bluuebunny/crossref_metadata_2025 huggingface dataset:
Serum fibroblast growth factor‐21 is responsive to a hypocaloric very low carbohydrate ketogenic diet but not a low fat diet
Authors: Kawiecki Diana M , Freidenreich Daniel J , Quann Erin E , Labonte Cherise C , Puglisi Michael J , Ballard Kevin D , Volk Brittanie M , Forsythe Cassandra E , Fernandez Maria‐Luz L , Kraemer William J , Volek Jeff S
Published: June 2021

Ketogenic diets were recently shown to induce hepatic expression and increase circulating levels of fibroblast growth factor 21 (FGF‐21), which coordinates lipid homeostasis by inducing hepatic lipid oxidation, ketogenesis, lipolysis, and triglyceride clearance. Very low carbohydrate ketogenic diets (VLCKD) in humans consistently improve features of metabolic syndrome. We hypothesized a VLCKD would increase FGF‐21 in adults. Fasting serum FGF‐21 concentrations were measured by RIA in 39 overweight men and women who consumed a VLCKD (1504 kcal:%CHO:fat:protein=12:59:28) or a low fat diet (LFD) (1478 kcal:%CHO:fat:protein=56:24:20) for 12 weeks. All subjects had detectable levels of FGF‐21 in serum with no gender differences. FGF‐21 was reduced (p<0.05) by the VLCKD (2.51 ± 0.75 to 2.19 ± 0.56) and unchanged after the LFD (3.02 ± 0.61 to 3.03 ± 0.69). Baseline concentrations and delta changes in FGF‐21 were not correlated with weight loss, body fat, glucose, insulin, lipoproteins, or other metabolic syndrome markers. Contrary to our expectations, these findings indicate that serum FGF‐21 concentrations decrease after a hypocaloric, ketogenic diet in overweight humans. The metabolic significance of this response is unclear, but may represent a down‐regulation in the need for lipolytic signaling as a consequence of the potent stimulatory effects of carbohydrate restriction on hepatic lipid oxidation.
Effects of natural plant extract on activities of enzymes related with fat metabolism in lean and fa/fa Zucker rats
Authors: Choe Myoen , Lee Hyeon‐Ju , Kim Dae‐Jung , You Jin‐Kyoun , Seo Dong‐Joo , Lee Won‐Kyoung , Park Eun‐Hee , Cho Jea‐Won , Kim Myoung‐Dong
Published: June 2021

Since obesity is well known as a potential risk factor for chronic diseases such as hyperlipidemia and diabetes, many researches have been focused on finding natural anti‐obesity compounds. This study was carried out to estimate anti‐obesity effects of plant extract on activities of enzymes related with fat metabolism. Changes in activities of acyl‐CoA synthetase, lipoprotein lipase and hormone‐sensitive lipase were compared for lean and fa/fa rats. While extracts from Plantago asiatica and Letinus edodes gave rise to an augmented acyl‐CoA synthetase activity in fa/fa rats, those from Semen plantaginis and Plantigo asiatica resulted in decreased lipoprotein lipase activity in adipose tissue of fa/fa rats. Abdominal hormone‐sensitive lipase activities in 10‐week old Zucker rats were significantly enhanced by Zea mays L. This study indicated that plant extract might be developed as a plausible anti‐obesity nutraceutical supplements.
Effect of endogenous n‐3 PUFA on inflammation and oxidative stress
Authors: Fernandes Gabriel , Rahman Md Mizanur , Bhattacharya Arunabh , Banu Jameela , Kang Jing X. , Halade Ganesh
Published: June 2021

Accumulated oxidative stress is now considered to be a key mechanism underlying the aging process. Calorie‐restriction (CR) is recognized to slow the rate of accrual of age‐related oxidative stress and the production of the overall inflammatory gene expression. Recently, several studies have demonstrated that n‐3 PUFA act as anti‐inflammatory and antioxidative agents. This study was undertaken to examine the synergistic role of n‐3 fatty acids and 40% CR on the activity of inflammatory cytokines and antioxidant enzymes in transgenic Fat‐1 mice. Thesetransgenic mice express a Caenorhabditis elegans desaturase, capable of converting n−6 to n−3PUFA endogenously and decrease the n−6/n−3 PUFA ratio in tissues which is considered to be very important determinant for good health. In this study, Fat‐1 transgenic mice and wild type control mice were fed 10% corn oil (CO) either AL or CR (40%) for 5 months. The n−6/n−3 PUFA ratio was significantly lower in serum, liver and muscle tissues of Fat‐1 transgenic mice as compared to control mice. Significantly lower serum pro‐inflammatory cytokines, TNF‐α, IL‐1β, IL‐6, CRP, higher muscle and liver antioxidant enzymes, SOD, CAT, GPX were observed in Fat‐1 transgenic mice as compared to wild type control. Also, a significant up‐regulation of the expression of life prolonging silent information regulator‐1 (SIRT1) was found in CR mice, particularly much higher levels in Fat‐1 CR mice. Interestingly, the PPAR‐γ expression was found to be reduced in Fat‐1 CR mice as compared to wild type control CR mice. Taken together, our studies suggest that n‐3 PUFA with moderate calorie restriction may extend survival and delay aging by modulating expression of cytokines, free‐radical scavenging enzymes, and life prolonging SIRT1 gene etc. Research support: NIH R01 AG030161‐A1.
”What do guys want?” Perceived and desired body shape: results from the Strong Bodies and Strong Minds Unite Sisters! Project
Authors: Kavanagh‐Prochaska Katherine , Wetherall Karen Balnicki , Kittle Leah , Jahns Lisa
Published: June 2021

Sorority women may be at risk for unhealthy eating or physical activity behaviors due to discordant body image perceptions. However there is little empirical evidence. The lack of data on perceived body shape ideals, especially among minority sororities, highlights the need for further research. Three hundred and forty of 1,994 members of 4 minority (n = 46 (72%)) and 13 non‐minority (n = 294 (15.6%)) sororities responded to a web‐based survey investigating body shape perception. Participants rated self‐perceived body shape (S), desired body shape (D), and body shape desired by males (DM) using the 9‐item Stunkard figure scale (1983). Thirty‐eight percent of the variability in S was explained by D (r = 0.614; p<0.05). However there was little correlation between S and DM (r = 0.28; p<0.05). Results did not differ by sorority type though the minority sorority sample size was small, possibly affecting power to detect differences. The strong relationship between S and D indicates that sorority women are generally satisfied with their body shape. However, the lack of relationship found between S and DM indicates that sorority women perceive their body shape to be different from what males desire.
Quality aspects in the determination of sodium in bread
Authors: Castanheira Isabel , Coelho Ines , Santiago Susana , André Catarina , Silva Ana Sanches
Published: June 2021

Objective: In this work was studied the impact of sampling plan measurement uncertainty and appropriate reference materials in the determination of assigned value of sodium content in several bread types consumed in Portugal.

Methods: The sampling plan was designed according to the national food balance sheets. Sodium was determined by flame photometry and measurement uncertainty was estimated in accordance with NIST guide.

Results: The highest sodium value determined was 650 mg/100 g. Relative measurement uncertainty reaches in some cases 10%, where the largest contribution comes from sampling. In all types of bread analysed, the sodium values are significantly different between bakeries under study (p<0.001).

Conclusions: Our study reveals that measurement uncertainty and the use of appropriated reference materials facilitates the evaluation of sodium content available in different bakeries. Sampling plan incorporated in quality assurance programme is a key criteria for extrapolate analytical into intake values and reduce the additional imprecision in nutrient intake estimation. The quality framework of sodium content in Portuguese bread can be applied in other countries across and beyond Europe.

This work was completed on behalf of Project LMARSA and funded under the Regional Programme for European Cohesion.
Anemia and Decline of CD4 Cell Count in HIV+ Drug Users in Miami
Authors: Sales Sabrina , Campa Adriana , Rafie Carlin , Le Katherine , Entengoff Kirstin , Page John B , Baum Marianna K
Published: June 2021

Objective:

This study investigates the association of anemia with HIV disease progression and survival over 30 months in HIV+ drug users in Miami.

Methods:

After obtaining consent, demographic and medical questionnaires were completed every 6 months for 30 months in HIV+ drug users. Blood was drawn for CD4 cell count, viral load, and CBC. Anemia was defined as hemoglobin <12g/dL in women and <14g/dL in men, and hematocrit<37% in women and <42% in men. Kaplan Meier Estimation was used to evaluate the rate of decline of CD4 cell count and mortality over 30 months.

Results:

Of 231 participants, 77% were black, and 73% were men. Mean age was 42 years ± 7.1. The prevalence of anemia was 46% (n=106). Of those with anemia, only 19% had macrocytic/normochromic anemia and 0.05% microcytic/hypochromic anemia, suggesting nutritional deficiency. Anemia was significantly associated with a more rapid decline of CD4 cell count to below 200 cells/μL (p<0.001) when compared to participants without anemia. There was also a trend in decreased survival overtime in those who had anemia compared to those without anemia, (80% vs. 94%, p=0.094).

Conclusions:

Nutritional etiologies of anemia do not seem to be the predominant cause of anemia in this cohort. Anemia in our cohort was strongly related to advanced HIV disease and efforts to identify the cause and provide treatment may help to delay disease progression.

Funded by NIDA
Regulation of RANK expression mediated by L‐ascorbic acid in RAW264.7 cells:involvement of cNOS, NF‐kB and Ikaros
Authors: Joo Haemi , Son Eunwha , Rhee Dong‐Kwon , Pyo Suhkneung
Published: June 2021

L‐ascorbic acid (AsA) has been known to induce osteoclastic differentiation through osteoclast differentiation factor in some osteogenic cells. However, the precise mechanism of AsA in osteoclastogenesis is not completely understood. This study investigated the effect of AsA on the expression of receptor activator of nuclear factor‐kappa B (RANK) which is indispensable for osteoclast differentiation. Our data showed that AsA treatment up‐regulated the expression of RANK mRNA and protein in RAW 264.7 cells, which was inhibited by DPI, cNOS inhibitor. AsA also induced the expression of cNOS protein, indicating that cNOS is involved in AsA‐induced RANK expression. The level of RANK expression was decreased in Ikaros dominant negative isoform‐overexpressed RAW 264.7 cells compared with that of control cells. In addition, AsA dose‐dependently increased the DNA binding affinity of the Ikaros transcription factor, suggesting that Ikaros acts as a positive regulator on RANK expression. DPI also inhibited the nuclear expression of NF‐kB p65 induced by AsA. Furthermore, DPI significantly decreased the DNA binding activity of NF‐kB induced by AsA, implicating that cNOS is clearly related to NF‐kB activity. Overall, the present data demonstrate that AsA might stimulate osteoclastic differentiation by enhancing RANK expression and this process might be related to cNOS, NF‐kB and ikaros activation.
Nutritional status of Korean hemodialysis patients with and without diabetes mellitus
Authors: Cho Ju‐Hyun , Jang Sang‐pil , Kim Wha‐Young
Published: June 2021

Prevalence of chronic renal failure with diabetic patients was increasing in Korea. The purpose of this study is to investigate the nutritional status of Korean hemodialysis patients with and without DM. Anthropometry, hematological variables and food intake were analyzed. Food intake was taken using 24hr recall for consecutive 2days including one dialysis and one non dialysis day. An average intake of 2days was considered as daily food intake. Urea reduction rate was significantly lower in DM than nonDM group, DM group showed lower adequacy of dialysis. BMI, triceps skin fold thickness were in normal range for both groups. However, BMI was significantly higher in DM than nonDM group. Serum protein, hemoglobin and hematocrit were below normal range for both groups. These values implied mild protein inadequacy and possible anemia in patients. Energy/IBW(kcal/kg IBW/d) intake was inadequate in DM and significantly lower compared to nonDM group. Intake of calcium, vitaminB2 were below Estimated Average Requirements in both groups. DM group was not differ from nonDM group in anthropometry, hematological variables and food intake. BMI was higher, even though their energy intake and URR was lower. In conclusion, our subjects regardless the presence of DM, nutrition intake status was not satisfactory, and more active nutrition monitoring is needed. (This work was supported by second stage of BK21 project in 2007)
Biochemical markers of nutritional assessment and inflammation in gastroenterological surgical patients
Authors: Zago Liliana , Infantino Carlos Gonzalez , Danguise Eduardo , Callegari Mariana , Morgan Lucila , Río María Esther
Published: June 2021

It is accepted that inflammation is a confusion factor for the interpretation of biochemical markers of nutritional status because result in modifications attributable to nutritional deficiencies. In order to evaluate the influence of inflammation on nutritional assessment of surgical patients with no infection, 45 patients from programmed gastroenterological surgeries were studied. Hemoglobin, erythrocyte protoporphyrin (EP), serum vitamins A (VA) and C (VC), total proteins (TP), albumin (Alb), transthyretin (TTR) and retinol binding protein (RBP) were evaluated in fasting samples taken before surgical procedure. Patients were classified by C‐Reactive Protein as a marker of inflammation (CRP positive when >0.6 mg/dl; I+). Patients with inflammation (I+; n=17) presented a decreased level of VA (X±SD: 35.6±16.1 vs 49.5±19.1 μg/dl; p=0.0162), Alb (3.9±0.7 vs 4.2±0.5 g/dl), TTR (17.7±7.4 vs 27.6±7.6 mg/dl) and RBP (3.1±1.1 vs 4.3±1.8 mg/dl) respect to that with no inflammation (I−; n=28). No changes of VC (0.69±0.45 vs 0.66±0.42 mg/dl), TP (6.1±0.6 vs 6.1±0.6 g/dl) and EP (43.6±22.3 vs 39.2±27.3 μg/dl) were founded. A higher number of patients I+ presented low hemoglobin (47% vs 32%). We conclude that chronic inflammation with no infection expose the patient to a higher chance of a subnormal nutritional profile that may compromise evolution by increasing the risk of complications. Supported by UBACyT B077.
Modulation of proteasome activity by vitamin E in THP‐1 monocytes
Authors: Munteanu Adelina , Ricciarelli Roberta , Massone Sara , Zingg Jean‐Marc
Published: June 2021

The anti‐retroviral protease inhibitor, ritonavir, stimulates CD36 scavenger receptor expression in THP‐1 monocytes, an event that is prevented by α‐tocopherol co‐treatment. Ritonavir furthermore leads to cellular proteasome inhibition, increasing the level of oxidative stress in these cells. Since α‐tocopherol can reduce oxidative stress, it appears likely that normalization of CD36 over‐expression is a consequence of scavenging free radicals produced after proteasome inhibition by ritonavir. Indeed, the proteasome inhibitor, ALLN, produced levels of oxidative stress similar to those induced by ritonavir; however, α‐tocopherol was able to reverse only the effect of ritonavir, and not that of ALLN. Accordingly, α‐tocopherol could normalize the amounts of 3‐nitrotyrosine‐modified proteins only after ritonavir treatment. In the absence of any proteasome inhibitor, intrinsic cellular proteasome activity was not modulated by α‐tocopherol; however, some natural vitamin E analogues and synthetic derivatives could significantly inhibit cellular proteasome activity. Because in our experimental system oxidative stress is reduced by α‐tocopherol only when the proteasome is inhibited by ritonavir and not by ALLN, it is concluded that α‐tocopherol normalizes proteasome activity through a mechanism that does not involve the antioxidant property of the vitamin.
Gamma Tocopherol alone and in Combination with Alpha Tocopherol Decreases Biomarkers of Oxidative Stress and Inflammation in Subjects with Metabolic Syndrome
Authors: Devaraj Sridevi , Leonard Scott , Traber Maret G , Jialal Ishwarlal
Published: June 2021

Metabolic syndrome (MetS) is associated with increased incidence of diabetes and cardiovascular disease (CVD). Prospective clinical trials with alpha tocopherol (AT) have been equivocal. AT supplementation decreases circulating gamma tocopherol (GT), thus, we tested supplementation with GT (800 mg/day), AT ( 800 mg/day), the combination or placebo for 6 weeks on AT and GT concentrations, biomarkers of oxidative stress and inflammation in MetS subjects (n=20/group). Plasma AT and GT levels increased following supplementation with AT alone, GT alone or in combination. AT supplementation significantly decreased GT levels. Urinary alpha and gamma CEHC, metabolites of the respective Ts also increased correspondingly, with the combination resulting in significant increases in both compared to placebo. HsCRP significantly decreased in the combined AT +GT group (p<0.05). There was a significant decrease in TNF with AT alone or in combination with GT. Plasma MDA/HNE and lipid peroxides were significantly decreased with AT, GT alone or in combination (p<0.01). Nitrotyrosine levels were significantly decreased only with GT or GT+AT but not with AT (p<0.01). Thus, the combination of AT and GT supplementation appears to be superior to either alone on biomarkers of oxidative stress and inflammation and needs to be tested in prospective clinical trials to elucidate its utility in CVD prevention.
Distribution of Biotin in Cerebrospinal Fluid of Children: Most of the Biotin is Bound to Protein
Authors: Bogusiewicz Anna , Stratton Shawna L , Ellison Dale A , Mock Donald M
Published: June 2021

Direct measurement of total avidin binding substances (TABS) in CSF underestimates the amount of true biotin as measured by HPLC/avidin‐binding assays. We investigated the mechanism of the underestimation as follows: 1) To evaluate whether detection of free biotin was masked by an unknown CSF component that blocks binding to avidin, biotin (independently quantified as 3H‐biotin) was added to CSF. TABS assay detected 100% of the added biotin, providing evidence against masking. 2) To determine if in vitro conversion of biotin to metabolites confounded detection, 3H‐biotin was incubated for 72 h in CSF at 4°C. No conversion of 3H‐biotin to biotin metabolites was observed by HPLC, providing evidence against conversion. 3) To evaluate the possibility that biotin is bound to a macromolecule and released during HPLC, CSF was acid hydrolyzed. Detectable biotin increased approximately 30 fold, providing evidence in favor of this mechanism. The 30 fold increase accounts for most, but not all, of the 90 fold increase observed for this sample after HPLC. Biotin‐binding proteins in CSF were further characterized by SDS‐PAGE, western blot, and streptavidin detection. Three bands were detected. These studies provide evidence that reversible and/or covalent binding to proteins causes substantial underestimation of CSF biotin. We speculate that this phenomenon may have confounded previous studies. NIH DK 36823
Effects of three doses of ascorbate on tyrosine hydroxylase immunoreactivities in the brain of paraquat‐injected C57BL/6 mice
Authors: Kang Min Jeong , Koh Hyun Chul , Lee Sang Sun
Published: June 2021

There has been increasing evidence that ascorbate is an important antioxidant, enzyme co‐factor and neuromodulator in the brain. Paraquat produces a selective and potentiated neurotoxicity to the nigrostriatal dopamine system. We investigated the effects of ascorbate on tyrosine hydroxylase immunoreactivities (TH‐ir) in substantia nigra (SN) and striatum of paraquat‐injected C57BL/6 mice. Male C57BL/6 mice (aged 7 week and 23–25g) were divided into five groups (n = 6). Ascorbate (4, 40, 400 mg/kg b.w., i.p.; LCPQ, ICPQ, HCPQ) treated daily for 4 weeks from 3 d before first paraquat injection. Paraqaut (10 mg/kg, i.p.) was administrated twice per week for 3 consecutive weeks. Saline was injected in intact mice (CON) and paraquat‐injected mice (CONPQ). The densities of TH‐ir neurons in the SN were in LCPQ and ICPQ groups higher than in CONPQ and HCPQ groups. The densities of TH‐ir fibers in the ventral subregion of rostral striatum and the dorsal subregion of caudal striatum were decreased approximately 20% in the CONPQ group relative to the CON group. In conclusion, these paraquat‐induced neurotoxicities in the brain may be prevented by 4 and 40 mg/kg dose of ascorbate. On the other hand, the 400 mg/kg dose of ascorbate did not show protective effects on paraquat‐injected mice. However, further studies are needed to understand the mechanism of ascorbate in the paraquat‐injected mice.
Vitamin K kinetics in humans after consumption of 13C‐labeled phylloquinone from kale
Authors: Novotny Janet A. , Baer David J. , Kurilich Anne C. , Britz Steven J. , Clevidence Beverly A.
Published: June 2021

The absorption and plasma elimination of vitamin K was investigated by uniformly labeling phylloquinone in kale with carbon‐13 and feeding the kale in a clinical study. Seven healthy volunteers ingested a single 400 g serving of kale with 30 g vegetable oil. The kale provided 156 nmol of phylloquinone. Serial plasma samples were collected and analyzed for the appearance of 13C‐phylloquinone by HPLC‐MS. Six of the subjects showed significant amounts of labeled phylloquinone in plasma, with labeled phylloquinone detectable for between 7 and 28 days following the dose. One subject's plasma was not consistently enriched above the detection limit, and this subject's baseline plasma was the lowest of the group. Average peak plasma concentration for the 6 subjects with detectable 13C‐phylloquinone was 2.1 nmolar. At the peak plasma concentration, the 13C‐phylloquinone was elevated between 5% and 34% above baseline (14% on average). The peak plasma 13C‐phylloquinone concentration represented 5±4% of the 13C‐phylloquinone in the kale dose. Carbon‐13 labeling of leafy vegetables for use in clinical nutrition studies is a feasible method for studying the absorption and metabolism of phylloquinone. Funded by USDA/ARS.
Inter‐relationship of fat‐soluble vitamins in progression of renal calcification
Authors: Fu Xueyan , Mernitz Heather , Wallin Reidar , Wang Xiang‐Dong , Booth Sarah L
Published: June 2021

Both 1alpha, 25 dihydroxyvitamin D3 (1,25D, 5.0 μg/kg diet) and 9‐cis retinoic acid (9cRA, 15 mg/kg diet) inhibited lung carcinogenesis in a tobacco carcinogen (NNK)‐initiated A/J mouse model, but 9cRA mitigated 1,25D‐induced renal calcification (Mernitz et al, 2007). Matrix gamma‐carboxyglutamic acid protein (MGP) requires vitamin K for gamma‐carboxylation, which confers its function as an inhibitor of soft tissue calcification. We therefore hypothesized that fat‐soluble vitamins may regulate MGP expression and function. RT‐PCR (n=10 per group) and western blotting (n=3 per group) were used to assess MGP expression, and gamma‐carboxylation in kidneys of 4 groups of mice: Control (NNK alone), 1,25D (NNK+1,25D), 9cRA (NNK+9cRA) and COMBO (NNK+1,25D+9cRA). Kidney MGP mRNA levels were significantly higher (~two‐fold) in 1,25D group compared to all other groups (p<0.01). However, there were less carboxylated MGP levels (relative density) in the 1,25D group [1.8±1.1 vs. 4.6±0.8 (Control); 5.2±0.7 (9cRA); 6.5±3.2 (COMBO); p<0.05]. In contrast, tissue vitamin K levels (pmol/g) were lowest in the 9cRA and COMBO groups [67±32 (Control); 63±13 (1,25D); 18±7 (9cRA); 33±14 (COMBO); p<0.01]. Although the role of vitamin K in this process is unclear, these data suggest that 1,25D can induce renal calcification through the impaired carboxylation of MGP, whereas 9cRA protects against it.
A comparison of hyperimmune egg protein and placebo for efficacy and safety among healthy young adults
Authors: Scheett Timothy P , Martin Tyler D. , Carr Benjamin M. , Koster Carolyn A. , Celmer Patricia A. , Whitener Jeffrey A. , Webster Michael J.
Published: June 2021

To evaluate the safety and efficacy of oral HIE supplementation during a sample training program. Twenty‐four recreationally active males (23.6 yrs, 176 cm, 69.2 kg and 17.1 % body fat) were randomly assigned to either HIE (n=12) or an egg protein placebo (PLA) group. Participants were supplemented with 4.5 g/d for 2 d, 9 g/d for 2 d and 13.5 g/d for 6 d. HIE and PLA supplements were identical in appearance and taste before and after mixing with 237 mL of milk. Subjects recorded duration and severity of adverse events in a daily log. HIE and PLA had a 100% compliance with the study protocol. 17% (n=2) of HIE and 25% (n=3) of PLA reported experiencing at least one adverse event. HIE reported experiencing adverse events for a total of 10 d as compared to 24 d for PLA. In HIE one subject reported experiencing stomach ache and diarrhea for 3 d (severity of 2 on a 10 pt scale) and another subject reported a skin rash lasting 4 d (severity of 6). In PLA one subject reported experiencing stomach ache and vomiting for 3 d and increased thirst and feeling tired/sleepy for 3 d (severity of 7 and 8, respectively). Subjects reported a runny nose (n=3) lasting 1–3 days (1–5 severity) and a cough (n=2) lasting 3 d (severity 1–5). HIE was associated with fewer adverse events of similar or lesser severity than PLA and all adverse events were minimal and transitory in nature with none requiring medical intervention.

Supported by Legacy for Life, LLC, Melbourne, FL
Multiple roles for Mcm10 during lagging strand synthesis
Authors: Bielinsky Anja‐Katrin , Das‐Bradoo Sapna , Haworth Justin , Chattopadhyay Sharbani
Published: June 2021

In lagging strand synthesis, Okazaki fragments are initiated by DNA polymerase‐alpha/primase, which produces a short RNA/DNA primer. Subsequently, a switch occurs from pol‐alpha to pol‐delta. We believe that minichromosome maintenance protein (Mcm) 10 coordinates this switch as it interacts with pol‐alpha and proliferating cell nuclear antigen (PCNA), the accessory factor for pol‐delta. This polymerase switch may be triggered by the non‐proteolytic ubiquitination of Mcm10, which is a pre‐requisite for Mcm10 to bind PCNA. Importantly, ubiquitinated Mcm10 can no longer interact with pol‐alpha. Because we have shown that the catalytic subunit of pol‐alpha is rapidly degraded in the absence of Mcm10, this modification might provide a mechanism to release pol‐alpha from the replication fork. We are currently trying to understand how Mcm10 is ubiquitinated and whether the Mcm10‐regulated degradation of pol‐alpha plays a role during replication.
Effects of polysaccharides from Salicornia herbacea on proliferation and apoptotic activity in mouse splenocytes
Authors: Kim Seonhee , Ryu Deokseon , Kim Yongho , Choi Hyunju , Lee Dongseok
Published: June 2021

This study investigated the proliferation and apoptotic activities of Salicornia herbacea in mouse splenocytes. Crude (CSP) and other kinds of fine polysaccharides (SPI and SPII) were prepared from Salicornia herbacea by ultrafiltration and gel filtration chromatography. Mouse splenocytes were treated with CSP, SPI or SPII (1, 2, 4, 8 mg/ml). The proliferation and apoptosis of the cells were used as indices for immunomodulating activity. The effect of proliferation was evaluated using MTS assay. Apoptosis of the cells was assayed quantitatively by Annexin V technique using a flow cytometry. In results, the proliferation was more increased in the SPI and SPII treated cells compared to the CSP treated cells. The most proliferation was shown in the splenocytes cultured with SPI at the concentration of 8 mg/ml for 24hr. The proliferation of splenocytes and separated T‐cells was higher (5.5 times and 3.7 times, repectively) than the control. Treatment of the mouse splenocytes with the SPI and SPII suppressed apoptosis of the cells. Moreover, those proliferation and suppression of the apoptotic death were in dose‐dependent manner. In conclusion, polysaccharides from Salicornia herbacea may have a beneficial effect on immunomodulating activity. (This work was supported by BK21, NURI, and BPRC)
The polycyclic flavonoid, trifolirhizin, inhibits proliferation in LNCaP and PC‐3 human prostate cancer cells by arresting cell cycle progression at G1/S and G2/M, respectively
Authors: Zhang Jason J.J. , Schoene Norberta W. , Cheng Zhihong , Yu Liangli Lucy , Cheng Wen Hsing , Lei Kai Y.
Published: June 2021

Trifolirhizin, found in the roots of Sophora flavescens, a traditional medicinal plant widely used in Asia, is a potent polycyclic flavonoid with anticarcinogenic effects. To elucidate the mechanisms underlying the anticancer activities of trifolirhizin, the effect of trifolirhizinon on the inhibition of cell cycle progression, induction of apoptosis and regulation of tumor suppressor gene expression was examined in human derived prostate cancer cells LNCaP and PC‐3 as well as normal human prostate epithelial cells (PrEc). The treatment of as low as 25 μM trifolirhizin in LNCaP cells for 6 days resulted in 40% inhibition of cell proliferation. Treatment of higher dosages caused the growth inhibition effect to be manifested in as early as 3 days. PC‐3 cells were not affected by trifolirhizin till the very high dosage of 125 μM that resulted in more than 50% reduction in cell proliferation after 4 to 6 days of treatment. Most importantly, for PrEc cells, trifolirhizin was effective only at the highest concentration of 200 μM. The growth inhibition in trifolirhizin treated LNCaP and PC‐3 cells was associated with cell cycle arrest at the G0/G1 and G2/M phase, respectively, in a dosage dependent manner. Moreover, the protein expression of cyclin E was unchanged but that of cyclin D1, p53 and p21 was suppressed in 50 μM trifolirhizin treated LNCaP cells, suggesting a p53 independent pathway.
Effects of red ginseng ethanol extract on lipid metabolism in C57BL/6J mice fed high‐fat diets
Authors: Mun Eun‐Gyeng , Kim Min‐A , Kim Young‐Sook , Wee Jae‐Joon , Cha Youn‐Soo
Published: June 2021

This study evaluated the effects of supplementation of red ginseng ethanol extract (RG‐EtOH; 0.1%, 0.5%, and 1% of high‐fat diet) in C57BL/6J mice fed high‐fat diet (60% fat kcal) for 8 weeks. The RG‐EtOH diet groups did not show any difference in food and energy intakes compared with the high‐fat diet group, but had lower final body weights, epididymal, and back fat pad weights. Triglyceride and total cholesterol concentrations of RG‐EtOH diet groups in serum were significantly lower than high‐fat diet group. Higher concentrations of RG‐EtOH, however, did not have greater effects on serum lipid profiles. Although the 1% concentration of RG‐EtOH group had lower lipid profiles and fat pad weight than high‐fat diet group, the effect was not as great as in the 0.5 percent of RG‐EtOH. These results were confirmed by mRNA expressions of enzymes (ACS, CPT‐I, PPAR‐α, PPAR‐γ, and ACC) which are related to lipid metabolism by reverse transcriptional‐PCR. We suggest that 0.5% RG‐EtOH is the most effective dose for improving lipid metabolism. Also, our study show that red ginseng ethanol extract reduced obesity induced by high‐fat diets. (This study was supported from the KT&G Central Research Institute.)
Effects of polysaccharides from Salicornia herbacea on anti‐diabetic activity in rats
Authors: Ryu Deokseon , Kim Seonhee , Kim Jiyeon , Lee Sangyeon , Choi Hyunju , Lee Dongseok
Published: June 2021

This study investigated the physiological activities of polysaccharides isolated from Salicornia herbacea. Polysaccharides were prepared by extraction with hot steamed water (CSP) and by further ultrafiltration (SPI). Male Spreque‐Dawley rats were divided into four groups which were normal control group (NC), diabetic control group (DC), diabetic CSP group (DCSP), and diabetic SPI group (DSPI). Diabetes was induced by injection of Streptozotocin (55mg/kg B.W. i.p.). CSP and SPI were administrated to the rats as drinks (2%) for 6 weeks. The glucose, triglyceride, total cholesterol, and HDL‐cholesterol concentrations were measured in the blood. In results, food intakes were markedly higher in the DC than those of the NC and were not significantly decreased in the DCSP and DSPI. Body weight, feed efficiency ratio (FER), blood glucose levels in the diabetic group were significantly improved by the treatment of the CSP and SPI (p<0.05). Increments of the serum total cholesterol and triglyceride levels in the DCSP and DSPI were smaller than those in the DC. Serum HDL‐cholesterol levels in the DCSP and DSPI were higher than those in the DC. Especially, the treatment of SPI showed the improved anti‐diabetic activity in the diabetic rats. It is suggested that polysaccharides from Salicornia herbacea could be useful as anti‐diabetic functional agent. (This work was supported by BK21, NURI, and BPRC)
Chronic effect of CLA isomers on bone mineral density, fat and lean mass in C57BL/6 female mice
Authors: Halade Ganesh Vasantrao , Rahman Md Mizanur , Banu Jameela , Fernandes Gabriel
Published: June 2021

Conjugated linoleic acids (CLA) are a class of positional, geometric, conjugated dienoic isomers of linoleic acid (LA). Dietary CLA supplementation results in a dramatic decrease in body fat mass and increase in lean mass as well as protects against bone loss both in mice and humans. However, little information is known of the long term effect of CLA isomers in animals as well as in humans. In order to investigate the isomer specific effects of CLA on fat and lean mass, the purified c9,t11‐CLA, (0.5%), t10,c12‐CLA (0.5%) and CLA‐mix (0.25%, c9,t11‐CLA + 0.25% t10,c12‐CLA) were fed along with 10% corn oil (CO) to 6 months old C57BL/6 female mice (n=8–10) for 6 months. All of the dietary interventions of CLA groups were compared with CO fed mice as a control group. After 6 months, mice were scanned using DEXA to measure the bone mineral density (BMD), lean and fat mass. The results revealed that, the body weights were significantly decreased in both t10,c12‐CLA, CLA‐mix groups compared to c9,t11‐CLA and 10% corn oil. Further, t10,c12‐CLA and CLA‐mix groups significantly decreased the total and abdominal fat mass compared to c9,t12‐CLA and CO group. In addition, both CLA‐mix and t10,c12‐CLA significantly increased the lean mass and also BMD in lumbar spine and distal femoral metaphysis compared to corn oil. The overall % fat of the body were decreased significantly in t10,c12‐CLA and CLA‐mix compared to c9,t11‐CLA and CO group. In conclusion, the specific effect of t10,c12‐CLA induces both significant fat mass reduction, maintains higher lean and bone mass compared to c9,t12‐CLA and CO.

Research support: NIH R21 AG027562
Probucol impairs cardiac function in apo E‐KO mice
Authors: Moghadasian Mohammed H , Othman Rgia , Jassal Davinder , Fang Tielan , Zhao Zhaohui , Kroeker Amy , Riediger Natalie , Xu Zuyuan , Walker Kristy , Le Khoung , Azordegan Nazila
Published: June 2021

Background: Despite beneficial cholesterol‐lowering and antioxidant properties of probucol, it paradoxically increases coronary atherogenesis in apo E‐KO mice. The aim of this study was to investigate the effects of probucol on cardiac function in apo E‐KO mice.

Materials and Methods: Male apo E‐KO and C57BL/6 mice were fed an atherogenic diet with (treated, n=8) or without (control, n=8) 1% probucol for 16 weeks. Plasma cholesterol levels were measured. Murine echocardiography was performed serially at 6 week intervals.

Results: Probucol treatment resulted in significant reductions in plasma cholesterol levels in both apo E‐KO (−30%, p<0.05) and C57BL/6 mice (−90%, p<0.01). Apo E‐KO mice treated with probucol showed early regional LV systolic dysfunction as early as 6 weeks with further decrease over time [endocardial velocity −40%; strain −40%, ejection fraction −10% (p<0.01)], with no impact on mortality. Such effects of probucol were not observed in C57BL/6 mice.

Conclusions: Probucol impairs cardiac function in apo E‐KO mice. This may be related to previously observed coronary atherogenesis due to probucol in this animal model.

Supported in part by URGP, HSF and NSERC.
Effect of nicotinic acid on plasma lipid and hepatic gene expression in obese insulin resistant dogs.
Authors: Le Bloc'h Jérôme , Leray Véronique , Zaman Muhamad‐Quaid , Serisier Samuel , Ouguerram Khadija , Nguyen Patrick
Published: June 2021

Nicotinic acid (NA) is well known for improving plasma lipids in man (decrease in triglycerides (TG), total (TC) and LDL‐cholesterol (LDL‐C) and increase in HDL‐cholesterol (HDL‐C) levels). The aim of this study was to assess the effects of NA in obese insulin resistant dogs.

Eight beagle dogs were alloted to 2 groups : treated (n=5) and control (n=3). Treated dogs were given 375mg/d of NA for one week and then 500mg/d for 3 weeks. Insulin sensitivity, plasma TG, TC, free fatty acids and insulinemia were assessed at the end of the experimental period. Plasma lipoproteins were separated, and LDL‐C or HDL‐C were measured. Hepatic expression of ABCA1 and SR‐BI was quantified using real‐time RT‐PCR.

NA treatment did not induce any change in either insulin sensitivity or glycemia, insulinemia and free fatty acid concentrations. In the treated group, TG were reduced by 30% (p < 0.05), and TC by 25% (p < 0.001). Analysis of lipoprotein cholesterol showed a reduction in LDL‐C (p < 0.05) whereas HDL‐C was unchanged. All these parameters were unchanged in the control group. No change in the ABCA1 and SR‐BI expression was observed in liver.

Our results confirm the effects of NA that have been described in humans, except that concerning the increase in HDL‐C. According to observations recently done in mice, as dogs are devoid of CETP activity, we can hypothesize that CETP might be the link between plasma TG decrease and change in HDL‐C.
Modulation of Platelet Function by Recombinant Thrombomodulin Hematologic Implications
Authors: Adiguzel Cafer , Iqbal Omer , Fareed Daniel , Hoppensteadt Debra , Jeske Walter , Simmikannu Michael , Fareed Jawed
Published: June 2021

While recombinant thrombomodulin's (rTM) effect on blood clotting are known, its effect on platelets is not clear. This study investigated the effect of rTM (ART 123, Asahi Pharmaceutical, Japan) on platelets using platelet aggregation, release and activation assays. In platelet aggregation studies, citrated whole blood was supplemented with 0–10 μg/ml rTM. Aggregation was induced by ADP (5 and 2.5 μM) or thrombin (0.5 U/ml). Platelet activation was measured by flow cytometry following stimulation with rTF or ADP. Platelet release was measured as 14C serotonin release (SRA) from washed platelets. In platelet aggregation assays, rTM did not inhibit ADP‐induced aggregation, but concentration‐dependently inhibited thrombin induced aggregation (IC50: 0.42 μg/ml). rTM enhanced microparticle generation at low concentrations, but at concentrations > 0.31 μg/ml rTM concentration dependently inhibited microparticle formation (IC50: 2.5 μg/ml). At concentrations >5.0 μg/ml, rTM completely blocked TF‐induced microparticle formation, but did not block P‐selectin expression. rTM inhibited thrombin‐induced serotonin release. These studies suggest that while rTM does not inhibit agonist induced platelet aggregation, it can inhibit TF induced microparticle formation. As rTM concentrations up to 10 μg/ml do not affect P‐selectin expression, it is unlikely to alter primary hemostasis at therapeutic levels.
Pharmacokinetics of Defibrotide in Non‐Human Primates is Dose Dependent.
Authors: Fareed Jawed , Hoppensteadt Debra , Neville Brian , McGeehan Elizabeth , Iacobelli Massimo , Jeske Walter
Published: June 2021

Defibrotide (DF) is a mammalian tissue derived polyelectrolyte which has been used for the treatment of arterial and venous thrombosis and microangiopathic disorders. Until recently indirect methods have been used to measure the levels of DF. More recently a colorimetric method, based on chromogenic coupling, is developed to measure the absolute amount of this agent. To investigate the absolute intravenous pharmacokinetics (PK) of DF in non‐human primates, graded dosages of defibrotide (1.0–5.0 mg/kg) were administered to individual groups of monkeys (n=6). Whole blood and plasma based clotting times were measured for up to 12 hours after the intravenous bolus at these dosages. In addition, tissue factor pathway inhibitor (TFPI) antigen levels and absolute levels of defibrotide were also measured using the colorimetric assay. The half life of 1.0, 2.5 and 5.0 mg/kg IV bolus ranged from 30–120 minutes. The peak concentration was also dependent on the dosage and ranged from 30±4 ug/ml suggesting a linear PK. DF produced minimal effects on the clotting assays, however, a dose dependent increase in TFPI antigen levels and defibrotide levels using a colorimetric method were observed. Repeated administration of defibrotide every 12 hours resulted in a progressive increase in AUC0–12, suggesting partial accumulation. In addition, at higher dose, DF resulted in an increased anti‐Xa activity which was dose independent. These studies clearly suggest that the pharmacologic actions of DF involve both the direct (plasmatic) and indirect pharmacodynamic components.
Suledoxide Mediated Inhibition of Tissue Factor Activation of Platelet. A Potential Mechanism for the Observed Therapeutic Effects in Diabetic Nephropathy
Authors: Adiguzel Cafer , Iqbal Omer , Debra Hoppensteadt , Cunanan Josephine , Fareed Jawed
Published: June 2021

Suledoxide (Alfa Wassermann, Bologna) is used for the management of cardiovascular disorders and has been developed in the United States for the treatment of diabetic nephropathy. Suledoxide is composed of 80% fast moving heparin and 20% dermatan sulfate resulting in a lower anticoagulant effect in comparison to unfractionated heparin. To investigate Suledoxide's effect on platelet function, studies were carried out to characterize its effects on agonist induced platelet aggregation and tissue factor (TF)‐induced activation of platelets. Supplementation of Suledoxide to platelet rich plasma did not alter ADP, epinephrine, and collagen‐induced platelet aggregation. However Suledoxide inhibited TF‐mediated microparticle formation (IC50=3.5 μg/ml), platelet aggregate formation (IC50=7.5 μg/ml), and P‐ selectin expression (IC50=4.0 μg/ml). Another heparinoid, Danaparoid, inhibited microparticle formation (IC50=3.5 μg/ml) but did not inhibit P‐selectin expression and platelet aggregate formation. These results suggest that while Suledoxide exhibits a lower anticoagulant effect, it still capable of inhibiting TF‐mediated platelet activation. This effect may contribute to the observed therapeutic effects of Suledoxide in diabetic nephropathy. These results also suggest that inhibition of TF activation of platelets by Suledoxide may be independent of its anticoagulant effects.
Effects of endurance exercise training on endothelial function and atherosclerosis in the hereditary hypercholesterolemic swine model
Authors: Newcomer Sean , Roseguini Bruno , Esquivel Arturo Arce , Bender Shawn , Turk James , Laughlin Harold
Published: June 2021

Hereditary hypercholesterolemic (HH) swine develop advanced atherosclerotic lesions throughout the coronary and peripheral vasculatures. The purpose of this study was to test the hypothesis that endurance exercise training would increase endothelial function and decrease the prevalence of atherosclerosis in this swine model. Ten adult pigs were randomly assigned into either exercise trained (EX) or sedentary groups (SED). Exercise training consisted of 16 weeks of treadmill running. Upon sacrifice the following arteries were harvested: thoracic aorta (TA), carotid (CA), brachial (BR), femoral (FEM) and coronary arteries (LAD, RCA, CFX). In vitro assessment of endothelium‐dependent dilation (EDD) was determined by administering increasing doses of acetylcholine to BR and FEM rings and bradykinin to CFX rings. Atherosclerotic lesion formation was quantified through Sudan IV staining in the TA, CA, BR, FEM, LAD and RCA. Endurance exercise training increased EDD in the BR and FEM but not in the CFX. The percentage of Sudan IV staining was not significantly different between EX and SED swine in the TA, CA, BR, FEM, RCA and LAD arteries. These findings demonstrate that endurance exercise training increases EDD in the BR and FEM arteries, but does not alter the development of atherosclerosis in HH swine.

Supported by NIH HL083597, HL36088 and HL52490
Effects of acute exercise on endothelial nitric oxide synthase (eNOS) in rat vasculature
Authors: McAllister Richard Murray , Weimer Cory Matthew
Published: June 2021

eNOS is phosphorylated by shear stress in cultured endothelial cells; whether eNOS phosphorylation occurs in vivo is unknown. We hypothesized that acute exercise induces phosphorylation of eNOS via the protein kinase Akt. Rats were either cage‐confined (Sed, n=4) or run on a treadmill for 30 min (Ex, n=4). Endothelial proteins were extracted from each rat aorta in a buffer containing the phosphatase inhibitor calyculin (100 nM). NOS activity ([3H]‐L‐arginine assay) did not differ (P>0.10) between Sed and Ex (Sed, 1001±200 fmol/mg/h; Ex, 1269±150). Phosphorylated eNOS (Ser1177; Western blotting) also did not differ between groups (Sed, 1017±141 units; Ex, 879±216). Phosphorylated Akt (Ser473) was not detected in most vessels; phosphorylated protein kinase A (PKA; Thr198) was detected (Sed, 886±242 unts; Ex, 1496±210; 0.05<P<0.10). These data suggest that levels of shear stress at rest are sufficient to phosphorylate eNOS at Ser1177. Further, they may indicate that PKA rather than Akt is the mediator of eNOS phosphorylation in vivo.

Supported by NIH RR1 8276 and a Faculty Research Award from the College of Veterinary Medicine, University of Missouri.
Interconnections between the transcription and processing of mRNA precursors
Authors: Manley James L.
Published: June 2021

My laboratory studies the mechanisms and regulation of pre‐mRNA splicing and polyadenylation in vertebrate organisms, with emphasis on understanding how these processes are connected to the transcription of mRNA precursors by RNA polymerase II and to other nuclear events. We use a variety of different methods, including biochemical, genetic and structural, to investigate these pathways and the protein factors involved, and to analyze the consequences on cell growth, development and viability when these processes are disrupted. I will present results from our most recent analyses of these problems.
Events leading to intestinal barrier failure and systemic inflammation following hemorrhagic shock
Authors: Thuijls Geertje , Derikx Joep P , de Haan Jacco‐Juri , Hadfoune M’hamed , Heineman Erik , Buurman Wim A
Published: June 2021

Hemorrhagic shock (HS) leads to systemic inflammatory response syndrome (SIRS), a potential deleterious complication. A current hypothesis states that gut barrier loss is crucial in the development of SIRS. We aimed at elucidating the sequence of events leading to gut barrier loss and SIRS following HS.

Rats were subjected to HS and sacrificed at 15, 30, 60, 90 min after HS, and compared with sham controls. Intestinal neutrophil influx (NI), tight junction (TJ) integrity, enterocyte integrity, bacterial translocation (BT), and SIRS were assessed.

HS leads to early NI from 15 min after HS, followed by a significant loss of TJ proteins Claudin‐3 and ZO‐1 from 30 and 60 min after HS, respectively (100% versus 71% and 100% versus 41%, respectively, P<0.05, students t‐test (stt)). Enterocyte integrity loss measured by plasma Ileal Lipid Binding Protein levels, is markedly increased from 60 min after HS (0.6 versus 11.8 ng/ml, P<0.0001, stt). Intestinal barrier function loss detectable from 60 min after HS is represented by BT (0.0 versus 47.9 CFU/g tissue, p<0.005, wilcoxon signed rank test (wsrt)). SIRS measured by plasma TNFα levels is significantly elevated from 60 min after HS (0.0 versus 26.1 pg/ml, p<0.05, wsrt).

HS leads to NI and gut barrier integrity loss, followed by BT and SIRS within 90 min after shock. Ongoing studies are directed at unraveling the causal relation between succeeding events.