bluuebunny/crossref_metadata_2025 huggingface dataset:
Population attributable risk for dementia in the Seattle region
Authors: Montine Thomas
Published: June 2021

Previously published community‐ or population‐based studies of brain aging and dementia with autopsy are limited to one gender, ethnic group, clergy, or focused assessments. Our goal is to determine the contributors to dementia in a typical U.S. population of older individuals. We considered autopsy data from the Adult Changes in Thought (ACT) study, an on‐going longitudinal population‐based study of brain aging and dementia. Analyses were based on data collected between 1994 and 2006, which included approximately 3,400 men and women who were 65 years or older and cognitively intact at the time they were sampled in King County, WA. All consecutive autopsies (n = 221, 20% of deaths) from the cohort were evaluated. A weighted multivariate analysis was employed to account for potential participation bias, and thereby facilitate interpretation of the results beyond autopsied subjects. After adjusting for age, gender, education, and APOE, independent correlates of dementia (relative risk, 95% CI; overall p‐value) included Alzheimer's disease (5.89, 1.62–17.60; p < 0.05), microvascular brain injury (4.80. 1.91–10.26; p < 0.001), and neocortical Lewy bodies (5.08, 1.37–18.96; p < 0.05). Estimated population attributable risk for the three processes for dementia was 45% for Braak stage, 33% for microvascular brain injury, and 10% for neocortical Lewy bodies. Our results underscore the therapeutic imperative for Alzheimer's and Lewy body diseases, and provide evidence for immediate use of strategies that target cerebral microinfarcts to prevent or delay onset of dementia in our progressively aging population.
Microprojections regulate proliferation and activity of cardiomyocytes derived from mouse embryonic stem cells
Authors: Biehl Jesse K. , Yamanaka Satoshi , Boheler Kenneth R. , Russell Brenda
Published: June 2021

The goal was to test if the physical properties of the microenvironment control proliferation and activity of the mouse embryonic stem cell (mESC) progeny. Microtopographic features were fabricated by photolithography to create 15μm high projections spaced 80 or 500μm apart tetragonally in poly‐dimethyl‐silicone (PDMS) membranes. mESC differentiation began in hanging drops, followed by suspension culture, before the resulting embryoid bodies were disassociated and plated on either flat or the microtextured surfaces. The number of heterogeneous SC derivatives observed with phase microscopy was 60 ± 20% (n=3) on the 80μm microprojections compared to flat PDMS. Similar results were seen for pure cardiomyocytes derived using a puromycin resistant cassette incorporated into the NCX1 promoter. Only 43±12% (n=3) and 75±16% (n=5) of the cardiomyocytes were found on the 80μm and 500μm spaced microprojections, respectively, compared to the flat PDMS. The beating rate per minute of the cardiomyocytes was recorded by video microscopy and was 1.8±0.4 fold higher on the microprojections compared to the flat with surprising changes in coefficients of variance of 0.50 and 0.21 respectively (n=5). Results suggest that microtopography affects both expansion and beating characteristics of mESC progeny. T32HL007692 and HL 62426.
Simultaneous permeation and metabolism of methyl nicotinate in human, snake, and shed snake skin
Authors: Ngawhirunpat Tanasait , Opanasopit Praneet , Rojanarata Theerasak , Panomsuk Suwannee , Chanchome Lawan
Published: June 2021

The transdermal permeation and metabolic characteristics of methyl nicotinate (MN) in stratum corneum and split‐thickness of human skin and three species of shed snake and snake skin (Elaphae obsoleta, Naja kaouthia and Python molurus bivittatus) were evaluated. The total flux from MN‐saturated solution through human skin was not significantly different with that through snake and shed snake skin of Elaphae obsoleta and Naja kaouthia, but significantly higher than that through snake and shed snake skin of Python molurus bivittatus (P<0.05). A great difference in skin esterase activity was observed between human and snake in both snake skin and shed snake skin of all species. NA flux increased with an increase in MN donor concentration and reached a plateau, suggesting that metabolic saturation was taking place in the skin. NA flux at the plateau and MN donor concentrations at which the NA flux reached a plateau also varied by species. These findings indicated that the discrepancy in transdermal profiles of MN among skins tested was predominantly due to the difference in the esterase activity in the skin. This research was supported by the Thailand Research Fund and the Ministry of Education, Thailand (RMU 4880011).
Characterization of stretch‐activated nonselective cation channels in pregnant rat uterine smooth muscle
Authors: Kim Young‐Hwan , Chung Seungsoo , Lee Young‐Ho , Ahn Duck‐Sun
Published: June 2021

Although the exact mechanisms that initiate parturition have been still uncertain, the stretching of the myometrium as a result of fetal growth has been considered as one of key event in the initiation of labor. However, the direct evidence for the presence of stretch‐activated non‐selective cation channels(SACC) in uterine smooth muscle has not yet been reported. In this study, the stretch can activate the SACC in pregnant rat myometrial cells using patch clamp technique. The application of negative pressure within a pipette in cell‐attached or inside‐out patch mode induced the opening of ion channels, which has single channel conductance of ~25pS, and activity of these ion channels are disappeared after removing of negative pressure. There was no significant difference in the stretch‐induced activation of ion channel between cell‐attached and inside‐out patch mode, which might exclude the possible involvement of secondary messenger systems in the activation of these channel by stretch. The channels shows similar permeability to both Na+ and Cs+ ions. Increasing [Ca2+]i in inside‐out patch mode significantly enhanced the channel activity at same negative pressure. From these result, it may be concluded that stretch can directly activate the SACC in pregnant rat uterine smooth muscle cells, and SACC might contribute to the spontaneous contraction during labor.
Effect of nifedipine, ketoprofen or the combination on the phenylephrine‐enhanced peristalsis in an ex vivo model of the isolated, intraluminally perfused porcine ureter
Authors: Purdy Ralph Earl , Shelkovnikov Stan , Summers Scott M. , Herz Jeffrey M. , Tedford Clark E. , Box Geoffrey N. , Louie Michael K. , Clayman Ralph V.
Published: June 2021

Intraluminal exposure of the ureter to nifedipine and/or ketoprofen could prove beneficial by relaxing smooth muscle and inhibiting peristalsis. Omeros’ proprietary combination product candidate, OMS201, includes nifedpine and ketoprofen as active pharmaceutical ingredients and is designed to provide clinical benefits during endoscopic urological procedures by reducing smooth muscle contractions and inhibiting inflammation and pain. To evaluate the single agents as well as the OMS201 combination product, isolated naïve porcine ureters were cannulated and subjected to antegrade perfusion. Ten μM phenylephrine was added to the external medium to induce ureteral peristalsis. Addition of 0.3 μM nifedipine to external bathing medium resulted in a rapid aperistalsis in less than 5 minutes. In contrast, intraluminal perfusion with 9 μM nifedipine required 14 ±2.6 minutes to cause aperistalsis. Intraluminal ketoprofen, 27 μM, was without significant effect on amplitude or rate. The combination of nifedipine (9 μM) and ketoprofen (27 μM) in OMS201 was effective in reducing peristalsis, demonstrating the contribution of nifedipine to smooth muscle relaxation and inhibition of peristalsis. It is concluded that while the urothelial lining is a drug barrier, intraluminal 9 μM nifedipine can penetrate to the ureteral smooth muscle and fully inhibit peristalsis. Funded by Omeros Corp.
STIM1 regulates store operated calcium entry (SOCE) in human airway smooth muscle
Authors: Thompson Michael A , Zhang David , Sieck Gary , Pabelick Christina , Prakash Y S
Published: June 2021

Intracellular Ca2+ regulation in human airway smooth muscle (hASM) involves plasma membrane (PM) Ca2+ influx, including that triggered by sarcoplasmic reticulum (SR) Ca2+ depletion, i.e. store‐operated Ca2+ entry (SOCE). While transient receptor potential channels (e.g. TRPC3, TRPC4) are thought to mediate SOCE, the mechanisms triggering or regulating SOCE are being actively investigated. Recent studies suggest that stromal interaction molecule 1 (STIM1) acts as a signaling mediator between PM and SR, potentially regulating Ca2+entry via SOCE. In hASM, fluorescence microscopy and western blot analyses were used to verify the presence of STIM1. SR Ca2+ depletion with 10μM cyclopiazonic acid (CPA) resulted in increased STIM1 expression in the PM fraction, compared to controls. Transfection with 50nM STIM1 siRNA for 48h significantly decreased STIM1 levels. In cells where [Ca2+]i was measured using fura 2‐AM, SOCE was significantly reduced in cells transfected with STIM1 siRNA. Green‐fluorescent protein (GFP)‐tagged STIM1 transfection showed intracellular localization in non‐stimulated cells, with increased PM staining following CPA exposure. Subsequent immunostaining revealed co‐localization of STIM1 with TRPC3 and TRPC4. These results indicate that STIM1 is present in hASM and suggest a role for this protein in regulating SOCE.

This work has been supported by NIH grants UL1RR024150‐01(CR to CMP, CR20 and Early Career Development Award to YSP), HL74309 (GCS), and the Department of Anesthesiology (CMP, YSP), Mayo Clinic College of Medicine, Rochester, MN
Purinergic regulation of ENaC in rat and mouse collecting duct
Authors: Bugaj Vladislav , Pochynyuk Oleh , Stockand James D
Published: June 2021

The epithelial sodium channel (ENaC) plays a critical role in setting systemic salt and water balance and thus, blood pressure. Activity of this channel is limitingfor Na+ reabsorption at the renal collecting duct. Cell signaling in response to purinergic stimulation decreases ENaC activity in models of polarized collecting duct epithelia. Moreover, targeted deletion of purinergic P2Y receptors causes hypertension in mice associated with dysfunctional salt handling at the collecting duct. Thus, we test here the hypothesis that the purinergic agonist, ATP, modulates the activity of ENaC in isolated, split‐open mouse and rat collecting ducts. In addition, we define the cellular signaling pathway underpinning such regulation. We find that ATP rapidly decreases ENaCopen probability in both the mouse and rat collecting duct. Dwell time analysis reveals that ATP forces ENaC toward a closed state. Inhibiting purinergic signaling withsuramin blocks the actions of ATP and rescues ENaC activity. The PLC inhibitor, U73122, recapitulates the actionof suramin. We conclude that ATP through metabotropic purinergic receptors coupled to PLC decreases ENaC activity by rapidly decreasing channel open probability. It is likely that PLC‐mediated reduction in apical membrane PI(4,5)P2 levels couples purinergic signaling to decreases in channel open probability in native collecting ducts. These results suggest that purinergic signaling is a major regulator of ENaC activity in renal epithelialcells.
Repeated exposures to hyperbaric air suppress neurite outgrowth in PC12 cells
Authors: Isada Tetsuro , Hirose Munetaka , Murata Eri , Shigemi Kenji
Published: June 2021

Objective: Scuba divers are at risk of central nervous system damage under hyperbaric exposure, which would directly causes lesions of neural cells.

Method: After NGF stimulation, PC12 cells were compressed for 30 minutes as single exposure, and repeated exposures for 30 minutes were applied 3 times with intervals 3,15,30 minutes. Then neurite outgrowth was observed. Downstream signals of NGF (TrkA and ERK) were also examined under single and repeated exposures.

Result: Repeated exposures to hyperbaric air with either 3 or 15 min interval suppresses NGF‐stimulated neurite outgrowth. Although TrkA showed no change in autophosphorylation, repeated exposures with 3 min interval reduced ERK protein expression significantly.

Conclusion: Direct effect of repeated exposures to hyperbaric air on neural cells likely causes CNS lesions in scuba divers.
Time‐dependent alterations in peripheral chemoreceptor activity during postnatal hyperoxia
Authors: Bavis Ryan W , Donnelly David F
Published: June 2021

Postnatal hyperoxia, as low as 30% O2 and lasting 1–4 weeks, attenuates the ventilatory response to acute hypoxia due to profound impairment of peripheral chemoreceptors. The present study investigated the time course of this impairment by recording single‐unit chemoreceptor action potentials (APs) following 1, 3, 5, 8 and 14 days of hyperoxia treatment. Rat pups were placed in hyperoxia (60% O2) at P7. Pups were then removed at P8, P10, P12, P15 and P21 and single‐unit activities were isolated and recorded in vitro from the soma of petrosal neurons with projections to the carotid body. Compared with chemoreceptors of normoxia‐reared pups, hyperoxia enhanced the peak‐discharge response to acute hypoxia after 1 day of hyperoxia. In contrast, the peak‐discharge response was significantly reduced by 5 days of hyperoxia and remained low for the duration of the treatment period. Conduction time for an AP to progress from the carotid body to petrosal ganglion was also lengthened by the fifth day of hyperoxia treatment. Because nerve conduction time lengthens with the same time course as changes in the acute response to hypoxia, it suggests that at least some of the response impairment is due to changes in afferent nerve excitability characteristics. Supported by NIH grants P20 RR‐016463 (Maine INBRE) and HL‐073500.
Metabolic syndrome abolishes A2A receptor and KATP channel involvement in coronary arteriolar dilation to adenosine in Ossabaw swine
Authors: Bender Shawn B. , Sturek Michael , Laughlin M. Harold
Published: June 2021

Reduced coronary flow reserve to adenosine and altered adenosine receptor expression/signaling are common in diabetes and metabolic syndrome (MetS). We hypothesized that reduced adenosine 2A (A2A), but not 2B (A2B), receptor and ATP‐sensitive (KATP) and voltage‐gated (Kv) potassium channel function contribute to reduced coronary arteriolar dilation to 2‐chloroadenosine (2‐CADO) in MetS. Swine were fed control (C) or excess kcal atherogenic diet for 9 wk to study early stage MetS. Dose‐ response curves to 2‐CADO in isolated coronary arterioles were performed in the absence or presence of ZM241385 and/or alloxazine to inhibit A2A or A2B receptors, respectively. Responses were also examined in the presence of glibenclamide or 4‐aminopyridine to inhibit KATP or Kv channels, respectively. Dilation to 2‐CADO was similar in C and MetS swine. A2A and A2A+2B, but not A2B, receptor blockade similarly reduced maximal 2‐CADO dilation in C arterioles. Conversely, A2A or A2B blockade had no effect while A2A+2B inhibition right shifted the curve in MetS arterioles. KATP and Kv inhibition right shifted the curve in C, while only Kv inhibition did so in MetS arterioles. Dilation to SNP was unchanged. These data suggest that MetS disrupts pathways involved in coronary dilation to adenosine, which are compensated for, thereby preserving arteriolar dilation to adenosine in MetS. Support: HL52490, AR048523, RR013223, HL062552.
Effects of metformin on endothelial glycocalyx properties in db/db mice
Authors: VanTeeffelen Jurgen Willem , Eskens Bart , Zuurbier Coert , Vink Hans
Published: June 2021

Recent studies show impaired glycocalyx properties during hyperglycemic conditions (1,2). The current aim was to determine the effects of the popular anti–diabetic drug metformin on glycocalyx properties. Db/db mice, 6–8 wks of age, were treated with metformin in their drinking‐water for 2 wks (MET, n=5); non‐treated db/db mice served as control (CON, n=5). At the experimental day, mice were infused with FITC–labeled 70 kDa dextrans (D70) and Texas Red‐labeled 40 kDa dextrans (D40), blood samples collected for 30 min, and plasma dextran concentrations determined using a spectrophotometer (2). Systemic glycocalyx D70 excluding volume was determined from the difference in initial distribution volume of D40 versus D70 (2,3), and was 0.009 ± 0.030 ml in CON and increased to 0.183 ± 0.057 ml in MET (p<0.05). The ratio of D70 to D40 clearance, reflecting glycocalyx barrier properties (2,3), was 0.619 ± 0.026 in CON and decreased to 0.530 ± 0.026 in MET (p<0.05). Dry to wet weight ratio, measured in heart and kidney, was 0.286 ±0.006 in CON and decreased to 0.257±0.009 in MET (p<0.05). Fasting blood glucose level was not changed in MET. These data show that MET treatment in db/db mice results in improved glycocalyx properties, irrespective of a change in fasting glucose. The glycocalyx improvement might underlie the observed restoration of organ fluid balance by metformin.

Supported by Established Investigatorship from Netherlands Heart Foundation (2005T037) and Dutch Diabetes Research Foundation (2006.00.027)
Irreversible damage to rostral pontine nuclei (RPN) in adult awake goats attenuates a secondary physiologic response to hypoxia
Authors: Bonis Joshua M , Davis Suzanne , Kiner Tom , Krause Katie , Qian Baogang , Pan Lawrence , Forster Hubert
Published: June 2021

We chronically implanted microtubules into the rostral pons of 6 goats to assess the response to hypoxia following irreversible damage to RPN. All goats recovered uneventfully after implantation. After breathing room air for 30 minutes (min), goats were subjected to 10.6% O2 for 30 min. PaO2 at min 6 and 26 of hypoxia was 28.41±0.57 and 26.87±0.58mmHg respectively. The normal response to hypoxia (Table) was a 1° (∼3 min) spike in pulmonary ventilation (VI) followed by a “roll‐off” in VI (∼6 min), and lastly a 2° increase in VI (within 20 min) associated with an increase in body temperature, shivering, VO2, and VCO2. Following irreversible damage to RPN, the 1° spike and “roll‐off” in VI were unchanged from pre‐damage conditions, but the 2° increase was attenuated (P<0.05). We conclude that damage to RPN does not alter the presumed peripheral chemoreceptor 1° response to hypoxia nor the VI “roll‐off”, but attenuates the 2° physiologic response.

Comparison of physiologic variables in response to hypoxia, before and after RPN damage.

VE(%)
PaCO2(Δ mmHg)
Temp(Δ °C)
VO2 (l/min)
VCO2 (l/min)

1° increase
Roll‐off
2° increase
6 min hypoxia
26 min hypoxia
Final
26 min hypoxia
26 min hypoxia

Pre‐damage
174
130
251
−5.47
−8.18
0.96
158.1
197.1

Post‐damage
176
128
195*
−5.31
−6.82
0.59*
8.9*
85.1*

Significant difference between pre‐ and post‐damage by one‐way ANOVA (P<0.05). Supported s.
Chemosensory function of amphibian skin epithelium: Amiloride inhibition of basolateral membrane depolarization in response to increased bath osmolality correlates with afferent neural activity and behavior.
Authors: Hillyard Stanley D , Willumsen Niels J
Published: June 2021

In dehydrated toads cutaneous water absorption is associated with initiation a behavior, the water absorption response (WR), in which the toads press the skin to a rehydration surface. WR initiation is suppressed in toads given a hyperosmotic (250 mM) NaCl solution. Associated with WR suppression the activity of spinal nerve #6 increased when the skin was exposed to this solution and the basolateral membrane potential (Vb) of principal cells in the isolated skin depolarized. These results support our hypothesis that the chemosensory properties of the skin are analogous to taste cells of the mammalian tongue where depolarization of Vb also stimulates neural activity. Here we show the frequency of WR initiation increased from 21% to 50% of trials in toads given 250 mM NaCl with 10 μM amiloride, that the time toads allow ventral skin contact is greater in the presence of amiloride, and that the depolarization of Vb is greatly reduced, relative to NaCl alone. The frequency of WR initiation was also greater with 250 mM Na gluconate as the rehydration solution (40% vs 21% with NaCl) and was increased to 80% of trials when 10 μM amiloride was added to the Na gluconate solution. Inhibition of the transcellular pathway with amiloride and the paracellular pathway with gluconate have similar behavioral responses, and variability in the behavioral responses correspond with variability in Vb and paracellular conductance.
Development of a robust, quantitative normal‐phase HPLC method for flavan‐3‐ols and procyanidin oligomers in chocolate and cocoa‐containing samples
Authors: Robbins Rebecca J. , Leonczak Jadwiga , Johnson J. Christopher , Li Julia , Kwik‐Uribe Catherine , Prior Ronald L , Gu Liwei
Published: June 2021

A novel normal‐phase HPLC method for the quantitative determination of flavan‐3‐ols and procyanidins in chocolate and cocoa‐containing food products has been developed. Chromatographic separation based on degree of polymerization (DP = 1–10) was achieved on a diol stationary phase column (normal phase) with a binary gradient of acetonitrile (CH3CN:HOAc) and methanol‐water (CH3OH:H2O:HOAc). This novel methodology employs more environmentally friendly solvents than previously published methods and demonstrates greater resolution for larger oligomers. Quantitation was achieved with fluorescent detection (FLD: ex 230 nm/em 321 nm) and the use of external standards. With these solvents, fluorescence quantum yields were significantly greater than observed in previous methods. To have oligomers (DP1 −10) within the detection range of the instrument in a single run, and behave linearly over a wide range of concentrations, an FLD photomultipler tube gain gradient was employed. Sample preparation investigations were conducted to eliminate common chromatographic interferences and assess extraction efficiencies. Method performance was examined on selected cocoa‐containing foods (including NIST reference materials) with multiple analysts and laboratories. This method demonstrated accurate and reproducible quantitation of the flavanols and procyanidins in chocolate and cocoa‐containing products
Proteomic analysis of factors secreted by glioblastoma that induce endothelial tissue factor
Authors: Brat Daniel J , Tucker‐Burden Carol , Rong Yuan , Chen Gang , Durden Don , Van Meir Erwin , Pohl Jan
Published: June 2021

We have proposed that intravascular thrombosis initiates or propagates necrosis in GBM, leading to hypoxia‐induced angiogenesis and rapid biological progression. We investigated factors secreted by gliomas following PTEN loss and under hypoxia that induce endothelial expression of tissue factor (TF), a potent procoagulant that could promote thrombosis. We used a PTEN null human GBM cell line (U87MG) with an inducible wt PTEN (23.11 cells) to model progression. Conditioned media (CM) from 23.11 +/− PTEN was collected after 48 hours of normoxia (21% O2) or hypoxia (1% O2). CM was tested on human brain derived microvascular endothelial cells (HBMEC) for its effect on proliferation, apoptosis, tube stability on Matrigel, or TF expression. Proteins from CM were precipitated, concentrated and labeled by Isotope Coded Affinity Tags (ICAT). Paired ICAT samples were separated and analyzed by tandem mass spectrometry (MS/MS). Proteins identified by MS/MS were measured in CM by ELISA and tested directly on HBMEC. We found that CM from PTEN− 23.11 cells strongly and consistently induced HBMEC TF expression compared to PTEN+ cells and that hypoxia potentiated this effect. CM from PTEN− and PTEN+ gliomas did not have differential effects on proliferation or apoptosis. ICAT coupled with MS/MS identified MMP‐2 (1.5‐fold) and macrophage migration inhibitory factor (MIF; 1.3‐fold) as proteins secreted by PTEN− compared to PTEN+ gliomas. IL‐8 (3.2‐fold) and insulin‐like growth factor binding protein 3 (IBP‐3; 3.2‐fold) were secreted at much higher levels in hypoxia. Both purified IL‐8 and MIF induced endothelial TF in the concentration range in CM. Thus, MIF and IL‐8 are secreted factors that induce endothelial TF and may be associated with the development of thrombosis in GBM.
Mechanical and chemical regulation of cells in a 3D environment
Authors: Chen Qian
Published: June 2021

Functional tissue engineering requires cells growing in a 3D environment that mimics in vivo conditions. First, mechanical signals are transduced from extracellular matrix to the cells. Pericellular matrix is at the ideal location to be involved in transmitting mechanical signals from the microenvironment to a cell. We found that changes of the content of matrilins that link various pericellular molecules surrounding chondrocytes affect mechanical stimulation of chondrocyte proliferation and gene expression. Thus, pericellular matrilins may play a role in chondrocyte mechanotransduction. Cells may convert mechanical signals into chemical signals. We found that the mechanoresponsive region in the promoter of the α1(X) gene, a mechanoresponsive gene in cartilage, coincided with the Bone Morporgenic Protein (BMP)‐responsive elements in the promoter. Mechanical loading stimulated production of BMP 2/4 by chondrocytes, and addition of BMP antagonist noggin abolished the mechanoresponsiveness of type X collagen promoter. This observation indicates that mechanical loading stimulated type X collagen transcription by activating BMP‐dependent signaling pathways. It suggests a novel mechanism by which mechanical loading regulates α1(X) gene transcription through activating BMP signaling. This mechanism may provide insight into mechanical and chemical regulation of cells in tissue engineering.
Disruption of BCATm in mice alters hypothalamic expression of genes involved in energy balance
Authors: Shen Li , Zhou Jun , Martin Roy J , Morrison Christopher D , Hutson Susan M
Published: June 2021

Leucine has been shown to act locally within the hypothalamus to regulate hypothalamic neuropeptide expression and feeding behavior. However, the long‐term in vivo consequences of leucine signaling and the role of branched‐chain amino acid (BCAA) metabolism in this signaling remain unclear. To investigate the role of BCAA metabolism in neuronal function we utilized mice deficient for BCATm, which is the enzyme catalyzing the first step in peripheral BCAA metabolism. BCATM−/− mice exhibit elevated plasma BCAAs and reduced adiposity and body weight despite eating more food, along with increased energy expenditure. Brains from fed mice were quickly removed, frozen on dry ice and stored at −70°C. The midbrain was dissected using a cryostat. Arcuate nucleus (ARC) micropunches were collected for five continuous coronal sections, and levels of agouti‐related protein (AgRP), neuropeptide Y (NPY) and proopiomelanocortin (POMC) mRNA in the ARC were determined using real‐timereverse transcriptase polymerase chain reaction. Compared with wildtype mice, POMC expression was significantly decreased and AgRP expression increased in the arcuate nucleus of BCATm−/− mice (p<0.05). These findings indicate that blocking BCAA metabolism alters the expression of hypothalamic genes regulating energy balance, and may provide a mechanism whereby dietary amino acids regulate energy homeostasis.
Transforming Growth Factor‐beta 1 in colorectal cancer: implications to treatment response, disease and nutritional outcomes
Authors: Ravasco Paula , Grillo Isabel Monteiro , Camilo Maria
Published: June 2021

Background: TGF‐beta 1 has been shown to contribute to breast & bladder cancer progression, suggested in colorectal cancer cell lines. This study investigated potential interactions between serum concentrations of TGF‐beta 1 & pro‐inflammatory cytokines, cancer characteristics, response to radiotherapy (RT) & metabolic/nutritional factors in 101 patients with colorectal cancer (CRC). Methods: Prospective longitudinal study, before & after neoadjuvant RT, evaluated: serum concentrations of TGF‐beta 1, TNF‐a, IL‐1ra, IL‐6 (ELISA, R&D Systems®, USA), cancer stage/histology, RT response, Resting Energy Expenditure (REE‐indirect calorimetry), %weight loss, usual/current diet (diet history+24hr recall). Results: Before & after RT, higher TGF‐beta 1 was associated with higher TNF‐a, IL‐1ra & IL‐6 (p<.01), stages III/IV (p<.02), histology grades 2/3 (p<.003), higher REE + weight loss >5% + intake reduction >25% (p<.01). Predictive value analyses, adjusted for stage/histology, showed that higher baseline TGF‐beta 1 predicted a poorer RT response (HR:8.25; 95% CI 2.74–26.47, p=.003), increased REE (HR:8.25; 95% CI 2.74–26.47, p<.002), greater weight loss (HR:8.15; 95% CI 2.22–25.40, p<.002) & intake reduction (HR:7.15; 95% CI 2.25–16.11, p<.004) after RT. Conclusions: Our results suggest an association between higher TGF‐beta 1 and poorer RT response, nutritional wasting & possibly prognosis.
OXIDATIVE STATUS IN HEAVY SMOKERS: EFFECT OF NUTRITION SUPPLEMENTS
Authors: Bamonti Fabrizia , Novembrino Cristina , Gregori Dario , Lonati Silvia , Schiraldi Gianfranco , Cighetti Giuliana
Published: June 2021

Long‐term cigarette smoking can cause oxygen free radical (ROS) formation and antioxidant capacity depletion, leading to oxidative stress involved in many disorders. Active ROS produces lipid peroxidation generating molecules such as oxidized LDL (oxLDL) and malondialdehyde (MDA) used as indexes of oxidative damage.

An adequate diet and/or supplementation may attenuate this condition.

To evaluate potential beneficial effects of supplementation with two different encapsulated formulas, consisting primarily of mixed juice powder concentrate, a placebo‐controlled double‐blind study was performed on 61 heavy smokers (>20 cigarettes/d, duration ≥10y, age 40–60y, 35 male). Subjects were randomized into 3 groups: A‐placebo; B‐fruit/vegetable; and, C‐fruit/vegetable/berry. We measured serum oxLDL and plasma free MDA (f‐MDA), the chemically active form, index of recent damage.

Values (mean±SD) were compared at baseline (T0) and after 3 months supplementation (T1).

Nutritional supplementation produced a decrease in oxidative damage indexes, more evident in group C than B, and seems to attenuate smoking effects.

Support: NSA, Collierville, TN
Perspective of Managers towards Breastfeeding Support in the Workplace
Authors: Chow Tan , Olson Beth H.
Published: June 2021

Managers influence new mother employees’ perceptions of breastfeeding support in the workplace. The objective of this study was to assess manager attitudes towards breastfeeding support. Five focus groups were conducted with managers mainly from the finance, education and community sectors, who had experience supervising non‐managerial female employees. The results were transcribed verbatim and analyzed using NVivo. Emerging themes were knowledge of and need for company policies, managers’ role in providing support to breastfeeding employees, workplace consequences such as decreased productivity and coworker conflict, and benefits both to the health of mothers and infants and to the company. Most managers did not know if a company policy existed and showed mixed attitudes towards the need for a policy. All managers felt they should accommodate breastfeeding employees, but the extent of accommodation might vary by job nature, workflow, and facilities. Managers concerned lower productivity and co‐worker jealously of breastfeeding time, but that benefits would include recruitment, employee retention and loyalty, and potential increased productivity. Managers showed some understanding of breastfeeding benefits compared to infant formula. This research will be used to develop an instrument to measure managers’ attitudes towards breastfeeding support in the workplace. Funded by: National Research Initiative of the USDA Cooperative State Research, Education and Extension Service, grant number 2006‐35215‐16703
Determination of carotenoids in yellow maize, the effects of saponification, and food preparations
Authors: Muzhingi Tawanda
Published: June 2021

Maize is an important staple food consumed by millions of people in many countries. Yellow maize naturally contains carotenoids which not only provide provitamin A carotenoids but also xanthophylls which are known to be important for eye health. This study was aimed at 1) evaluating the effect of saponification during extraction of yellow maize carotenoids, 2) determining the major carotenoids in 36 genotypes of yellow maize by high performance liquid chromatography with a C30 column, and 3) determining the effect of cooking on the carotenoid content of yellow maize. The major carotenoids in yellow maize were identified as all‐trans lutein, cis‐ isomers of lutein, all‐trans‐zeaxanthin, α‐ and β‐cryptoxanthin, all‐trans β‐carotene, 9‐cis β‐carotene and 13‐cis β‐carotene. Extraction procedures without saponification showed a significantly higher yield than that obtained using saponification. In addition, cooking by boiling yellow maize at 100Â°C for 30 minutes increased the carotenoid concentration, while baking at 450Â°C for 25 min decreased the carotenoid concentrations by almost 70% as compared to the uncooked yellow maize flour. Therefore, yellow maize is a good source of provitamin A carotenoids and xanthophylls, the stability of maize carotenoids depends on cooking methods and no saponification is needed for analysis of maize carotenoids
The maternal transfer of lutein and zeaxanthin into the fetus during pregnancy
Authors: Connor Sonja L. , Connor William E. , Bezzerides Eugenia A. , Wang Yingming
Published: June 2021

Mammal and avian embryos obtain lutein and zeaxanthin from maternal sources. To quantify this transfer in humans, we analyzed maternal and newborn plasma for lutein and zeaxanthin content at delivery. Newborn venous blood was obtained from the umbilical cord. For 15 term births, maternal plasma lutein was 17.6 ± 7.9 mcg/dL vs. newborn lutein of 3.9 ± 2.1, and maternal zeaxanthin was 3.7 ± 1.9 mcg/dL vs. newborn zeaxanthin of 0.9 ± 0.5. Newborn levels were 22 percent lower for lutein and 24 percent lower for zeaxanthin than respective maternal values. In six pre‐term births, values for lutein and zeaxanthin were much lower for both mothers and infants than in full terms: for lutein, maternal 8.5 ± 4.4 and pre‐term 2.0 ± 1.0; and for zeaxanthin, maternal 2.0 ± 1.0 and preterm 0.6 ± 0.3 mg/dL. The newborn plasma values of lutein and zeaxanthin were only 20 to 24 percent of maternal values. One reason for lower fetal values might be lower HDL levels in the fetus. HDL in cord plasma has been shown to be only 27 percent of the maternal HDL, the major transporter of lutein and zeaxanthin. Perhaps HDL can only carry so much lutein and zeaxanthin. These data indicate placental transfer of lutein and zeaxanthin. More transfer probably occurred later in pregnancy. Women who had higher plasma lutein and zeaxanthin delivered infants whose plasmas also had higher lutein and zeaxanthin levels. USDA grant
Fetal growth only partially explains the increased risk for obesity in children of women who gain excessive weight during pregnancy: A large multi‐center, multi‐ethnic cohort study
Authors: Stettler Nicolas , Wrotniak Brian H. , Shults Justine , Butts Samantha
Published: June 2021

OBJECTIVE:

To evaluate if fetal growth (assess by birth weight) explains the association of excessive pregnancy weight gain with obesity in the offspring at age 7 years.

METHODS:

Retrospective cohort study of 10,228 subjects of the Collaborative Perinatal Project (1959–72). Logistic regression was used after adjustment for important confounding factors, then, after further adjustment for birth weight.

RESULTS:

Pregnancy weight gain was excessive in 11% of women and 5.7% of offspring became overweight at age 7 years. Birth weight was higher after excessive vs. appropriate pregnancy weight gain and among overweight vs. non‐overweight children. Children of mothers who gained excessive vs. appropriate weight were more likely to be overweight: OR= 1.48, 95%CI: 1.06–2.06, p=0.02. After adjusting for birth weight, the association magnitude relative decrease was 5%, but it remained significant: 1.40, 1.00–1.95, 0.05.

CONCLUSIONS:

Fetal growth only partially explains the increased risk for obesity in children of women who gain excessive weight during pregnancy, perhaps due to fetal programming expressed after the neonatal period. Improved adherence to pregnancy weight gain recommendations may be a new and effective way to prevent childhood obesity, independent of fetal growth, since, currently, 46% of U.S. women exceed these recommendations. Support: Nutrition Center of The Children's Hospital of Philadelphia
Nutrient adequacies in mexican children
Authors: Rosas Verónica Mundo , Ramírez Sonia Rodríguez , Levy Teresa Shamah
Published: June 2021

The aim of this study was to describe the reported energy, nutrient intake and adequacies in mexican children. We studied a national probabilistic sample of 3705 from 12 to 59 months of age in the National Survey of Health and Nutrition 2006 in Mexico (ENSANUT‐2006). A 7‐days semi‐quantitative food frequency questionnaire was used. Nutrient adequacies were estimated using the Dietary Reference Intakes and stratified according to area (urban or rural) and socioeconomic status (SES). SES was determined by household characteristics and possessions of goods and divided into tertiles (low, middle and high). Intake and dietary inadequacy risk (adequacy <50%) were estimated.

Our results revealed that energy, macro and micro‐nutrient adequacies were lower in rural (RA) than in urban areas (UA). The risk of inadequacy (RI) in rural areas was: Energy: 16.5% RA vs 7.4% UA, vitamin A: 21.4% RA vs 10.6% UA, folate:17% RA vs 13.8 UA and calcium: 18.2% RA vs 11.2% UA.

The RI was higher in the low SES: Energy: 14.9%, vitamin A: 18.4%, folate:17.7% and calcium: 18.3%. The RI iron was lower (<5%) in all areas and SES. However 1.6% of the median national total iron intake provides from to heme iron.

Mexico is experiencing a inequalities across areas and socioeconomic status. This information can contribute to the decision making to focus the beneficiaries of food programs, using indicators of available and access to foods.
Talin isoforms in cardiac development and adult heart
Authors: Manso Ana Maria , Ong Shannon , Padilla Steve , Lo Su Hao , Ross Robert S
Published: June 2021

Talin (tln) is a cytoskeletal linker protein. Tln has two genes: tln1 and tln2. The aim of this study was to evaluate the expression pattern of tln in mouse heart. For this we used tln2 gene‐trapped mice. This trap lead to expression of a chimeric protein with the N‐terminal portion of tln2 fused to b‐galactosidase (b‐gal). Tln2 expression was detected by monitoring b‐gal activity. Tln2 was detected in heart at early stages in cardiac development (E8 and E9) and its expression was more intense in heart compared to other organs at E10, 11, 12, and 15, as well as in the neonatal and adult heart. Due to the high similarity of tln2 and tln1, there are no specific antibodies to differentiate these two proteins. Our gene‐trapped mice allowed us to use antibodies against the C‐terminal domain of talin to distinguish tln1 from tln2 expression. Inmunofluorescence analysis of adult heart and isolated cells showed that tln2 is mainly expressed in cardiomyocytes, whereas tln1 is localized predominantly in endothelial cells and cardiac fibroblasts. In cardiomyocytes, tln2 was localized specifically in costameres. These results suggest that: (a) tln2 expression in the heart vary depending on the stage of development and (b) tln2 may have functions distinct from tln1 in heart due to its preferential expression in distinct cell types and locations.Supported by grants HL073393 and HL046345 from the NHBLI/ NIH
Research of algae at waters of river Morava Binqës
Authors: kurteshi kemajl , Hafner Dubravka , Vehapi Idriz , Gecaj Abdulla , Ismaili Muharrem , Salihu Dilaver , Ibrahimi Haqif , Sahiti Naim
Published: June 2021

During the summer 2006 we investigate the algae of the upstream part of the river Morava Binqes. We taken the samples from 4 locality. We determinate the composition of the algae: 6 species belong divission Cyanophyta, 28 species belongs Bacillariophyta, 3 species Euglenophyta and 6 species Chlorophyta, altogether 43 species. According the bioindicator species we are done the qualitative estimation of waters of river Morava Binqes. Based on the presence of species as saprobiologic bioindicator and saprobic level , the waters of river Morava Binqes belongs the second level of bonity respectively beta‐ mesosaprob
T cells shoot from the waist: low dose IL‐15 induces snap arming of CD44low T lymphocytes in the absence of antigen.
Authors: Tamang David L , Alves Bryce , Elliot Viki , Fraiser Stephanie , Redelman Doug , Hudig Dorothy
Published: June 2021

The current paradigm for lymphocyte activation is that T cells require two signals, antigen recognition and co‐stimulation, to become cytotoxic over the course of 3–5 days. This report describes a novel observation whereby freshly isolated splenocytes without prior exposure to specific antigen became functionally cytotoxic within 24 hours after culture with low‐dose IL‐15 (7x10‐10 M). The observed cytotoxicity was great enough so that one P815 target was killed per CD8+ T cell. Cytotoxicity was T cell dependent, as measured by anti‐CD3 redirected 51Cr release to detect the activity of all T cells. Furthermore, the effector T cell population killed P815 targets even when CD44hi (memory effector‐associated phenotype) CD8+ T cells were depleted. Cytotoxic capacity was gained before granzyme B could be detectable by flow cytometry. Phenotypic markers for the T cell population resembled that of resting T cells. Since IL‐15 is physiologically bound to IL‐15 receptors of dendritic cells and IL‐15 message is produced by a variety of peripheral tissues, this work suggests that effector T cells may gain cytotoxic potential earlier than currently thought.
Anti‐RNA Antibodies with Differential Specificity towards Polyribonucleotides from a Murine Model of Lupus
Authors: Kole Hemanta , Bolland Silvia
Published: June 2021

Autoantibodies against nucleic acids were found in a lupus prone FcγRIIB−/‐ mice sera in B6 background. With the addition of the yaa (Y‐chromosome autoimmune accelerator) gene, pathogenecity in these male mice was significantly enhanced and autoantibodies switch from anti‐DNA to anti‐RNA specificity. Here we have characterized anti‐RNA antibodies from a hybridoma clone H526 that was produced by the fusion of R2yaa spleenocytes with myeloma cells. Hybridoma H526 produced anti‐RNA IgG2a‐λ antibodies. Sequence analysis showed only a few amino acid changes in the VH and VL regions compared to B6 germ‐line Ig variable regions. H526 antibody was found to deposit in the mouse kidney after 24 hrs of injection. When R2yaa mouse was immunized with H526 antibody, we found a marked reduction of anti‐nucleolar antibodies in the serum indicating neutralization of pathogenic antibodies by anti‐H526 antibody. For the antigenic epitope characterization for the anti‐RNA H526 antibody, we examined many polyribonucleotides of various lengths and found that synthetic polyribonucleotides, Poly (G, C) of 40 mers is optimal for the antibody binding in an ELISA. The Poly (G, C)40 also selectively binds anti‐RNA antibodies from R2yaa serum. Further characterization of Poly (G, C) in pathogenic B cell binding, detection, and cellular localization and antigen presentation are in progress.
The Role of ICOS in Peripheral Inflammation in Lupus
Authors: Eardley Leah Diane , Odegard Jared , Craft Joseph
Published: June 2021

T and B cells, dendritic cells (DC) and macrophages infiltrate the kidneys of lupus‐prone mice, with analogous infiltrates found in humans with lupus nephritis. These cells, as well as circulating autoantibodies, can produce severe renal disease in lupus. We are interested in the role of CD4+ T cells in promoting such inflammation. We have also utilized the costimulatory molecule ICOS to investigate the mechanisms by which CD4+ T cells contribute to peripheral inflammation in lupus.

Our lab and others have demonstrated that the costimulatory molecule ICOS is necessary for cellular infiltration into the kidneys of MRL/Faslpr mice, a murine model of lupus. ICOS−/− MRL/Faslpr mice have greatly reduced renal cellular infiltrates. The infiltrates of kidneys were characterized by FACS, immunofluorescence, and histology. ICOS is expressed on the majority of CD4+ T cells found in the kidney. Ex vivo stimulated ICOS−/− CD4+ T cells from MRL/Faslpr mice produce significantly less quantities of effector cytokines, such as IFNγ, TNFα, IL‐10, and IL‐5, compared to ICOS+/+ CD4+ T cells from MRL/Faslpr mice. However, similar levels of certain chemokines, including MIG, are found in ICOS−/− MRL/Faslpr kidneys, compared to ICOS+/+ MRL/Faslpr kidneys. In addition, ICOS+/+ and ICOS−/− MRL/Faslpr CD4+ T cells expressed similar amounts of some chemokine receptors (CXCR3, the receptor for MIG), but altered amounts of other chemokine receptors (CCR6). Although the implications for CD4+ expression of CCR6 is unclear, as less than 10% of kidney infiltrating CD4+ T cells express CCR6. Taken together, these data demonstrate ICOS dependent and independent cytokine and chemokine production in lupus mice, as well as chemokine receptor expression by CD4+ T cells in MRL/Faslpr mice.
Mechanical injury by tape stripping causes Th2 responses in mice
Authors: Oyoshi Michiko K , Geha Raif S
Published: June 2021

Atopic dermatitis (AD) is a chronic inflammatory skin disease associated with cutaneous hyperreactivity to environmental allergens. In AD, skin injury caused by scratching and subsequent entry of antigen in the skin is critical to disease development. We hypothesize that DCs that emigrate from mechanically injured skin polarize the T cell response towards Th2. We used the hapten FITC as a marker to isolate DCs that have recently migrated from skin to draining lymph node (DLN). FITC was applied to shaved and tape‐stripped mouse skin, a surrogate for scratching, or to shaved skin without tape stripping. After 24 hrs, CD11cFITChi cells were enumerated in DLN, and examined for their capacity to cause proliferation and cytokine secretion in T cells. In each of the two groups CD11c+FITC+ constituted ~10% of DCs in DLNs and supported comparable proliferation of naïve DO11.10 CD4+T cells to OVA323–339 peptide. In contrast, FITC+ DCs from DLN of shaved/stripped skin induced significantly greater secretion of the Th2 cytokines IL‐4, IL‐5, IL‐13 and IL‐10, and less production of the Th1 cytokine IFN‐γ. IL‐10 is released in skin following mechanical injury. FITC+ DCs from DLN of IL‐10−/− mice were impaired in their ability to support Th2 cytokine secretion, consistent with our previous finding that the Th2 response to EC sensitization was impaired in these mice J Clin Invest. 114:399–407, 2004). The results demonstrate that mechanical skin injury polarizes skin DCs to support Th2 differentiation of naïve T cells; this is in part dependent on IL‐10.
An essential role of the G protein‐coupled formylpeptide receptor mRPR2 in the development of allergic airway inflammation
Authors: Chen Keqiang , Le Yingying , Hu Jinyue , Huang Jiang , Gong Wanghua , Bauchiero Samantha , Howard OM Zack , Tessarollo Lino , Wang Ji Ming
Published: June 2021

The human G protein‐coupled formylpeptide receptor‐like 1 (FPRL1) and its mouse homologue mFPR2 mediate the chemotactic activity of a variety of polypeptides associated with inflammation and microbial infection. In order to elucidate the role of mFPR2 in vivo, we established mice with disrupted mFPR2 gene. Although mFPR2−/− mice developed normally, they showed significantly reduced cellularity in the bone marrow and lymphoid organs, as well as defects in the maturation of bone marrow‐derived myeloid dendritic cells in vitro. In a model of oval‐albumin‐induced allergic airway inflammation, mFPR2−/− mice showed severely reduced infiltration of leukocytes into the lung and the bronchial lumen, with decreased production of key immunoglobulins involved in type II allergic responses. Further studies revealed significantly reduced recruitment of CD11c+ antigen presenting cells in the spleen and lymph nodes of oval‐albumin immunized mFPR2−/− mice. These results indicate a key role of mFPR2 in the development of allergic airway inflammation requiring dendritic cell activation and trafficking into the lymphoid organs.