 1.8 antagonist. We also recorded the responses of spindles to succinylcholine (200μg) and stretch before and after injecting A803467. In “freely perfused” rats (n=6), A803467 (1mg) significantly reduced the pressor response to lactic acid (16±5 to 4±4ΔmmHg) and capsaicin (47±7 to 26±8ΔmmHg). In “ligated” rats, 1 mg of A803467 significantly reduced the pressor response to lactic acid (32±13 to 3±3ΔmmHg, n=6) but not to capsaicin, which required a dose of 2mg (66±11 to 37±17ΔmmHg, n=4). Surprisingly, we found that A803467 (1mg) reduced the responses of 10 spindles to succinylcholine (34±11 to 4±3 Δ imp/s p=0.04) and stretch (83±17 to 0.4±1 Δ imp/s; p<0.01). We conclude that A803467 reduces the reflex response to lactic acid and capsaicin; however, it may be working in part on channels other than NaV 1.8. One possibility is the T type calcium channel, whose blockade with Mibefradil also attenuated the responses of spindles to stretch and succinylcholine.
Inflammation reduces the response to forskolin and expression of the NaHCO3 cotransporter, NBCe1, in the proximal colon of IL10−/− mice
Authors: Fan Shujun , Schultz Michael , Butt Grant
Published: June 2021

The cause of the reduced anion secretion in the inflamed intestine is unknown. In this study, the effect of proximal colonic (PC) inflammation on electrogenic anion secretion and expression of associated transport proteins were investigated. Interleukin10‐knockout (IL10−/−) mice infected with Helicobacter typhlonius were used as a model of inflammation and uninfected, asymptomatic IL10−/− mice served as controls. Electrogenic anion secretion was measured in the Ussing chamber and protein expression determined with immunohistochemistry and Western blotting. The short circuit current (Isc) response to forskolin was reduced in the inflamed PC (ΔIsccon=99.4±13.5μAcm−2; ΔIsc inflamed=60.3±10.9μAcm−2, X±SEM, n=8, P<0.05). This was due to a decrease in the 4,4′‐diisothiocyanatostilbene‐2,2′‐disulfonic acid‐sensitive (500 μM serosal [s]) component of the response, with no change in the bumetanide‐sensitive (100 μM s) component. Consistent with this, NKCC1 expression was not altered by inflammation, whereas NBCe1 expression was markedly (P<0.001, n=6) reduced, particularly in cells in the upper half of the crypts. This suggests inflammation reduces colonic HCO3− secretion, which may affect luminal pH regulation and growth of luminal bacteria, as well as modifying mucus hydration. The resultant dysbiosis and altered intestinal barrier is likely to exacerbate the inflammation. Supported by the OMRF.
Klotho Gene Deficiency Exacerbates Early Diabetic Nephropathy
Authors: Lin Yi , Suh John , Sun Zhongjie
Published: June 2021

Diabetic nephropathy is the most common cause of end stage renal disease. Previously, we reported that when challenged with streptozotocin (STZ), Klotho mutant (KL+/−) mice had higher blood glucose levels compared with wild type counterparts. Here, we explored the role of Klotho, anti‐aging protein, in a mouse model of STZ‐induced early diabetic nephropathy. Male KL+/− mice (6 to 8 weeks) were injected with multiple low doses of STZ. Urinary excretions of albumin and blood glucose levels were measured. Kidneys were collected for histological examination and molecular assays of transforming growth factor‐β1 (TGFβ1) and mammalian targets of rapamicin (mTOR) signaling. Deficiency of Klotho in KL+/− mice exacerbated STZ‐induced increases in urine albumin, blood urea nitrogen levels, expansion of mesangial matrix in glomeruli, and kidney hypertrophy, suggesting a protective role of Klotho in kidney functions and structures. Furthermore, we found that deficiency of Klotho significantly enhanced STZ‐induced TGFβ1 signaling and mTOR signaling in kidneys. These data suggested deficiency of Klotho may exacerbate diabetic nephropathy via enhancing both TGFβ1 and mTOR signaling in kidneys. Thus, Klotho gene or protein delivery may have therapeutic potential for early diabetic nephropathy.
Types of tracheal submucosal glands: implications of literature controversy
Authors: Barros Mirna Duarte , Landim Amanda P , Dobre Ana Clara M , Andreotti Joao Duvilio Biazzi , Curcio Daniella Franco , dos Anjos Duarte Ana Lúcia
Published: June 2021

In what concerns the morphological description of tissues and organs there are some conceptual discrepancies in the classical literature used as the knowledge foundation of basic sciences in health and medical colleges. Such controversies may create conflicts and jeopardize the relationship of trust between students and faculty members. This paper compares basic histology books and atlases with respect to the main information on tracheal subucosal glands. We analyzed 23 books and 21 atlases (from which 17 were virtual atlases), focusing information regarding presence and classification of tracheal submucosal glands. All references agreed that exocrine glands were present in the tracheal submucosa, however we found controversy about its types: presence of “seromucous glands” were referred by 39.1% of the references; presence of “mucous and serous glands”, by 26.1%; “mucous glands with serous components”, 17.4%; presence of both “mucous” and “seromucous”, 4.3%; “mixed glands” 4.3%; mucous, serous and mixed glands, 2.2%; mucous only, 2.2%; serous only, 2.2%. No information on gland type could be found in 2.2% of the references.

CONCLUSION

There is no consensus in what concerns the classification of the glands present in the trachea submucosa. The lack of consistence in basic information and terms may be a source of problems in standardization of histological terminology. Furthermore, such controversy may affect student's confidence in the subject and the staff, since the reliability of the theoretical basic knowledge is questioned.
Comparison between Student Desired Grades and Actual Final Grades in Undergraduate Human Anatomy
Authors: Dunham Stacey
Published: June 2021

Anat A215, Basic Human Anatomy, is a large enrollment (400+) lecture/lab course offered each fall and spring semester at Indiana University. Each semester students complete a short survey in which they indicate name, major, standing in school, previous anatomy coursework, and the lowest grade they would want for the course.

In this retrospective study the information provided by students in these surveys was compared to their final course grade earned in Anat A215, as recorded in the instructor grade files. The first goal of this study was to identify how accurately students are predicting their own grades. Second, this study intended to identify any associations between ability to accurately predict course grades and the variables included in the survey, including demographic information and students’ reported desired grade and corresponding rationale for earning that grade (to maintain GPA, to raise GPA, to get into specific undergraduate or graduate programs). Results show that of 350 students, 40% scored lower than their stated “lowest” desired grade. Further analysis will look for trends impacting both successful and failing grade predictions by students.

Grant Funding Source: None
Learning human neuroanatomical structure and function using a computer‐based learning tool.
Authors: Severson Arlen R , Forbes Donna J
Published: June 2021

An interactive computer‐based neuroanatomical learning tool is being used in an integrated neuroscience course to facilitate learning neuroanatomical structures, relationships, and significant morphological and functional features of the human central nervous system. Gross images of intact and dissected brains, sections of the whole brain, brain stem and spinal cord, and MR images are included. Neural structures to be identified are listed on the right‐hand side of the computer screen. Clicking on the name of a specific structure produces a transparent overlay of that structure. Structures can be identified also by moving the pointer over the image and clicking on a specific structure. With either approach, the structure is identified by a colored overlay and a collapsible dialog box at the bottom on the screen that provides a brief description of the structure and its functional role in the nervous system. The learning tool is also available for the iPad. Data from student surveys and comments indicate that this is an effective way to learn the anatomical and functional aspects of the nervous system that is available on a 24/7 basis. With less instructional laboratory time, limited laboratory materials, and fewer faculty available for laboratory instruction, the computer‐assisted learning tool accomplishes our educational goals and is enthusiastically accepted by the students. (Supported by MMF)
Coordinated Degradation of Replisome Components Ensures Genome Stability Upon Replication Stress
Authors: Noguchi Eishi , Roseaurlin Laura , Toda Takashi
Published: June 2021

The stabilization of the replisome complex is essential in order to achieve highly processive DNA replication and preserve genomic integrity. Conversely, it would also be advantageous for the cell to abrogate replisome functions to prevent inappropriate replication when fork progression is adversely perturbed. However, such mechanisms remain elusive. Here we report that replicative DNA polymerases and helicases, the major components of the replisome, are degraded in concert in the absence of Swi1, a subunit of the replication fork protection complex. In sharp contrast, ORC and PCNA, which are also required for DNA replication, were stably maintained. We demonstrate that this degradation of DNA polymerases and helicases is dependent on the ubiquitin‐proteasome system, in which the SCF‐Pof3 ubiquitin ligase is involved. Consistently, we show that Pof3 interacts with DNA polymerase ε. Remarkably, forced accumulation of replisome components leads to abnormal DNA replication and mitotic catastrophes in the absence of Swi1. Swi1 is known to prevent fork collapse at natural replication block sites throughout the genome. Therefore, our results suggest that the cell elicits a program to degrade replisomes upon replication stress in the absence of Swi1. We also suggest that this program prevents inappropriate duplication of the genome, which in turn contributes to the preservation of genomic integrity.
Vaccination of BALB/c Mice Against a Murine Hookworm Model
Authors: Schlotterback Mark William , Comella Phillip , Horien Corey , Neutzling Kari , Luk Kevin , Richards Elizabeth , Pearson Jonah
Published: June 2021

Intestinal hookworm infections are a significant medical problem affecting nearly 600 million people worldwide. Our laboratory is investigating the use of a secreted parasitic protein as a potential vaccine against human hookworm. This project demonstrates a pre‐clinical trial designed to examine the safety, tolerability and efficacy of five novel, different peptide and recombinant protein vaccines. Peptide design relied heavily on bioinformatics analysis of protein conservation across human‐parasitizing species of hookworm, as well as antigenicity, hydrophilicity and flexibility of potential peptide sequences. These sequences were prepared as vaccines with KLH‐conjugation and the recombinant protein vaccine was isolated by nickel affinity chromatography of its His‐tag. Each vaccine was administered to seven age and sex‐matched Balb/c mice, followed by identical boosts 14 and 28 days later, with serum samples collected each day of immunization. Challenge infections were administered 14 days after the last boost and efficacy was assessed based on adult worm burden levels 14 days after infection. Of the five unique constructed vaccines, one repeatedly yielded statistically significant protection and demonstrated unique serological profiles of antigenicity and immunogenicity, exhibiting pre‐clinical protection against the mouse model of this chronic and debilitating disease.
Beta‐arc 2 hydrogen bond network within hemolysin A facilitates hemolytic activation
Authors: Glasgow Evan , Weaver Todd
Published: June 2021

In the current study, the hemolytic exoprotein from Proteus mirabilis, termed HpmA, was cloned in a truncated form (HpmA265) and assayed to characterize its biological activity and thermal stability. A template‐assisted hemolytic assay measured the ability of native HpmA265 and six mutant variants to facilitate the refolding of denatured full‐length HpmA into its active conformation, allowing it to lyse RBCs. Circular dichroism spectroscopy was employed to estimate the denaturation temperature (Tm) of native HpmA265 and the six mutant variants (each of which harbored one or more serine substitutions). The mutations were located in the region of the protein called beta‐arc 2, which forms the base of the beta‐helical architecture of the protein, a structure common to many proteins secreted via the two‐partner secretion pathway in Gram‐negative bacteria. Tm changes ranged from −4.0 to −12.8°C among the mutants, and hemolytic activation (by template‐assisted refolding of full‐length HpmA) was markedly decreased in the triple and quadruple mutants, but activity remained near the native parameters for the single and double mutants. This suggests that an integrated network of hydrogen bonds in beta‐arc 2, with cooperative interactions in the hydrophilic pocket and the exterior carboxyamide ladder, lend substantial structural stability to HpmA265. This research is supported in part by NSF‐RUI 1050435 (to TW).
Kinetic mechanism of human DNA ligase I
Authors: O'Brien Patrick J
Published: June 2021

DNA ligase I (Lig1) is essential for DNA replication and repair. It catalyzes the joining of single‐strand DNA breaks via three chemical reactions. In the first step the enzyme is activated by transferring the adenylyl group from ATP to an active site lysine; in the second step the adenylyl group is transferred to the 5′‐phosphate at the nick; in the third step the new phosphodiester bond is formed and AMP is released. We used transient kinetic approaches to determine the minimal kinetic mechanism for human Lig1 in the presence of Mg2+ and other divalent metal ions. Although all three reactions catalyzed by Lig1 require divalent metal ions, individual steps exhibit different affinity for metal and are susceptible to inhibition by different metal ions. These different patterns of inhibition suggest that there are multiple divalent metal ion binding sites and that the metal sites change throughout the reaction cycle. This work provides deeper insight into the mechanism of DNA ligation and illustrates how divalent metal ions can inhibit a single enzyme via very different mechanisms. The results have implications for abortive ligation, which generates blocked (adenylylated) 5′ends and for the inhibition of DNA ligation by transition metals.
Association of gestational diabetes mellitus with occurrence of Autism: A Cohort Study
Authors: Al‐Farsi Yahya M , Al‐Khaduri Maha , Al‐Sumri Hanaa , Al‐Farsi Omar , Al‐Sharbati Marwan , Waly Mostafa , Ouhtit Allal , Al‐Shafaee Mohammed
Published: June 2021

Background

Several antenatal and perinatal adverse events have been reported to be potentially associated with increased risk of occurrence of autism spectrum disorder (ASD). Among these is the gestational diabetes mellitus (GDM).

Objectives

To evaluate the potential association between GDM and ASD.

Methods

A retrospective cohort study has been nested on a community trial titled as “Delaying the Development of Diabetes Mellitus type 2 (AMAL study)” in Oman. The study population included 532 women with a total of 3,196 pregnancies. Multi‐variate Cox proportional hazard regression analyses were conducted to explorer the association between GDM and ASD, adjusting for important confounders. The main outcome measure was hazard ratio (HR) of the effect of GDM on ASD.

Results

A total of 42 ASD cases have been enumerated over 3714 person‐years of follow up. In the crude model, pregnancies complicated with GDM carried a higher risk of ASD children than pregnancies without GDM (HR = 2.3; 95% CI = 1.5, 3.4). the adjusted model, the association persisted even with control of the confounding effect of maternal age (HR = 1.71; 95% CI = 1.1, 2.3).

Conclusion

This study provides a supportive evidence that GDM may carry an increased risk of occurrence of ASD. It calls for more strict control of GDm, especially that diabetes mellitus and obesity have reached alarming epidemiologic rates in Oman and the region.
Cytotoxic properties of alpha‐synuclein/fatty acid complex
Authors: Kim Doohun
Published: June 2021

Neurodegenerative diseases such as Alzheimer's disease (AD) and Parkinson's disease (PD) can be attributed to the specific degeneration of neuronal cells in the brain. However, the natures and action modes of toxic species remain largely unknown. Here, we present a simple and fast method for the preparation of neurotoxic complex with á‐synuclein, which is implicated in PD.
The Multiple Solvent Crystal Structures Method of P22TSP
Authors: Smith Mychal D , Mattos Carla , Barbirz Stefanie
Published: June 2021

The multiple solvent crystal structures (MSCS) method uses organic solvents as molecular probes to map the binding surfaces of proteins. MSCS was used to map the surface of tailspike protein from Salmonella phage P22 (P22TSP). P22TSP is a viral adhesion protein with receptor binding and destroying functions. P22TSP structures were solved to 1.9 angstroms resolution or better. Crystals of P22TSP were soaked in solvent/aqueous buffer mixtures for data collection with the following organic solvent contents: 50% glycerol; 60% acetone; 60% isopropanol; 50% 2‐ methyl‐2,4 pentanediol; 50% tert‐butanol; 50% dimethylsulfoxide; 50% dimethylformamide; 1M trimethylamine‐N‐oxide; 1M glucose; and neat hexanediol. Structures were solved using the Crystallographic Object—Oriented Toolkit (COOT) and Pythonbased Hierarchical ENvironment for Integrated Xtallography (Phenix) for refinement. The MSCS map depicts hydrophobic pockets not only within P22TSP's known active site, but also at a remote location that constitutes the active site found in evolutionarily related tailspike family members.
Physical Model of the Staphylococcus aureus Transpeptidase PBP2a in Complex with an Anti‐Methicillin‐Resistant Staphylococcus aureus Cephalosporin
Authors: Assad Fatima , Bhiki K. , Eusebio D. , Flores E. , Paulino M. , Ramirez A. , Ramirez H. , Valentin T. , Zaman M. , Sussman J. L. , Lovering A. L. , Westblade L. F. , Granberry A.
Published: June 2021

Transpeptidases, also known as penicillin‐binding proteins (PBP), catalyze cross‐linking of peptidoglycan polymers during bacterial cell wall synthesis. The natural transpeptidase substrate is the D‐Ala‐ D‐Ala peptidoglycan side chain terminus. Beta‐lactam (β‐lactam) antibiotics, which include the monobactams, penicillins, cephalosporins and carbapenems, bind and irreversibly inhibit transpeptidases by mimicking the D‐Ala‐D‐Ala substrate, resulting in the inhibition of cell wall synthesis and ultimately bacterial cell growth. Overuse and misuse of β‐lactams has led to the generation of methicillin‐resistant Staphylococcus aureus (MRSA) isolates that have acquired an alternative transpeptidase, PBP2a, which is neither bound nor inhibited by β‐lactams. MRSA isolates are resistant to all β‐lactams, can be hospital‐ or community‐acquired, and are the cause of significant morbidity and mortality. Furthermore, they are often only susceptible to “last resort antibiotics”, such as vancomycin. Recently, two cephalosporins ‐ ceftobiprole and ceftaroline ‐ that bind and inhibit PBP2a have been developed. The Hostos‐Lincoln Academy SMART Team (Students Modeling A Research Topic) has generated a model of the PBP2a/ceftobiprole complex (PDB 4DKI) using 3D printing technology to illustrate the mechanism of action of ceftobiprole. Supported by a grant from the Camille and Henry Dreyfus Foundation
Analyses of Patatin‐like Phospholipase Domain‐containing 2 (PNPLA2) Transcripts: Discrepancies between Expressed Sequence Tag (EST) and Experimental Data
Authors: DesJardin Jacqueline Talea , Notari Luigi , Becerra S. Patricia , Subramanian Preeti
Published: June 2021

Pigment epithelium‐derived factor‐receptor (PEDF‐R), a PNPLA2 protein found in the retina, has phospholipase A2 (PLA2) activity that is enhanced upon binding PEDF, a retina survival factor. Amino acid D166 forms part of the PLA2 catalytic dyad required for activity and is encoded on PEDF‐R exon 4a (E4a). Interestingly, EST databases predict that PEDF‐R has multiple isoforms: one with an E4a deletion in mouse sequence, and another with an exon 6 (E6) deletion in mouse and rat sequences. It is unknown whether alternatively spliced PEDF‐R transcripts exist in the retina. Here, we explored the proposed PEDF‐R isoforms in cell lines of retinal origin (R28, 661W) and in various mouse tissues. We designed PCR primer pairs that encompassed the predicted E4a deletion and/or E6 deletion to amplify the full length and predicted splice variants. The PCR products resolved by gel electrophoresis showed a single band consistent with full length PEDF‐R rather than splice variants. Additionally, we performed western blots of R28 and 661W cell extracts and found that multiple PEDF‐R antibodies detected only a single band migrating as expected for full length PEDF‐R. The data point to a single transcript and no PEDF‐R alternative spliced variants. Considering ESTs offer partial gene coverage and possible sequence errors, our results underlie the importance of validating conclusions drawn from EST databases. Supported by NIH‐NEI‐IRP.
Translocation of Mitochondrial HSP60 Precedes Cytochrome C Release Following Cerebral Hypoxic‐Ischemic Injury in the Neonatal Rat: A Role in Apoptosis Induction?
Authors: Reynolds Christian , Jones Michelle , Przyklenk Karin , Sanderson Thomas
Published: June 2021

The mitochondrial HSP60 is unique among the family of heat shock proteins in that it has pro‐survival and pro‐apoptotic functions. Here we demonstrate that translocation of HSP60 occurs during/immediately following hypoxic‐ischemic (HI) insults to the neonatal rat brain. We employed a rat model of neonatal Encephalopathy, in which 7day old pups underwent unilateral common carotid artery ligation followed by 120min of hypoxia (8% O2). Injury was assessed using triphenyltetrazolium chloride staining 24hour after HI. The hemisphere ipsilateral to the ligation (hypoxic‐ischemic) exhibits a robust injury comprising 30% of the hemisphere with primarily cortical involvement. Notably, the hemisphere contralateral to the ligation (hypoxic) is uninjured. To examine the effects of cerebral HI on cytosolic accumulation HSP60 and cytochrome C (cytoC), mitochondrial and cytosolic fractions were isolated via differential centrifugation immediately following the insult and after 4hrs of recovery. Samples were analysed via western blot. Normoxic, uninjured littermates were used as controls. HI results in a 2× increase of HSP60 immunoreactivity in the cytosol immediately following the insult. Furthermore, cytoC release was not observed at this early time point. These data suggest that translocation of HSP60 may play a role in the induction of apoptosis following cerebral HI in the neonatal rat.
Comparing endocranial ontogenetic trajectories in extant great and lesser apes
Authors: Scott Nadia , Neubauer Simon , Gunz Philipp , Hublin Jean‐Jacques
Published: June 2021

Differences in adult morphology arise as a result of modifications to ancestral developmental patterns. Studies of evolutionary changes to brain development in primates can provide important clues about the evolution of human cognition, but are hindered by the lack of preserved neural tissue in the fossil record. As brain structure is mirrored in the endocranium, we have used endocasts (virtual casts of the endocranial cavity) to capture brain shape in a geometric morphometrics analysis. Here, we compare the postnatal developmental changes of the endocasts of modern humans (n=93), chimpanzees (n=61), gorillas (n=64), orangutans (n=72) and hylobatids (n=21) to assess evolutionary changes in the tempo and mode of brain development. We tested whether ontogenetic trajectories based on Procrustes shape coordinates and centroid sizes overlapped in shape‐space and form‐space and differed in their direction. In the subspace of the first three principal components, explaining >;71% of the variance in shape‐space, our results show a conserved pattern of endocranial shape development in hominids. Hominids and hylobatids have similar ontogenetic trajectories in shape‐space, yet have distinct shapes at all stages of development. These findings indicate that differences in ontogenetic shape changes are observable from the first stage of postnatal development, suggesting that divergences from an ancestral pattern of primate development occur at an earlier prenatal stage. This research was supported by the Max Planck Society and by the Natural Sciences and Engineering Research Council of Canada (NS).
Community acceptability and utilization of micronutrient powders in Bihar, India
Authors: Young Melissa Fox , Kekre Priya , Verma Pankaj , Srikantiah Sridhar , Majumdar Amlan , Trehan Shamik , Das Arup Kumar , Chaudhuri Indrajit , Sheth Meggha , Webb‐Girard Amy , Ramakrishnan Usha , Martorell Reynaldo
Published: June 2021

Multiple micronutrient powders (MMPs) represent an innovative solution to reduce anemia and perhaps improve complementary feeding (CF) practices. The objective of this pilot was to examine community acceptability and utilization of MMPs to inform the design of a randomized‐control trial in Bihar, India. A 10 day pilot study of acceptability was conducted in 27 households with children ages 6–24 months. Households were counseled on MMP product use and age appropriate CF. In‐depth interviews with caregivers were conducted on three occasions (day 0, 5, and 10). MMP use was high regardless of caste, religion or age of child (100% tried product at least once, 84% had complete compliance). Caregivers reported overall ease of use of product and agreed there was a need for MMPs in the community. Caregiver's perceptions of MMPs were generally favorable and many even reported improved child appetite, CF practices and well‐being of the children. Two households felt MMP causes loose stools and reported negative experiences. Potential barriers identified were misconceptions that MMP would have immediate benefits in reducing illness. Formative research informed the development of locally appropriate visual aids/counseling materials. MMPs integrated with CF counseling are a promising locally acceptable intervention to improve child nutrition in Bihar.

Funding provided by a BMGF grant through a subcontract with CARE‐India.
Application of Urine Metabolomics to Probe the Effects of Long‐Term Bed Rest as a Model for Spaceflight
Authors: Ericson Karen Louise , Cooper B. R. , Peels M. E. , Deng Y. , Smith S. M. , Coburn S. P.
Published: June 2021

This project aims to determine if urine metabolomics can be a meaningful approach to assess spaceflight health effects. Muscle loss in spaceflight might be caused by decreased muscle synthesis or increased muscle breakdown, and identification of an appropriate biomarker might suggest appropriate countermeasures. In this study, urine samples from 8 subjects in a bed rest study (paired before and during bed rest) were analyzed using hydrophilic interaction liquid chromatography (HILIC) and positive ionization mode time‐of‐flight mass spectrometry. Data analysis indicated measurable differences in urinary compounds of the paired groups. Of 1873 unique compounds found, 20 were identified as having high significance (consistent increase or decrease during the sample period) and were thus considered possible biomarkers. Comparison of the bioinformatics programs used here, Agilent Mass Profiler Professional and Purdue's in‐house Omics Discovery Pipeline, could be valuable for future metabolomic studies. Identification of high‐significance compounds is continuing. Support: Grant 10465 – Indiana Space Grant Consortium & Indiana University‐Purdue University Fort Wayne Office of Research & External Support (KLE).
Long‐term effects of dried plum consumption on insulin and glucose levels in postmenopausal women
Authors: Hooshmand Shirin , Garcia Sofia , Metti Dina , Vereda Yenina , Chai Shea C. , Arjmandi Bahram H.
Published: June 2021

Insulin and glucose play a crucial role in glycemic control. Postmenopausal women have an increased risk of imbalances in maintaining stable blood sugar levels as a result of changes in body composition. Dried plums are a good source of nutrients including fiber, potassium, magnesium, phosphorus, iron and boron. Nonetheless, the consumption of dried plums is often avoided due to their high sugar content (51.5%). However, the sugar in dried plums is composed of 28.5% sorbitol, which is not readily absorbed, and 25.4% fructose, which promotes a stable insulin response. Although short‐term studies evaluating the effects of dried plum consumption on serum insulin levels have been conducted, there is a lack of long‐term studies. Therefore, the objective of the present study was to identify the long‐term effects of consuming dried plum on insulin and glucose levels. One hundred women, 1–10 years postmenopausal, were randomly assigned to one of the two treatment groups: dried plum (100 g/d) or dried apple (comparative control). Blood samples were collected at baseline and 12 months to assess insulin and glucose levels. No significant changes were found in insulin and glucose levels between dried plum and control groups (p=0.6720 and 0.7855, respectively). Therefore, long‐term consumption of dried plums can be recommended to postmenopausal women for their health benefits without negatively affecting glycemic control.
Effect of lycopene on adipose tissue mass and inflammatory factor in high fat diet‐induced obese
Authors: Choi Soo‐Kyong , Ryu Hae‐Jeong , Seo Jung‐Sook
Published: June 2021

The present study was performed to investigate the effects of lycopene on adipose tissue mass and inflammatory factor in high fat diet‐induced obese gerbil. Gerbils were randomly divided into a normal diet group, a high fat diet group, lycopene‐supplemented normal diet group, and lycopene‐supplemented high fat group. Epididymal adipose tissue mass higher in high fat diet group than in normal diet group, but it was not affected by lycopene supplementation. Mesenteric fat tissue and brown adipose tissue mass increased by high fat diet but decreased significantly by lycopene supplementation. Concentration of adiponectin was increased in lycopene treatment group when compared with HF group, but the change was not statistically significant. There were no statistically significant differences in plasma level of CRP among the experimental groups. The level of oxidized LDL was increased in high fat diet group compared with in the normal diet group, but it was reduced by lycopene feeding. A significant negative correlation was observed between plasma adiponectin and mesenteric fat tissue. Plasma concentration of ox‐LDL was significantly correlated with epididylmal adipose tissue, brown adipose tissue and mesenteric fat tissue mass. These results suggest that dietary supplementation of lycopene may prevent the chronic diseases related to lipid metabolism through suppression of inflammatory factor.
Validation of the Dietary Inflammatory Index in the Women's Health Initiative
Authors: Tabung Fred , Steck S E , Ma Y , Ockene J K , Shivappa N , Hurley T , Liese A D , Zhang J , Shikany J M , Park H L , Millen A E , Martin L W , Jiao Li , Hou L , Agalliu I , Hingle M , Liu S , Rosal M C , Hebert J R
Published: June 2021

We previously developed a dietary inflammatory index (DII) from extensive literature review to assess the inflammatory potential of diet. The purpose of this study was to validate the novel DII based on responses to a food frequency questionnaire (FFQ), using data from three 24‐hour dietary recalls (24HR), as well as inflammation biomarkers in the Women's Health Initiative Observational Study. Two separate DII scores were calculated: one from three 24HR and another from an FFQ (n=461). Pearson correlation and cross‐classification of quintiles were used to compare the 24HR‐ with the FFQ‐derived DII. In a separate analysis of baseline data (n= 2899), the FFQ‐derived DII was used to predict plasma levels of hs‐CRP, IL‐6, and TNFα‐R2 using multiple linear regression models.

The correlation coefficient between the FFQ‐ and 24HR‐derived DII was 0.64 (95%CI 0.58, 0.69). The FFQ‐derived DII was positively associated with hs‐CRP (β=1.02, P=0.03) in all race/ethnicities; IL‐6 in African Americans (β=1.10 P<0.0001) and European Americans (EA) (β=1.07 P=0.002); and TNFα‐R2 in EA (β=76.09 P=0.001).

The FFQ‐derived DII appears to provide valid estimates of the inflammatory potential of diet. The DII represents a new tool for assessing the relationship between diet and chronic diseases such as cancer.

Funding: The WHI was funded by the NHLBI and Dr. Hebert by the NCI (K05 CA136975).
Dietary patterns of Korean older adults and cognitive function
Authors: Kim Jihye , Yu Areum , Choi Bo Youl , Nam Jung Hyun , Kim Mi Kyung , Oh Dong Hoon , Yang Yoon Jung
Published: June 2021

The objective of the study was to investigate the association between dietary patterns and cognitive function in Korean older adults. The subjects were 808 older adults participating in the Yangpyeong Cohort Study between July 2009 and August 2010. Dietary intake was estimated by using a 106‐items food frequency questionnaire. Cognitive function was assessed by Korean version Mini‐Mental State Examination (MMSE‐KC). Factor analysis using the principal component analysis method was applied to identify major dietary patterns. Four major dietary patterns were derived: 1) vegetables, seaweeds, fruits, mushroom, potatoes, fish, seafood, nuts, and green tea (healthy pattern without dairy foods); 2) meat, chicken, eggs, bread, flour based food, and coffee (western pattern); 3) dairy foods, soybean milk, seafood, fruits, mushroom, and snack (healthy pattern with dairy foods); 4) white rice (white rice pattern). Healthy pattern with dairy foods was inversely related to the risk of cognitive impairment after adjusting for confounders (4th v. 1st quartile, OR 0.57, 95% CI 0.38–0.87, P for trend=0.010). White rice pattern was positively associated with the risk of cognitive impairment after adjusting for confounders (4th v. 1st quartile, OR 2.14, 95% CI 1.37–3.33, P for trend=0.011). This study suggests that a healthy diet with dairy foods may have a beneficial effect on preventing or postponing cognitive impairment. Whereas, eating mainly white rice without other balanced side dishes may increase the risk of cognitive impairment.

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2012R1A1A1041792).
Exercise ameliorates high fat diet induced intestinal inflammation
Authors: Campbell Sara Chelland , Wisniewski Paul J , Aydin Christopher , Lightfoot Stanley A
Published: June 2021

Oxidative stress and inflammation have emerged over the past decade as key features in chronic disease. Research has consistently supported the role of regular exercise in decreasing pro‐inflammatory responses thereby reducing the risk of developing these chronic diseases. The overall objective of this study was to identify if exercise attenuates the intestinal inflammatory response in an animal model of diet‐induced obesity. The intestine has received wide‐spread attention recently due to changes in permeability associated with high fat diets (HFD). Twenty‐four C57BL/6NTac mice (n=6/group) were randomly assigned to one of 4 groups: 1) normal (10% fat)‐sedentary; 2) normal‐exercise; 3) HFD (45% fat)‐sedentary; and 4) HFD‐exercise for 12 weeks. Animals were gradually acclimated to motorized treadmill running which culminated with the animals running 4 days/week for 40 minutes per day at 15m/min. Upon sacrifice duodenal sections were harvested and fixed in 10%NBF. The duodenums were cut longitudinally so 90+% of each duodenum could be seen and all cuts were analyzed by Dr. Stanley Lightfoot in duplicate. Results showed that HFD doubled the width of duodenal villi compared to lean animals (95μm vs. 44 μm), resulting in thinner spaces between the villi due to increased inflammatory and fat cells present in the villi. Exercise caused enhanced vessel dilation in lean animals and eliminated inflammatory and fat infiltrate in HFD animals resulting in protecting the villi from widening. These results suggest that exercise can ameliorate intestinal inflammation and could contribute to alleviating chronic diseases of intestinal origin, like obesity.