bluuebunny/crossref_metadata_2025 huggingface dataset:
The role of central memory CD8 T cells in the kidney and the role of these cells in genesis of hypertension
Authors: Zhang Feng , Xiao Liang , Harrison David G
Published: June 2021

Immunological memory is an important function of the adaptive immunity in response to pathogenic stimuli. Effector memory T cells, which are CD44 high CD62L low, reside in various non‐lymphoid tissues and serve as a first line defense against foreign antigens. We have previously shown that T cells are important in hypertension, but the role of memory T cells has not been defined. Here, we show that hypertension also causes immunological memory. The presences of CD8+/ CD62Lhi/CC44hi, CCR7hi central memory‐like cells in the kidney are increased 5 to 10‐fold compared to sham infused mice. Moreover, the central memory CD8 T cells produce IFN‐r and up‐regulate IL‐18R. To examine a potential role of these cells in hypertension, we re‐infused a low dose of angiotensin II that minimally raises blood pressure in naïve mice (200 ng/kg/min) 4 weeks after stopping an initial infusion of either high dose angiotensin II or vehicle. This low‐dose angiotensin II increased blood pressure to 137±6mmHg in the animals that had previously received a vehicle infusion, but to 172±12mmHg in mice that had received angiotensin II. Thus, these studies show that hypertension is associated with the development of central memory CD8+ T cells and that these cells may promote a severe hypertensive response to subsequent exposure to angiotensin II. This memory model allows us to investigate the role of memory CD8 T cells in genesis of hypertension.
Estrogen Downregulates NaPi‐IIa in the Kidney Through the Activation of Both Estrogen Receptors ER alpha and ER beta
Authors: Amlal Hassane , Burris Dara , Matu Joseph , Levi Moshe , Siddiqui Farraz
Published: June 2021

In these studies, we examined the effect of estrogen (EST) on phosphate transporters in the kidney and determined the respective role of estrogen receptors (ER) isoforms (ERα and ERβ) in this effect. We first demonstrated by RT‐PCR that both ERα and ERβ are expressed in the proximal tubule (PT) cells. Next, OVX rats were injected with EST or its vehicle for 3 days and the expression of Pi transporters in the PT was examined in the kidney cortex. Molecular studies demonstrated that NaPi‐IIa mRNA and protein were significantly reduced by EST in a dose‐dependent manner, whereas the expression levels of NaPi‐IIc and Pit2 were unchanged. Using specific agonists of either ERα (4,4′,4″‐(4‐ Propyl‐[1H]‐pyrazole‐1,3,5‐triyl)trisphenol or PPT) or ERβ (2,3‐ bis(4‐Hydroxyphenyl)‐propionitrile or DPN) or both (PPT+DPN), we demonstrated that food intake was decreased in PPT‐ and PPT+DPN− but not in DPN‐treated rats. PPT and DPN alone did not affect NaPi‐IIa protein abundance and did not cause phosphaturia despite a decrease in the expression of its mRNA levels. However, PPT+DPN caused a sharp downregulation of NaPi‐IIa along with significant urinary Pi wasting. These studies demonstrate that the phosphaturic effect of EST is mediated specifically through downregulation of NaPi‐IIa and that this effect is mediated via a complex mechanism involving the activation of both ERα and ERβ isoforms. NIH/NIDDK 5RO1DK83582
Stretch‐sensitive microRNA expression and signaling events in vascular smooth muscle
Authors: Turczynska Karolina , Swärd Karl , Hellstrand Per , Albinsson Sebastian
Published: June 2021

Vascular smooth muscle cells are constantly exposed to mechanical force by the blood pressure, which is thought to regulate smooth muscle growth, differentiation and contractile function. We have previously shown that expression of small non‐coding RNAs, microRNAs (miRNAs), is essential for regulation of smooth muscle phenotype including stretch‐dependent contractile differentiation.

In this study we have investigated the effect of mechanical stretch on miRNA expression and the role of stretch‐sensitive miRNAs for intracellular signaling in smooth muscle.

Mechanical stretch was applied to mouse portal veins in organ culture ex vivo by attaching a gold weight at one end of the vessel for 24h. MiScript whole genome miRNA PCR arrays analysis, comparing miRNA levels in stretched versus non‐stretched portal veins, revealed a dramatic decrease in the miR‐144/451 cluster level with no change in other active miRNA strands. Furthermore, we could confirm that these miRNAs target the AMPK signaling pathway, which correlates with an increased AMPK activation in stretched portal veins.

In conclusion, our results suggest that reduced expression of miR‐144/451 in stretched portal veins could participate in the regulation of AMPK signaling. This effect may represent a negative feedback mechanism, limiting stretch‐induced growth in vascular smooth muscle.
Unilateral renal denervation (DNx) improves autonomic balance in conscious rabbits with chronic heart failure (CHF)
Authors: Schiller Alicia , Haack Karla , Curry Pamela , Zucker Irving
Published: June 2021

A hallmark of CHF is autonomic imbalance with increased sympathetic tone. Renal DNx in CHF patient trials has proven beneficial, but the protective mechanisms remain unclear. We hypothesized that in CHF, unilateral renal DNx would improve cardiac autonomic balance. New Zealand White rabbits (n= 14) were instrumented with ventricular pacing leads to induce CHF and placed into one of four groups: non‐pace non‐DNx (Sham‐INV), non pace DNx (Sham‐DNx), pace non‐DNx (CHF‐INV) or pace DNx (CHF‐DNx). Two weeks following DNx (stripping the left renal artery of all visible fibers)/sham surgery, animals were paced at 380 BPM and CHF was confirmed by echocardiography. Heart rate (HR) and variability (HRV) were calculated from the ECG signal. CHF‐INV rabbits showed a significant decrease in HRV compared to Sham‐INV (p < .03) and this was attenuated in CHF‐DNx rabbits (7.2 ± 1.0 ms vs 13.6 ± 0.4 ms, p < .001). In CHF‐INV rabbits, bradycardia induced by IV metoprolol to assess sympathetic tone was exacerbated compared to sham (−24.0 ± 3.4 bpm vs −12.4 ± 2.4 bpm, p < .05) but was normalized in CHF‐DNx (−14.5 ± 1.4 bpm, p < .05), indicating a reduction in sympathetic tone by DNx. Conversely, the tachycardia in response to IV atropine as an assessment of vagal tone was not improved in CHFDNx vs CHF‐INV animals. These results suggest renal DNx improves cardiac autonomic balance in CHF by a reduction of cardiac sympathetic tone. Supported by PO1HL62222
The PreBötzinger Complex (preBC) Partially Mediates Opioid‐Induced Respiratory Depression in Young but not in Adult Rabbits
Authors: Stucke Astrid G , Zuperku Edward J , Prkic Ivana , Hopp Francis A , Stuth Eckehard AE
Published: June 2021

It is yet unclear which brainstem areas are involved in respiratory depression by clinical doses of opioids. This study investigated whether the effects of the intravenous mu‐opioid agonist remifentanil could be reversed by local injection of naloxone (NAL) into the preBC and whether this reversal was different in young and adult animals.

The study was approved by the local Animal Care Committee and conformed to NIH standards. 12 adult (3–4kg) and 15 young rabbits (13–23d, 110–550g) were anesthetized with 1–1.5 MAC sevoflurane, tracheotomized, ventilated and decerebrated. The location of the preBC was identified by neuronal recordings and the phrenic response to injection of D,L‐homocysteic acid. Remifentanil (0.08–0.5 mcg/kg/min) was infused intravenously until significant respiratory slowing and depression of peak phrenic activity (PPA) occurred. NAL (1mM, 70nl in young, 280nl in adults) was injected locally into the area of the preBC bilaterally. Local NAL partially reversed the depression of PPA (p=0.02) and the lengthening of expiratory time (p=0.01) in young but not in adult rabbits. Respiratory drive (PPA/inspiratory time) was unchanged. This is the first study to show that the preBC partially mediated the opioid‐induced changes in respiratory timing, though not in respiratory drive, in young rabbits but played no role in adult animals.

Supported by FAER grant MRTG‐BS‐02/15/2010‐Stucke
Processing of vestibular inputs by the medullary lateral tegmental field of conscious cats: implications for generation of motion sickness
Authors: McCall Andrew A , Moy Jennifer D , DeMayo William M , Puterbaugh Sonya R , Miller Daniel J , Catanzaro Michael F , Yates Bill
Published: June 2021

The dorsolateral reticular formation of the caudal medulla, the lateral tegmental field (LTF), participates in generating vomiting. LTF neurons exhibited complex responses to vestibular stimulation in decerebrate cats appropriate for a brain region that generates motion sickness. Since responses of brainstem neurons to vestibular stimulation can differ between decerebrate and conscious animals, the current study examined the effects of vestibular stimulation on the activity of LTF neurons in conscious felines. We examined whether LTF neurons had spatial‐temporal convergence (STC) behavior, which is due to the convergence of vestibular inputs with different spatial and temporal properties. The proportion of LTF neurons with STC behavior in conscious animals (25%) was similar to that in decerebrate cats. Far fewer neurons in other regions of the feline brainstem had STC behavior. However, responses to vertical plane vestibular stimulation were considerably different in decerebrate and conscious felines for LTF neurons lacking STC behavior. In decerebrate cats, most LTF neurons had responses similar to those of otolith organ afferents; in conscious animals, the response properties were similar to those of semicircular canal afferents. These differences show that higher centers of the brain regulate the labyrinthine inputs relayed to the LTF.
Evidence that 5‐HT neurotransmission within the nucleus tractus solitarii (NTS) differentially modulates glutamate release via 5‐HT3 and 5‐HT1A receptors
Authors: Hosford Patrick S , Ramage Andrew G , Mifflin Steve
Published: June 2021

The effects of spontaneously released 5‐HT were investigated in NTS containing slices (300 μm). Recordings were made of glutamatergic mEPSCs (abolished by CNQX) from voltage clamped (−60 mv) NTS neurones in the presence of gabazine (25 μmol) and TTX (1 μmol). Low doses (300μM; n=5) of 5‐HT inhibited (frequency decreased by 53±12%), while high doses 10μM increased mEPSC frequency (1285±276%) and amplitude (21±5 %). In presence of the 5‐HT3 antagonist granisetron (3μM) 5‐HT now only caused a decrease (31±11%) in frequency. The 5‐HT3 receptor agonist PBG (1μM) increased mEPSC frequency by 497±112% and amplitude by 18±4%, which was also blocked by granisetron. Granisetron (1μM; n=6) alone decreased mEPSC frequency by 35±4% as did the SSRI citalopram (0.5μM, n=5; 54±5%) and the OCT3 inhibitor decynium 22 (1μM; n=4; 27±2). The effects of the reuptake inhibitors were blocked by WAY‐100635 (1μM). The data indicates that the spontaneous release of 5‐HT in the NTS only activates presynaptically located 5‐HT3 and 5‐HT1A receptors. However, the ability of 5‐HT uptake blockade to only increase 5‐HT1A receptor activation suggests that the ability of spontaneously released 5‐HT to activate presynaptic 5‐HT3 receptors is not subject to modulation by reuptake. This would be consistent with these 5‐HT3 receptors being heteroreceptors. These data shows that endogenous 5‐HT has two distinct modulatory actions on glutamate release.
Myocardial AT1 gene expression is increased in a model of left ventricular hypertrophy induced by high salt intake.
Authors: Pereira Rafael Canavel , Katayama Isis Akemi , Heimann Joel Claudio
Published: June 2021

Objective

To evaluate gene expression of the renin‐angiotensin system components in a model of left ventricular hypertrophy induced by high salt intake demonstrated in a previous study.

Methods

Wistar rats were fed normal (NS1.3% NaCl) or high (HS 8%) salt diet since weaning. From the 7th week of age, two HS subgroups received hydralazine or losartan (HZ 15 and LOS 20mg/kg/day). Angiotensinogen, renin, ACE, ACE2, AT1 and AT2 gene expression in left ventricle were measured.

Results (n=8/group)

NS
HS
HS+HZ
HS+LOS

Angiotensinogen/GAPDH (IDV)
0.21±0.017a
0.19±0.017a
0.21±0.013a
0.19±0.024a

Renin/GAPDH (IDV)
0.71±0.02a
0.68±0.02a
0.67±0.02a
0.67±0.02a

ACE/GAPDH (IDV)
0.87±0.03a
0.83±0.05a
0.97±0.06a
097±0.05 a

ACE2/GAPDH (IDV)
0.41±0.02a
0.43±0.02a
0.37±0.03a
0.34±0.02a

AT1/GAPDH (IDV)
0.23±0.00a
0.46±0.06b
0.45±0.06b
0.22±0.00a

AT2/GAPDH (IDV)
0.52±0.04a
0.51±0.03a
0.45±0.03a
0.44±0.02a

Data are reported as mean±SEM. Means in row with superscripts without a common letter differ, P<0.05

Conclusion

High salt intake increases AT1 gene expression, however it is prevented by losartan treatment. Supported by FAPESP and CNPq.
AMP‐activated protein kinase enhances myogenin expression and myogenesis
Authors: Fu Xing , Zhao Junxing , Zhu Mei‐Jun , Du Min
Published: June 2021

AMP‐activated protein kinase (AMPK) is a master regulator of energy metabolism. We previously demonstrated that AMPK activity is correlated with skeletal muscle development, but the underlying mechanisms are unclear. Myogenin regulates myoblast fusion, a critical step of myogenesis. We hypothesized that AMPK regulates myogenesis through controlling myogenin expression. In C2C12 cells, AMPKα1 but not AMPKα2 knockdown by shRNA down‐regulated myogenin mRNA expression and its protein content, as well as myosin heavy chain (MHC) protein content. AMPKα1 knockout (KO) primary myoblasts but not AMPKα2 KO myoblasts showed reduced myogenin expression and reduced MHC protein content as compared to wild‐type (WT) myoblasts. AMPKα1 was found to be expressed at a much higher level than AMPKα2 in myoblasts. AMPK activation by AICAR promoted myogenin expression in WT, AMPKα1 KO, and AMPKα2 KO primary myoblasts. Histone deacetylase (HDAC) 5 knockdown by shRNA increased myogenin mRNA and protein contents, together with enhanced MHC expression and myotube formation in C2C12 cells. Mutation of two phosphorylation sites in HDAC5, Ser 259 and 498, abolished the regulatory role of AMPK on myogenin expression in both C2C12 cells and primary myoblasts. In conclusion, AMPK activity promotes myotube formation through enhancing myogenin expression, which is mainly mediated by AMPKα1 via HDAC5 phosphorylation. NIH R01HD067449
Fucosylated TLR‐4 signaling mediates microbial induction of intestinal fut2 expression
Authors: Nanthakumar Nanda N , Meng Di , Newburg David S.
Published: June 2021

Toll‐like‐receptor (TLR) binding by specific microbial pattern‐recognition molecule leads to a cascade of innate immune response by activating NF‐κB and/or IRF‐dependent inflammatory signaling pathways. Bacterial‐colonization is also known to stimulate mucosal fucosylation in the gut without activating inflammatory responses, however, it is not known whether this non‐inflammatory response is mediated by a specific TLR. The goal of this study is to determine, if a specific TLR mediates colonization‐dependent intestinal fucosylation. Induction of high fucosylated phenotype of the mature gut occurs in the wildtype mice, but not in the TLR4−/− mice. In the absence of microbiota, a novel and previously unreported form of fucosylated‐TLR4 is generated in the colonocytes. A TLR4 specific ligand, LPS, or monocolonization by a gram‐negative bacterium, Bacteroides fragilis, is sufficient to induce intestinal fucosylation without activating inflammatory responses. This novel fucosylated TLR4 is also activated by a specific α1,2‐fucosylated epitope binding ligand, UEA1, that generated a signaling pathway that is non‐inflammatory and NF‐κB and IRF3 independent. The ERK and JNK‐dependent signaling by the gut microbiota by activating this novel fucosylated TLR4 result in transcriptional induction of the secretor (fucosyltransferase 2; fut2) gene in the epithelium that leads to elevation of fut2 mRNA and FucT II activity. The resultant mucosal fucosylation creates a niche that facilitates colonization by a distinct fucose‐utilizing microbiota and this type of interkingdom communication between the microbiome and the epithelium seems to be central to establishing a protective microbial ecosystem in the gut. This may help explain of a significant association between lack of secretor gene expression and intestinal inflammatory diseases such as necrotizing enterocolitis and Crohn's disease.

Supported by HD013021, AI075563, and DK070260.
Sulforaphane protects small intestinal mucosa from indomethacin‐induced injury in mice via nrf2‐dependent mechanisms
Authors: Sato Junya , Yanaka Akinori
Published: June 2021

Background

NSAIDs cause GI mucosal injury by generating reactive oxygen species. Sulforaphane (SFN), an isothiocyanate compound rich in broccoli sprouts, up‐regulates nrf2‐dependent antioxidant enzymes. We have previously shown that SFN up‐regulates hemoxygenase‐1 (HO‐1) expression in intestinal epithelial cells in vitro, and protects small intestine from indomethacin (IND)‐induced injury in vivo. In the present study, we examined if nrf2 plays a role in the protection by SFN from IND‐induced small intestinal injury using nrf2‐deficient mice in vivo.

Methods

IND‐induced small intestinal injury was induced by injection of IND in nrf2+/+ and nrf2−/− C57/BL6 mice. Sulforaphane glucosinolate (SGS), a precursor of SFN, was orally administered before injection of IND. Mucosal injury was evaluated by lesion score, Evans blue exudation, myeloperoxidase activity, expression of TNF‐α and IL‐6. Mucosal invasion of anaerobic enterobacteria was quantitated by incubating the mucosa in Gifu anaerobic medium agar.

Results

1. IND caused small intestinal injury, effects demonstrated more in nrf2−/−, than in nrf2+/+ mice. 2. SGS attenuated the IND‐induced injury in nrf2+/+, but not in nrf2−/− mice. 3. SGS inhibited invasion of anaerobic bacteria into the mucosa in both types of mice.

Conclusions

1. Nrf2 contributes to protection of small intestinal mucosa from IND‐induced injury, 2. SFN affords protection against IND‐induced small intestinal injury, at least in part by up‐regulating nrf2‐dependent antioxidant enzymes.
What do medical students learn from the process of dissection?
Authors: Nicholson Helen Diana , Barrett Anthony , Martyn Helen
Published: June 2021

Dissection has been argued by Anatomists to be a useful tool for learning Anatomy. The aim of this study was to investigate what students perceive they learn from dissection. Medical students were invited to take part in three electronic surveys. These included both quantitative questions and free text responses and were delivered at the beginning of the programme, soon after their first experience of dissecting and during the last semester of Anatomy teaching. After nearly 2 years >; 80% of students felt that working in the dissecting room had been a positive experience and the level of stress that students experienced working in the dissecting room had decreased. Students identified the hardest parts of dissection as being the smell and cutting the face and the best parts as learning about the body and seeing pathology and evidence of previous surgery in the cadavers. Students reported that dissection had helped them develop a 3D appreciation of the body and an understanding of the importance of biological variation. When asked what they had learnt about themselves the common themes were that they had learnt to work as part of a team, come to terms with death and had increased their self‐confidence by learning to deal with difficult situations. This study provides data from the students’ perspective that dissection is a valuable experience that contributes to both their learning of Anatomy and professional development.
Advancing the Playing Field. A look at how virtual reality can change the future of anatomy teaching learning process
Authors: Duarte Jhonathan , Vilá Cristina Vanesa , Vilaró José , Veras Wilson R
Published: June 2021

Play combines all three aspects of learning: visual, auditory, and kinesthetic. Visual learners have a preference for seeing. Auditory learners’ best learn through listening. kinesthetic learners prefer to learn via experience—moving, touching, and doing. Through play, children learn a set of skills such as social skills, creativity, handto‐eye coordination, problem solving, and imagination. It is argued that these skills are better learned through play rather than through flashcards or academic drills. An element of “play” is an essential condition for perennial learning to occur in all individuals. So why should we leave this tool behind while learning medicine? Commonly, lectures are designed to favor one or maybe two types of learning styles, usually leaving gaps within medical education. Play is more than simply having a good time; it is about working out rules, understanding constraints, being absorbed by novelty, using imagination and innovating. It is also about extending, modifying and augmenting learning activities. This project brings an overt element of play to the representation of fundamental anatomical concepts and thus engages students more strongly in the learning process. The “Microsoft Kinect™ gesture recognition device” enables users to simultaneously stimulate all three learning styles at once leading to the most profound learning experience. With the Kinect you experience an immersive 3D environment, with auditory commands coupled with kinesthetic control. Theoretically providing a learning machine that can be utilized in labs, tutorials, lectures, and even at home. This project brings the element of play to the representation of fundamental anatomical concepts, and thus engages students more strongly in the learning process.
Learning by teaching: service learning in anatomy
Authors: Montante James Michael , Nazar Rami , Bee Mary
Published: June 2021

Service learning enhances a student's assimilation and integration of the course material in a way that is not possible in the classroom. Our service‐learning project involved having university students presenting anatomy material from their lecture course to urban secondary school students, and assessing the impact on both student groups. University students gained an appreciation for teaching in the urban environment and the depth of knowledge that only comes from teaching. Students in the client institutions were able to see that the material taught in their course is relevant and valued outside of their classroom. Secondary students scored significantly higher (p<0.001) on post‐tests relative to pre‐tests. University students scored significantly higher (p<.005) on exam questions related to the organ system that they presented relative to the students that did not present on that system. Comments from instructors and students were overwhelmingly positive. This service‐learning project was an effective learning strategy for both the secondary and university students. Sponsored by the Ford Motor Company through a Ford Community Corps Partnership Grant.
Understanding craniofacial variability caused by loss of Gata3 function
Authors: Eberhart Johann K. , Swartz Mary E.
Published: June 2021

Craniofacial malformations are extremely variable and we seek to understand the causes of this variability. We identified a zebrafish gata3 mutant with defects to the palatal skeleton; these defects are highly variable dependant upon the genetic background. Mutants in one inbred genetic background almost completely lose the trabeculae, a portion of the zebrafish palate. Lineage tracing of the trabeculae precursors shows that these cells become mislocalized anterior to their normal location. In a second genetic background, the trabeculae are present but the chondrocytes are not stacked out as a single row of flattened cells, as is found in wild‐type embryos. This failure of stacking results in shortened trabeculae and is consistent with a defect in cell intercalations. In order to characterize these background‐dependent phenotypic differences, we are seeking to understand the signaling pathways regulating gata3‐mediated cellular behaviors. We have found neural crest cells express gata3 and require the function of Gata3 for proper palatogenesis. Loss of Smad5 function causes loss of gata3 expression in the neural crest, consistent with the more severe mutant phenotype resembling Bmp loss of function phenotypes. We are currently examining neural crest cell movements and the expression of members of the planar cell polarity pathway in gata3 mutants in both genetic backgrounds. This work is supported by NIH R00DE018088.
Ser/Thr/Tyr (S/T/Y) phosphorylation of flagellar motility protein (FliC) affects elastase secretion in Pseudomonas aeruginosa (PA01)
Authors: Suriyanarayanan Tanujaa , Ravichandran Ayshwarya , Ishihama Yasushi , Swarup Sanjay
Published: June 2021

Pathogenic bacteria are replete with strategies to interact, colonize and establish infection in host cells, including phosphorylation/dephosphorylation of S/T/Y residues at different stages of pathogenesis. We have previously reported the S/T/Y phosphoproteome of Pseodomonas. The objective of this work is to understand the significance of FliC phosphorylation in affecting its surface‐associated phenotype. This study focuses on a phosphosite of flagellin type B (FliC) identified from a pool of 57 phosphopeptides in the PAO1 proteome. Removal of this phosphosite shows significant effects in extracellular virulence factor secretion through Type 2 Secretion System (T2SS) but not swimming motility, contrary to expectation. The FliC phosphosite mutation leads to 35–50% higher levels of elastase secretion via T2SS. Intracellular levels of the secreted proteins and the T2SS machinery levels remain unaffected by the FliC mutation, suggesting an increase in T2SS secretion efficiency. The aim of this study is to provide a link between flagellin phosphorylation and virulence factor secretion and improve our understanding of coordinated control of virulence strategies using PAO1 as a model bacterial pathogen. Funding source: Mechanobiology Institute, National University of Singapore, Singapore, SCELSE, Nanyang Technological University
Application of two‐dimensional electrophoresis for plasma of normal and diabetic rats
Authors: Braga Camila Pereira , Pozzi Carla Mariane Costa , Vieira José Cavalcante Souza , do Carmo Federici Padilha Cilene , Okoshi Marina Politi , Fernandes Ana Angélica Henrique , de Magalhães Padilha Pedro
Published: June 2021

The aim of this study was to compare plasma gels of normal and diabetic rats by two‐dimensional electrophoresis. Experimental animals were randomly divided into two groups (n = 8): G1 = control and G2 = diabetic rats (induced by intraperitoneal administration of streptozotocin in a single dose concentration of 60 mg/kg body weight). A pool of plasma samples were diluted in a specific buffer and applied to IEF (isoelectric focusing) strips with pH in the range of 4–7. Once hydrated for 12h, the strips were brought to the IEF system for running the first dimension. After this step, the strips were equilibrated in a specific buffer for 15 minutes and applied onto polyacrylamide gels at 10% w/v for running the second dimension. At the end of the separation steps (performed in duplicate), the protein spots were fixed in the gel and then stained with Coomassie Blue. The gels were scanned and analyzed using an image processor. For the image analysis, it was found that the protein spots for both experimental groups were more concentrated in the range of 24–76 kDa; 416 spots were found in G1 and 513 in G2, with a percentual matching of 53 and 43% for G1 and G2 respectively. These matching values show that there was some difference between the treatments. There is great interest in the plasma proteome as a discovery source of new biomarkers in diabetes as it is thought to contain subsets of other tissue proteomes.

Financial Support: FAPESP 2011/21672–1
Method for Improved Thermal and Chemical Stability of Carotenoids in MonaVie Juice Products
Authors: Quan Danyi , Xiong Maxwell , Pugh Mike , Talbott Shawn
Published: June 2021

BACKGROUND

Carotenoids, fat‐soluble potent antioxidants, are found in yellow, orange and red fruits and vegetables, and provide protection from free radical damage. However, formulating with carotenoids is difficult because they are susceptible to oxidation and degradation if exposed to oxygen and heat. Ensuring the stability of carotenoids in juices, is a big challenge. To combat this, an effective delivery system is required to ensure the integrity and potency.

OBJECTIVE

We used a proprietary microencapsulated carotenoid blend (natural beta‐carotene, lycopene, and lutein). The advantages include the elimination of ingredient cross‐interaction, increased heat stability, and improved shelf‐life and potency.

METHODS

The juices were placed on accelerated and long term stability studies. At each time point, the juice samples were extracted and assayed by validated extraction and HPLC methods.

RESULTS

The MonaVie Juices with the carotenoid microencapsulated powder blend has an excellent stability profile, suggesting that the carotenoid potency is maintained at least 85% at one year shelf life; compared to only 30% potency of juices containing a commercial granulated beta‐carotene over the same time period.

CONCLUSION

The results show that a microencapsulated powder blend significantly increases the thermal and chemical stabilities of carotenoids in juices, combats the oxidation of carotenoids, and help the carotenoids maintain the potency during the shelf life.
Biomarkers Reflecting Inflammation and Nutrition Determinants of Anemia (BRINDA): Project Overview
Authors: Namaste Sorrel , Suchdev Parminder , Aaron Grant
Published: June 2021

There is a need to further understand the relationship between inflammation and nutrition biomarkers and refine the relative contribution of nutritional and other risk factors for anemia across settings. The CDC, GAIN, and NICHD formed a collaborative research group called Biomarkers Reflecting Inflammation and Nutrition Determinants of Anemia (BRINDA). We identified data from nationally and regionally representative surveys that included preschool children (PSC, 6–59 mo), school‐age children (SAC, 5–14 yr) and women of reproductive age (WRA, 15–49 yr) that at minimum measured hemoglobin, inflammation (C‐reactive protein and/or alpha1‐acid glycoprotein), and iron status. Of 23 datasets identified, permission to date was received for 15. Individual country and aggregated analyses will be performed. Priority research topics generated include: 1) association among inflammation, anemia, iron, and vitamin A, as well as and approaches to account for the effect to adjust for the effects of inflammation on nutrient interpretation; and 2) risk factors and their relative contribution to anemia. BRINDA results will inform guidelines on the measurement and interpretation of anemia and micronutrients. Findings will also guide the development of a research agenda for future longitudinal studies. Research support: Bill & Melinda Gates Foundation, CDC, GAIN, NICHD
Protective effects of Fuzhuan Tea against high saturated fat diet‐induced inflammation and liver injury in Wistar rats
Authors: Tracey Ashley , Wei Yuren , Wang Dong , Magnuson Aaron , Gentile Christopher , Foster Michelle , Pagliassotti Michael , Weir Tiffany
Published: June 2021

Fuzhuan tea is a fermented Chinese tea with a unique phytochemical profile which has purported benefits for regulating lipid and glucose metabolism. The mechanism of action by which fuzhuan tea mediates beneficial effects is unknown. Our preliminary studies demonstrate that fuzhuan tea reduced inflammation and liver dysfunction in rats fed a high saturated fat diet (HSFD). Male Wistar rats consumed either control (12% kcal fat) or high saturated fat diets (45% kcal fat with 56% saturated fat), with or without fuzhuan tea supplementation (1400 mg/kg body weight per week) for 8 weeks. Animals given a high saturated fat diet had begun to show increased insulin resistance and markers of inflammation and ER stress in the liver were significantly elevated (p<0.05). In contrast, these effects were attenuated in tea‐fed rats consuming HSFD. Furthermore, HSFD resulted in elevated systemic endotoxin levels (1.7 EU/ml compared to 0.7 EU/ml in control diet fed animals) whereas no endotoxin was detected in rats supplemented with fuzhuan tea. Tea‐consuming animals showed ~4‐fold higher cecal levels of Lactobacillus spp., which modulate intestinal inflammation for improved gut barrier function. In summary, our data suggests that fuzhuan tea may provide protective effects against systemic inflammation and liver injury via effects mediated in the gastrointestinal tract. Funded by: DK072017 CAHS minigrant
Parasitic infection among young children in Mwami catchment area, Chipata District, Eastern Zambia
Authors: Mukaire Pamela E , Katuli Sozina , Ormsby Gail M , Cordero‐MacIntyre Zaida
Published: June 2021

Objective

To assess the prevalence of malaria and intestinal parasitic infections, as well as hemoglobin levels in a sample of children.

Methods

We screened for P. falciparum, Ascaris lumbricoides, helminthiasis, hookworms and hemoglobin levels from blood and stool samples collected among 138 children aged 2–7 years from five rural villages. Blood was obtained by finger prick or drawn from the arm. Blood slides were assessed using Field's Stain A&B staining and screened with an electric microscope for malaria species identification. Fifty fields were examined on each slide. Grade of malaria parasitemia was reported based on the average number of parasites seen per field. Stool was examined using direct wet mount method and the formalin‐ethyl acetate concentration method in the laboratory. Hemoglobin (Hb) levels were diagnosed using a hemoglobimeter. We used Pearson correlation to compare Hb level to malaria status.

Results

Of this study population, 52.8% tested positive for malaria; all cases were identified to be caused by P. falciparum. The mean level of hemoglobin was 10.0 ± =1.3 mg/dl. Pearson correlation comparing Hb level to malaria status was not significant (−0.068). Hookworm infection was found in less than 5%; Ascaris lumbricoides was found in only 2% of the children.

Conclusion

Over half of the children tested positive for malaria. Findings for helminthiasis were lower than expected.
NEUROPROTECTOR ROLE OF PIOGLITAZONE ON EXPERIMENTAL MODEL OF CEREBRAL MALARIA IMPROVES SURVIVAL AND REDUCES CEREBRAL LEUKOCYTE RECRUITMENT
Authors: Limborço‐Filho Marcelo , Oliveira‐Lima Onésia Cristina , Kangussu Lucas Miranda , Marins Fernanda Ribeiro , Oliveira Antônio Carlos , Carvalho‐Tavares Juliana
Published: June 2021

Pioglitazone(PIO) acts as an anti‐inflammatory drug, and therefore, we sought to investigate its role on experimental cerebral malaria(CM). PIO treated C57BL/6 mice(TM) showed protection against infection compared to non‐treated mice, as evidenced by delay in mortality and clinical improvement without altering the parasitemia. TM showed a decrease in rolling(1,6±0,5 vs 4,9±0,8 cels/min) and adhesion(3,4±0,8 vs 11,1±1,6 cels/100um) leukocytary process on pia mater endothelium(5dpi), as well a reduction on cerebral myeloperoxidase activity(1,1±0,2 vs 2,5±0,44; 1,95±0,25 vs 3,4±0,4 UR/brain 100mg; 5–6 dpi, respectively). By histopathological analysis, TM show a reduction on vascular obstruction and bleeding on brain parenchyma(6 dpi). This partial neuroprotection(5–6 dpi) was confirmed with reduction on Evan's blue dye leakage into the brain parenchyma of TM(8,32±2,08 vs 13,82±1,14; 10,61±1,29 vs 16,13±1,07 dye ug/brain 100mg), as well a reduction of brain hemorrhage(21,3±1,8 vs 31,8±1,7;26,7±2,8 vs 37,7±3,9 Hb ug/brain 100mg). This therapeutic strategy suggests that PIO is relevant to delay CM pathogenesis probably due its property of inhibiting gen transcription of proinflammatory mediators. CAPES/CNPq
An entire, isolated liver in papier mâché by Auzoux
Authors: LE FLOCH‐PRIGENT Patrice Pierre , GILLOT Jean Bernard
Published: June 2021

A complete, isolated liver in papier mâché made by Auzoux was studied. It was of natural size. Very realistic and painted with natural colors, this anatomical model was very representative of the production by Dr Auzoux during the 19th century in Paris. It can not be dated with certainty and it belonged very probably to the “real‐size human model”, an iconic collection. The numbers inscribed on the model were identified. The anatomical veracity was excellent.
Ablation of Neurogenesis in α1A‐Adrenergic Receptor Activated Mice
Authors: Goldenstein Brianna , Perez Diane , Doze Van
Published: June 2021

It is unclear what role α1A‐adrenergic receptors (ARs) have on neurogenesis, cognition and mood. The goal of this study is to examine how the α1A‐ARs relationship to neurogenesis influences cognition and mood. Mice over‐expressing the α1A‐AR showed increased cognition and neurogenesis over normal mice. We hypothesize activation of the α1A‐AR alone is not responsible for improvements in cognition and mood, but it is the receptor‐stimulated neurogenesis. To test this hypothesis, mice are treated with selective α1A‐AR agonist, cirazoline (CRZ). A cannula and osmotic pump is surgically inserted into mice with either artificial cerebral spinal fluid or anti‐mitotic agent cytosine arabinoside (AraC). Mice undergo behavioral testing (Morris water maze, novel object recognition, forced swim, tail suspension, and open field), are sacrificed, brains fixed, sectioned and stained with cell markers for immature and dividing neurons. Stereology was used to estimate cell populations. Preliminary results show AraC mice are significantly slower in solving the Morris Water Maze than AraC‐CRZ mice. The results of this study may lead to development of therapeutic strategies for neurodegenerative diseases. Support: NSF Graduate Research Fellowship Program Grant, NSF Grant EPS‐0447679, NSF CAREER Grant 0347259, NIH Grant 5P20RR017699.
Theranostic Nanoparticles for Targeting Drug Resistant Triple Negative Breast Cancer
Authors: Miller‐Kleinhenz Jasmine , Qian Weiping , Zhou Hongyu , Wang Y Andrew , Mao Hui , Yang Lily
Published: June 2021

Triple negative breast cancer (TNBC) is the most aggressive breast cancer subtype and more than 60% of TNBCs are resistant to chemotherapy. To effectively treat chemoresistant TNBC we have developed uPAR‐targeted magnetic iron oxide nanoparticles (IONPs) carrying therapeutic agents. uPAR is highly expressed in aggressive and invasive breast cancer tissues. Our goal is to determine whether uPAR–targeted theranostic IONPs selectively deliver drugs into CD44+/CD24‐cancer stem cells (CSC) and enhance therapeutic effects in those cells. Using an orthotopic primary human TNBC xenograft model derived from surgically resected drug resistant TNBC tissues in nude mice, we have demonstrated the effect of tumor growth inhibition following systemic delivery of the targeted IONPs. Histological analysis of post‐chemotherapy TNBC and primary tumor xenograft tissues with or without uPAR‐targeted IONP‐Doxorubicin (Dox) treatment shows high expression levels of CD44 and uPAR in primary TNBC tissues and human tumor xenografts. Primary TNBC xenografts treated with conventional Dox have increased levels of CD44 and uPAR. Conversely, the tumor tissues from the uPAR‐targeted IONP‐Dox treated group show decreased levels of CD44 and uPAR expression. These results support further investigation of theranostic nanoparticles as an approach to overcome drug resistance in TNBC.

Research supported by NIH grant R01 CA154129A‐01
Use of a Recombinant Fluorescent MMP‐2 as a Reporter for Tracking the Involvement of This Protease in Target Organ Damage Found in Cardiovascular Diseases
Authors: do Prado Alejandro Ferraz , Azevedo Aline , Castro Myrella Lessio , Gerlach Raquel Fernanda
Published: June 2021

Matrix Metalloproteinase 2 (MMP‐2) is elevated in hypertensive heart and vessel remodeling, being also highly expressed in organs that show function change and structural damage due to hypertension and atherosclerosis. Expression of MMP‐2 alone caused structural and functional changes in heart and kidney. It is not known yet which are the consequences of increased circulating MMP‐2 in vivo. To investigate this, we constructed a clone containing the MMP‐2 cDNA encoding MMP‐2's catalytic domain fused to the green fluorescent protein (GFP). Recombinant protein was expressed in bacteria, purified and checked for integrity and activity. Zymography, SDS‐PAGE and western blotting (WB) analyses were used for characterization. The purified protein was injected into ApoE Knockout mice (ApoE−/−) and controls (C57BL6) for four weeks. Whole organs (arteries, brain, heart, and kidneys) were excised to check for the presence of MMP‐2 according to the level of fluorescence using the IVIS spectrum imaging system. Homogenized organs levels of MMP‐2 were checked in a fluorimeter, being those further confirmed by Western Blot against the GFP protein. Results suggest spread of MMP‐2 via circulation to organs that display funtional and structural damage in cardiovascular diseases.

This study was supported by The State of Sao Paulo Research Foundation (FAPESP)
Perioperative Administration of Atorvastatin and Simvastatin has Cardioprotective Effects in Type 1 Diabetic Rats
Authors: Crespo Maria J. , Cruz Nildris , Quidgley Jose , Creagh Osmar , Torres Hector , Rivera Karines , Hernandez Cesar , Casiano Hector
Published: June 2021

Perioperative statin administration has been suggested to improve postoperative cardiovascular outcomes in patients undergoing surgery, but the optimal administration time and the best statin suited to improve postoperative outcomes in diabetics is unknown. In this study, we evaluate by echocardiography the effects of simvastatin (SV, 10 mg/kg/d), pravastatin (PV, 10 mg/kg/d), and atorvastatin (AV, 10 mg/kg/d) on the cardiovascular status of anesthetized streptozotocin (STZ)‐induced diabetic rats with compromised cardiac function. Echocardiographic measurements revealed that 24 hours after administering statins, cardiovascular parameters did not improve. In contrast, daily administration of SV and AV over a one‐week period significantly improved ejection fraction from 44% to 63% with AV, and to 65% with SV (N=9, P<0.05). The cardiac output index also was significantly improved by AV and SV. Only AV increased stroke volume from 0.50±0.08 ml to 0.83±0.13 ml (N=9, P<0.05). Unlike the other statins tested, PV provided no beneficial effects after either 24 hours or one‐week administration. Our results indicate that daily administration of atorvastatin and simvastatin for one week improves cardiac performance in fully anesthetized diabetic rats. This study has strong clinical implications for the improvement of postoperative outcomes in diabetic patients. MBRS‐RISE Grant R25‐GM061838.
Using Project‐Based Learning to Develop a Creative High School Course in Pharmacology
Authors: Eberle‐Wang Kim
Published: June 2021

At Springside Chestnut Hill Academy (SCH) we educate innovators, break‐through thinkers and problem solvers. To that end, the science department has developed “Introduction to Pharmacology” to expose students to some of the principles of drug mechanisms of action. For example, students form drug discovery teams to test the chemical properties of aspirin formulations, and to measure the antibiotic activity of natural compounds such as honey, and tea tree oil. They create films of their research using iMovie and Garageband. A field trip to Pennsylvania Hospital and to The Mutter Museum introduce the history of medicine in Philadelphia. Dose response curves are analyzed in the “Drunken Fly Lab” and the “Worm Cardiovascular Lab”. Guest pharmacologists visit to discuss their research and to mentor student research projects. The goal of this popular elective course, which is in its sixth year at SCH, is to inspire students to consider taking future courses in the fields of pharmacy, pharmacology, nursing, and medicine.
Combination of Anti Diabetic and Anti‐Obesity Drugs Prevents Tumor Progression
Authors: Bravo Claudio , Carlini Maria Jose , Saggar Neha , Sokol Michael , Puricelli Lydia Ines , De Lorenzo Mariana Silvia
Published: June 2021

Obesity causes subclinical inflammation in adipose tissue that contributes to insulin resistance and cancer progression. Chronic inflammation predisposes to different forms of cancer, and diabetes is correlated with increased risk of breast cancer. Caloric restriction (CR) reduces insulin resistance, adiposity and inflammation and the anti‐diabetic drug metformin (MET) is a CR mimetic agent. In this study, we evaluated whether the combination of MET with the anti‐obesity drug orlistat (OR) enhances the anti‐cancer properties of MET on 4T1 cells, a highly metastatic breast cancer model. Combination of MET (5mM) + OR (10μM) significantly decreased in vitro cell proliferation (p<0.01), adhesion (p<0.05) and migration (p<0.01) compared to vehicle and each drug alone. 8‐weeks old female BALB/c mice received vehicle, MET (in drinking water, 3mg/ml), OR (240mg/kg/d) or MET + OR. After 3 weeks on drugs, 4T1 cells (105) were injected into mice and drugs continued till the end of the experiment. At 30 days, the MET + OR treatment induced a 50% reduction in tumor volume (p<0.01) and a significantly lower average wet tumor weight (p<0.01). Also, MET + OR treated mice had lower total vessel length (p<0.01) and a reduced number and size of spontaneous lung metastases (p<0.01). Our results provide a new rationale and experimental basis for the use of metformin plus orlistat as cancer therapy.
EFFECTS OF EXERCISE TRAINING ON CARDIAC AND RENAL OXIDATIVE PROFILE IN AN EXPERIMENTAL MODEL OF RISK FACTORS ASSOCIATION
Authors: de Oliveira Brito Janaina , Bernardes Nathalia , Conti Filipe Fernandes , da Palma Renata Kelly , Lemos Guilherme , Irigoyen Maria Claudia , De Angelis Kátia
Published: June 2021

The aim of study was to verify the effects of exercise training (ET) on hemodynamic parameters and on cardiac and renal oxidative stress (OE) in hypertensive ovariectomized rats submitted to fructose overload. Females Wistar (n=8) and SHR (n=32) rats were divided into: normotensive (N), hypertensive (H), hypertensive ovariectomized (HO), hypertensive ovariectomized submitted to fructose overload (HOF) and trained hypertensive ovariectomized submitted to fructose overload (THOF). The ET was performed on treadmill (1h/day;5d/wk;8wks). The arterial pressure (AP) was directly recorded. The OE profile was verified by thiobarbituric acid reactive substances (TBARS) protein carbonylation (CARB) and antioxidant enzymes activities. AP was increased by association of risk factors, and was reduced by ET. Tachycardia was observed in the HFO group compared to THOF group (343±16 vs. 398±14 bpm). The hypertension increased TBARS (H:2.78±0.39;HO:4.78±0.4;HFO:3.87±0.23 vs. C:1.69±0.09 umol/mg prot) and CARB in renal tissue when compared to C group, which were attenuated by ET. The cardiac catalase and superoxide dismutase were reduced in the hypertensive groups, but the ET normalized the catalase. The superoxide dismutase activity was higher in the THOF group in renal tissue. Concluding, the ET attenuated hemodynamic dysfuctions and cardiac and renal oxidative stress caused by the association of risk factors, suggesting benefits of this non‐pharmacology approach after ovarian hormones deprivation.

Financial Support:

FAPESP (2010/17188–4; 2007/57595–5; 2011/15828–9).