bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tracing studies in rat provide sensitive gene expression data for neurons innervating the bladder and urethra
Authors: Brinkmann Hannah , Miner Wesley Dennis , McMurray Gordon , Walsh Roddy , Gardner Clare Evelyn , Ward Sari Annelie , Christy Clare 
Published: June 2021

The aim of the study was to explore novel tissue based techniques to generate gene expression profiles of both afferent (sensory) and efferent (motor) neurons that specifically innervate the rat bladder and urethra.Retrograde neuronal tracing studies in combination with laser capture microdissection (LCM) and microarray analysis were used to generate gene expression profiles of both afferent and efferent neurons that specifically innervate the rat bladder and urethra. A complete rat study (n=12) was performed in which fluorescently labelled cholera toxin B was injected into the bladder and urethra. Traced neurons were isolated by LCM, RNA was extracted, amplified and analysed by microarray.Extensive analysis of all the data derived from this study give very sensitive and powerful gene expression data from only 50–100 microdissected neurons. Expression of sensory neuron specific markers could be confirmed in dorsal root ganglion neurons whereas expression of genes involved in motor function of the lower urinary tract could be confirmed in efferent neurons e.g. Onuf's Nucleus.The resulting gene expression profile will increase our understanding of signalling pathways involved in the control of the lower urinary tract. Data derived from this study will not only be relevant for safety and efficacy but will also enable us to identify targets for the treatment of urinary frequency and urge incontinence (UFUI).
How to build a signalling organelle: A molecular pathway for biogenesis of the ciliary membrane
Authors: Nachury Maxence 
Published: June 2021

The primary cilium is a hair‐like projection present at the surface of nearly every vertebrate cell. How such a complex organelle is assembled remains an open question that has become all the more relevant with the emergence of primary cilium dysfunction as the central pathomechanism in a range of inherited disorders characterized by obesity, brain malformations, retinal degeneration, kidney cysts and skeletal abnormalities.Strikingly, all these symptoms coincide in Bardet‐Biedl Syndrome (BBS). However, despite the identification of twelve BBS genes, the molecular basis of BBS had remained elusive. Last year, the discovery of a stable complex of seven BBS proteins (the BBSome) considerably simplified the apparent molecular complexity of BBS. Most tellingly, the BBSome associates the ciliary membrane and with Rabin8, the guanosyl exchange factor for the small GTPase Rab8 and Rab8‐GTP enters the primary cilium to promote extension of the ciliary membrane.Thus, BBS is likely caused by defects in vesicular transport to the primary cilium. With this realization comes a flurry of questions that will be addressed in my presentation: What is the precise molecular activity of the BBSome and Rab8 in this process? What are the effectors of Rab8 involved in ciliogenesis? Does the ARF‐like GTPase ARL6/BBS3 also function in vesicular transport to the cilium?
Antioxidation effects of polysaccharide fucoidan extracted from seaweeds in skin photoaging
Authors: Ku Mi‐Jeong , Lee Myeong‐ Sook , Moon Hee‐Jung , Lee Yong‐Hwan 
Published: June 2021

The skin aging can be divided into two areas, intrinsic aging and photoaging. Damage to human skin is due to repeated exposure to ultraviolet (UV) radiation. Especially UVB causes cutaneous changes such as sunburn, skin aging and skin cancer. UVB irradiation induces formation of reactive oxygen species (ROS) and depletes cellular antioxidant stores. In the present study, using the human skin fibroblast HS68 cell line, the author investigated antioxidation effects of fucoidan through UVB‐induced glutathione (GSH), malondialdehyde (MDA), ROS and DNA fragmentation. Fucoidan, with doses of 10 μg/mL, inhibited (8.4%) the production of MDA. GSH level in HS68 cells was increased (5.8%) at pretreated with fucoidan 1 μg/mL. The levels of intracellular reactive oxygen species (ROS) were reduced by dose‐dependently.Based upon these results, it was suggested that fucoidan might play an important role in the celluar antioxidation effects against UVB irradiation‐induced photoaging.*This work was supported by the grant of KICOS
Induction of differentiation in K‐562 leukemia cells by PMA and AD 198
Authors: Barham Whitney Jo , Lothstein Leonard 
Published: June 2021

Both the phorbol ester phorbol‐12‐myristate‐13‐acetate (PMA) and the anthracycline N‐benzyladriamycin‐14‐valerate (AD 198) are protein kinase C (PKC) activators. Both act by binding to the C1b domain of PKC. PMA and AD 198 promote apoptosis in cells exposed to their respective cytotoxic doses. PMA promotes differentiation when administered at lower than cytotoxic levels. AD 198 may too induce differentiation when administered at non‐cytotoxic levels.K‐562 chronic myeloid leukemia cells were exposed to AD 198 at a non‐cytotoxic dose. Other cells were exposed to a non‐cytotoxic dose of PMA. Western blot analysis showed the level of expression of p21 in these samples, revealing significant p21 expression in the PMA treated cells, and no expression in the AD 198 treated cells. Reverse transcriptase polymerase chain reaction (RT‐PCR) was used to probe for differentiated cell specific RNAs including CD61, pdgf, and c‐myc. Significant amounts of both CD61 and pdgf were found in the PMA treated cells, but were not found in those treated with AD 198. C‐myc was absent in all samples. It was concluded that AD 198 does not induce differentiation as PMA does. Since AD 198 and PMA both increase PKC‐ ε expression, but only PMA promotes PKC‐ ε activation and nuclear localization (both implicated in differentiation), differential PKC‐ ε expression may account for this functional difference. Research funded by NIH NCI #CA 100093.
Antagonists of ribonuclease inhibitor: Small molecules, dendrimers, and peptides
Authors: Ellis Gregory A , Lavis Luke D. , Johnson R. Jeremy , Raines Ronald T. 
Published: June 2021

Recent successes have indicated the feasibility of disrupting protein–protein interactions (PPIs) with small molecules, peptidomimetics, and peptides to modulate cellular processes. Among the tightest of all known PPIs are the interactions between ribonuclease inhibitor (RI) and its ribonuclease ligands, which bind with femtomolar affinity even though these ribonucleases share only ∼30% amino‐acid sequence identity. Due to this high affinity, identifying antagonists of this interaction would be a notable advancement in the area of PPI inhibition.Multiple strategies are currently being used to identify antagonists of RI. Several preliminary antagonists have been identified through screening of small‐molecule libraries. In addition, poly(amidoamine) dendrimers were screened as ribonuclease mimics and demonstrated moderate affinity for RI. Finally, phage display is being conducted to identify peptide antagonists.Ribonucleases that evade RI exhibit selective toxicity for cancer cells, and the lack of affinity for RI is considered a primary determinant of that toxicity. Therefore, it is hypothesized that inhibiting the RI–ribonuclease interaction with antagonists will potentiate the cytotoxic activity of modestly RI‐evasive ribonucleases, possibly leading to extremely potent chemotherapeutics.This work was supported by Grant CA73808 (NIH) and the Wisconsin Alumni Research Foundation.
Applications of Chemotaxis in the Classroom
Authors: Burke Teresa Rose , Cruikshank William 
Published: June 2021

Cells of the immune system migrate toward specific chemical signals generated by an inflammatory response. During inflammation, specific cells are responsible for secreting chemokines. Chemokines are small cytokines that promote the chemotaxis (cell migration) of specific target cells that display the corresponding protein receptors. Many of these target cells regulate the immune system by either enhancing or suppressing the reaction.In this laboratory experience, mouse models were used to demonstrate chemotaxis of human T‐cells toward two specific chemokines, IL‐16 and MIP. Human primary lymphocytes were labeled with an internal fluorescent marker and injected into immunocompromised SCID mice. The chemoattractants were then intratracheally instilled into the lungs of the mice. After 2–4 days, lung and lymph node tissue was extracted and cell migration was analyzed using a flow cytometer.The curriculum designed from this experiment is geared towards an Advanced Placement Biology course. The main concepts that this unit will cover are cell communication, membrane structure, protein receptors, and immune system response. Students will use column chromatography to separate an “unknown” lymphocyte concentration from an imitation human blood suspension. The Ochterlony technique will be used to analyze the composition and the chemotaxis of the “unknown” lymphocytes towards a specific chemokine, either IL‐16 or MIP. Results will be analyzed based on the formation of precipitates in the agar.
Endothelial CD47 interaction with SIRP‐γ is essential for T cell transendothelial migration in vitro
Authors: Stefanidakis Michael , Parkos Charles A , Luscinskas Francis W 
Published: June 2021

6006‐AAILeukocyte transendothelial migration (TEM) is a critical event in inflammation and requires sequential action of several families of adhesion molecules. CD47 or Integrin‐Associated Protein (IAP) is a 50kD transmembrane protein that belongs to the IgSF and is highly expressed on platelets, leukocytes, endothelial and epithelial cells, and neurons. CD47 is known to bind two members of the signal regulatory protein family, SIRPα and SIRPγ. So far, little is know about the role of endothelial CD47 in T cell TEM under physiological flow conditions in vitro or in vivo. FACS and Western blot analysis show that CD3+ T cells express SIRPγ but not SIRPα. Confocal microscopy showed that CD47 was enriched at endothelial junctions, suggesting a role for CD47 in T cell TEM. To determine the role of endothelial CD47 in T cell TEM, function blocking mAb to CD47 or SIRPγ were used in an in vitro flow model of TEM. mAb to CD47 or SIRPγ inhibited T cell TEM by 65 % and 75 % (n=4), respectively, but had no effect on adhesion. Both blocking mAb also decreased T cell apical migration velocity. Strikingly, the TEM of murine T helper 1 cells through TNF‐α activated murine CD47 deficient endothelium was inhibited by 80 % (n=2). In summary, these data indicate that interactions between endothelial CD47 and SIRPγ play an important role in T cell TEM under flow conditions. Support by grants from the American Heart Association and NIH HL36028.
Characterization of Islet Infiltrating Lymphocytes in NOD mice
Authors: Young Ellen , Hess Paul , Arnold Larry , Tisch Roland , Frelinger Jeffrey 
Published: June 2021

Nonobese diabetic (NOD) mice are a major animal model for the study of Type 1 diabetes (TID). However, the lymphocytes infiltrating the islets are not well characterized because of low cell numbers. A detailed analysis of the islet infiltrating lymphocytes was carried out using 9 color flow cytometry. Interestingly, up to 30% of the CD4+ T cells and 40% of the CD8+ T cells infiltrating the islets were CD62Lhi and CD44lo indicating a naïve phenotype. In contrast, islet infiltrating CD8+ T cells with specificity for islet‐specific glucose‐6‐phosphatase catalytic subunit‐related protein (IGRP) had an effector phenotype. As the number of CD4+T cells per islet increased the relative number of CD8+ T cells also increased. The percentage of naïve CD4+T cells increased from less than 10% to 30% of the CD4+ T cell population and the percentage of IL‐4+ CD4+ T cells fell, although the number per islet increased linearly with the increase in CD4+ T cells per islet. We concluded that infiltrates with the same number of cells per islet had the same composition regardless of source, i.e. male or female NOD mice. There was a significant difference in the time to reach the same cell number, with females accumulating cells at an earlier age.Research support ‐ NIAID
Different clinical course of experimental autoimmune encephalomyelitis (EAE) in neurointermediate pituitary lobectomized (NIL) plus adrenalectomized (ADX) Lewis and Wistar rats
Authors: Quintanar‐Stephano Andrés , Organista‐Esparza Alejandro , González‐Delgado Evangelina , de Jesús García‐Gallegos Diego , Campos‐Tenorio Beatríz , Reyes‐Echeverría María Cruz , Kovacs Kalman , Berczi Istvan 
Published: June 2021

We have described that NIL Lewis rats develop mild signs of acute EAE with no changes in the HPA axis activity. Lewis and Wistar rats develop EAE, but differences in the HPA axis response to EAE exist between them. This suggests a different strain‐specific role of corticosterone (CO) in the immunoregulation of the EAE. Here we study the effects of CO and neurohypophyseal (NH) hormones deficiency (NIL) on the EAE. Rats from both strains were sham operated (SHAM) or NIL. Three weeks after surgeries rats were immunized with an encephalitogenic emulsion, and clinical course of EAE registered before and after ADX (performed at 27 day post‐immunization (DPI)). SHAM animals of both strains developed similar transient paralytic scores of EAE (10–20 DPI). In both strains, NIL induced decreased scores of the EAE. In Wistar SHAM ADX and NIL ADX groups no relapses of EAE occurred, whereas in Lewis SHAM ADX and NIL ADX groups a prompt and worse relapse of the EAE occurred, similar to the observed in the acute paralysis.Conclusions: 1) In NIL Wistar and Lewis groups, decreased severity of the EAE is under the NH hormones control, 2) In Wistar rats CO‐independent mechanisms protect against the EAE relapses, 3) In Lewis rats CO is required to protect against the EAE relapses, 4) Although NH hormones and CO participate in the immunopathology of the EAE it seems that they work through different mechanisms.
Brief exercise increases circulating myeloperoxidase in boys with maturational influences observed at baseline
Authors: Schwindt Christina Danell , Zaldivar Frank , Cooper Dan 
Published: June 2021

Rationale: Circulating myeloperoxidase (cMPO) is elevated in asthmatics suggesting leukocytes are inordinately activated in these individuals and may play a role in wheezing. Intriguingly, MPO is increased in response to exercise in non‐asthmatic adults, and exercise is a major trigger for bronchoconstriction in asthmatics. We hypothesized that exercise would increase cMPO in children and the response to exercise would be greater in asthmatics.Methods: Thirty‐one boys [9‐asthmatic late pubertal (AL), mean age 16.1 years; 8‐asthmatic early pubertal (AE), mean age 10.6 years; and 14 healthy control late pubertal (CL), mean age 17.0 years] performed 6–8 minutes of moderate exercise on a cycle ergometer. Blood was sampled at pre‐exercise (PE) and end‐exercise (EE).Results: CMPO did not differ between groups at PE (AL 8.0±1.6 ng/mL, AE 9.8±1.7 ng/mL, and CL 5.8±1.3 ng/mL); however, in 3 asthmatics PE cMPO was >2SD above normal. All groups demonstrated a significant increase from PE to EE [AE Δ =3.9±1.5 ng/mL, p=0.013; AL Δ =4.5±1.4 ng/mL, p=0.0032; and CL Δ =5.0±1.1 ng/mL, p=0.0001] without a significant difference among groups. There was no correlation (|r|<0.4) in PE cMPO or ΔcMPO to leukocytes, lung function, or fitness (BMI%, max VO2, VO2/kg). There was a significant negative correlation (r=0.54, p<0.02) between PE cMPO and age in controls.Conclusions: Baseline cMPO may be influenced by maturational status. Brief exercise led to significant increases in cMPO in all children. The magnitude of difference was not related to asthma. We speculate that exercise‐induced asthma may result from an exaggerated response to leukocyte activation that occurs normally in children even with brief exercise.
Dendritic cells as anti‐inflammatory regulators of extra‐pulmonary acute lung injury
Authors: Venet Fabienne , Lomas‐Neira Joanne , Chen Yaping , Chung Chun‐Shiang , Ayala Alfred 
Published: June 2021

Acute Lung injury (ALI) is one of the most common organ dysfunctions in critically ill patients and is associated with lung inflammation and apoptosis. However, the cellular interactions in the lung contributing to its pathophysiology are unclear. The objective of this study was to determine the contribution of dendritic cells (DCs) to lung apoptosis, inflammation and neutrophil recruitment during ALI resultant from hemorrhagic shock (Hem)/sepsis (CLP).Using a dual insult model of Hem followed 24h later by CLP, we measured, 24h after CLP, caspase 3 activity, lung myeloperoxidase (MPO) and pro‐inflammatory cytokine production in the lung and plasma of CD11c‐Diphteria Toxin (DT) Receptor conjugated vs background mice injected intraperitoneally (IP) or intratracheally (IT) with DT prior to shock.Systemic DC depletion (IP DT) was associated with a marked increase in IL‐6, TNF‐α and MCP‐1 concentrations after Hem/CLP, whereas lung apoptosis and neutrophil recruitment (MPO) were not modified in comparison with background mice. Similar results were observed after local lung DCs depletion (IT DT). Preliminary experiments suggest that this DC effect is mediated through the recruitment and/or activation of macrophage to/in the lung. Thus the resident and possibly recruited DCs may play here‐to‐for unappreciated role in suppressing lung inflammation and promoting the resolution of ALI. (NIH‐HL73525)
TLR‐2 Dependency of IL‐6 Induction by PG‐PS in Bone‐Marrow Derived Dendritic Cells
Authors: Kannan Krishnaswamy , Sisk Bryan , Kimpel Donald , Ortmann Robert 
Published: June 2021

Peptidoglycan‐polysaccharide (PG‐PS 10S), a purified streptococcal cell wall preparation, has been widely used as an antigen/adjuvant to induce arthritis in female Lewis rats. We earlier demonstrated that PG‐PS can induce IL‐1β, IL‐6 and TNF‐α in bulk cultures of rat splenocytes and in human PBMC. In this study, we investigated the cellular source of these pro‐inflammatory cytokines using naïve bone‐marrow derived dendritic cells (BM‐DCs) obtained from C57BL/6 wt, TLR2‐KO and TLR9‐KO mice. Our results show a dose‐dependent increase of IL‐6 production with the incubation of PG‐PS at 0–30 μg/ml for 24 and 48h. BM‐DCs derived from B6 and TLR9‐KO mice produced significantly higher quantities of IL‐6 over media controls. However, TLR2‐KO BM‐DCs produced no detectable IL‐6, thus suggesting a TLR2 dependency of this cytokine induction.It was also noteworthy that TLR9‐KO dendritic cells produced five fold more IL‐6 as compared to B6 wt DCs suggesting a regulatory control between TLR9 and TLR2 in the induction of IL‐6. We extended these studies to human PBMCs, peripheral blood monocyte‐derived DCs and purified CD4+ T‐cells and found a similar trend in IL‐6 induction in a dose‐dependent manner. Taken together, our results suggest that PG‐PS induction of IL‐6 in dendritic cells is a TLR‐2 dependent process.
The CD3 ζ Subunit of the T Cell Receptor is Required for an Efficient Immune Response During Bacterial Infection
Authors: Becker Amy M , Young Jennifer A. , Forman James , van Oers Nicolai S.C. 
Published: June 2021

The T cell receptor (TCR) complex is comprised of a ligand binding αβ heterodimer and two signaling modules, CD3 ζζ and CD3 γε/δε. Each signaling subunit has at least one copy of an ITAM signaling motif. TCR engagement results in bi‐phosphorylation of the ITAM on tyrosine residues, allowing ZAP‐70 to complex. This initiates an intracellular signaling cascade, leading to T cell activation and effector function.We generated CD3 ζ transgenic mice (YF1‐6) lacking all forms of phospho‐ζ. Previous data have shown that phospho‐ζ is not required for T cell development or signaling. Conversely, after TCR stimulation, CD8+ T cells from YF1‐6 mice produce 2‐fold less IFN‐γ than C57BL/6 mice. To determine the impact of this difference, we established a Listeria monocytogenes (LM) infection model. We found that YF1‐6 mice could not clear primary LM infections and had 5 to 50‐fold higher CFUs in the liver and spleen compared to control mice. Yet, after secondary infection, YF1‐6 and C57BL/6 mice cleared the bacteria. This suggests that phospho‐ζ is required for innate cell function during LM infection. We are examining whether this results from aberrant CD8+ T or NK cell function, as both populations are thought to be important for early IFN‐γ production and LM clearance. Defining the role of phospho‐ζ during innate immune responses to infections may reveal new therapeutic targets. Supported by NIH T32 AI005284‐28, AI042953, AI45764
Effects of the Mexican program Oportunidades on the dietary intake of energy, iron, zinc and vitamin A in children 1 to 4 years old in rural areas
Authors: Ramírez‐Silva Ivonne , Rivera Juan , Leroy Jef 
Published: June 2021

Introduction: Mexico's “Oportunidades” program provides conditional cash transfers to poor families and fortified food supplements to their young children for improving human capital.Objective: Assess the program's as well as the food supplement effects on daily dietary intake of energy, iron, zinc and vitamin A.Methodology: Secondary data analysis of a randomized effectiveness study in children 12–59 mo. Diet was evaluated using a 24‐hour recall questionnaire. Fffects were estimated using a random‐effects multiple linear regression model. The control group was compared with the overall program's beneficiaries (PB) as well as with the subgroups of consumers (SC) and non‐consumers (SNC) of fortified supplements.Results: Overall PB consumed more iron, zinc and vitamin A than the control group at all ages, while greater energy intake was found only in children 12–23 months (65 kcal; p<0.05). SC group consumed greater amounts of iron (7.7 mg/day), zinc (7.6 mg/day), and vitamina A (398.5 mcgRE/day) (all p<0.05) than the control group. No differences in intake were found between SNC and SC (p>0.05).Conclusion: Positive impacts of the program on energy and micronutrient intakes were documented. These dietary effects were due to the intake of the fortified food supplement and not to cash transfers.
Dietary intake in children consuming three supplements with identical micronutrient content
Authors: Rodriguez Fabiola Mejia , Islas Clara Dominguez , Neufeld Lynnette M , Feregrino Raquel Garcia , Guerra Armando Garcia , Lopez Lourdes Flores 
Published: June 2021

The objective of this analysis was to assess intake of energy (E) and protein (P) from diet only in children consuming three nutritional supplements. Communities (18 per supplement) were randomly assigned to receive a fortified food (FF), syrup (SY) or Sprinkles (SK). Children 6 to 12 mo of age, beneficiaries of the Oportunidades program in Mexico were eligible to participate. A food frequency questionnaire (FFQ) referring to the previous month was applied periodically; for this analysis we assess intake after 10 m supplementation. Appropriate statistical analyses for cluster randomized trials were used. 927 children were assigned to treatment (FF n=265; SY n=323; SK n=339). E and P intake were significantly (p<0.05) lower in breastfed than non‐breastfed children and in girls than boys. Among breastfed children, median (interquartile range) intake of E and P were significantly (p<0.05) lower in FF group (1038, 791–1414 kcal; 31, 22–48 g) than in SY group (1291, 946–1672 kcal; 39, 28–52 g) but not the SK group (1119, 812–1692 kcal; 34, 21–50 g). E intake is high and likely overestimated by our FFQ. Nonetheless, we believe that this does not impede comparisons between supplementation groups. The results of this study suggest that consumption of the FF may result in some displacement of energy and protein from the usual home diet.The Oportunidades program financed this study and holds the rites to the data presented.