bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Vitamin B6 Deficiency on Lipid Metabolism in HepG2 Cells
Authors: Zhao Mei , da Silva Vanessa , Ralat Maria , Gregory Jesse F
Published: June 2021

We previously observed a decrease of plasma arachidonate (C20:4 n‐6) and an increase of erythrocyte oleate (C18:1 n‐9) in human volunteers with marginal vitamin B6 (VB6) deficiency. The objective of our current study was to investigate whether VB6 deficiency affects lipid metabolism in HepG2 cells and mechanisms involved. HepG2 cells were cultured for 6 wk in regular minimal essential medium (RMEM) containing 2.2 μM pyridoxal (PL) or VB6 – free MEM containing 10 nM, 20 nM, and 50 nM PL. After 6 wk, the intracellular pyridoxal 5′ – phosphate was 73 % lower for cells cultured in MEM containing 10 nM PL versus 2.2 μM PL. Total fatty acids increased 21%, the proportion of total arachidonate decreased from 4.3% to 3.6% (p < 0.05), and that of total oleate increased from 32% to 35% (p < 0.05) in VB6 –deficient cells cultured in medium with 10 nM PL compared to cells cultured in RMEM. Moreover, membrane lipid analysis showed that the proportion of arachidonate and dihomo‐γ‐linoleate (C20:3 n‐6) decreased respectively from 5.1% to 4.5% (p = 0.01) and from 0.9% to 0.6% (p < 0.001) in cells cultured in 10nM – PL medium compared to cells in RMEM. Our study demonstrated that VB6 deficiency moderately affects fatty acid profiles in HepG2 cells. Further investigation will focus on fatty acid metabolic kinetics in HepG2 cells with VB6 deficiency. Supported by NIH grant DK072398.

Grant Funding Source: NIH grant DK072398
Lipids Profile After Consumption of Different Types of Vegetable Oils: A Pilot Test in Rats
Authors: Gaona Gabriela , Reyes Patricia , Delgado Rubí , Montañez Alvaro , Ferriz Roberto A , del Carmen Caamaño María , Anaya Miriam A , García Olga P , Rosado Jorge L , García Teresa , de la Torre‐Carbot Karina
Published: June 2021

Coronary Heart Disease (CHD) is one of the major causes of death worldwide. Its development is strongly related to intake and type of fat more important than total amount of fat. The objective of the present study was to evaluate the effect of the consumption of canola, high oleic safflower, grape seed, soybean and avocado oil on lipid profile. An experimental controlled study was carried out for 6 weeks. Groups of 3 wistar rats were formed. Six groups were fed with a different type of vegetable oil added to standard rat chow. There were 2 control groups: one with no oil added and other who received partially hydrogenated oil. Hydrogenated oil group showed the highest levels on all lipid profile parameters. Avocado (45.83), canola (51.67) and soybean (48.10) oil group, presented significantly lower total cholesterol concentration mean (mg/dL) compared with hydrogenated oil group (70.27). Tryglycerides concentrations mean (mg/dL) were singnificantly lower in the canola (25.07), safflower (18.73), and grape seed (18.83) oil groups, compared with hydrogenated oil group (39.73). Soya oil group presented a better LDL/HDL ratio mean (2.27) in comparison with grape seed (3.00), hydrogenated (3.58), canola (2.90), and safflower (3.39) oil groups (P=0.05). The type of vegetable oil consumed in the diet modifies the lipid profile.
Low density lipoprotein quantification: a new look
Authors: Schlegl‐Zawadzka Malgorzata , Hartwich Jadwiga
Published: June 2021

Routine lab test for quantification of LDL plasma level is regarded as LDL cholesterol assessment. LDL is not a homogenous entity. The relative content of cholesterol decreased from large more buoyant to small, dense (sdLDL) particles (Swinkels 1987). LDL phenotype B i.e. the predominance of sdLDL in LDL plasma pool was significantly associated with a threefold increased risk of myocardial infarction (Austin 1988). People with low or normal cholesterol may have a high number of LDL particles, even if their cholesterol is being lowered by diet, exercise, or medication (Cromwell 2006). The need of therapeutical modulation of small, dense LDL particles content in LDL plasma pool is widely discussed (Rizzo 2007). Dietary effects differ in subject with LDL subclass phenotypes A and B (Krauss 2005). Diet‐induced favorable changes in plasma lipid and lipoprotein profile were noted only in the ‘post‐dietary LDL density responders’ (Hartwich 2009). Qualitative and quantitative measurements of LDL have been shown to be of benefit in cardiovascular risk assessment, so the classic lipoprotein analysis should be improved by high separation techniques.
Incorporation profiles of isomers of conjugated linoleic acids (CLA) into raft and non‐raft lipids of CHO cells.
Authors: Subbaiah Papasani Venkata , Gould Ian Gopal , Lal Samanta
Published: June 2021

The aim of this study was to determine whether part of the biological effects of CLA is due to their incorporation into membrane lipids, and consequent modification of membrane raft function. The incorporation of four CLA isomers, trans 10‐cis 12 (A), trans 9‐trans 11 (B), cis 9‐trans 11 (C), and cis 9‐cis 11 (D), and of linoleic acid (LA) into raft lipids, and their positional distribution in phospholipids was studied. In all cases, more than 60% of the cellular CLA was present in the raft fraction. Part of the CLA was converted to the 18:3 and 20:2 derivatives by the cells. Of the total CLA incorporated, 60% was present in phosphatidylcholine (PC), and 30% in phosphatidylethanolamine (PE). Unlike LA, which was present almost exclusively in the sn‐2 position of phospholipids, 30–40% of A, C, and D, and up to 70% of B were found in the sn‐1 position of PC. About 65% of the total CLA in PE was present in sn‐1 position for all the isomers. In all cases endogenous 18:1 was the predominant fatty acid replaced by CLA, but saturated fatty acids were also replaced by B. Furthermore, A, C, and D inhibited the Δ9 desaturase activity, as evident from the decreased ratios of monunsaturates/saturates. CLA‐B, however, increased this ratio, indicating no inhibition of the enzyme by it. These results show that CLA incorporate preferably into raft lipids, with a large percentage in sn‐1 position of phospholipids, which may affect the membrane functions.
Stearidonic acid‐enriched soybean oil as an effective source of long‐chain polyunsaturated fat in foods
Authors: Lemke Shawna , Wilkes Richard , Welsby David , Goldstein Daniel , Krul Elaine , Mukherjea Ratna , Harris William
Published: June 2021

Health benefits of the long chain polyunsaturated fatty acids (LCPUFAs), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are well documented. To meet intake needs, plant‐based sources are being developed that will augment and provide an alternative to fish sources that are limited in sustainable supply. A soybean that produces 20–30% w/w stearidonic acid (SDA) has been developed through biotechnology. SDA is the delta‐6 desaturase product of alpha‐linolenic acid (ALA) and clinical data show that it readily enriches blood tissues with EPA. SDA soybean oil (SBO) is processed similar to commodity SBO, is similarly bland in flavor and has been successfully added to a range of foods resulting in acceptable flavor and shelf life. Fruit and nut granola bars were developed by incorporating SDA SBO and other omega‐3 oils and evaluated throughout 12 months of shelf life. Bars made with SDA SBO were similar to control bars in flavor, off flavor, and consumer acceptance for overall liking and flavor throughout the 12 month shelf life. Modeling consumption of SDA SBO in a variety of processed foods using NHANES 1999–2002 and at level that is achievable in foods, i.e., 375 mg SDA/serving, resulted in an estimated mean intake of 2.2 g SDA/day. The results demonstrate that SDA‐enriched SBO has the potential of being formulated into shelf‐stable products with consumer appeal that can increase the effective intake of LCPUFAs.
Fatty acids of plasma phospholipids and erythrocytes are reliable biomarkers of n‐3 polyunsaturated fatty acid supplementation
Authors: Rudkowska Iwona , Plourde Mélanie , Julien Pierre , Lemieux Simone , Couture Patrick , Vohl Marie‐Claude
Published: June 2021

The availability of reliable biomarkers to assess the efficacy of dietary interventions is a prerequisite of meaningful clinical studies with n‐3 polyunsaturated fatty acids (PUFA).

Objective

To determine the suitability of using either plasma phopholipids (PL) or erythrocytes fatty acids (FA) composition as indicators of n‐3 PUFA status.

Methods

We measured FA % in erythrocytes and plasma PL obtained from 28 men supplemented daily with 3g of n‐3 PUFA (1.9 g eicosapentaenoic acid (EPA) and 1.1 g docosahexaenoic acid (DHA)) during 6 weeks.

Results

After supplementation, EPA, DHA and EPA+DHA in plasma PL or erythrocytes increased by 334%, 43%, 174% and 195%, 16%, 29%, respectively (p<0.05 versus baseline for all analyses). EPA in plasma PL was correlated to EPA in erythrocytes at baseline (r=0.77, p<0.01) and after supplementation (r=0.63, p<0.01). In addition, a correlation (r=0.47, p=0.02) was seen between the combined EPA+DHA in plasma PL and erythrocytes after supplementation.

Conclusion

Even tough differences do exist in the FA composition of plasma PL and erythrocytes, both biomarkers significantly increased with n‐3 PUFA supplementation over 6 weeks suggesting that they are both reliable biomarkers of n‐3 PUFA status after supplementation. Funding provided by a CIHR Operating Grant.
Lipidemic and cholesterolemic effects of feeding an algal source of docosahexaenoic acid to mice
Authors: Pietrofesa Ralph A , Barnes Kimberly M
Published: June 2021

Algal sources of ω‐3 polyunsaturated fatty acids are abundant in docosahexaenoic acid (DHA) but have little to no eicosapentaenoic acid. Our objectives were to evaluate algal sources of DHA vs fish oil on body fat and serum lipids. Male mice (n=100, 10 wk old) were fed 12% lipid diets containing either 0 (soy oil; SO), 5, or 10 g/kg DHA from fish oil (FO5; FO10), algal oil (AO5; AO10), or dried algal biomass (AB5; AB10) for either 2 or 4 wk. A body fat index was calculated and serum lipids were determined. Body weight and feed intake did not differ. At 2 wk, FO‐fed mice were leaner than AO‐fed mice (P < 0.01), but only tended (P = 0.09) to be leaner than SO‐fed mice. AO10‐fed mice were fatter (P < 0.01) than SO‐fed mice. At 4 wk, there were no significant differences in body fat index, but FO‐fed mice tended (P = 0.06) to be leaner than all other groups. Serum triacylglycerol (TAG), total cholesterol (TC), and LDL cholesterol (LDLC) were reduced only in the FO10‐fed mice at 2 wk (P < 0.01) but at 4 wk, FO5 and AO10‐fed mice also had lower serum TAG, TC, and LDLC (P < 0.01). In conclusion, algal DHA does not appear to be as effective as fish oil at reducing body fat or serum lipid parameters.
Iodine intake from dietary supplements among pregnant females in the US: National Health and Nutrition Examination Survey 1999–2006
Authors: Gahche Jaime Jacqueline , Bailey Regan , Mirel Lisa , Dwyer Johanna , Russell Robert , Sempos Christopher , Picciano Mary Frances
Published: June 2021

During pregnancy, there is an increased need for iodine because iodine is essential for adequate neural development and growth of the fetus. Therefore adequate iodine nutriture is critical at that time. The Estimated Average Requirement (EAR) is 160 μg. Previous reports indicate that iodine is available in only 50% of prenatal dietary supplements (DS) sold in the US. The purpose of this study was to characterize use and the amount of iodine in DS used by pregnant women in a population‐based survey, the NHANES during 1999–2006. The oversample of pregnant women 15–39 years of age during that period permitted reliable estimates on this nationally‐representative sample of 1234 pregnant women. While 78%(SE 2) of these pregnant women reported taking one or more DS in the past 30 days, only 21%(SE 2) of them reported consuming one that contained iodine. The most common amount reported on the label was 150 μg iodine/serving. Average daily iodine intake was estimated using the amount of iodine from the DS facts label, frequency of use over the past 30 days, and the amount consumed on the days the DS was taken. Mean iodine intake for iodine supplement users (e.g. pregnant women taking one or more DS containing iodine) was 125 μg (SE 6) and the median was144 μg. 76% (SE 5) of the pregnant females using a DS containing iodine met or exceeded the EAR of 160 μg. However, intakes of women using a DS not containing iodine or not using a DS should be further investigated.
Dietary patterns affected the increase of Bone Mineral Content in Korean girls.
Authors: Noh Hwa Young , Song YoonJu , Park Min Kyung , Li Shan Ji , Joung Hyojee , Paik Hee Young
Published: June 2021

The aim of this study is to identify the dietary patterns among Korean girls and explore its relationships with change of body mass index (BMI), body fat, and bone mineral density (BMD) and bone mineral contents (BMC) during 22 months. Girls aged 9–11 yr were recruited and 3‐day dietary data was collected four times and anthropometric values were measured baseline and 20 months later. Reduced rank regression (RRR) was used to derive dietary pattern using the change of BMI, body fat, BMD and BMC as response variables. Two dietary patterns were identified and the first was characterized by low intake of nuts & seeds, processed meat, potatoes, and the eastern grains and the second was characterized by high intake of fruits, nuts & seeds and milk & dairy products and low intake of vegetables, mushrooms, and kimchi. In the change of BMI and body fat, two dietary patterns had similar trend, but the change of BMC was different by dietary patterns. First dietary pattern was negatively correlated with the change of BMC (r=−0.49, p<0.01), while second dietary pattern was positively correlated with the change of BMC (r=0.25, p<0.01). The higher RRR score of the second dietary pattern was associated the larger change of BMC. Our results suggest that high intake of fruits, nuts & seeds, and milk & dairy products may contribute to increasing the peak bone mass among young girls. This work was supported by the Korean Science & Engineering Foundation.

Grant Funding Source: the Korean Science & Engineering Foundation
Effects of duration of proton pump inhibitor (PPI) therapy on markers of bone health in men and post‐menopausal women
Authors: Pabin Zarina , Newell Brian E , Richards Rickelle , Franz Kay B , Davidson Robert T
Published: June 2021

Because PPI therapy reduces stomach acid which theoretically could reduce mineral absorption, this observational study compared measures of bone health biomarkers, bone mineral density (BMD), dietary habits, and physical activity levels of men (n=31) and non‐estrogen supplementing post‐menopausal women (n=23). Participants were further divided according to duration of proton pump inhibitor (PPI) therapy; more than 5 years (n=16), less than 5 years (n=15), and no PPI therapy (n=23). The shortest duration of PPI therapy was 2 months and the longest duration of PPI therapy was 25 years with a mean duration of 7.5 years. No significant differences were found between measures of spine BMD, urinary deoxypyridinoline (i.e. bone resorption), urinary calcium and magnesium, serum osteocalcin (i.e. bone formation), serum parathyroid hormone, serum magnesium, serum 25 hydroxyvitamin D, dietary and supplement intake, or physical activity levels. However, mean hip BMD was higher in females than in males in participants who took PPI therapy for any duration. In the no‐PPI therapy group, hip BMD was not significantly different between genders. This study suggests that there may be no measurable or clinically significant effects of long term PPI therapy on bone health.
Effects of Astragalus membranaceus supplementation with calcium on the bone mineral density and bone metabolism in Ca‐deficient ovariectomized rats
Authors: Kim Hee Jung , Kang Se‐Chan , Kim Mi‐Hyun
Published: June 2021

Recently, it was reported that traditional herbs, Astragalus membranaceus (A. membranaceus) had estrogenic effect in vitro. To examine the possible role of A. membranaceus supplementation with calcium on bone status in Ca‐deficit ovariectomized rats, a total of 48 female rats were used and divided into 6 group, and A. membranaceus ethanol extract (500 mg/kg) or Ca (800 mg/kg) were administered orally for 8 weeks with feeding Ca‐deficient diet. The groups were as follow: (1) Normal control, (2) Sham control, (3) OVX control, (4) A. membranaceus administration after OVX (OVXM), (5) Ca administration after OVX (OVXCa), (6) A. membranaceus and Ca administration after OVX (OVXMCa). A. membranaceus supplementation significantly increased the ash contents of tibia (p<0.001) and femur (p<0.001) in OVX rats. Also, A. membranaceus alone or combined with Ca administration significantly decreased the urinary Ca excretion (p<0.001). BMDs and bone strengths of tibia and femur in OVXCa and OVXMCa were significantly higher than those in OVX control or OVXM group. Uterine weight was significantly increased by A. membranaceus administration in OVX rats. These results suggest the possibility of using the A. membranaceus as functional material being able to augment calcium's effect on bone health in postmenopausal women.
Anti‐osteoporotic effects of silibinin and silymarin and on osteoblast differentiation and RANKL‐dependent osteoclast differentiation
Authors: Kim Jung‐Lye , Choi Jung‐Suk , Oh Seon‐Mi , Kang Young‐Hee
Published: June 2021

Bone‐remodeling imbalance induced by increased bone resorption and osteoclast formation is known to cause skeletal diseases such as osteoporosis. Silibinin (SB) is the major active constituent of silymarin (SM), the mixture of flavonolignans extracted from blessed milk thistle. Several studies suggest that SB is a powerful antioxidant and has antihepatotoxic properties and anti‐cancer effects against carcinoma cells. This study investigated that SB and SM had anti‐osteoporotic effects. Murine macrophages were pre‐incubated with 10 μM SB and SM for 5 d in presence of RANKL. Non‐toxic SB inhibited RANKL‐mediated expression of TRAP in mature osteoclasts. In addition, SB markedly attenuated gelatinolytic MMP‐9 activity elevated by RANKL. SB suppressed the formation of multi‐nucleated osteoclasts, which was also observed by SM. Furthermore, MC3T3‐E1 mouse cells were treated with SB and SM for 14 d in a differentiate medium. It was found that SB and SM had a significant induction of osteoblast proliferation and differentiation. Additionally, alkaline phosphatase activity and matrix mineralization were enhanced with SB and SM, as determined by von Kossa and Alizarin red stainings. These results demonstrate that SB and SM were potential therapeutic agents for the prevention osteoporosis. Supported by the Regional Research Universities Program/Medical & Bio‐Materials Research Center.
Effect of Natural Plant Water Extracts Suppressed Melanin Production In Human skin cell(CCD‐986sk)
Authors: Jang Jung Yeong , Chung Mi Ja , You Jin kyoun , Kim Dae Jung , Seo Dong Joo , Kim Tae Hyuk , Choe Myeon
Published: June 2021

Melanin is the pigment responsible for the color of human skin. Tyrosinase is known to be a key enzyme that catalyzes the synthesis of melanin in melanocytes. Therefore, we intend to verify the effect of whitening using these natural plant water extract (MH: Ephedrae, Santalum album, BDH: Agrimonia pilosa ledeb,SHC: Houttuyniae herbal, ESC: Houttuynia cordata Thunberg, JGP: Sorbus amurensis Koehne, TSJ: Coscuta japonica chois ). The effect of natural water extracts on antioxidant activities, tyrosinase activity, melanin biosynthesis have been studied in a human skin cells or in a cell‐free system. The tyrosinase activity and melanin content was assessed using the RT‐RCR, western blotting and spectrophotometric methods. The results show that the SHC has a greater ability to inhibit tyrosinase activity than tested five extracts. MH, SHC, and JGP significantly suppressed melanin production in a human skin cells.

(Following are results of a study on the “Human Resource Development Center for Economic Region Leading Industry” Project, supported by the Ministry of Education, Science & Technology(MEST) and the National Research Foundation of Korea(NRF).)
Do organic snack foods taste healthier because of their label?
Authors: Lee Jenny Wan‐Chen
Published: June 2021

In contrast with past ‘health halo’ studies, which focused on the perceptions associated with organic labels, this study investigated the influence of organic labels on behavior and post‐consumption evaluations. Fifty‐four subjects, ages 20 to 28, were presented with cookies and potato chips labeled either ‘organic’ or ‘non‐organic.’ After consuming them, subjects rated the snacks based on a number of sensory, nutritional, and value dimensions. Participants consuming snacks labeled ‘organic’ reported higher levels of taste experience (p=.011), appearance ratings (p=.023), and estimated nutritional values (p=.006) than those who consumed snacks without the label. By generating a more positive consumption experience, such labels may stimulate greater consumption of organic foods overall—including foods which are not necessarily healthy or nutritious. Interestingly, the participants most influenced by such labels also self‐reported higher preferences of recycling (r=.477, p=.01) and a greater appreciation of nature (r=.413, p=.036). Identifying those segments particularly susceptible to these labels may serve as accurate predictors of post‐consumptive behavior.

Grant Funding Source: none
Has the obesity epidemic leveled off in US children? What do recent data tell us?
Authors: Wang Youfa , Beydoun May , Caballero Benjamin
Published: June 2021

Recent data suggest that childhood obesity has leveled off in the US. This is encouraging, though problematic if inaccurate. To address this important but complicated public health problem, we systematically examined recent trends in obesity and various adiposity measures and energy balance‐related behaviors in US children using published findings and analysis of recent nationally representative data from National Health and Nutrition Examination Survey (NHANES) and Youth Risk Behavior Survey (YRBS). We found complex patterns in adiposity measures and across population groups. Results also varied by dataset and data analysis method. In general, obesity/overweight rates have increased during 1988–2008 and since 1999 in various US children sex‐, age‐, and ethnic groups using NHANES. In 1988–94, 1999–04, and 2005–08, obesity (%) in boys aged 2–19 y was 11.7, 15.9, 18.4; in girls, 11.2, 14.2, 16.7. There are signs of a plateau in the prevalence in recent years for some groups. YRBS showed obesity (%) was 10.7 in 1999, 10.5 in 2001, 13.1 in 2005–07. The data did not show material improvements in US young people's diet and physical activity in recent years. At least some observed plateau in obesity observed in recent NHANES is due to sampling and analysis issues and short survey periods. More data are needed to monitor the trends. Vigorous effort still urgently needs to promote healthy lifestyle to fight obesity epidemic.
Bovine pituitary extract interferes with SARS‐CoV replication
Authors: Jelesijevic Tomislav , Uhl Elizabeth , Michel Frank , Hogan Robert Jeffrey
Published: June 2021

Severe acute respiratory syndrome (SARS) was the first pandemic of the 21st century, and the causative agent was found to be a novel RNA coronavirus virus (SARS‐CoV). Unlike other human coronaviruses, which induce mild respiratory disease, SARS‐CoV infection was characterized by age‐dependent differences in mortality rates. The mortality rate for people over 60 was around 50%, while in those under 24 years of age it was under 1%. With this in mind, we hypothesized that the observed differences in mortality between young versus elderly people may be due to differences in the secretion of pituitary gland bioactive molecules. To test this hypothesis, the effects of bovine pituitary extract (BPE) on SARS‐CoV replication were examined in vitro by adding various amounts of BPE to African green monkey kidney epithelial cells (Vero E6). Interference with SARS‐CoV replication was observed via TCID50 assays and plaque assays. Results of both assays indicated significant, BPE dose‐dependent inhibition of SARS CoV replication. Future studies are aimed at identifying the molecules that mediate the observed responses and their mechanism(s) of action.

Research is supported by NIH grant 1R21AI071109‐01A1 (RJH).
Effects of claudin overexpression in carcinoma cell lines
Authors: Zavala‐Zendejas Veronica Elizabeth , Torres‐Martínez Ana Cristina , Salas‐Morales Berenice , Rendon‐Huerta Erika Patricia
Published: June 2021

Claudins are proteins that preserve cell polarity and determine the size and charge of the molecules that pass through the paracellular space in epithelial tissues. Altered claudin expression in early and late stages of several cancers has been related to increased invasiveness, metastatic potential, poor prognosis or tumor recurrence. We analyzed if claudin‐2, ‐6, ‐7 or ‐9 overexpression confered changes in cell motility, invasion and proliferation in a human gastric adenocarcinoma cell line or in a breast cancer cell line. MCF‐7 and AGS cells were transfected with these claudins and proliferation rate, Matrigel invasion and claudin‐1 expression were determined. While overexpression of claudin‐2, ‐6, ‐7 and ‐9 in AGS cells enhanced their invasive potential, in MCF‐7 cell line only claudin ‐7 and ‐9 had a similar effect. Proliferation rate of claudin‐2, ‐6, ‐7 and ‐9 transfected AGS cells increased, however claudin‐2 overexpression in MCF‐7 cells had the opposite effect. Our results show that the induced modifications in proliferation or invasive potential might be associated with changes in claudin‐1, expression.
Galectin‐3 mediates chronic airway allergic inflammation and airway remodeling
Authors: Ge Xiao Na , Bahaie Nooshin S , Kang Bit Na , Hosseinkhani Reza M , Ha Sung Gil , Frenzel Elizabeth M , Liu Fu‐Tong , Rao Savita P , Sriramarao P
Published: June 2021

The role played by Galectin‐3 (Gal‐3) in airway remodeling, a feature of chronic asthma that leads to airway dysfunction and poor clinical outcome in humans, was investigated in a murine model of asthma. Wild‐type (WT) and Gal‐3 knock‐out (KO) mice were subjected to repetitive allergen challenge with ovalbumin (OVA) up to 12 weeks and bronchoalveolar lavage fluid (BALF) and lung tissue collected after the last challenge were evaluated. Chronic OVA challenge induced Gal‐3 expression in the lungs of WT mice. Cell counts in BALF and lung immunohistology demonstrated that eosinophil infiltration in OVA‐challenged Gal‐3 KO mice is significantly reduced compared to WT mice. Also, eotaxin‐1, IL‐2, IL‐4 and TGF‐β expression in these mice was substantially lower than in OVA‐challenged WT mice. Decreased eosinophil recruitment in Gal‐3 KO mice was associated with diminished remodeling of the airways with significantly reduced mucus secretion, sub‐epithelial fibrosis, smooth muscle thickness, and peribronchial angiogenesis. Finally, leukocytes from Gal‐3 KO mice demonstrated decreased trafficking on endothelial adhesion molecules compared to WT cells. Overall, blockade of Gal‐3 in a chronic setting leads to diminished airway inflammation and remodeling potentially due to reduced trafficking of inflammatory cells to sites of inflammation. 1National Institutes of Health grants AI35796, U19‐AI70535, HL0793041.
Toll‐like receptor 2 signaling pathway in lung injury
Authors: Jezierska Agnieszka , Kolosova Irina A , Verin Alexander D
Published: June 2021

Endothelial cell (EC) barrier is critical for regulation of fluids and gas exchange through an alveolar space. Disruption of this barrier leads to pulmonary edema and lung inflammation [hallmarks of acute lung injury (ALI)], often followed by multi‐organ failure or even death. Based on data of literature, we hypothesize that in development of this pathological conditions Toll‐like receptor 2 (TLR2) signaling is involved. To test this hypothesis we employed several methods including measurements of transendothelial electrical resistance, fluorescent imaging and western blotting. Our data demonstrate that TLR2 specific agonist, Zymosan and TLR2 specific antibody activate pro‐contractile pathways including increased in myosin light chain (MLC) phosphorylation, p38 and ERK MAPK activation, leading to increase in actin stress fibers, gap formation and EC permeability increase. This was accompanied by active beta‐catenins liberation from endothelial junctional complexes with VE‐cadherin. Based on our data, we strongly believe that targeting and inhibiting TLR2 or TLR2 related proteins with specific agonists may benefit patients with ALI or even more severe form acute respiratory distress syndrome (ARDS).
Characterization of chondrocyte differentiation of ATDC5 cell line induced by Ascorbic acid
Authors: Temu Tecla Mtui , Phornphutkul Chanika , Gruppuso Philip
Published: June 2021

The ATDC5 cell line exhibits a multistep of chondrogenic differentiation observed during endochondral bone formation. ATDC5 cell line is routinely induced to differentiate by exposure to insulin at high concentration thus limiting the ability to study nutrients and growth factors signal mechanism. We previously observed spontaneous induction of differentiation upon exposure to ascorbic acid containing alpha MEM medium. In this study, we first characterize the process of ATDC5 cells differentiation using ascorbic acid containing alpha MEM medium. Compared to high dose insulin, ascorbic acid induced mRNA expression of Collagen X by more than 2 folds. Ascorbic acid had no significant effect on the IGF‐1 or insulin receptors while high dose insulin downregulated both receptors respectively. Our findings suggest that we can employ ascorbic acid containing alpha MEM medium as a condition of choice for studying ATDC5 cells differentiation. We investigated the mechanism whereby ascorbic acid induced the differentiation of ATDC5 cells. Inhibitor of collagen synthesis completely blocked the effects of ascorbic acid on ATDC5 cells. When plates containing ascorbic acid‐condition chondrocyte matrix were used as substrate for prechondrocyte cells were able to differentiate in the absence of ascorbic acid an indication that collageneous matrix formation is essential for the induction of ATDC5 cells differentiation.
Changes in protein profile composition of coronary luminal membrane induced by ischemia and ischemia/reperfusion
Authors: Rubio Rafael , Arroyo‐Flores Blanca , de la Rosa Ana Barba , Knabb Maureen
Published: June 2021

Endothelial luminal glycocalyx is a layer of membrane bound and unbound molecules complexes and it is a poorly understood dynamic organelle. Ischemia‐reperfusion (I/R) injury disrupts the glycocalyx gross structure causing microvasculature dysfunctions not fully understood. We hypothesized that I/R injury might cause chemical changes in the constituents of glycocalyx and an associated decrease in mass. In the present study, during control conditions, isolated rat hearts (n=5/group) were perfused (37°C) with sulfo‐NHS‐LC‐biotin to biotinylate luminal endothelial cell proteins. Only biotin‐label proteins were isolated by passing the protein membrane extract through a streptavidin‐affinity column. The groups of hearts were: control (C), ischemia (I; ischemia 30min), and ischemia‐reperfusion (IR; reperfusion 30 min). The biotinylated membrane proteins were examined by 2‐D electrophoresis. The protein profiles comparative analysis between three conditions in gel 2‐D showed that the proteins profiles of C, I and I/R are different. Molecular weight (MW) population analysis show profiles were: MWC >MWI >MWI/R. Similar analysis for isoelectric points (pI) show profiles were: pIC >pII ≤pII/R. The results indicate that I and I/R injuries reduce MW of the luminal glycocalyx proteins and alter their pI. Funded by CONACyT SEP‐42567, CONACyT‐SALUD 2004‐C01‐156 and Fulbright Award #9583
Rac1‐mediated H2O2 generation suppresses MMP‐9 expression via ERK
Authors: Murthy Shubha , Carter A Brent
Published: June 2021

Our previous observations demonstrate that deletion of Rac1 from cells of the myeloid lineage protects mice from asbestos‐induced pulmonary fibrosis due to attenuated H2O2 generation. To address whether this was due to increased matrix degradation, we investigated the expression of matrix metalloproteinase (MMP‐9) in Rac1 null macrophages. Compared to wild type (WT) macrophages, MMP‐9 gene expression was significantly greater in Rac1 null cells. Site directed mutagenesis of the MMP‐9 promoter revealed that SP‐1 and AP‐1 were necessary for MMP‐9 expression. As these transcription factors are activated by MAP kinases, we examined the role of ERK in Rac1‐mediated MMP‐9 gene transcription. ERK activation was dramatically increased in Rac1 null cells and transfection with dominant‐negative ERK significantly decreased MMP‐9 promoter activity. Exogenous H2O2 inhibited MMP‐9 promoter activity in Rac1 null cells whereas over expressed catalase in WT cells enhanced ERK activation and MMP‐9 expression. Thus Rac1‐mediated H2O2 production suppresses MMP‐9 expression via inhibition of ERK. In vivo, MMP‐9 mRNA was enhanced in lung macrophages of Rac1 null mice and in WT mice administered catalase after asbestos exposure. These data suggest that enhanced MMP‐9 expression attenuates, in part, the development of asbestos‐induced pulmonary fibrosis. This work was supported by NIH grant ES‐015981.
Low immunohistochemical expression of MGMT in ACTH secreting pituitary tumors of patients with Nelson syndrome
Authors: Salehi Fateme , Scheithauer Bernd W , Moyes Veronica J , Drake William M , Syro Luis , Manoranjan Branavan , Sharma Soniya , Horvath Eva , Kovacs Kalman
Published: June 2021

Objective

MGMT expression in brain and pituitary tumors has been correlated with temozolomide treatment. Few medical therapies are available in patients with Nelson Syndrome. The aim of the present study was to assess immunohistochemical expression of MGMT in ACTH‐secreting pituitary adenomas in patients with Nelson Syndrome.

Methods

Our material consisted of specimens from ACTH‐secreting pituitary adenomas from patients with Nelson Syndrome. Immunohistochemical staining for MGMT was performed using the streptavidin‐biotin‐peroxidase complex method. MGMT immunoreactivity was assessed microscopically and recorded as an estimated percentage of nuclear MGMT immunopositivity. (0=none, 1=<10%, 3=<50%, 4=>50%)

Results

Male:Female ratio was 3:5, with average patient age being 62 (range 57–66). Five of the eight specimens (65%) exhibited no MGMT immunoreactivity, with two out of eight cases (25%) showing slight MGMT immunopositivity and one out of eight cases (12%) demonstrating moderate MGMT immunopositivity (<25%).

Conclusions

Temozolomide therapy may be useful in patients with Nelson Syndrome. Absent or low MGMT staining in brain and other neoplasms has been shown to correlate with successful treatment with temozolomide, and recent reports assessing aggressive pituitary adenomas suggest similar outcome.
Sensory Dysesthesia: Rare Presentation of Subependymoma
Authors: KURUKUMBI MOHANKUMAR , Ramidi Ganga , SAMINENI SWETHA , JAYAM‐TROUTH ANNAPURNI , DABI ALOK
Published: June 2021

Background

Subependymomas are uncommonly encountered ependymal tumors, accounting for less than 1% of all intracranial neoplasms and distinguished from ordinary ependymomas because of their generally better prognosis. Here we present a case of subependymoma with rare clinical presentation.

Case presentation

19 year old male student presented to us with complaints of persistent left sided pain and numbness for one month duration following herpes zoster over left face. Clinical examination was normal except decreased perception of pain and touch on the whole left side of the body.

MRI with gadolinium of the brain revealed a posterior fossa mass almost occluding the fourth ventricle with infiltration to the right side immediately behind the pontine tegmentum probably impinging on the right spinothalamic tract causing the patient symptoms. Post operative tumor histopathology revealed the classical appearance of subependymoma with characteristic clusters of nuclei in a dense fibrillary background.

Conclusion

Post operatively patient remained pain free at six month follow up. Supependymoma is generally asymptomatic. In this patient whole left sided dysesthesia raised the suspicion of central pathology than unlikely post herpetic etiology. This case is reported for the interesting atypical presentation and the importance of diagnosing benign tumor which can change the prognosis.
Modern methods in histo‐ and cytopathology: a view from Russia
Authors: Jargin Sergei V.
Published: June 2021

Immunohistochemistry, in situ hybridization, molecular and other modern methods are used in Russia mainly in research and are hardly available for routine histological and cytological diagnostics even in case of direct indications. These methods, requiring meticulous work with pipettes and test‐tubes, have never been primarily developed in Russia; the reagents, equipment and knowhow were imported from abroad. The following can be heard in support of such attitude: essential diagnostic questions can be answered on the basis of sections stained with hematoxylin‐eosin or Romanovsky‐Giemsa smears in most cases; information obtained by means of sophisticated methods is usually irrelevant for praxis or useful only for palliative care prolonging poor‐quality life of the terminally ill: precise classification of malignant neoplasms, primary localization of a metastasizing tumor etc. Obviously for that reason modern methods used in histo‐ and cytopathology are not covered by medical insurance. Last time, in view of the rapidly improving economy, new arguments are voiced: now we can purchase modern equipment and ready‐made kits; and the broadening international cooperation will make foreign knowledge and skills flow massively into our country. It is said, in other words, that modern methods will be brought to Russia on a gold‐rimmed saucer. After that, this abstract will be only of historical interest.
Sphingosine 1‐phosphate‐induced autophagy is mediated through activating endoplasmic reticulum stress response in human prostate cancer PC‐3 cells
Authors: Shih Chih‐Jen , Huang Yuan‐Li , Chang Chi‐Lun , Huang Wei‐Pang , Lee Hsinyu
Published: June 2021

Sphingosine 1‐phosphate (S1P) is a bioactive lysophospholipid that binds to a family of G protein‐coupled receptors (GPCRs), termed S1P1~S1P5. Our previous study has reported that S1P inhibits PC‐3, a human prostate cancer cell, proliferation through activation of autophagy. In addition, S1P‐induced autophagy plays a prosurvival role in PC‐3 cells. Accumulating evidences have studied that the cytoprotective effect of autophagy can be triggered by ER stress induction. Thus, we attempted to investigate whether S1P‐induced autophagy is resulted from triggering ER stress in PC‐3 cells. By monitoring XBP‐1 mRNA splicing, a characteristics of ER stress, we demonstrated that S1P triggers ER stress in a concentration and a time‐dependent manner. In addition, we also showed that S1P1 and S1P5 are required for S1P‐induced ER stress by using RNA interference experiments. Furthermore, signaling analyses revealed that PI3K, PLC, and ROS production were involved in S1P's effects on ER stress induction. On the other hand, knockdown of XBP‐1 significantly abolished S1P‐induced autophagy. In summary, our results demonstrated for the first time that S1P‐triggered ER stress is responsible for autophagy in PC‐3 cells.

Support by: Institute of Zoology
Beneficial Vascular Effect of A Non‐selective PPAR Activator In Aorta of Spontaneously Hypertensive Rats
Authors: Qu Chen , Leung Susan W.S. , Vanhoutte Paul M. , Man Ricky Y.K.
Published: June 2021

Wy14643 is a potent activator of peroxisome proliferator‐activated receptor (PPAR) α and γ. The present study aimed to determine whether or not Wy14643 improved endothelial dysfunction in hypertension, and if so, the mechanism involved. Isometric tension in isolated thoracic aortic rings of spontaneously hypertensive rats was recorded. Wy14643 caused greater relaxations than fenofibrate (PPARα activator) or rosiglitazone (PPARγ activator). L‐NAME (nitric oxide synthase inhibitor) and ODQ (soluble guanylyl cyclase inhibitor) alone and in combination inhibited these relaxations to the same extent. Compound C (AMP‐activated protein kinase (AMPK) inhibitor) reduced Wy14643‐induced relaxations to the same extent as L‐NAME. Endothelium‐dependent contractions evoked by acetylcholine in the presence of L‐NAME were reduced by Wy14643, fenofibrate but not rosiglitazone. Moreover, Wy14643 and fenofibrate inhibited acetylcholine‐induced prostanoids release to the same extent. Our data suggests that the three PPAR activators induce nitric oxide‐mediated relaxation through activation of AMPK. This relaxing effect is more prominent with Wy14643. Together with the ability to reduce the release of endothelium‐dependent contracting factor, it appears that Wy14643 produces better protection against vascular diseases in spontaneous hypertension.
Role of TLR4 expression in experimental hypertension
Authors: Bomfim Gisele Facholi , Tostes Rita , Fortes Zuleica , Carvalho Maria Helena Catelli
Published: June 2021

Cardiovascular system is exposed to pathogens and danger signals (endogenous ligands), resulting in activation of pattern‐recognition receptors including Toll like receptors (TLR). TLR4 have been shown in experimental models of atherosclerosis and heart failure, however it has not been described in hypertension. Our purpose was to investigate the gene expression of TLR4 in aorta, mesenteric arterioles, heart and kidney of spontaneously hypertensive rats (SHR) in phases: pre‐hypertensive (5 weeks) and hypertensive (10 and 15 week) and in this last the effect of treatment with anti‐hypertensive. A gradual increase in TLR4 mRNA in mesenteric arterioles, heart and kidney was observed in SHR according to age. At the age of 15 weeks, the gene expression of TLR4 in aorta of SHR was reduced while in the kidney it was increased when compared with WT rats. The treatment for 15 days with losartan and amlodipine decreased BP and TLR4 expression in mesenteric arterioles, heart and kidney. On the other hand, both treatments increased the TLR4 expression in aorta. We suggest that TLR4 has a role in the chronic inflammatory process present in hypertension.
High molecular weight kininogen regulates endothelial function
Authors: Kolte Dhaval Sanjeev , Osman Noah , Yang Jia , Shariat‐Madar Zia
Published: June 2021

This study demonstrates that the bradykinin (BK) precursor, non‐enzymatic cofactor high molecular weight kininogen (HK), regulates endothelial function. HK circulates in complex with two different zymogens, namely, prekallikrein (PK) and factor XI (FXI). Both PK and FXI participate in the contact phase of blood coagulation through their binding to HK. The assembly and activation of the HK‐PK complex on endothelial cells results in the generation of BK and nitric oxide (NO). Since no clinical abnormality is apparent in patients with PK deficiency, investigations were performed to determine the functional role of HK on endothelial cells using a flow‐based assay. HK positively regulated endothelial cell function by modulating Ca2+‐dependent process and endothelial nitric oxide synthase (eNOS) function. The inhibitors of BK‐dependent signaling pathway impaired HK‐mediated signal transduction in endothelial cells. HKH20 and HOE140 significantly reduced HK‐ or HK/PK‐induced intracellular [Ca2+]i changes. This is a novel finding indicating that HK contributes to the maintenance of endothelial barrier function in the absence of PK. Taken together, these findings highlight a novel potential function for HK‐receptor‐mediated crosstalk in regulating endothelial cells and suggest that HK may serve as a cardioprotective peptide. This work was supported by NSF MRI 0619774, AHA and NCRR/NIH P20RR021929 to ZSM.
Role of A1 adenosine receptors in vascular reactivity and inflammation in a murine model of allergic asthma
Authors: Ponnoth Dovenia S , Nadeem Ahmed , Tilley Stephen , Mustafa S. Jamal
Published: June 2021

We reported earlier that allergic mice have altered adenosine (AD) mediated vascular reactivity and systemic inflammation. This study was done to determine the role of A1 adenosine receptors (A1AR) in inflammation, vascular and airway reactivity using A1 WT and KO mice. Mice were divided into 4 groups: Control (Con), Con+AD, allergen challenge (Sen), Sen+AD and were sensitized on days 1, 6 with 200 μg ragweed (RW) i.p followed by 1% RW aerosol challenges (days 11–13). Con+AD and Sen+AD groups received one AD aerosol challenge (6 mg/ml) on day 14. Precontracted aortae (10−7M phenylephrine) were used in organ bath studies. WT Sen and Sen+AD groups had significantly lower relaxation (22.8±4.9% & 17.4±7.8% vs 48.64±1.93% in Con at 10−5M, p<0.05) to NECA (nonselective AD analog) and higher contraction (44.44±4.1& 46.2±5.4% vs 10.62±3.39 in Con at 10−6M, p<0.05) to CCPA (A1 selective agonist) than WT Con. All KO groups had a concentration dependent relaxation to NECA. ICAM‐1 and IL‐5 (2.01±0.76 & 1.9±0.5 vs 1.03±0.4 & 1.05±0.48 in Con, p<0.05) were higher in allergic WT aorta (undetected in KO). WT Sen and Sen+AD mice had higher airway hyperreactivity (AHR) to NECA (699.6±117.7% & 1312.7±116.6% vs 285.05±65.4% in Con, p<0.05). Thus, allergic WT mice had altered vascular reactivity, AHR and vascular inflammation which suggest that the A1 AR is proinflammatory systemically in this model. Supported by HL027339, HL04447 & HL071802
Investigation of fourth‐grade children's dietary recall accuracy and their achievement test scores (ATS)
Authors: Baxter Suzanne Domel , Guinn Caroline H , Royer Julie A , Hardin James W , Mackelprang Alyssa J , Smith Albert F
Published: June 2021

Each of 374 children (96% Black) was observed eating school breakfast and school lunch, and then interviewed to obtain a 24‐hour dietary recall that included the 2 observed meals. The school district provided ATS. Regression analysis showed that ATS were negatively correlated with omission rates (standardized regression coefficient [SRC] = −0.256; p < 0.001) and intrusion rates (SRC = −0.118; p = 0.015), 2 measures of reporting error for food items, and total inaccuracy (SRC = −0.120; p = 0.019), a measure that combined reporting errors for food items and amounts, and positively correlated with correspondence rates (SRC = 0.234; p < 0.001), a measure of reporting accuracy for energy. Effect sizes were small to medium for intrusion rate and total inaccuracy, and medium to large for omission rate and correspondence rate. Knowing that children's ATS are related to their dietary recall accuracy could be helpful for future school‐based nutrition studies which use dietary recalls provided by children. As achievement testing is routine in school districts, researchers could obtain ATS (for individual children) from districts and consider them when using dietary recalls to assess, for example, the relative validity of food frequency questionnaires or the effectiveness of school‐based interventions to improve children's dietary intake. Funded by R01HL074358.