bluuebunny/crossref_metadata_2025 huggingface dataset:
Methyl gallate protects against oxidative stress‐induced apoptosis in PC12 cells
Authors: Crispo James , Piché Matthew , Ansell Dominique , Eibl Joseph , Tai Isabella , Ross Gregory , Tai T.C.
Published: June 2021

Recent studies demonstrate that polyphenols reduce oxidative damage by increasing cell viability, decreasing intracellular ROS and altering GSH metabolism. In addition, polyphenols such as epigallocatechin gallate (EGCG) have been reported to inhibit apoptotic signaling and increase neural cell survival. In an effort to better understand the benefits associated with polyphenol consumption, the aim of this study was to investigate potential anti‐apoptotic effects of EGCG, methyl gallate (MG), gallic acid (GA) and N‐acetylcysteine (NAC) on H2O2‐induced apoptosis in PC12 cells and elucidate protective mechanisms. Pre‐treatment with MG and NAC significantly increase viability of H2O2‐stressed cells, while pre‐treatments with EGCG and GA exacerbates stress. Analyses of apoptosis and mitochondrial membrane potential reveals that MG pre‐treatment prevents mitochondria depolarization, however does not inhibit apoptosis. Further, analysis of DNA degradation and caspase cleavage shows that MG inhibits activation of caspase 9 and has a partial inhibitory effect on DNA degradation. These findings suggest that MG may have potential therapeutic properties against mitochondria‐mediated apoptosis.

Research funded by the Northern Ontario School of Medicine and Natural Sciences and Engineering Research Council of Canada.
Novel Pyrimidin‐2‐yl‐benzylidene imines and 2‐[(pyrimidin‐ylimino)‐methyl]‐phenoles and their Antibacterial Activity
Authors: Hoti Ramiz , Kalaj Vuksan , Vehapi Idriz , Ismaili Hamit , Thaçi Veprim , Bicaj Muhamet
Published: June 2021

Some new Pyrimidin‐2‐yl‐benzylidene‐imines are synthesized by catalytic reactions of aromatic aldehydes and corresponding 2‐Aminopyrimidines. Condensation of Benzaldehyde 1a and 2‐Aminopyrimidines 2(a,b) gave pyrimidin‐2‐yl‐benzylidene imines 3(a,b). By condensation of salycilaldehyde 1b and 2‐Aminopyridines 2(a,b), corresponding 2‐[(pyrimidin‐ylimino)‐methyl]‐phenoles 3(c,d) were synthesized. The antibacterial activity of products 3(a–d) were investigated and results were submitted for their activities against Escherichia coli, Clebsiella and Staphylococcus aureus.

Compound 3a showed emphatic bactericide activity against Staphylococcus aureus. Compounds were more active against. Light activity against Escherichia coli exhibited compounds 3a and 3c, whereas 3d exhibited the strongest activity against Clebsiella. In general there was a direct positive correlation between increasing concentration of the compound and antibacterial activity
Differential effects of flaxseed oil (Linum usitatissimum) on cell growth between malignant and non‐malignant cell lines
Authors: Buckner Alison Linda , Buckner Carly , Lombardo Domenic , Abou‐Zaid Mamdouh , Lafrenie Robert
Published: June 2021

Many cancer patients take vitamins and supplements as potential treatments and therefore many researchers have begun to systematically evaluate the potential of these “functional foods”. Flax, in the form of seeds or seed‐derived oil, is recognized for its exceptional nutritional value due to its high concentration of fiber‐based lignans and large amounts of omega fatty acids. We have examined the anti‐cancer effects of flaxseed oil by studying its direct effects on cancer cell growth in vitro. A total of seven different oils, including flaxseed oil, were characterized by HPLC and GC/MS analysis and used to treat cells. Treatment with flaxseed oil was associated with a rapid slowing of growth by the aggressive murine melanoma cell line B16‐BL6. Treatment of B16‐BL6 with each of the other characterized oils, showed no significant change in cell growth. Interestingly, non‐malignant cell lines such as HSG cells, showed an increase in cell growth following flaxseed treatment. Treatment with flaxseed oil inhibited growth of all 6 tested malignant cells in a dose‐dependent manner and did not inhibit growth of non‐malignant cell lines. Therefore, in addition to improving the efficacy with which cancer treatments selectively target and destroy neoplastic cells, current efforts in the development of novel therapeutic agents seek to minimize aversive side effects to improve patient‐well being and quality of life.
Reduction effect of neutropenia by herbal medicine treatment to chemotherapy
Authors: Choi Han‐Seok , Choi Hyeong‐Sim , Shin Yong‐Cheol , Ko Seong‐Gyu
Published: June 2021

It has been reported that there could be a possibility of side effects on neutropenia while chemotherapy is applied to treat prostate or breast cancer; however, these side effects could be reduced or even eliminated via combining with herbal medications. In this study, we had how made neutropenia induced model. First, we have injected cyclophosphamide(150mg/kg, every 2 days) into a targeted mice to drive neutropenia. Next, to two different group, we have injected G‐CSF(0.1mg/kg, every 2 days) to the one side and herbal medicine to another. After the treatment, we have measured the quantity of neutrophils from its blood cells; moreover, we have then measured level of cytokines secreted from bone marrow cells and spleenocytes. From the acquired information, we observed that the levels of both cytokines and neutrophils are recovered from both groups, G‐CSF and herbal medicated. It is shown that herbal medicine could possibly used as a substitute of G‐CSF as a treatment for neutropenia. As a consequence, herbal medications could benefit chemotherapy.
Anti‐allergic effect of Polar Lichen Extract in animal models
Authors: Choi Ko Woon , Byeon Hye‐Eun , Yim Joung‐Han , Lee Hong Kum , Pyo Suhkneung
Published: June 2021

Allergies are immediate hypersensitive responses to antigens and IL‐4 is involved in the initiation and development of allergic responses. Lichen‐derived compounds have been known to have a variety of biological activities including anti‐inflammatory activity, but the effect of polar lichen extract (PLE) on allergic responses is not known yet. The present study was undertaken to examine whether PLE has an inhibitory effect on allergic response in mouse splenocytes and mouse model. Our results showed that PLE suppressed both T and B cells proliferation induced by mitogens. RT‐PCR analysis also revealed that mRNA levels of IL‐4 were in vitro suppressed in the presence of diosgenin. In allergic mouse model, application of PLE on atopic dermatitis like skin lesions improved skin condition and inhibited starching behaviors. In addition, PLE application suppressed IL‐4 mRNA levels and its secretion. Moreover, PLE suppressed foot swelling in rats. Taken together, these results suggest that PLE has a potent inhibitory role in T cell activation and may be a candidate for therapeutic agent in allergy.
The extract of Pharbitis nil Choisy has bone regenerative potential in the healing process of calvarial defects in a rabbit model
Authors: Lee Kang Jin , Baek Eun Ji , Park Sang Yong , Shin Heonsub , Park Jeonghyeok , Shin Yeongjun , Han Chang‐Kyun , Yi Tae‐Hoo
Published: June 2021

The purpose of this study was to evaluate the regenerative potential of Pharbitis nill Choisy in the healing process of calvarial defects in rabbits. Calvarial defects were surgically created through an 8 mm trephine bur at the rabbit calvarium. In the experimental group, mixture of beta‐tricalcium phosphate and BuOH fraction of Pharbitis nil Choisy was applied at the rabbit calvarial defects. In the control group, beta‐tricalcium phosphate was only applied. Animals were sacrificed at healing periods of 4, 8, and 12 weeks. Data analysis performed through computer tomography analysis, and bone mineral density measurement and histophathological analysis. The computed tomographic analysis performed at 4 to 12 weeks after operation and mixture of beta‐TCP and BuOH fraction of Pharbitis nil Choisy was almost absorbed and the amount of osteogenesis was increased. Four, 8, and 12 weeks after the operation, Bone mineral density analysis was 0.1821 g/cm2, 0.1715 g/cm2, and 0.2125 g/cm2, respectively in the control group. while the experimental group were 0.2468 g/cm2, 0.3619 g/cm2, and 0.3217 g/cm2. Accordingly, bone mineral density appeared higher in the experimental group than in the control group. Therefore, we suggest that mixture of beta‐tricalcium phosphate and BuOH fraction of Pharbitis nil Choisy have effect on effective osteogenesis in the defective region of the rabbits calvarial bone.
The Herpes simplex virus‐1 DNA polymerase exhibits 3′ phosphatase activity
Authors: Ravi Rupesh Kanchi , Boehmer Paul E.
Published: June 2021

Herpes simplex virus‐1 (HSV‐1) is a large dsDNA virus that encodes its own DNA replication machinery, including enzymes involved in nucleotide metabolism. We recently demonstrated that the HSV‐1 DNA polymerase (UL30) exhibits AP and 5′ dRP lyase activities. Moreover, UL30 in conjunction with the viral uracil DNA glycosylase as well as cellular AP endonuclease and DNA ligase IIIα/XRCC1, perform uracil‐initiated Base Excision Repair (BER). As an alternative route to repairing base adducts and abasic sites, and as a mechanism to process ssDNA breaks we have investigated the ability of UL30 to process various 3′ ends. Interestingly, we find that UL30 can degrade 3′ sugar phosphate as well as 3′ phosphate terminated DNA. Moreover, UL30 exhibits a 3′ phosphatase activity that removes 3′ phosphate prior to 3′–5′ exonuclease action. These novel activities would allow UL30 to process AP sites following β‐elimination though incision by its inherent AP lyase activity or DNA breaks that result in 3′ phosphate terminated DNA following βδ‐elimination or X‐ray irradiation. Further studies will investigate how UL30 can repair this damage in conjunction with other factors. These findings further highlight the role of DNA repair in the maintenance of a viral genome with implications on both lytic replication and reactivation from latency.

This work was supported by grant GM 062643 from the NIH.
CML‐08‐62, a novel NF‐kB inhibitor, inhibits the growth of human fibrosacoma cells through the induction of apoptosis
Authors: Lee Chang Woo , Kim Jang Hyun , Kang Moo Rim , Lee Heesoon , Lee Ki Hoon , Lee Kiho , Kang Jong Soon , Kim Hwanmook , Park Song‐Kyu
Published: June 2021

NF‐κB, an inducible transcription factor, regulates the immune response and plays critical roles in the pathogenesis of chronic inflammatory diseases and a variety of human cancers. It has been reported that suppression of NF‐κB limits the proliferation of cancer cells. In this study, we investigated the anti‐cancer activity of CML‐08‐62, a novel NF‐κB inhibitor. This compound significantly suppressed the proliferation of 16 human cancer cell lines tested. CML‐08‐62 most effectively inhibited the growth of HT‐1080 cells, the human fibrosarcoma cell line, among the cell lines. Furthermore, it was shown that CML‐08‐62 was able to inhibit the NF‐κB expression in HT‐1080 cells. Flow cytometry analysis showed that the apoptotic death of HT‐1080 cells. Cleaved caspase 3 and cleaved caspase 9, which were active forms involved in apoptotic signaling pathway, and cleaved PARP were also detected in HT‐1080 cells treated with CML‐08‐62. In addition, fluorescence imaging analysis showed that the growth of the HT‐1080‐RFP tumors expressing fluorescence signal decreased dose‐dependently. The tumor inhibitory rates of CML‐08‐62 at doses of 30 and 50 mg/kg were 20% and 47%, respectively. Taken together, our results indicate that CML‐08‐62 exerts an anti‐cancer activity in vitro and in vivo and may be a promising therapeutic agent.
A study of the role of GATA4 gene polymorphism in myocardial infarction
Authors: Bakhit Dana Mohammed , Muiya Paul , Gueco Daisy , Meyer Brian F , Al‐Mohanna Futwan , Alshahid Maie , Dzimiri Nduna
Published: June 2021

The role of GATA4 in cardiovascular disease remain to be elucidated. We initially sequenced the GATA4 coding regions to identify variants of interest in 250 individuals, and investigated the role of 5 SNPs, rs3729855 (C>T), rs3729856 (A>G), rs1062219 (C>T), rs11785481 (C>T) and rs3203358 (C>G) in predisposing individuals to myocardial infarction (MI) by the real‐time PCR in 3803 individuals, all of Saudi origin. We then tested the impact of congenital heart disease (CHD) by comparing the CHD patients with the rest. The results pointed to a significant association for the rs3729856 (p.S377G) with CHD [Odds ratio(95% Confidence Interval) = [1.48(1.06–2.08); p=0.023], and protective effect for rs3729855 [0.96 (0.42–2.19); p=0.005. In order to rule out the influence of CHD on other relationships, we excluded CHDs from further analyses involving 2713 MI patients and 986 controls. Interestingly, the rs3729856 [1.16(1.01–1.34); p=0.034] also conferred risk for MI, while the rs3729855 [0.68 (0.52–0.89); p=0.005] showed protective properties. Furthermore, both the rs3729856 [1.14(0.99–1.31); p=0.076] and rs3729855 [1.37(1.00–1.88); p=0.052] were also associated with hypertension, but not with type 2 diabetes mellitus. These observations implicate the GATA4 as a susceptibility gene for both MI and CHD.

This work was supported through the Royal Cardiovascular Research Grant of the King Faisal Specialist Hospital and Research Centre
Caffeic acid disturbs adipokine resistin‐induced adhesion molecule upregulation in cultured human endothelial cells
Authors: Lee Eun‐Sook , Gong Juhyeon , Kang Young‐Hee
Published: June 2021

Adipokines has been implicated in the pathogenesis of atherosclerosis. Recent studies suggested that resistin is associated with endothelium dysfunction and proinflammation, which plays a major role in triggering and sustaining atherogenesis. This study investigated that resistin induced expression of cell adhesion molecules of E‐selectin, VCAM‐1 and ICAM‐1 in human endothelial cells(HUVEC) and that such induction was suppressed by 1–20μM caffeic acid, a naturally occurring phenolic compound. We found that resistin induced the expression of all three adhesion molecules within 8 h and hence enhanced THP‐1 monocyte adhesion on activated endothelium. Non‐toxic caffeic acid at ≥ 1μM attenuated expression of E‐selectin and VCAM‐1 of resistin‐inflamed HUVEC. In addition, the monocyte adhesion to resistin‐activated endothelial cells was diminished by adding 10μM caffeic acid. However, the elevated expression of another adhesion molecule PECAM‐1 was not influenced even by 20μM caffeic acid. These results demonstrate that caffeic acid blocked monocyte trafficking to resistin‐activated endothelium via retarding expression of cell adhesion molecules. Thus, caffeic acid may have a therapeutic potential disturbing obesity‐triggered atherosclerosis. This research was financially supported by the Ministry of Knowledge Economy through the Technological Development Project for Regional Strategic Plan.
Sympathetic nerve‐mediated nitrergic dilation of basilar arteries of the rat in vivo
Authors: Chang Hsi‐Hsien , Lee Hui‐Chao , Chen Po‐Yi , Lee Yuan‐Chieh , Chen Mei‐Fang , Kuo Jon‐Son , Lee Tony Jer‐Fu
Published: June 2021

Brain arteries receive sympathetic nerves originating in superior cervical ganglion (SCG), and norepinephrine (NE) is thought the transmitter for vasoconstriction. The exact role of the sympathetic nerve in regulating brain circulation, however, remains unclear. Results of studies in isolated cerebral arteries indicate that activation by nicotine of cerebral perivascular sympathetic nerves induces NE release. NE then diffuses to act on β2‐adrenoceptors located on nitrergic nerve terminals to release nitric oxide (NO) that relaxes the smooth muscle. This axo‐axonal interaction in regulating cerebral blood flow, therefore, was examined in the rat in vivo. Electrical stimulation of SCG caused dilation of basilar arteries in anesthetized normotensive Wistar‐Kyoto rats (WKY). The vasodilation was blocked by inhibitors for NO synthesis and beta2‐adrenoceptors. In spontaneously hypertensive rats (SHRs), electrical stimulation‐of‐SCG‐induced nitrergic dilation of basilar arteries, however, was diminished. This is consistent with reported findings of decreased parasympathetic nerve terminals in cerebral arteries of SHR. These findings indicate that activation of cerebral perivascular sympathetic nerves does not cause significant vasoconstriction, but predominant nitrergic dilation of the basilar arteries in normotensive animals. This effect of sympathetic nerves may change in hypertension.
Generation of Transgenic Mice Overexpressing a Hybrid Enzyme Up‐regulating Biosynthesis of Vascular Protector Prostacyclin
Authors: Mohite Anita J , Chillar Annirudha J , So Shui‐Ping , Ruan Ke‐He
Published: June 2021

Prostacyclin (PGI2) is a potent vascular protector through its anti‐platelet aggregation and vasodilation properties. Non‐steroid anti‐inflammatory drugs (NSAIDs) by inhibiting PGI2 production promote heart diseases. Recently, we have successfully engineered a hybrid enzyme (COX‐1‐10aa‐PGIS, by linking cyclooxygenase isoform 1(COX‐1) with PGI2 synthase (PGIS) through a 10 amino acids (10aa) linker) with triple catalytic functions directly converting arachidonic acid into PGI2. PGI2 biosynthesis was upregulated in the mammalian cells, transfected with the hybrid enzyme cDNA. Thus, with the objective of translating these in vitro results into in vivo, we generated transgenic mice overexpressing COX‐1‐10aa‐PGIS. PCR using mouse tail tissue confirmed the incorporation of COX‐1‐10aa‐PGIS gene in the transgenic mouse chromosome. Permanent expression of the hybrid enzyme was identified by Western blot. The catalytic activity of the hybrid enzyme to form PGI2 was determined by enzyme activity assay and LC/MS analysis of transgenic mouse tail and urine, respectively. In conclusion, a transgenic mouse overexpressing the hybrid enzyme with upregulated PGI2 biosynthesis was successfully obtained. The mice can be used to further uncover the mechanisms of the vascular protection of PGI2 and test the side effects of NSAIDs.

Funding: R01 HL56712 (for KHR); RO1HL079389 (for KHR) and RC1 HL100807 (for RD and KHR)
Transdermal dl‐methylphenidate potentiates the pharmacokinetic interaction with ethanol relative to oral dosing in a mouse model
Authors: Bell Guinevere Hannah , Novak Andrew , Griffin William , Patrick Kennerly
Published: June 2021

Objective

Test the hypothesis that transdermal dl‐methylphenidate (MPH) potentiates the pharmacokinetic interaction with ethanol relative to oral MPH dosing; thus resulting in a greater elevation of d‐MPH and the metabolite l‐ethylphenidate (EPH) in blood, brain and urine.

Methods

A novel murine model of transdermal MPH was developed using ¼ of a 12.5cm2 MPH patch (Daytrana®) with/without ethanol (3 g/kg po). Used MPH patches were analyzed to determine the average dose delivered (7.5 mg/kg) and this dose was used for oral MPH‐ethanol comparisons. MPH and EPH were enantioselectively quantified by GC‐MS of prolyl derivatized piperidyl fragments.

Results

The concentration of d‐MPH was significantly elevated by ethanol, especially following transdermal dosing. Approximately 10X more l‐MPH reached the blood with transdermal delivery vs. oral. In the ethanol‐transdermal MPH group, the concentration of l‐EPH was significantly higher than d‐EPH.

Conclusions

The greater ethanol‐induced elevation of the “active” d‐MPH isomer when using transdermal vs. oral dosing appears to be mediated by the higher circulating levels of l‐MPH which competitively inhibit carboxylesterase 1. These findings have implications for abuse liability and cardiovascular toxicity of concomitant transdermal MPH and ethanol should this animal model hold for ADHD patients who consume ethanol. Supported by RO1AA16707.
INVOLVEMENT OF PRELIMBIC BUT NOT INFRALIMBIC CORTEX ON THE PRESSOR RESPONSE TO CHEMOREFLEX ACTIVATION IN UNANESTHETIZED RATS
Authors: Granjeiro Érica Maria , Scopinho América , de Aguiar Corrêa Fernando Morgan , Resstel Leonardo Barbosa
Published: June 2021

INTRODUCTION AND OBJECTIVES

The ventral portion medial prefrontal cortex (vMPFC) comprises the prelimbic cortex (PL) and the infralimbic cortex (IL). It has been reported an important role of vMPFC on cardiovascular reflexes modulation. However, there is no study showing the role of this area on cardiovascular responses to chemoreflex activation. We report here a comparison between the effects of pharmacological inhibition of PL or IL neurotransmission on the pressor and bradycardic responses to chemoreflex activation evoked by KCN, i.v. administration.

RESULTS AND CONCLUSION

Bilateral microinjections of 200 nl of the unspecific synaptic blocker CoCl(2) (1 mM) into the PL significantly reduced the pressor (60 ± 3 vs 44 ± 3 mmHg, n=9, p<0.01) but had not effect on the magnitude of bradycardic response to chemoreflex activation (−219 ± 24 vs −201 ± 16 bpm, n=9, p>0.05). However, when local synapses in the IL were inhibited by bilateral injections of CoCl(2) into that area, no significant changes were observed on cardiovascular responses evoked by chemoreflex (n=6, p>0.05). These data indicate the involvement of the vMPFC PL area in modulation of the pressor response induced by chemoreflex activation but not in the decrease in heart rate evoked by this reflex. They also suggest that the neurons located on PL area do not play a major role in the neurotransmission of the autonomic responses to chemoreflex activation.
Vasoconstriction‐induced inward remodeling of isolated arterioles involves reactive oxygen species‐dependent activation of matrix metalloproteinases
Authors: Martinez‐Lemus Luis A. , Zhao Guiling
Published: June 2021

We previously demonstrated that inward remodeling induced by prolonged agonist‐induced vasoconstriction in isolated arterioles requires reactive oxygen species (ROS) generation and activation of matrix metalloproteinases (MMPs). We hypothesize that ROS participate in the vasoconstriction‐induce inward remodeling process in part by promoting MMP activation. This hypothesis was tested in isolated and pressurized (60 mmHg) rat cremaster arterioles exposed to norepinephrine (NE, 10−5.5 M) plus angiotensin‐II (Ang‐II, 10−7 M) for 4 hours. Prolonged vasoconstriction induced inward remodeling and increased the gelatinolytic activity of arterioles (P<0.05). ROS‐inhibition with apocynin, tempol, or the Rac‐inhibitor, NSC, significantly affected the development of inward remodeling and reduced the arteriolar gelatinolytic activity induced by NE and Ang‐II. These results indicate that during prolonged vasoconstriction MMPs are in part activated through a pathway dependent on the generation of ROS, and that ROS are required for the acute remodeling of the vascular wall. As MMP‐inhibition prevents the development of inward remodeling, these results suggest that vasoconstriction‐induced inward remodeling in resistance vessels is associated with ROS‐dependent activation of MMPs.

Funded by AHA and NIH.
The Effect of Chronic Kidney Disease on Endothelial‐Dependent Dilation and Progenitor Cells
Authors: Kuczmarski James Matthew , Darocki Mark , Sikes Robert A. , Cooper Carlton R. , Farquhar William B. , Edwards David G.
Published: June 2021

The purpose of this study was to determine if circulating hematopoietic and myeloid progenitor cell subpopulations are reduced, colony forming ability (CFA) of peripheral blood mononuclear cells (PBMNC) is impaired, and endothelial‐dependent dilation (EDD) is attenuated in chronic kidney disease (CKD) patients as compared to healthy controls. Ten patients with stage 3–5 CKD and ten healthy controls were studied. Brachial artery flow‐mediated dilation was used to assess EDD. Circulating progenitor cells (CPC) were quantified in venous blood samples via flow cytometry and PBMNC were cultured to assess CFA. EDD was reduced in CKD (5.39 ± 1 vs. 10.12 ± 1.4; p = 0.007) as were hematopoietic CPC subpopulations CD34+/KDR+ (276.9 ± 65.9 vs. 475.2 ± 93.2, p = 0.046), CD34+/CD45‐ (131.8 ± 30.4 vs. 428.4 ± 100.7, p = 0.004), and CD34+/KDR+/CD45‐ (107 ± 2.8 vs. 298.5 ± 41.2, p < 0.001). Furthermore, myeloid CPCs CD45+ (9.9 × 104 ± 1.3 × 104vs. 1.32 × 105 ± 1.6 × 104, p = 0.07) and CD45+/KDR+ (1.1 × 104 ± 5.9 × 103vs. 4.1 × 104 ± 1.8 × 104, p = 0.07) were lower and CFA (2.64± .502 vs. 4.3 ± .953, p = 0.065) was impaired, approaching significance. Reductions in CPC populations may be a potential mechanism by which vascular integrity is compromised, leading to endothelial dysfunction, ultimately increasing the risk of cardiovascular disease and contributing to renal disease progression in CKD patients.

Supported by Grants DK077306 and P20 RR016472‐09.
The novel non‐depolarizing, normokalemic cardioplegia formulation based on the combination of adenosine‐lidocaine (Adenocaine™) exerts superior anti‐neutrophil effects by synergistic actions of its components
Authors: SHI WEIWEI , JIANG RONG , Dobson Geoffrey P , Vinten‐Johansen Jakob
Published: June 2021

Objectives

We tested the hypothesis that the combination of adenosine and lidocaine (AL) synergistically attenuates neutrophil (PMN) functions.

Methods

1. Superoxide anion (O2‐) generation, 2. PMN adherence, 3. Transmigration. Group differences (one‐way ANOVA) were accepted at P value<0.05.

Results

The isolated porcine PMNs released superoxide dismutase (0.1mg/ml)‐sensitive O2‐ in a burst pattern upon activation. Both adenosine and lidocaine alone inhibited O2‐ production in a dose‐dependent pattern. AL further inhibited O2‐ generation in a synergistic manner. In addition, adenosine (0.1–10 μM) inhibited O2‐ generation by cytochalasin B (priming), whereas lidocaine (1–100 μM) inhibited O2‐ release stimulated by both cytochalasin B and fMLP (activation). Both adenosine and lidocaine alone inhibited PAF‐induced PMN adherence to endothelial cells. Adenosine (50μM) and lidocaine (100 μM) decreased adherence to 71.6±5.4% (n=8, P<0.05) and 73.2±8.1% (n=7, P<0.05) of stimulated control, respectively. AL further suppressed adherence to 47.2±4.3% (n=7, P<0.05). Adenosine (100μM) reduced PMN transendothelial migration to 80.1±6.7% of control (n=6, P<0.05), whereas lidocaine (100μM) showed no significant inhibition. AL did not further decrease transmigration.

Conclusions

AL suppresses PMN O2‐ generation and adherence in a synergistic manner relative to adenosine or lidocaine alone.
Glucose‐6‐Phosphate Dehydrogenase Interacts with Cav1.2 and Regulates [Ca2+]i in Coronary Artery Smooth Muscle
Authors: Gupte Rakhee S , Kurebayashi Naomi , Gupte Sachin A
Published: June 2021

G6PD plays a role in promoting contraction of coronary artery (CA); however, the molecular mechanism underlying this observation is still elusive. We have investigated the mechanism through which G6PD may regulate [Ca2+]i in CA. We performed Western blot analysis of different cell fractions of CA and found that G6PD and Cav1.2 co‐eluted with caveolin 1, suggesting that G6PD and Cav1.2 co‐localized in the caveolae fraction. Hence, we hypothesized that G6PD may be involved in regulating [Ca2+]i by interacting with Cav1.2. We performed co‐immunoprecipitation experiments and found that G6PD interacted with Cav1.2 in resting coronary artery smooth muscle cells (CASMC) and CA. Interestingly, stimulation of CASMC and CA with KCl (30 mM), U46619 (100 nM) or PDBu (10 μM) increased G6PD‐Cav1.2 complex. Furthermore, we co‐expressed human cardiac GFP‐fused‐Cav1.2 and RFP‐fused‐G6PD in HEK 293T17 cells. FRET analysis confirmed their interaction in vivo with an efficiency of 0.47±0.05. To determine the implication of their interaction on CA function, we silenced G6PD in CA by siRNA resulting in decreased KCl‐induced increases in [Ca2+]i and contraction of CA. Moreover, ectopic expression of G6PD in CA enhanced KCl (30 mM)‐evoked increases in [Ca2+]i and contraction by 2‐ or 3‐fold as compared to control CA. These results, therefore, provide novel findings that G6PD interacts with Cav1.2, and suggest that G6PD & NADPH redox promotes CA contraction by elevating [Ca2+]i, at least partly, through regulating L‐type Ca2+ channel function. (Support by NIH grant HL085352)
In vitro and in vivo analysis of macelignan induced toxicity on heart
Authors: Javaregowda Palaksha Kanive , Hong Yunkyung , Kim Sunmi , Hong Yonggeun
Published: June 2021

Macelignan which isolated from Myristica fragrans Houtt has been suggested that it acts as PPAR α/γ dual agonist for diabetic patients. However, macelignan contains a small amount of safrole considering as carcinogen. Here we investigated that its toxic effect on heart in vitro and in vivo. The systolic blood pressure, pulse and the component of blood serum in spontaneously hypertensive rats (SHR) and control rats were measured. The expression of cell death markers were analyzed by RT‐PCR and immunoblot. We confirmed that PPAR α/γ was expressed on H9c2 cells, neonatal cardiomyocytes and heart. Compared to control, macelignan significantly reduced the beating number and frequency as a concentration dependent manner (p<.05). Cell viability was strikingly decreased at 16 h after treatment. And the expression of autophagic markers as well as apoptotic molecules was maximized at 16 h (p<.05). A significant reduction of the systolic blood pressure was shown in both strains, and pulse was also strikingly decreased after its injection (p<.05). However, the component of serum was not changed by macelignan. Here, we found a cytotoxicity induced by macelignan in vitro and in vivo. Therefore, the use of macelignan as an anti‐cancer or anti‐inflammatory agent should be carefully considered.

Funding: BioGreen21 Program (Code No. 20070401‐034‐006‐009‐02‐00), Rural Development Administration, Korea
IRF2BP2 is a new NFAT1 partner acting as a repressor protein
Authors: Carneiro Flavia Raquel Gonçalves , Ramalho‐Oliveira Renata , Mognol Giuliana P. , de Araujo‐Souza Patrícia S. , Viola João P.B.
Published: June 2021

NFAT family of transcription factors encodes 5 members and is present in a wide range of cell types, regulating genes involved in cell proliferation, angiogenesis, cell cycle progression and apoptosis. NFAT family has two well conserved regions: the regulatory domain and DNA binding domain. N and C‐terminal ends are transactivation sites and show less sequence similarity. To understand the molecular mechanisms related to NFAT functions we performed a yeast two hybrid screening using the less conserved NFAT1 C‐terminal end as bait. We identified that NFAT1 interacts with IRF2BP2 transcriptional repressor by its RING domain. This interaction seems to be NFAT1 specific, since the other conserved regions of NFAT1 do not interact with IRF2BP2. Furthermore, NFAT1 and IRF2BP2 co‐localize in the nucleus in activated cells and luciferase report assays demonstrate that IRF2BP2 represses the NFAT1‐dependent activation of IL‐2 and IL‐4 promoters and TNF‐alfa k3 element. We also identified a nuclear localization signal in IRF2BP2 and mutation of this signal abolishes the repression function of IRF2BP2. Our data demonstrated that IRF2BP2 interacts with NFAT1 and repress its transcriptional activity, suggesting that IRF2BP2 can be a new regulator of NFAT1 transcriptional activity.

Financial support: CNPq, FAPERJ, ICGEB
Circulating hepcidin at term pregnancy and in cord blood independently reflects maternal and fetal iron status
Authors: Westerman Mark , Ervasti Mari , Punnonen Kari , Olbina Gordana , Ostland Vaughn , Luukkonen Susanna , Heinonen Seppo , Pulkki Kari , Sankilampi Ulla
Published: June 2021

Background

The aim of this study was to determine serum hepcidin concentrations and its associations with iron status in pregnant women and cord blood. 191 full‐term pregnant women and newborns were studied. Blood from pregnant women was obtained within 24h of delivery, and venous cord blood shortly after birth. Serum hepcidin was measured by C‐ELISA and compared to hematological and iron indices, including ferritin, TfR, %TSat, hsCRP, and EPO.

Results

Hepcidin in pregnant women was significantly lower than in the cord blood [GMC; 13.5 vs. 72.9 ng/mL, respectively]. Serum hepcidin was <5 ng/ml in 12% of pregnant women and in 2% of newborns. Hepcidin was significantly higher in vaginal and emergency section deliveries, compared to elective Cesarean. In pregnant women at term, 7–8% were iron deficient (Hb<110 g/L or %HYPOm >3.4%). Hepcidin was correlated with hsCRP and gestational weeks. 20% of newborns had Hb <145 g/L and 5% had TSat <30%, and were classified as iron deficient. Reference ranges for serum hepcidin at term pregnancy was 1.4 – 70.6 ng/mL (n=126; median 11.9 ng/mL) and for cord blood, 20.1 – 231.2 ng/mL (n=142; median 77.45 ng/mL).

Conclusions

Maternal hepcidin levels are suppressed at term facilitating iron transfer to the fetus, while newborn hepcidin levels are normal for the existing iron stores. Research funded by a grant to Kupio University Hospital and by Intrinsic LifeSciences.
YB‐1 mediates fetal gene reprogramming during cardiac stress
Authors: Strauch Arthur Roger , Willis William L. , David Jason J.
Published: June 2021

Fetal smooth muscle alpha‐actin (SMA) is re‐expressed in adult cardiomyocytes in response to biomechanical stress. Transglutaminase 2 (TG2) is associated with myocardial fibrosis and catalyzes gamma‐glutamyl‐peptide linkages via a transamidation reaction in response to reactive oxygen. TG2 was expressed in ventricular cardiomyocytes after ischemia/reperfusion (I/R) injury. Amino‐ and carboxyl‐terminal sequences in the mRNA‐binding, stress granule protein YB‐1 are homologous to TG2 consensus crosslinking motifs. Myocytes exposed to a biotin‐pentylamine TG2 substrate formed biotin‐tagged 50 and 100 kDa YB‐1 variants in the presence of TG2‐activating hydrogen peroxide. Studies using recombinant YB‐1 (rYB‐1) as a TG2 substrate revealed substantial formation of YB‐1 oligomers in the range of 150–250 kDa which varied as a function of rYB‐1 concentration and enhanced in the presence of SMA mRNA. Endomyocardial biopsies collected from patients more than 9 years after heart transplant showed elevated expression of both fetal SMA and low (50 kDa) and high (180 kDa) molecular weight YB‐1:SMA mRNA complexes. YB‐1 deposition also was noted at cardiac intercalated discs after I/R injury. We conclude that TG2‐mediated transamidation of YB‐1 monomers may be potentially important for regulating fetal gene mRNA stability and translational efficiency in the stress‐injured heart. Supported by NIH HL085109.
Altered calcineurin‐NFAT signaling in the hearts of NFATc2 null mice
Authors: Sin‐Chan Patrick , Michel Robin N.
Published: June 2021

The Ca2+‐ dependent phosphatase, calcineurin, and its downstream transcriptional effectors, nuclear factor of activated T‐cells (NFAT), are major intracellular modulators of cardiac hypertrophy. Transcription factor NFATc2 has recently been identified as the major NFAT isoform responsible for calcineurin‐mediated cardiac hypertrophy. We thus investigated the role of each NFAT isoform (c1–c4), the NFATc2 binding partner GATA‐4, and NFAT/GATA‐4 kinases in regulating the cardiac phenotype by assessing their expression at the transcript and protein levels in NFATc2 −/− mice. Cardiac mRNA and protein expressions of other members of the NFAT family were similar in wild type and NFATc2−/− mice, suggesting that individual NFAT isoforms are independently regulated. Expression of cytoplasmic GATA‐4 was unchanged whereas nuclear GATA‐4 was higher in hearts from NFATc2 −/− mice, indicating increased nuclear transit of this transcription factor in the absence of NFATc2. In support of this, the expression of MAPK, responsible for GATA‐4 DNA binding ability, remained unchanged whereas GSK3‐β, which promotes GATA‐4 nuclear export, was lower in NFATc2 −/− hearts. Taken together, our data identify molecular partners that may compensate for the lack of NFATc2, thereby providing further insight towards understanding calcineurin‐mediated cardiomyopathy. Supported by CIHR, NSERC and CRC to RNM.
THE VENOUS DRAINAGE OF THE CAUDATE LOBE. STUDY ON CORROSION CASTS
Authors: MATUSZ PETRU L , PUSZTAI Agneta Maria , HORDOVAN Eniko‐Christine , PURECA Ana , DAESCU Ecaterina , PRILIPCEANU Georgeana
Published: June 2021

The group of small hepatic veins form the lower group of hepatic veins ‐ participating in the venous drainage (VD) of the caudate lobe (CL) ‐ and the lower portions of right and left hepatic lobes. VDCL was examined by a number of 150 hepatic corrosion casts; the pieces were made by injecting plastic mass in the vascularductal elements of the liver, followed by corrosion of the parenchyma with hydrochloric acid. The number of the CL veins varies from four to fifteen (average of 8.77 veins/piece). The analysis of VDCL depending collector hepatic veins (HV) demonstrates that: the left HV participates in VDCL 17.33% by the tail vein 1.3 (mean 1.80 veins/piece); the middle HV participates in VDCL 100% through the tail vein 4.8 (mean 5.2 veins/piece); the common trunk formed from the middle and left HV participates in the VDCL as a percentage of 11.11%; the right superior HV participates in VDCL as a percentage of 62.67% through the tail vein 2.4 (mean 2.75 veins/piece); the right inferior HV participates in VDCL as a percentage of 12% CL by the tail vein 1.3 (mean 2.22 veins/piece); the Inferior Vena Cava (IVC) venous drainage involved in the CL in 100% of cases, 1.4 veins (veins averaging 1.77 veine/piece). CL main venous drainage is done mainly to the Middle Hepatic Vein (98.46% of the parts) and to the retrohepatic portion of the IVC (84.09%) (Supported by CNMP 41092/2007).
Increased dopaminergic tone in the hippocampus by repeated fasting/refeeding in adolescent rats that experienced neonatal maternal separation
Authors: Jahng Jeong Won , Yoo Sang Bae , Kim Jin Young , Lee Jong‐Ho
Published: June 2021

We have reported that neonatal maternal separation (MS) leads to the development of anxiety/depression‐like behaviors with altered activity of the hypothalamic‐pituitary‐adrenal (HPA) axis in offspring. Adolescent MS rats showed sustained hyperphagia with increased HPA activity during repeated fasting/refeeding (RF) cycles. In this study, SD pups were separated from dam for 3 h daily during PND 2‐14 (MS) or left undisturbed (NH). Half of MS pups were deprived from food every other day, otherwise had free access to food from PND 28 (MS/RF). NH/C or MS/C had free access to chow throughout the experimental period. Rats were subjected to behavioral sessions or sacrificed on PND 40 after 6th refeeding day, brain tissues were collected for dopamine analysis by HPLC. The hippocampal contents of dopamine increased significantly in MS/RF, compared to NH/C or MS/C, and the striatal dopamine to NH/C. Metabolic rate of dopamine to DOPAC increased in hippocampus, but decreased in striatum, of MS/RF compared to NH/C. Metabolic rate to HVA did not change in hippocampus, but decreased in striatum, of MS/RF compared to NH/C. MS/RF showed increased ambulatory activity and anxiety‐like behavior compared to NH/C. Results reveal that RF cycles increases dopaminergic tones in the hippocampus and striatum of adolescent MS rats, via increasing dopamine neurotransmission and degradation at pre‐synaptic level and decreasing degradation both at pre‐ and post‐synaptic levels, respectively. Together with our previous reports, it is suggested that increased dopaminergic tones in the hippocampus and striatum may mediate the increased HPA axis activity, ambulatory activity and anxiety‐like behavior, perhaps, leading to a sustained hyperphagia during repeated fasting/refeeding cyles in adolescent MS rats.
Neuroprotective effects of treadmill exercise on brain damage in spontaneously hypertensive rats with focal cerebral ischemia
Authors: Shin Jinhee , Lee Seunghoon , Park Sookyoung , Chang Kyu‐tae , Hong Yonggeun
Published: June 2021

To determine the neuroprotective effects of treadmill exercise, normotensive Wistar Kyoto (WKY) rats and SHR were subjected to middle cerebral artery occlusion‐reperfusion (MCAo), and then subjected to treadmill exercise for 4‐weeks. In gait analysis, the stride length of the hindlimb increased continuously up to 72 h due to motor dysfunction. Brain tissues from the rats were stained with TTC to estimate the infarct volume. The infarct volume of the SHR was increased significantly following MCAo, as compared to that for the WKY rats (p<.05), which suggests that hypertension is a risk factor for cerebral vascular disorders caused by FCI. Moreover, the gene expression of BDNF, Trk, and nNOS were decreased, whereas iNOS gene expression was increased in MCAo‐brain tissue (p<.05). Also, the expression of BDNF, Trk, and nNOS was increased, whereas the iNOS level was decreased in the MCAo+exercise group (p<.05). These results suggest that MCAo is a strong causative factor for impairment of the brain, and that physical therapeutic intervention is beneficial for the functional rescue of neuromuscular communication in cases of brain damage, including those caused by FCI.

Funding: BioGreen21 Program (Code No. 20070401‐034‐006‐009‐02‐00), Rural Development Administration, Korea
Oral bezafibrates induce long and spontaneous torpor bouts in mice
Authors: Chu Lisa Pei , Swoap Steven John
Published: June 2021

FGF21 is a hormone released from the liver into the blood that mediates many of the physiological responses of fasting, such as lipolysis and ketogenesis. FGF21 is activated by the nuclear receptor PPARα when bound to its endogenous agonist, circulating free fatty acids, or to the synthetic agonist, bezafibrate. This study tested whether PPARα agonists enhance the metabolic suppression and accompanying fall in body temperature that occurs in mice in response to fasting. C57BL/6J mice were implanted with temperature telemeters, fed a normal diet for 2 weeks, fasted for 23 hours for a baseline torpor response and then randomly assigned to either a 1) ketogenic diet, to elevate circulating free fatty acids or 2) diet containing 0.5% bezafibrate for 2 weeks. Those fed the bezafibrate experimental diet entered spontaneous torpor bouts without caloric restriction towards the end of the two weeks, whereas those fed the KD diet showed no spontaneous torpor bouts. Moreover, mice on the bezafibrate diet had longer bouts of fasting‐induced torpor than either their pre‐diet status or when fed the ketogenic diet (403±71 minutes, 129±34 minutes, 90±35 mintues, respectively). Thus collectively, these findings suggest that while the ketogenic diet did not change torpor behavior, bezafibrate makes it more likely to for mice to enter torpor which may involve the PPARα pathway,
Effect of Treatment with Peptidase Inhibitors on Vasoactive Peptide Response in Renal Arteries from Diabetic Rats
Authors: Oltman Christine L. , Kleinschmidt Travis L. , Coppey Lawrence J. , Davidson Eric P. , Yorek Mark A.
Published: June 2021

The purpose of this study was to examine the effect of peptidase inhibitors on peptide induced relaxation of renal arteries from diabetic rats. Rats were injected with 60 mg/kg streptozotocin (IP), and treated with α‐lipoic acid (2.5g/kg), Enalapril (500mg/kg), Candoxatril (300mg/kg), AVE7688 (500mg/kg), or Alogliptin (200mg/kg) in the diet for 12 weeks. Tension studies were performed on tertiary branches of renal arteries Acetylcholine and sodium nitroprusside responses were not altered in phenylephrine constricted arteries. Atrial natruretic peptide (ANP) and calcitonin gene related peptide (CGRP) responses were attenuated in arteries from diabetic rats. Responses were improved when rats received the peptidase inhibitors, but not when rats were treated with α‐lipoic acid. Thus, responses to vasoactive peptides are reduced in diabetic rats, and treatment with peptidase inhibitors, improves dilation to these peptides. Supported by VA merits to Drs. Oltman and Yorek.

Blood glucose and response to ANP and CGRP (% Relaxation)

Group
n
Glucose (mg/dl)
ANP(10nM)
CGRP(10nM)

Control
11
110 ± 5
97.7 ± 1.8
79.4 ± 5.5

Diabetic
12
578±13*
77.1 ± 4.9*
47.1 ± 5.9*

α‐lipoic acid
8
501 ± 33
79. 6 ± 4.9
54.1 ± 6.9

Enalapril
6
570 ± 22
94.8 ± 3.7#
87.5 ± 4.6#

Candoxatril
8
546 ± 14
96.7 ± 1.7#
80.4 ± 4.1#

Alogliptin
7
536 ± 22
96.2 ± 2.1#
72.3 ± 8.8#

AVE7688
7
575 ± 22
94.8 ± 3.3#
77.1 ± 5.9#

Control vs Diabetic P ≤ 0.05

Diabetic vs treatment, p ≤ 0.05.
Gene networking and canonical pathway analysis in a JMJD3 knockdown human monocytic cell line
Authors: Das Nando Dulal , Jung Kyoung Hwa , Choi Mi Ran , Chai Young Gyu
Published: June 2021

JMJD3, a JmjC family histone demethylase, can be induced by the transcription factor NF‐kB in response to various stimuli. JMJD3 is crucial for erasing H3K27me3, a histone modification associated with transcriptional repression, and is responsible for the activation of a diverse set of genes. The objectives of this study were to identify the genes in THP‐1 human monocytic cells that are significantly affected by the knockdown of JMJD3, particularly in expression levels and in downstream effects on signaling pathways. THP‐1 cells were subjected to lentiviral JMJD3 shRNA transduction followed by analysis of gene expression by qRT‐PCR. Global gene expression levels were detected in JMJD3 knockdown (kd) THP‐1 human monocytic cells and in TNF‐stimulated JMJD3‐kd THP‐1 cells using a 44‐k human whole genome array. Finally, datasets were analyzed using Ingenuity pathway analysis (IPA) software. IPA analysis reveals that JMJD3 attenuation down‐regulates various key genes in NF‐kB, chemokine and CD40 signaling and mostly affects inflammatory disease response molecules. Moreover, this study significantly highlights the connection of NF‐kB with JMJD3, which suggests an epigenetic regulation of the inflammatory genes.

Funded by NRF‐2009‐0077023; Korea.
Effect of exercise training on anaphylaxis in perfused rat liver and in conscious rat
Authors: Shibamoto Toshishige
Published: June 2021

Hepatic venoconstriction is involved in rat anaphylactic hypotension. We determined effects of exercise training (ET; four week treadmill running) on both anaphylaxis‐induced segmental venoconstriction in perfused rat livers and systemic anaphylaxis in conscious rats sensitized with the antigen of ovalbumin. Roles of nitric oxide (NO) in the effect of ET on venoconstriction of perfused livers were also examined. In the isolated livers perfused portally with blood, the portal venous pressure and the sinusoidal pressure were measured to determine the pre‐ (Rpre) and post‐sinusoidal (Rpost) resistances. In conscious rats, systemic arterial pressure and portal venous pressure were measured. Antigen caused 4.6‐ and 1.7‐fold increases in Rpre and Rpost, respectively, in the perfused livers of sedentary rats. Anaphylaxis‐induced increase in Rpre was significantly attenuated by 24% by ET. Inhibition of NO with L‐NAME (100 μM) enhanced anaphylactic venoconstriction, but not alter the effect of ET on the Rpre increase. In contrast, ET attenuated neither anaphylactic hypotension nor portal hypertension in conscious rats. In conclusion, ET attenuated the anaphylaxis‐induced presinusoidal constriction in perfused rat livers, independently of NO production. However, this action was not evident in conscious rats and ET did not inhibit anaphylactic hypotension in conscious rats.