bluuebunny/crossref_metadata_2025 huggingface dataset:
Magnesium Glycinate Supplementation in Bariatric Surgery Patients and Physically Fit Young Adults
Authors: DiSilvestro Robert A , Joseph Elizabeth , Starkoff Brooke E , Devor Steven T
Published: June 2021

Moderate deficiencies of magnesium could occur in diverse groups of people. Three questions that need to be answered are: One, what forms of magnesium supplements can correct the problem? Two, what types of people are prone to this problem? Three, what benefits are derived from correcting this problem? Some recent pilot data from our laboratory has been directed toward these questions. In one study, 6 week supplementation with 400 mg of magnesium as magnesium glycinate was effective at raising plasma magnesium readings in people who have had bariatric surgery (comparison made pre‐surgery to 6 weeks post‐surgery). Magnesium oxide did not produce this effect in these same type people, albeit at a slightly lower dose. In physically fit young adults, supplementation for 4 weeks with 400 mg of magnesium as magnesium glycinate, but not placebo, raised erythrocyte potassium content (a reflection of magnesium's role in body potassium retention and distribution). Magnesium supplementation also improved two measures of exercise performance (pre‐supplementation vs post‐supplementation). Interestingly, plasma magnesium was not increased significantly, though this does not provide the most sensitive indicator of altered magnesium status. In conclusion, the positive results obtained in this pilot work suggests that young fit adults, as well as people who have had bariatric surgery, tend to have less than optimal magnesium intake. The glycinate form of magnesium can improve magnesium status. In addition, in the pilot work, no reports were given of diarrhea, which can be a problem with some types of magnesium supplements.
Knock‐Downof Atp7a Expression Reduces Hephaestin Expression and Activity in A Rat Intestinal Epithelial Cell Line (IEC‐6)
Authors: Gulec Sukru , Doguer Caglar , Collins James F
Published: June 2021

Iron homeostasis is determined by intestinal absorption as no active excretory mechanisms exist. Recent studies demonstrated that copper content and expression of a copper transporter (the Menke'scopper‐transporting ATPase; Atp7a) increasein duodenal enterocytes during iron deprivation. Atp7adelivers copper to the trans‐Golgi for cuproenzyme synthesis and exports copper from enterocytes. Hephaestin(Heph), also expressed in enterocytes, is a multi‐copper ferroxidase which oxidizes exported iron to promote binding to transferrin. Heph represents the best characterized link between intestinal iron and copper homeostasis. Whether the copper transport function of Atp7a is necessary for Heph biosynthesis was tested in this study. Studies were performed in IEC‐6 cells in which Atp7a expression was silenced (>;90%) by siRNA. Metallothionein mRNA expression was induced in Atp7a knockdown (KD) cells, likely reflecting increased copper content. Moreover, Heph mRNA expression was reduced ~80% in the KD cells. It was subsequently shown that ferroxidaseactivity (measured by a spectrophotometric transferrin assay) in pure membrane and cytosolic fractions was reduced 50 and 35%, respectively (p<0.001), in KD cells as compared to negative control siRNA‐transfected cells. This investigation thus suggests that Atp7a influences Hephbiosynthesis in enterocytes and provides further mechanistic details regarding the influence of copper on iron homeostasis.
Secreted products from human adipose tissue decrease mitochondrial function of colon cancer cells
Authors: Schwartz Betty , Yehuda‐Shnaidman Einav , Nimri Lili
Published: June 2021

Obesity is linked to increased risk of colon cancer, the third most common cancer. Adipose tissue (AT) from obese differs from AT of lean in their immunogenic profile, body fat distribution and metabolic profile. AT from obese release free fatty acids (FFAs), and many pro‐inflammatory chemokines, factors known to play a key role in regulating malignant transformation or cancer progression.

Mitochondria are important organelles in cellular processes, including cellular respiration and energy expenditure, and programmed cell death (apoptosis). The metabolic networks that confer tumor cells oncogenic and metastatic properties, such as increased proliferation and the ability to avoid apoptosis, are still not well understood. Our goal in this study is to identify key molecular signals and interactions between adipocytes and colon cancer cells that may foster the genesis and growth of the latter.

We tested the effect of 24 hours exposure of conditioned media (CM) isolated from AT of obese vs. non obese subjects on the metabolic profile of colon cancer cell lines. We found that CM from obese subjects led to a reduction of basal and maximal oxygen consumption rates and to changes in the gene expression of mitochondrial proteins, with no effect on glycolytic proteins. This metabolic approach may lead to a more complete picture relating obesity to colon cancer.
Influence of dietary protein upon the development of obesity, insulin resistance (IR) and energy metabolism (EM) in malnourished mice fed a high fat diet (HFD)
Authors: Camargo Rafael Ludemann , Batista Thiago Martins , Branco Renato Chaves Souto , Gonçalves Vanessa dos Santos , Boschero Antônio Carlos , Carneiro Everardo Magalhães
Published: June 2021

Aim

To determine the influence of the dietary protein content upon obesity, IR and EM in malnourished mice fed a normal‐protein (NP‐HFD) or a low‐protein HFD diet (LP‐HFD).

Methods

Weaned male C57BL/6 mice were fed a protein‐restricted diet (6% protein ‐ R). After 6 weeks one group received a NP‐HFD (35% fat and 14% protein – RH) or LP‐HFD (35% fat and 6% protein – RRH).

Results

RH group had increased body weight (BW) (R=21.4±0.8; RH=30.7±1.0g) and perigonadal fat pad (R=1.6±0.08; RH=3.0±0.3%BW). RRH mice had lower BW (RRH=27.1±0.9g). RRH mice were protected from the hyperglycemia (R=65.8±3.1; RH=83.4±4.3; RRH=77.5±6.9mg/dL) and hyperinsulinemia (R=0.1±0.02; RH=0.3±0.03; RRH=0.1±0.03ng/mL) observed in RH mice. RH mice became glucose and insulin‐intolerant, which was improved in RRH mice (R=25.1±1.8; RH=40.7±3.2; RRH=29.9±3.1mg/dL.min/103/R=3.0±0.2; RH=4.3±0.3; RRH=3.3±0.3 mg/dL.min/103, respectively). Calorie intake was increased in HFD mice (R=10.1±0.79; RH=13.2±0.9; RRH=12.4±0.6kcal/day). During the dark period RH mice showed decreased energy expenditure which was prevented in RRH mice (R=136.7±4.0; RH=107.3±5.5; RRH=120.9±6.5 kcal/day/kg^0.75/103), despite the decreased ambulatory activity (R=24.3±2.8; RRH=16.2±0.9 arbitrary number/103) in this group.

Conclusion

RH mice became obese, hyperglycemic and IR, which was attenuated in RRH mice, possibly by regulation of the basal EM. Financial Support: FAPESP
Eight‐hr postprandial glycemic profiles after consumption of starch versus maltodextrin
Authors: Pelkman Christine , Lawton Clare , Croden Fiona , Camidge Diana , Finocchairo E Terrance , Dye Louise
Published: June 2021

A pilot study was conducted comparing glycemic responses to two starch ingredients vs maltodextrin (MD). A secondary objective was to compare two methods of blood glucose (BG) sampling – continuous interstitial glucose using an ambulatory glucose‐monitoring device (AGD), and capillary blood using a finger‐prick (FP) method. On three occasions, subjects consumed a breakfast of yogurt and 50 g of carbohydrate from a low‐fiber raw starch, a high‐fiber starch derived from high‐amylose maize (HI‐MAIZE ©260) or MD. Twelve men were tested with the FP method, and 8 with the AGD. For FP BG, a significant time*ingredient interaction was observed (p = 0.017). Higher values were observed for MD vs the two starches in the 90‐min period after breakfast. Over the remainder of the day, FP BG for MD tended to fall below the two starch conditions, but differences were small (~ 6 mg/dL, p = 0.07). In contrast, the AGD showed more differentiated profiles. In particular, a steep fall in BG was detected 3 hr after MD ingestion (~ 30% below baseline) whereas the FP method showed no fall. Both starches clearly mitigated this precipitous fall, with values at 3‐hr ~ 25% higher than MD (AGD), whereas the FP method showed little differences (~ 5%) between means. These results show that starch can prevent postprandial glycemic lows and that the FP method of sampling may lack the sensitivity needed to detect this effect. Supported by Ingredion Inc.
Longitudinal evaluation of estimated calorie and protein intake in patients on ketogenic therapy for seizures
Authors: Lennon Dylan J , Allen Hannah J , Giurcanu Mihai C , Borum Peggy R
Published: June 2021

Background

Ketogenic therapy (KT) is prescribed as a ratio of grams of fat to carbohydrate plus protein. Prescribed Calories (Cals) and protein (Pro) describe total dietary energy recommendations (Recs) and should support each patient's nutritional state.

Objectives

To compare Cal and Pro intake with prescribed Cal and Pro and with national Recs for healthy children.

Methods

24 hour diet records were entered into Nutrition Data System for Research. Cal (kcal/day) and Pro (g/day) intake was determined up to the first year of available data. Cal Recs were calculated from Estimated Energy Requirements for actual height, weight, age, gender, and physical activity. Pro Recs were calculated from DRIs for age and gender.

Results

Diet records were evaluated for intake as percent of prescribed Cals, Pro, recommended Cals, and DRI for Pro.

Intake of Prescribed (%)
Intake of Recommended (%)

Cal
Pro
Cal
Pro

(M=mean, SD=Stdev)

Month
n
M±SD
M±SD
M±SD
M±SD

1–3
16
97±8
99±8
95±17
124±22

4–6
15
98±17
99±17
93±15
120±24

7–9
14
96±10
99±10
91±18
119±16

10–12
14
95±5
97±5
87±16
113±14

Conclusion

When monitored regularly, parameters of diet intake reflect adherence to KT diet prescription, which is altered based on nutritional needs that may differ from those expected for a population of healthy children.

Supported in part by the National Institutes of Health (NIH) and National Center for Research Resources (NCRR) CTSA grant 1UL1RR029890
Metabolomic and genomic profiling of n‐3 polyunsaturated fatty acid effects on muscle metabolism in mice fed a high fat diet
Authors: Aljawadi Arwa , LeMieux Monique , Kalupahana Nishan S , Siriwardhana Nalin , Claycombe Kate , Moustaid‐Moussa Naima
Published: June 2021

We previously reported that feeding mice high‐fat (HF) diets enriched with eicosapentaenoic acid (EPA) decreased inflammation, adiposity and insulin resistance. In the current study, we used skeletal muscle from mice fed HF or HF‐EPA for 11 weeks to further dissect mechanisms mediating EPA effects on soleus muscle metabolism that may be responsible for improved insulin resistance and inflammation. We conducted metabolomic, energetic, gene and protein expression studies to determine global metabolic changes in this tissue. Our analyses showed that fatty acid and amino acid profiles were altered by the EPA diet. Both saturated fatty acids and docosahexaenoate (DHA) were elevated in muscle from the EPA group. Furthermore, essential amino acids for muscle protein synthesis and growth, namely branched chain amino acids (BCAAs) increased by the EPA diet. Surprisingly, we did not observe any significant changes in fatty acid oxidation between HF and HF‐EPA fed animals. These metabolomics findings were consistent with analyses of expression of lipid metabolizing genes in these muscles. From these data, we conclude that HF diets rich in EPA increased synthesis of both muscle fatty acid and essential amino acids but did not improve fatty acid oxidation in skeletal muscle. Further studies are needed to dissect the role of muscle metabolism in improving diet‐induced obesity, inflammation and insulin resistance
Adherence to a high protein and low fat energy‐restricted diet while participating in a circuit resistance‐exercise program promotes fat loss with no loss in fat free mass in postmenopausal women
Authors: Jung YP , Byrd M , Baetge C , Lockard B , Levers K , Galvan E , Jagim A , Simbo S , Oliver JM , Koozehchian M , Dalton R , Khanna D , Sanchez B , Kresta JY , Horrell K , Leopold T , Cho M , Springer S , Rivera A , Cerda C , Rasmussen C , Kreider R
Published: June 2021

41 sedentary women (55.3±10 yr, 45.0±4% body fat, 33.7±5 kg/m2) were randomized to participate in a control group (C) or the Curves Complete® (CC) weight loss and circuit resistance‐exercise program for 12‐wks. Participants in the CC program followed an energy‐restricted diet (30% C, 45% P, and 25% F) while participating in a circuit resistance‐training (3 d/wk) and walking (30 min, 3/d wk) program. Body mass, DEXA body composition, and resting energy expenditure (REE) measurements were obtained at 0, 4, 8, & 12 wks and analyzed by MANOVA. Data are presented as changes from baseline after 4, 8, and 12 wks for the C and CC groups, respectively. Participants in the CC program lost significant amounts of body mass (C −0.05±1.6, −0.1±2.2, 0.1±3.1; CC −2.1±1.7, −3.1±2.6, −3.9±3.2, kg, p=0.001), fat mass (C − 0.0±2.2, 0.2±2.3, −0.0±2.0; CC −2.2±1.4, −2.8±2.0, −4.1±2.4 kg, p=0.001) and body fat (C 0.1±1.7, 0.3±1.7, −0.1±1.7; CC −1.8±2.1, −2.2±2.3, −3.5±2.5 %, p=0.001) than controls with no time (p=0.54) or group × time effects on FFM (C −0.1±1.4, −0.1±1.7, 0.1±2.3; CC 0.2±1.8, 0.1±1.9, 0.5±1.6 kg, p=0.89). REE tended to differ among groups (C −20±149, 17±180, 39±190; CC −30±140, −65±169, − 85±130, kcal/d, p=0.03). Results indicate that post‐menopausal women who participate in an energy deficit higher protein and low fat diet with resistance‐exercise can promote fat loss without loss of FFM.
Reversal of Progress against Malnutrition in Egypt
Authors: Zaghloul Sahar
Published: June 2021

Objective

To assess trends and determinants of stunting, a monitoring indicator for economic and human development in Egypt, where marked economic and social changes have taken place.

Methods

WHO global database, Food and Agriculture Organization Food balance sheet and World Bank indicators were used to describe trends in childhood growth, food availability and economic development.

Results

Neonatal, infant and under five mortality rates decreased between 1995 and 2010. Prematurity and congenital anomalies among children < 5 years of age increased (20% and 30% respectively). Stunting decreased from 35% to 20.3% between 1990 and 2003 and bounced back to 30.7% by 2010. Moreover, the onset of stunting is now earlier (< 6 month of age). Iron deficiency anemia increased from 30 to 60% and from 30 to 46% among preschool girls and boys respectively. Gross domestic product per capita increased and inflation rate decreased between 1990 and 2011. A reverse pattern occurred between 2003 and 2004.

Daily calorie availability per capita did not change between 1995 and 2009 while calories from animal sources increased from 6.6% to 8.6%. Per capita protein from milk, eggs, poultry and pulses revealed a marked shortage compared to recommendations for 2–5 year old children.

Conclusion

Stunting reflects the cumulative effects of inadequate food intake, poor health conditions and projects low levels of human development and economic growth.

Congenital anomalies and prematurity may explain the early onset of stunting predicated by maternal poor health. Egypt, post Arab Spring, has a window of opportunity to reform its subsidy food and social programs to target improvement of maternal nutrition, strengthen breastfeeding and develop complementary feeding.
Adding access to online meal plans and monitoring to a structured weight loss program with resistance‐exercise promotes more positive changes in triglycerides
Authors: Levers K , Lockard B , Baetge C , Galvan E , Jagim A , Simbo S , Byrd M , Jung YP , Oliver JM , Koozehchian M , Dalton R , Khanna D , Sanchez B , Kresta JY , Horrell K , Leopold T , Cho M , Springer S , Rivera A , Cerda C , Rasmussen C , Kreider R
Published: June 2021

48 sedentary women (44.8±10 yr, 44.8±4% body fat, 33.9±6 kg/m2) were randomized to participate in the Curves Complete® weight loss and exercise program for 12‐wks with (CC‐OS) and without (CC‐NS) online support that provided access to meal plans and progress monitoring. Fasting blood samples were obtained at 0, 4, 8, & 12 wks and analyzed by MANOVA. Data are presented as changes from baseline after 4, 8, and 12 wks for the CC‐OS and CC‐NS groups, respectively. Overall MANOVA analysis revealed a significant time (p=0.001) difference with no significant group × time effects (p=0.57). Univariate analysis revealed significant time effects for TG, CHL, HDL, and LDL and that participants in the CC‐OS group experienced greater changes in TG (CC‐OS − 27.1±21, −24.2±24, −23.2±20; CC‐NS 0.9±29, −6.1±22, −6.7±26 %, p=0.001) with no differences observed among groups in total CHL (CC‐OS −11.8±11, −9.1±9, −7.5±11; CC‐NS −6.0±10, −4.0±11, − 3.6±10 %, p=0.17), HDLc (CC‐OS −0.8±11, 0.3±11, 5.0±15; CCNS −3.4±9, −0.1±13, 2.5±15 %, p=0.78), LDLc (CC‐OS −11.1±15, − 9.2±13, −7.8±19; CC‐NS −7.4±15, −6.6±16, −4.7±11 %, p=0.83) or blood glucose (CC‐OS −2.2±10, −1.9±8, −2.4±12; CC‐NS 1.2±14, 1.6±12, −0.6±15 %, p=0.75). Results indicate that adding online access to meal plans and monitoring can promote more favorable changes in TG during a structured weight loss program with resistance‐based exercise.
Barriers to engage parents in child obesity prevention and control: a multiple stakeholder analysis using the Family Ecological Model
Authors: Gehre Claudia , Chuang Emmeline , Blaine Rachel E. , Frankle Rebecca L. , Aftosmes Alyssa , Smith Lauren A. , Davison Kirsten K.
Published: June 2021

Barriers to engage parents in child obesity prevention are not well understood. We addressed this gap by utilizing the Family Ecological Model to identify and characterize barriers experienced by stakeholders in five sectors: WIC, health care, school, afterschool, child care. We conducted semi‐structured interviews with 40 stakeholders to identify factors influencing child obesity within their community and barriers to engage parents in prevention. Data were drawn at baseline from a multi‐sector study designed to reduce child obesity in two communities in eastern Massachusetts. Interviews were transcribed, coded, and summarized using NVivo 10. Stakeholders reported a wide range of barriers affecting parent engagement in child obesity prevention and control. Five main themes across sectors were identified, including family cultural beliefs (e.g. “chubby is healthy”), limited community resources, parents’ lack of education/awareness, lack of time, lack of money. Stakeholders also described poor cross‐sector communication and parents’ perceived lack of control over children's food intake as negatively affecting parental engagement. No obvious contradictory themes across sectors were identified and the results did not differ substantially between communities, suggesting that community stakeholders could share resources and work collectively to support parent engagement in obesity prevention.
Diet quality of rural breast cancer survivors as measured by the Healthy Eating Index‐2005
Authors: Christifano Danielle Nicole , Sullivan Debra K. , Befort Christie A.
Published: June 2021

Rural women are at increased risk for obesity compared to urban women. Obesity is a risk factor for breast cancer recurrence, but little is known about diet quality of breast cancer (BrCa) survivors in rural America. The objective of this study was to determine diet quality of rural BrCa survivors using the Healthy Eating Index‐ 2005 (HEI). Baseline data were obtained from a 2 year weight management trial for rural BrCa survivors in the Midwest. Participants (n=128; age 57.8 ± 7.8 years, BMI = 34.4 ± 4.3 kg/m2) underwent two baseline 24‐hour multiple‐pass dietary recalls (one weekday, one weekend). The recalls were entered into NDSR software and HEI scores were determined. Possible HEI scores range from 0–100, with higher scores indicating better diet quality. Mean HEI score was 56 ± 11, similar to the national average for adult women of 59 ± 4. Fruit and vegetable intake was 1.2 ± 1.1 and 2.9 ± 1.6 servings per day, respectively. Macronutrient profiles were 36 ± 7% fat, 45.8 ± 7.6% carbohydrate, and 16.9 ± 4.1% protein. Age was significantly correlated with total HEI score (p≤0.05). Specifically, older women had higher fruit (p≤0.01) intake, and lower sodium (p≤0.05) and saturated fat (p≤0.05) intake. Total HEI score did not differ across degree of rurality. Rural BrCa survivors are not meeting diet quality recommendations, and younger survivors may be in greater need of diet quality education than older survivors. Support of research by NIH R01 CA155014
Current consumption status of health functional food and influencing factors among office workers in Korea
Authors: Lee Soo‐Kyung , Lee Hee‐Won
Published: June 2021

As the life expectancy is getting longer and interests in health is increasing, functional foods have been gaining attention from consumers and producers alike. This study aimed to examine functional food consumption status. A pre‐tested questionnaire was self‐administered by a total of 496 office workers. Only 19.4% reported not having consumed any functional foods. A total of 52% answered current consumption and 28.6% past consumption. Among those who have consumed functional foods, 32.5% felt that the functional foods were effective, although 63.3% said they expected health improvement and maintenance from taking functional foods. Contrary to common beliefs, only 20% reported consuming functional foods for disease prevention. A total of 39.3% were consuming functional foods for relieving fatigue. Those who were either past or current functional foods tended to be heavier (p<0.05), to be more health conscious (p<0.001), to be more physically active only in men (p<0.05), compared to non‐consumers. It appears those who were more interested in health were more likely to invest in functional foods. This information would be useful to dietitians, public health workers, and functional food manufactures.
Interpretation of the IgG test‐based elimination diet
Authors: Shaidarov Mazhit , Tell Leonid
Published: June 2021

The ImmunoHealth Company's experience shows that elimination diet gives 82–86% of success rate. We claim that the IgG test results should be evaluated individually for each person. We chose several individuals and took blood samples four times for a one‐month period. We expected to see some variations as IgG reactions depend on myriad factors, including person's diet, heath or weakness during the day. We received adequate correlations not only within each group of foods, but also, when comparing the ratios in between a group of maximal and minimal IgG responses. This suggests that method is reproducible for an individual if the same test‐plates and conditions. Based on our computerized IgG‐test interpretation, we then suggested volunteers to undertake elimination diet program. During next months we monitored IgG levels and suggest correction diet to low down IgG response whenever possible. By the end of the 3 month program, more than 80% of individuals revealed weight loss and there healthy status was significantly improved. We consider this method to be well working when proper conditions are met.
Osteogenic activity of phloretin promoting osteoclast apoptosis in murine macrophages and inhibiting estrogen deficiency‐induced osteoporosis in mice
Authors: Lee Eun‐Jung , Cho In‐Hee , Kim Jung‐Lye , Kim Yunho , Kang Young‐Hee
Published: June 2021

Bone‐remodeling imbalance is known to cause skeletal diseases such as osteoporosis. Phloretin, a dihydrochalcone and a type of natural phenols, can be found in apple tree leaves. We have previously shown that phloretin inhibits osteoclast differentiation and bone resorption through disturbing TRAF6‐NFATc1‐NF‐κB pathway. RAW 264.7 macrophages incubated with 20μM phloretin and 17β‐estradiol for 5 d in the presence of RANKL stimulated capase‐3 activation, promoting osteoclast apoptosis. RANKL activated Akt and ERK via inducing c‐Src expression, which was attenuated by 1–20 μM phloretin. In addition, phloretin suppressed actin ring formation elevated by RANKL, indicating its inhibition of osteoclast fusion and resorption. This study further investigated inhibitory effects of phloretin on bone loss in ovariectomized (OVX) C57BL/6 female mice. OVX demoted bone mineral density (BMD) of mouse femoral bone tissues, reduced serum 17ßestradiol level and enhanced serum RANKL/osteoprotegerin ratio. The oral administration of 10 mg/kg phloretin to OVX mice for 8 weeks improved such effects, compared to sham‐operated mice. Phloretin attenuated TRAP activity and ß3 integrin expression and carbonic anhydrase II augmented in femoral bone tissues of OVX mice. Therefore, phloretin is effective in antagonizing osteoclastogenic resorption and preventing estrogen deficiency‐induced bone loss.
The Fanconi Anemia Pathway Induces Senescence and Suppresses Tumorigenesis in Vivo
Authors: Park Eunmi , D'Andrea Alan D
Published: June 2021

Fanconi Anemia (FA) is a rare human genetic disease characterized by bone marrow failure, congenital abnormalities, and a high incidence of cancer. The FA pathway regulates the synthesis of monoubiquitinated FANCD2, which is believed to function as a tumor suppressor in vivo. Disruption of the FA pathway, in human FA patients or in FA mouse models, results in an increased incidence of Squamous Cell Carcinomas (SCCs). Consistent with the role of FANCD2 as a tumor suppressor, Fancd2(−/−) murine embryo fibroblasts (MEFs) exhibit decreased RAS‐induced senescence and increased proliferation. We have also recently identified USP1, a cysteine protease which deubiquitinates FANCD2. Interestingly, Usp1(−/−) MEFs have elevated levels of monoubiquitinated Fancd2, and they exhibit increased RAS‐induced senescence and decreased proliferation. Their increase in senescence correlates with their increased cellular expression of TAp63 and p21. Moreover, Usp1(+/−) mice have a decreased incidence of H‐Ras induced squamous cell carcinomas in vivo compared to wild‐type sibling control mice. Taken together, our results indicate that downregulation of USP1 results in increased cellular levels of FANCD2‐Ub and an increased tumor suppressor activity in vivo. Pharmacologic inhibitors of USP1 may therefore function as cancer chemoprevention agents.
Effect of ketogenic therapy on seizure load score, medication load score, and response score in patients with intractable epilepsy
Authors: Allen Hannah J , Lennon Dylan J , Giurcanu Mihai C , Borum Peggy R
Published: June 2021

Background

The goal of ketogenic therapy (KT) for intractable epilepsy (IE) is to eliminate seizures and the need for medication.

Objective

Use Seizure Load Score (SLS), Medication Load Score (MLS), and Response Score (RS) to determine response to KT of patients with IE.

Methods

Seizure variables were ranked (1–5) to determine the Seizure Load (SL). Prescribed antiepileptic drug (AED) dose/kg was divided by the minimum recommended AED dose/kg to determine Medication Load (ML). SLS and MLS represent a % change in SL or ML from baseline (BL) at 100%. RS is the mean of SLS and MLS for specified time periods on KT. SLS, MLS, and RS were determined for patients on KT for the following time periods: 1–3, 3–6, and 6–12 months.

Results

Patients were stratified using SLS in the table below: Group 1 (SLS 50% better than BL), Group 2 (SLS better than BL, not 50% better), and Group 3 (SLS worse than BL). MLS and RS were determined for each group and time period on KT.

Conclusion

Patients can be stratified into response groups using SLS. MLS and RS provide additional information about the response to KT so that progress toward eliminating seizures and the need for medication can be assessed.

Supported in part by the National Institutes of Health (NIH) and National Center for Research Resources (NCRR) CTSA grant 1UL1RR029890
Ionizing radiation differentially modulates Wnt pathway components through ILK in normal and tumor bone cell lineages
Authors: Naves Marcelo Andery , Nogueira‐Pedro Amanda , Peres Giovani B. , Giordani Adelmo , Segreto Helena , Teixeira Vicente P.C.
Published: June 2021

Osteosarcoma (OS) frequently invades, mestastasizes, relapses and is characteristically radioresistent. Integrin‐linked kinase (ILK) modulates cell survival, death and radiosensibility. It can also interact with Wnt signaling pathway that is essential to skeletal development and may also play an important role in OS progression.

Murine pre‐osteoblast (MC3T3‐E1), rat OS (UMR‐106) and human OS (MG63) lineages were irradiated using a 60Co radiation source (2 and 6 Gy for 24 e 72 hr). We analyzed cell viability (MTT assay), adhesion (crystal violet assay), apoptosis, ILK, βcatenin, p‐GSK‐3β and p‐Akt protein expression (flow cytometry) and gelatinolytic activity (zymography).

Osteoblastic cells presented a substantial decrease in viability and cell adhesion. Apoptosis was reduced in MC3T3 groups irradiated with 6Gy dose. OS cells were less viable (except MG63 cells at 24 hr) and less adherent. Apoptosis increased in irradiated groups. ILK, p‐GSK‐3β, β‐catenin and p‐Akt expressions were significantly increased in all irradiated cell lineages, except in MC3T3 cells treated for 72 hr that showed a decrease in those protein expressions. Irradiation didn't modify cell gelatinolytic activity.

Ionizing radiation differentially modulated the cellular biology aspects of osteoblasts and OS lineages. This was probably accomplished by modulation of canonical Wnt signaling pathway through ILK.
Angiotensin II induced skin fibrosis in mice is associated with vascular injury
Authors: Stawski Lukasz , Haines Paul , Trojanowska Maria
Published: June 2021

We recently reported that Ang II induces skin fibrosis in mice (Stawski et al, 2012). The goal of this study was to further investigate the mechanisms involved in skin fibrosis induced by Ang II as a model for the pathogenic mechanisms of Scleroderma. Ang II was administered to mice for 7 and 14 days by subcutaneous osmotic pump. TUNEL and cleaved Caspase 3 staining showed increased number of apoptotic cells present in the vessels and in the connective tissue in Ang II treated mice. IHC staining showed increased number of Mmp12‐positive cells, including ECs in Ang II infused mice. MMP12 protein level was increased in Ang II treated human MVECs in a dose dependant manner. Pdgfrb‐positive cells were observed around the vessels in control mice, but were increased in all skin layers in Ang II mice. Real‐time PCR analysis showed a statistically significant increase in the levels of vascular injury markers: Angpt2 at day 7, Mmp12 and Thbs1 at day 14 and pericyte marker Adam12 at day 7 in the skin of Ang II infused mice.

These observations together with the previously reported data strongly suggest that the Ang II model closely reproduces one of the key features of SSc ‐ vascular injury. Moreover, persistent defect of endothelial cells in this model may lead to development of a chronic fibrogenic environment resulting in activation of peri‐endothelial cells that leads to fibrosis.
NFR2 EXPRESSION IN APAP ACUTE INTOXICATION
Authors: Cardoso João Felipe Rito , Cohen Célia , Vannucchi Hélio
Published: June 2021

Acetaminophen (APAP) overdose promotes the formation of NAPQI, the depletion of liver glutathione and oxidative stress. N‐acetilcysteine (NAC) is a cysteine donator that is a precursor for the synthesis of GSH. Nrf2 is a transcription factor, important in antioxidant response pathway, promoting the expression of phase II enzyme and the regulation of liver GSH. Male C57 mices, were fasted for 16h prior to the application of APAP (250mg/kg). The treatment with NAC (204mg/kg) occurred 1h after APAP challenge. The collection of liver tissue occurred 24h after treatment. Analysis: Histopathology by score of necrosis fields; hepatic levels of GSH by spectrophotometry and hepatic levels of Vitamin E (VE) by HPLC; the gene expression of NFR2 by RT‐qPCR (Taqman probes). Statistic Analysis: Student. t test (Graphpad Prism 5.0). The group (ANAC) that received treatment with NAC 1h after APAP challenge, showed a reduction of 28% of necrosis (p <0.05). ANAC GSH levels were 1.5 higher than the APAP levels (p <0.05), although VE presents no differences between the groups. The Nfr2 gene expression fold change had almost doubled compared with APAP group (p <0.05). The results show that after the treatment with NAC, the increase of NFR2 gene expression promotes the increase of GSH levels, saving the reduction of VE and consequently, the oxidative stress, protecting the liver from necrosis. Support by FAPESP (2011/12749–0 & 2011/17616–9)
BMI‐1 and homologous recombination markers in human breast cancer
Authors: Silveira Giorgia Gobbi , Celes Mara Rubia Nunes , Coutinho‐Camillo Claudia Malheiros , Soares Fernando Augusto , Rossi Marcos Antonio , Ribeiro‐Silva Alfredo
Published: June 2021

Camargo Cancer Hospital, Sao Paulo, Brazil The role of proteins involved in homologous recombination in breast carcinomas that express BMI‐1 still poorly understood. Our objective was to evaluate the relationship between BMI‐1 and regulatory proteins of homologous recombination. Using tissue microarrays containing 239 cases of primary breast tumors, protein expression was analyzed by immunohistochemistry. For gene expression, we performed Real‐Time PCR reactions in MCF‐7 cell line. BMI‐1 immunohistochemistry overexpression was related to p53 (p=0,003), BRCA‐1 (p=0,003), H2AX (p=0,024) and TopIIIβ (p≤0,001). Real‐Time PCR assay showed that BMI positive cells have low expression of H2AX and P53. Our results point to a relationship between BMI‐1 and homologous recombination markers, suggesting that the presence of BMI‐1 is an important event in breast cancer homologous recombination. Research support: FAPESP 2011/22849–2; FAPESP 2011/23042–5
The Role of the Protein C and Urokinase Systems in Prostate Cancer
Authors: Williams Ryan , Glasscock Laura , Pardhanani Pooja , Brunson Lacey
Published: June 2021

The objective of this study was to determine how the protein C and urokinase systems regulate prostate tumor cell migration. Prostate cancer (CaP) is the most frequently diagnosed cancer among males, with an estimated 238,590 new cases arising in the United States during 2013. Urokinase plasminogen activator (uPA) and its inhibitor, plasminogen activator inhibitor‐1 (PAI‐1), are regulators of blood coagulation and normal cell migration. Abnormal secretion of these two proteins by neoplastic cells in the tumor microenvironment has been linked to an increased likelihood of (CaP) invasion. Using a modified Boyden chamber, we tested the ability of activated protein C (APC), uPA, PAI‐1 and vitronectin (VN), an ECM protein, to mediate uPA and PAI‐1 dependent migration of the DU‐145 CaP cell line through Matrigel. As has been shown for ovarian cancer, in the presence of uPA and PAI‐1, APC competed with uPA for binding to PAI‐1, enabling uPA to increase the invasiveness of DU‐145 cells. We also determined that VN acts as a cofactor that accelerates APC‐PAI‐1 interaction, potentially allowing once inhibited uPA to degrade ECM and give neoplastic cells room to invade through tissue. We conclude that although protein C and APC alone do not affect CaP cell invasion, in the presence of uPA and PAI‐1, APC binds to PAI‐1, freeing uPA to facilitate CaP tumor cell invasion.
Gene expression patterns through oral squamous cell carcinoma development
Authors: Oliveira‐Costa Joao Paulo , Carvalho Alex Fiorini , Silveira Giorgia Gobbi , Carraro Dirce Maria , Soares Fernando Augusto , Ribeiro‐Silva Alfredo
Published: June 2021

Oral squamous cell carcinoma (OSCC) development is still a poorly understood phenomenon, in terms of gene expression profiles which might be involved in alterations leading to high invasiveness and aggressive behavior. Using 36 samples of OSCC cases, we performed cDNA and lincRNA microarrays, comprising whole human genome, in order to identify gene profiles with similar behavior throughout T1 to T4 tumor staging. We found 4 different gene expression profiles, either with a rise in expression levels, or decrease in expression levels, between T1, T2, T3 and T4 TNM staging. The four different gene expression profiles showed 58 differentially expressed genes, including an increased expression of RUNX family genes, and decreased expression of zinc finger proteins. Our study shows the existence of several genes with similar expression patterns during OSCC development, and also points to novel genes, which has not yet been implicated in oral carcinogenesis, providing new potential targets in oral cancer research.

Sponsored by FAPESP (2010/08637–0 and 2010/08400–0)
Absence of secondary phrenic motoneuron (PhMN) loss following lateralized cervical contusion
Authors: Little Lydia Nicole , Hussey Shaunn , Gonzalez‐Rothi Elisa J , Mercier Lynne M , Sanchez Deevee , Osteen Barbara , Fuller David , Lane Michael , Reier Paul
Published: June 2021

Cervical spinal cord injuries (cSCI) are often accompanied by respiratory complications, due to compromised integrity of the phrenic nucleus (C3–C5/6) and loss of diaphragm function. This study utilized a lateralized C3/4 contusion to characterize temporal changes in diaphragm activity (EMG) and ventilatory function following cSCI, pairing EMG recordings with whole‐body plethysmography. A second goal was to determine secondary PhMN loss to guide future neuroprotective strategies. Initial studies demonstrated diaphragm electromyographic (diaEMG) activity was immediately and markedly reduced, but showed recovery under baseline conditions within 2 days post‐injury. In contrast, diaEMG was persistently attenuated during hypercapnia. Conversely, ventilation was unaffected at baseline and only transiently affected by hypercapnic challenge. Other preliminary results indicate 63% initial loss of PhMNs after injury without evidence of further loss at 4 weeks post‐injury. This suggests persistence of normal ventilatory patterns most likely reflects compensatory plasticity in other spinal respiratory circuits. The absence of secondary PhMN loss challenges the concept of neuroprotection at least in this motor system. Whether impaired diaphragm activity is due to primary PhMN loss, reduced innervation of PhMNs caudal to the injury, or loss of other phrenic circuit constituents remains unknown.
Circulating 15‐hydroxyeicosatetraenoic acid levels are Elevated in a Mouse Model of Inflammatory disorders following High fat diet Administration.
Authors: Kaimi Nika , Vakili Ladan , Haghnegahdar Maryam , Farahmand foozhan , Sedigh farhad , Safar sheila , Meshkat arash , Niknam Daniel
Published: June 2021

Objective

To determine the plasma levels of the inflammatory marker 15‐hydroxyeicosatetraenoic acid in ApoE deficient mouse treated wit high fat diet.

Introduction

15‐hydroperoxyeicosatetraenoic acid (15‐HPETE) is a product of oxidation of arachidonic acid (C20:4). HPETEs are potent prooxidant molecules that can lead to stimulation of several metabolic pathways resulting in an inflammatory condition in many organs and systems. Fortunately our organism has evolved to use mechanisms to convert the potent 15‐HPETE to a lesser active molecule 15‐ hydroxyeicosatetraenoic acid, 15‐HETE. Still elevated levels of 15‐HETE are undesirable in most circumstances. We sought to determine the effect of a high fat diet in an animal model of inflammation namely ApoE deficient mouse. Using SepPak columns and a Q 4000 mass spectrometer, the plasma levels of 15‐HETE were determined. The data was analyzed using the software supplied by the manufacturer and statistical significance determined using ANOVA through Prism application.

Results

feeding a high fat diet amplified the circulating levels of 15‐HETE (p = 0.021 vs. the control group) which in turn can activate pathways that produce inflammatory molecules.

Conclusion

reducing fat consumption can lead to reduced level of plasma oxidized fatty acids which in turn can help alleviate the inflammatory pressure.
Effect of acute hypoxia on phenotype and plasticity of M1 and M2 macrophages
Authors: Malyshev Igor , Kalish Sergey , Mantulin Nikita
Published: June 2021

Federation Pathogenetic roles of hypoxia may be related with influences on immunity. The aim of study was to investigate the effect of hypoxia on the macrophage phenotype and plasticity in C57/BL mice, which have the pro‐inflammatory M1 macrophage phenotype, and in BALB/c mice, which have the anti‐inflammatory M2 one.

Methods

Hypoxia was produced in an altitude chamber. After hypoxia, macrophages were isolated from peritoneal lavage. NO production, inducible NO synthase content and cell shape were used as phenotype markers. To determine the phenotypic plasticity, we quantified the capability of macrophages for changing the phenotype to M1 and to M2.

Results

1) The response of macrophages to hypoxia has two successive phases, the immediate, anti‐inflammatory phase, and the delayed, pro‐inflammatory phase. These phases were more pronounced in Ñ57/BL6 M1 macrophages than in ÂÀLB/c M2 macrophages; 2) the effect of hypoxia on macrophage phenotypic plasticity depends on the genetically predetermined macrophage phenotype. Hypoxia increased the capability of macrophages for changing to the pro‐inflammatory M1 phenotype, while their capability for changing to the anti‐inflammatory M2 phenotype remained virtually unaffected.

Conclusion

Effects of hypoxia involve immune responses and depend on the genetically predetermined macrophage phenotype. This study was supported by the RFBR (12–04‐01191‐a)
Microparticles bearing RGD peptide promote integrin αvβ3‐dependent platelet adhesion in isolated, pressurized cerebral artery
Authors: Bagi Zsolt , Perez‐Balderas Francisco , Broskova Zuzana , Davis Benjamin , Anthony Daniel
Published: June 2021

Platelet‐derived microvesicles (PMVs, <500 nm) play a role in thrombotic diseases. How PMVs promote platelet adhesion is poorly understood. To dissect mechanisms we established a method for real‐time imaging of microparticle (MP) and platelet adhesion in intact, perfused arteries. Platelets, with or without prior administration of arginine‐glycine‐aspartate (RGD)‐coated, ironoxide MPs (RGD‐MP, ~700nm, Alexa‐488‐enriched) were delivered intraluminally into isolated, pressurized rat cerebral artery. Individual binding events were observed with high‐speed fluorescence imaging under constant flow (10 μl/min). We detected firm binding of RGD‐MPs on endothelium surface (130±51/mm2), which was reduced by adhesion‐blocking integrin αv antibody. RGD‐MP binding was enhanced (290±73/mm2) by pro‐inflammatory cytokine TNF, which also increased vascular expression of integrin αv. Endothelial adhesion of platelets was 14±3 event/mm2 in naïve arteries, which significantly increased by TNF (48±10/mm2). Prior administration of RGD‐MPs induced greater adhesion of platelets in naïve arteries (78±10/mm2), whereas RGD peptide solution or scrambled RDG peptide‐coated MPs had no effect. Thus, we demonstrate the multivalent RGD‐MP ‐ compared to free ligand in form of RGD peptide ‐ as a more effective construct to promote αvβ3‐dependent adhesion of platelets in the intact cerebral artery.
Oral Infection augments the production of Inflammatory Markers of Atherogenesis.
Authors: Meshkat Arash , Hamidi Homa , Mobayen Shirin , Vakili Ladan , Karimi Nika , Navab Kaveh , Kalantari Ryan , Farazmand Maryam , Haghnegahdar Maryam
Published: June 2021

Objective

To determine if oral infection will result in an increase in markers of inflammation that are known to augment atherogenesis.

Introduction

It is known that patients with inflammation or infection of the oral cavity have a higher risk of acquiring susceptibility and being in a state of systemic inflammation increasing their risk of having vascular abnormalities. In the present study we sought to examine if infection of the oral cavity would result in an increase in the levels of markers of atherogenesis.

Methods

Four month old female Apo E mice were treated with P. Gingivitis or with vehicle for 10 days. Plasma was prepared using heparinized capillary tubes and plasma separators and was cryopreserved. Circulating levels of interleukin 6 and 13‐HODE were determined using a commercially available ELISA kit and LC‐ESIMS/MS employing a Q4000 Quadrupole system. The data were analyzed using one way ANOVA taking advantage of GraphPad Prism application.

Results

In mice receiving the P. Gingivitis treatment the levels of circulating IL‐6 and 13‐HODE were significantly higher (p=0.011 and p=0.031, respectively). There was no difference in the levels of 20‐ HETE which is known not to increase in inflammation.

Conclusion

The present study indicates that oral infection with P. Gingivitis results in increases in the levels of circulating inflammatory molecules that are known to contribute to vascular dysfunction and increased vulnerability to atherogenic reactions.
Endothelium Specific Up Regulation of HO‐1 Gene by Lentiviral Transfection Promotes Release of Positive Regulators of Adipoctye Function
Authors: Getty Morghan Schuyler , Kim Dong Hyun , Abraham Nadar G.
Published: June 2021

We aimed to characterize the physiological effects of up regulating endothelial heme oxygenase‐1 (HO‐1) on adipogenesis in human mesenchymal stem cells (MSCs). Microvessel endothelial cells (HMEC‐1) were cultured along with a lentiviral (lenti) construct expressing human HO‐1 under the control of endothelium specific promoter vascular endothelium cadherin heme oxygenase (VECAD‐HO‐1) and Lenti‐VECAD‐GFP used as the control. In complementary experiments HMEC‐1 cells were cultured in the presence or absence of tin‐mesoporphrin (5 μM). Conditioned media (CM) was harvested and tested at 5%, 10% and 20%. The 10% CM from HMEC cells was used for ideal effect, along with adipogenic media, to measure paracrine effect on adipogenesis in MSCs. The MSCs exposed to CM from VECAD‐HO‐1 transfected cells demonstrated reduced (10.2 ± 1 at 490nm) adipogenesis (lipid droplets) as compared to MSCs exposed to CM from Lenti‐VECAD‐GFP cells (17.0±2, p>;0.05). CM from HMEC‐1 cells treated with inhibitor of HO‐1 activity promoted higher adipogenesis with increased adipocyte hypertrophy as opposed to HMEC‐1 treated activity transfected with the VECAD‐GFP control ( 24.5±1, p<0.01). These observations suggest that EC under stressed conditions releases a negative regulator as seen in VECAD‐HO‐1 in presence of HO‐1 inhibitor and increases adipocyte dysfunction.
Multifaceted regulation of Ca2+ efflux via the plasma membrane Ca2+‐ATPase by 17β‐estradiol in the vascular endothelium
Authors: Giles Jennifer , TRAN KIM , Firkins Rachel , VerMeer Mark
Published: June 2021

Many of the beneficial effects of estrogen in the vasculature involve Ca2+‐dependent activities. Ca2+ efflux via the plasma membrane Ca2+‐ATPase (PMCA) represents a major component of the Ca2+ signaling machinery in cells. We observed in vascular endothelial cells that chronic treatment with 17β‐estradiol dose‐dependently decreases PMCA activity without affecting PMCA expression levels. Treatment with 17β‐estradiol promotes tyrosine phosphorylation at both the 135‐kDa (PMCA) and 60‐kDa levels on PMCA immunoprecipitate. We identified this 60‐kDa protein to be a glycosylated form of the novel G protein‐coupled estrogen receptor 1 (GPER). Inhibition of Src kinase activity suppresses estrogen‐induced tyrosine phosphorylation on PMCA and GPER. Functionally, Src inhibition not only restores PMCA activity in cells treated with 17β‐estradiol, but increases it by 50%. Consistent with the 50% increase in PMCA activity upon Src inhibition in estrogen‐treated cells, the interaction between PMCA and calmodulin is increased by 50%, an effect that occurs independently of Src kinase activity. These results indicate that estrogen inhibits PMCA activity by promoting Src‐dependent tyrosine phosphorylation and interaction with GPER, effects that mask the stimulatory effect of enhanced CaM binding.