bluuebunny/crossref_metadata_2025 huggingface dataset:
A113: TCR‐independent Activation of T‐cells Through CD31 Triggering as an Etiology of Cytokine Production in Juvenile Idiopathic Arthritis
Authors: Ferguson Ian D. , Griffin Patricia , Michel Joshua , Kietz Daniel , Rosenkranz Margalit , Vallejo Abbe N.
Published: March 2014

Background/Purpose:

JIA is the banner diagnosis for a heterogenous group of rheumatic conditions characterized by local inflammation and destruction of the joint space and systemic manifestations. Pro‐inflammatory cytokines derived from T cells are considered effectors of disease. Recent data has shown that some of these cytokines come directly from a distinct subset of T cells expressing CD31, an adhesion molecule expressed by granulocytes and endothelial cells. In the present work, we examined whether the array of cytokines in blood and synovial fluid of patients are the same array produced by T cells activation through CD31 ligation independent of the TCR. We propose that CD31‐driven cytokine production by T cells is a self‐perpetuating mechanism of local and systemic inflammation in JIA.

Methods:

Following informed consent/assent, blood and/or synovial fluid were collected from patients with oligoarticular and polyarticular JIA. Similar blood samples were also collected from healthy subjects. Cytokine composition of the plasma and/or fluid were subjected to multiplex analysis using the Luminex platform. Phenotypes of mononuclear cells in blood and synovial fluid were examined by multicolor flow cytometry. Drawing from the results of the Luminex assay, receptor crosslinking bioassays using primary CD31+ T cells (in blood and/or synovial fluid) and CD31+ T cell lines were conducted. Cytokine production by these cells were assessed by flow cytometry and/or by Luminex.

Results:

The overall cytokine signature of JIA characterized by the dominance of IL‐6, IL‐10 and TNFα. Unexpectedly, patients with oligoarticular disease showed higher levels of IFNγ and IL‐17 family cytokines than those with polyarticular disease. Both types of patients however showed a similar cellular signature characterized by the preponderance of T cells deficient in CD4, CD8, and CD28, but expressing CD31. Results of CD31 cross‐linking bioassays, using either specific antibody or its recombinant ligand CD38, showed high levels of induction of IL‐2, IL‐17 and IFNγ. Specificity of CD31‐driven, TCR‐dependent cytokine production was verified by similar bioassays using somatic somatic T cell line mutants Loucy and JRT3 that lack TCR and TCR/CD3, respectively, but are both CD31+. The observed cytokine production was associated with the phosphorylation of the signaling substrates ZAP70, cAbl, and p65‐NFKB.

Conclusion:

Our data show that the cytokine signature of oligoarticular JIA appears distinct with the dominance of IFNγ and IL17. TCR independent, CD31‐driven induction of these cytokines suggests maladaptive T cell function in JIA. Further investigation on the relevance of IFNγ and IL‐17 to disease activity/clinical outcomes, and the CD31 signaling pathway will pave way to innovations in immunotherapy in JIA.
A125: Immunomodulatory Factors Produced by Mesenchymal Stem Cells After in vitro Priming with Danger Signals
Authors: Bukulmez Hulya , Bilgin Asuman , Bebek Gurkan , Caplan Arnold I. , Jones Olcay
Published: March 2014

Background/Purpose:

We have previously shown that pre‐exposure to IL‐1β augments the immunosuppressive activity of MSC using the mixed lymphocyte reaction as a readout assay. In this project, we studied the profiles of inflammatory cytokines secreted by MSCs at the protein level to identify biomarkers in in vitroprimed MSCs. The objective of this study was to determine the MSC response to “danger signals.”

Methods:

MSCs were isolated from a healthy donor and cultured as per a protocol published earlier by the Skeletal Research Center. After three passages, 200,000 cells per 35 mm culture dish were incubated in 2 ml DMEM with 10% fetal bovine serum at 37°C for 5 days until 90% confluent. The MSCs were then treated with LPS (10 μg/ml), IL‐1β (25 ng/ml), TNF‐α (50 ng/ml), or poly (I:C) (30 ng/ml) overnight (18 hours). Control and treatment groups were done in triplicate. The culture media were then collected and analyzed for secreted mediators using the RayBiotech® Human Cytokine Array (2000), as per the manufacturer's protocol. Chemiluminescense was detected using Kodak BioMax MR film, and was analyzed using the ImageJ program (rsbweb.nih.gov). Normalization and fold‐difference computations were then performed in MS Excel. Pathway analysis for biomarker detection was performed using the Ingenuity (ingenuity.com) program by the CWRU proteomics center.

Results:

There were clear distinctions in the patterns of inflammatory mediators produced by MSC upon exposure to LPS versus poly (I:C). LPS stimulated the production of mediators that affect mostly neutrophil chemotaxis and activation, including IL‐8 (1.8 fold), CXCL7 (2 fold), CXCL5 (2.4 fold), CXCL1 (1.6 fold). However, poly (I:C) induced mediators with activities on T cells and NK cells including IL‐15 (18 fold), IL‐4 (23 fold), TGF‐β3 (9.7 fold), CCL‐18 (4.6 fold). While poly (I:C) stimulated IL‐1β production (2.6 fold), LPS did not (0.7 fold), and neither agent increased TNF‐α significantly (1.3 and 2.1 fold). Interestingly, there was marked production of CCL1 (12 fold) upon exposure of MSC to IL‐1β but not to LPS (0.6 fold), TNF‐α (1.3) or poly (I:C) (2.1 fold).

Conclusion:

Priming MSC by TLRs ligands triggered different cytokine and chemokine expression profiles. These profile patterns may signify framework of the upcoming immune responses in the tissue. Accordingly, the candidate biomarkers for ongoing MSC immunomodulatory activities should include those for neutrophils versus those for T/NK cells. Furthermore, our results strongly suggest involvement of IL‐1β in MSCmediated activation of T‐regulatory cells by induction of the CCR‐8 ligand, CCL‐1. CCR‐8 is a known biomarker identifying CD4+ memory T‐cells enriched for FoxP3+ T‐regulatory cells and is involved in delivery of immunosuppression.
A137: Appropriateness of Rheumatology Lab Studies and Referrals among General Pediatricians
Authors: Correll Colleen K. , Spector Logan G. , Zhang Lei , Binstadt Bryce A. , Vehe Richard K.
Published: March 2014

Background/Purpose:

Rheumatology lab tests are often of low diagnostic utility when ordered by primary care physicians, and may lead to erroneous referrals. No prior studies have investigated pediatricians' reasons for ordering these tests. We surveyed pediatricians in three states of the Upper Midwest regarding their ordering of rheumatology lab tests and reasons for referral to pediatric rheumatology (PR).

Methods:

A web‐based survey was emailed to general pediatricians practicing in Minnesota, North Dakota, and South Dakota. The rheumatology tests queried included antinuclear antibody (ANA), anti‐double‐stranded DNA antibody (anti‐dsDNA), rheumatoid factor (RF), anti‐citrullinated peptide antibodies (anti‐CCP), and HLA‐B27. Basic descriptive analysis was performed. Subgroup analysis was performed to assess the relationship between the pediatrician's experience with PR (defined as years since completion of residency training, whether or not the training program had a PR division, time spent in a PR clinic during training and completion of continued medical education focused on PR in the last three years) and the appropriateness of lab tests and PR referrals.

Results:

The response rate was 15.3% (93/609). Eighty‐nine percent (83/93) of the respondents had ordered ANA, for reasons including abnormal joint exam (72%), musculoskeletal (MSK) pain with a normal exam (52%), concern for juvenile idiopathic arthritis (JIA) (47%), rash (41%), concern for systemic lupus erythematosus (39%), fatigue (35%), and fever (35%). Fifty‐nine percent (54/92) had ordered anti‐dsDNA, and the distribution of reasons paralleled that of ANA. Seventy‐three percent (67/92) had ever ordered RF, most commonly for abnormal joint exam (67%), joint pain with a normal exam (60%), concern for JIA (42%), and fever (30%). Only 8% (7/92) had ordered anti‐CCP. Forty‐two percent (39/92) had ordered HLA‐B27, primarily for back pain (59%). Of those who had ever referred a patient to a pediatric rheumatologist, 44% (37/85) did so for a positive ANA of unclear significance, 14% (12/85) for positive RF of unclear significance, and 9% (8/85) for anti‐dsDNA of unclear significance. Positive HLA‐B27 and anti‐CCP were less common referral reasons. Subgroup analysis did not show significant differences between experience level and appropriateness of lab tests and referrals to PR.

Conclusion:

Inappropriate ordering of rheumatology lab tests with subsequent referral to PR is common among pediatricians in the Upper Midwest and likely elsewhere. The correctness of lab test utilization and referral to PR did not obviously improve with increased experience in PR. New educational interventions are needed to improve general pediatricians' understanding and utilization of rheumatology lab tests and to decrease unnecessary referrals to PR.
A139: Body Mass Index and Height Change Over Time in Childhood‐Onset Systemic Lupus Erythematosus Cohort
Authors: Harris Julia G. , Maletta Kristyn I. , Kuhn Evelyn M. , Olson Judyann C.
Published: March 2014

Background/Purpose:

Systemic lupus erythematosus (SLE) can lead to significant morbidity and mortality related to coronary artery disease. Obesity is an additional risk factor for cardiovascular disease that appears to be more prevalent in this population. Obesity in SLE has been linked to decreased health‐related quality of life, functional limitations, pain, fatigue, diabetes mellitus, and increased malignancies. In addition, growth retardation is an important aspect to consider in a child or adolescent that is exposed to corticosteroids. Growth failure has been a proposed addition to the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SDI) to modify this validated scale for childhood‐onset SLE (cSLE). There is limited data discussing change in growth parameters over time in cSLE patients, which our study sought to do.

Methods:

A retrospective chart review was performed on patients diagnosed with SLE prior to age 18 who were followed in the Children's Hospital of Wisconsin Rheumatology Clinic for at least one year between January 1999 and August 2012. Weight and height were collected two times per year. Body mass index (BMI; kg/m2) and percentiles of measurements for age were calculated based on sex‐specific growth references from the Centers for Disease Control and Prevention for patients 2 to 20 years old. Additional information collected included age of diagnosis, medication use, presence of lupus nephritis, and items of disease damage based on the SDI.

Results:

Analysis was performed on 75 patients with mean disease duration of 4.55 years. There were 685 growth for age percentile values recorded. Mean BMI percentage per age at diagnosis was 64.81 compared to 76.31 at the 610 follow‐up visits, which was statistically significant (p < 0.001). At diagnosis, 33.3% of patients were overweight (≥85–95% BMI for age), obese (≥95–99% BMI for age), or severely obese (≥99% BMI for age) versus 49.3% at last available follow‐up visit. Sixty‐eight percent of patients had at least one BMI measurement out of the normal range. Mean percent height for age at diagnosis was 64.21 versus 46.58 at follow‐up visits (p < 0.001). Eight of the seventy‐five patients had growth percentiles <5% at last follow‐up visit.

image

Conclusion:

A majority of our cSLE population was overweight or obese during at least one time period. In addition, BMI statistically increased from diagnosis to follow‐up visits. Growth failure was present in greater than 10% of the population, and percent height for age statistically decreased after diagnosis as well. Interventions related to healthy living habits and medication adjustments should be made to address these important growth issues that can have long‐standing complications.
A146: Strategies for Assessment and Management of Severe Environmental Allergies at “Camp JRA”
Authors: Ostrov Barbara E. , Robbins Lisa
Published: March 2014

Background/Purpose:

Camp JRA (Juveniles Reaching Achievement) has been offering a week of fun, education, friendship, independence and support for 17 years to children with arthritis and related diseases. The number of campers has grown from 28 the first year to 120 maximum capacity for the past few years.

Experienced Pediatric Rheumatology nurses and physicians provide hands‐on care in the “Med Shed” at Camp JRA. Medical information is completed by parents, pediatric rheumatologists and primary care providers. The senior medical staff review and verify the information. Over the past 5 years we identified that coping with environmental allergies consumed a disproportionate amount of time for the staff and we felt unprepared to optimally address these issues.

We wish to share our strategy in addressing the growing concern about environmental sensitivities in a residential week long camp for children with rheumatic diseases.

Mindfulness of drug allergies is standard of care in any environment where medications are given. Coping with food allergies has assumed a significant position in our hierarchy of medical responsibilities in the Med Shed, as well. We found ourselves struggling to manage the increasing complexity of this issue. We found that some campers had multiple allergies listed that had not been reported, and no details were provided. For many years, we have run a very organized medical camp, however, this issue confounded our care of these complex children.

Methods:

The emergence of the multiple allergies issue was a barrier to providing a confidently safe camp environment. New approaches were developed and implemented.

1. We developed new allergy history questionaire that triggered progression to further detailed information.

2. We consulted with our Pediatric Allergist to develop an algorithm for assessment of symptoms (Attached to poster).

3. We developed an Allergy Action Plan specific to each camper with allergy issues, approved in advance by parent and signed.

4. We developed a system for Epi‐pen storage: over the door holder with multiple clear pockets, with the Epi‐pen, the child's photo, the signed Allergy Action Plan.

Results:

We oriented all medical staff who worked in the Med‐Shed to the new system. When implemented at Camp JRA in July 2013 and we experienced no allergy response problems. Increasing allergy sensitivities in patients requires a proactive plan for residential camps. We have developed a model for assessing allergies and reaction preparedness. This plan has improved our confidence that we can provide safe rheumatologic AND allergy response care.

Conclusion:

The development of increasing allergy sensitivities and concerns by families and patients requires a proactive plan for residential camps of children with rheumatic disease. We hve developed a model for assessing allergies and reaction preparedness.
A150: Control of Cell Proliferation in Lupus Nephritis: The Role of miRNAs and HER2
Authors: Costa Reis Ana Patrícia , Russo Pierre , Gallucci Stefania , Sullivan Kathleen E.
Published: March 2014

Background/Purpose:

miRNAS are responsible for post‐transcriptional gene silencing and typically regulate multiple targets. Our goal was to study the miRNA pattern of lupus nephritis (LN) in order to better understand its pathogenesis.

Methods:

A retrospective cohort study was performed on a large single‐center cohort of children exposed to anti‐Ro and/or anti‐La antibodies on whom prospective data has been collected since 1984. Inclusion criteria were: 1) first child born from a woman positive for anti‐Ro and/or anti‐La antibodies with a diagnosis of cutaneous lupus, systemic lupus erythematosus, Sjogren's syndrome, dermatomyositis or rheumatoid arthritis, 2) the mother underwent fetal echocardiography screening during pregnancy and/or the child had a postnatal ECG and 3) the child was ≥6 months old as of October 2013. We used Bayesian analysis for the association between prenatal use of AM and NLE.

Results:

A kidney LN miRNA signature was identified, which, according to Ingenuity pathway analysis, mainly reflected cell proliferation (p = 3.3E‐23‐0.049). miR‐26a, miR‐30b and miR‐4286 were significantly decreased in LN (p = 0.002; p = 0.005; p = 0.002). The levels of miR‐26a and miR‐30b were also decreased in the urine of LN patients when compared to controls (p = 0.03; p = 0.02). The genes significantly up‐regulated in the knockdowns of miR‐26a, miR‐30b and miR‐4286 when compared to controls were related to mitosis (p = 1.06E‐10). These results were validated by qRT‐PCR, including the increase on the expression of genes associated with cell cycle (CCNE2, E2F8, MAD2L1, MYBL1 and POLQ).

Based on these results, we examined HER2 expression, a protein that regulates miR‐26a and miR‐30b levels in breast cancer cell lines. We found that it was highly increased in the glomeruli and tubular compartment of LN patients and not in other mesangioproliferative diseases. Moreover, the average of HER2+ cells in the glomeruli of NZM2410 mice was higher than in B6 or Balb/c mice (p < 0.0001; p = 0.009) and it was associated with proteinuria (p = 0.001). Finally, HER2 expression was increased in mesangial cells by α Ninterferon exposure (p = 0.02) and IRF‐1 transfection (p = 0.0009).

Conclusion:

The kidneys of LN patients have a miRNA pattern characterized by a significant decrease of miR‐26a, miR‐30b and miR‐4286, which is predicted to be associated with increased proliferation. In addition, miR‐26a and miR‐30b are also decreased in the urine of LN patients, which makes them interesting candidates as biomarkers. It was previously shown that trastuzumab, an antibody against HER2, produces therapeutic actions by up‐regulating miR‐26a and miR‐30b in breast cancer cell lines. Since we demonstrated that HER2 was highly increased in the kidneys of LN patients and NZM2410 mice, blocking HER2 may be a new promising pathway to decrease cell proliferation and damage in this disease.
A151: Pediatric Rheumatology Care and Outcomes Improvement Network Demonstrates Performance Improvement on Juvenile Idiopathic Arthritis Quality Measures
Authors: Harris Julia G. , DeWitt Esi Morgan , Laxer Ronald M. , Ardoin Stacy P. , Gottlieb Beth S. , Olson Judyann C. , Passo Murray H. , Weiss Jennifer E. , Lovell Daniel J. , Lee Tzielan C. , Vora Sheetal S. , Griffin Nancy , Stock Jason A. , Darbie Lynn M. , Bingham Catherine A.
Published: March 2014

Background/Purpose:

Pediatric Rheumatology Care and Outcomes Improvement Network (PR‐COIN) is a multi‐site learning network designed to improve outcomes of juvenile idiopathic arthritis (JIA) care. Teams collect point of care data on measures of process of care and outcomes of care for the purposes of analysis to guide improvement activities. Eleven North American pediatric rheumatology centers participate. This report illustrates our improvement in several JIA process quality measures (QMs).

Methods:

Process of care QMs targeted for improvement include measurement of: arthritis‐related pain, physician global assessment, joint count, health‐related quality of life, physical function, as well as screening for uveitis, medication toxicity, and tuberculosis per guidelines. Outcome measures for JIA include clinical inactive disease, clinical remission on and off medications, no or mild pain level, and optimal physical functioning. Network goals were determined for each process and outcome measure. Data are collected with IRB approval and informed consent, and the shared registry for data entry is the ACR's Rheumatology Clinical Registry. Site‐specific and aggregate data are analyzed and displayed monthly via statistical process control charts allowing PR‐COIN to track performance over time. Individual centers use established quality improvement methodology to reach and exceed pre‐determined goals.

Results:

Data from 5112 encounters for 1134 JIA patients have been collected since April 2011. QMs with performance meeting or exceeding initial goals include documentation of complete joint count and measurement of arthritis‐related pain. For PR‐COIN network as a collective unit, QMs improved in six processes—measurement of functional ability, completion of ongoing medication toxicity labs, documentation of complete joint count, medication counseling for newly prescribed DMARDs, documentation of annual medication counseling, and measurement of health‐related quality of life. All of these measures had a shift above the baseline mean, demonstrating special cause. In addition, five sites have demonstrated individual improvement in at least one process QM.

Conclusion:

PR‐COIN sites are collectively and individually demonstrating significant improvements in JIA process of care QMs. Quality improvement efforts in PR‐COIN are ongoing with the goal of improving the outcome for patients with JIA.
A177: Program Evaluation of the ACR/CARRA Inter‐Institutional Mentoring Program (AMIGO) in Pediatric Rheumatology
Authors: Muscal Eyal , Moorthy Lakshmi N. , Riebschleger Meredith P. , Eberhard Anne , Huttenlocher Anna , Klein‐Gitelman Marisa S. , Prahalad Sampath , Stevens Kelly Rouster , Schneider Rayfel , Nigrovic Peter A.
Published: March 2014

Background/Purpose:

In pediatric rheumatology, the small size of many academic programs translates into limited mentoring options for early career physicians. To address this “mentorship gap,” in 2011 the American College of Rheumatology (ACR) and the Childhood Arthritis and Rheumatology Research Alliance (CARRA) joined together to develop AMIGO, the ACR/CARRA Mentoring Interest Group, that now includes greater than half of pediatric rheumatology fellows and junior faculty in the US and Canada. We report ongoing program evaluation encompassing pilot (2011) and full roll‐out (2012, 2013) phases of the AMIGO project.

Methods:

Mentees and mentors participating in the AMIGO project were surveyed via online questionnaire to determine dynamics of contact and perceived benefit. The entire pediatric rheumatology community was surveyed in 2011 to determine the state of mentoring and career development, with a repeat survey planned for early 2014. We tabulated the active dyads as of January 2014.

Results:

As recently reported, detailed evaluation of the pilot phase 17 months after initial roll‐out found that 19 of 20 pilot AMIGO dyads were still functioning, with substantial benefit noted by mentees in career guidance, scholarship, and job satisfaction. Benefits reported by mentors included improvement of their mentoring skills and development of their academic portfolios. Both mentees and mentors reported improved connectedness to the wider pediatric rheumatology community. 83 additional dyads were matched prior to 2012 and 2013 ACR meetings. Comparison of these 17‐month pilot results to a December 2013/January 2014 survey of all active AMIGO participants is ongoing. A questionnaire administered to the whole pediatric rheumatology community in 2011 (n = 135 respondents, including an estimated 64–70% response rate among fellows and junior faculty) found that approximately 60% of fellows and junior faculty had a career development mentor. We will assess whether the AMIGO program has had a global impact upon the mentoring culture in pediatric rheumatology by collecting community‐wide survey data in 2014 and then comparing to 2011 data.

Conclusion:

The 2011 AMIGO pilot program has confirmed the feasibility of a North American inter‐institutional mentoring program in pediatric rheumatology. Participants identified benefits to both mentees and mentors in multiple domains, most prominently in career guidance, a core goal of the program. Ongoing program evaluation will determine how much of this benefit has been sustained after participation was opened to the whole community, and whether the program has had a measureable impact on the overall state of mentoring in pediatric rheumatology.
Tendon‐to‐bone attachment: From development to maturity
Authors: Zelzer Elazar , Blitz Einat , Killian Megan L. , Thomopoulos Stavros
Published: March 2014

The attachment between tendon and bone occurs across a complex transitional tissue that minimizes stress concentrations and allows for load transfer between muscles and skeleton. This unique tissue cannot be reconstructed following injury, leading to high incidence of recurrent failure and stressing the need for new clinical approaches. This review describes the current understanding of the development and function of the attachment site between tendon and bone. The embryonic attachment unit, namely, the tip of the tendon and the bone eminence into which it is inserted, was recently shown to develop modularly from a unique population of Sox9‐ and Scx‐positive cells, which are distinct from tendon fibroblasts and chondrocytes. The fate and differentiation of these cells is regulated by transforming growth factor beta and bone morphogenetic protein signaling, respectively. Muscle loads are then necessary for the tissue to mature and mineralize. Mineralization of the attachment unit, which occurs postnatally at most sites, is largely controlled by an Indian hedgehog/parathyroid hormone‐related protein feedback loop. A number of fundamental questions regarding the development of this remarkable attachment system require further study. These relate to the signaling mechanism that facilitates the formation of an interface with a gradient of cellular and extracellular phenotypes, as well as to the interactions between tendon and bone at the point of attachment. Birth Defects Research (Part C) 102:101–112, 2014. © 2014 Wiley Periodicals, Inc.
The chondrocytic journey in endochondral bone growth and skeletal dysplasia
Authors: Tsang Kwok Yeung , Tsang Shun Wa , Chan Danny , Cheah Kathryn S.E.
Published: March 2014

The endochondral bones of the skeleton develop from a cartilage template and grow via a process involving a cascade of chondrocyte differentiation steps culminating in formation of a growth plate and the replacement of cartilage by bone. This process of endochondral ossification, driven by the generation of chondrocytes and their subsequent proliferation, differentiation, and production of extracellular matrix constitute a journey, deviation from which inevitably disrupts bone growth and development, and is the basis of human skeletal dysplasias with a wide range of phenotypic severity, from perinatal lethality to progressively deforming. This highly coordinated journey of chondrocyte specification and fate determination is controlled by a myriad of intrinsic and extrinsic factors. SOX9 is the master transcription factor that, in concert with varying partners along the way, directs the different phases of the journey from mesenchymal condensation, chondrogenesis, differentiation, proliferation, and maturation. Extracellular signals, including bone morphogenetic proteins, wingless‐related MMTV integration site (WNT), fibroblast growth factor, Indian hedgehog, and parathyroid hormone‐related peptide, are all indispensable for growth plate chondrocytes to align and organize into the appropriate columnar architecture and controls their maturation and transition to hypertrophy. Chondrocyte hypertrophy, marked by dramatic volume increase in phases, is controlled by transcription factors SOX9, Runt‐related transcription factor, and FOXA2. Hypertrophic chondrocytes mediate the cartilage to bone transition and concomitantly face a live‐or‐die situation, a subject of much debate. We review recent insights into the coordination of the phases of the chondrocyte journey, and highlight the need for a systems level understanding of the regulatory networks that will facilitate the development of therapeutic approaches for skeletal dysplasia. Birth Defects Research (Part C) 102:52–73, 2014. © 2014 Wiley Periodicals, Inc.
Engineering of a bispecific affibody molecule towards HER2 and HER3 by addition of an albumin‐binding domain allows for affinity purification and in vivo half‐life extension
Authors: Malm Magdalena , Bass Tarek , Gudmundsdotter Lindvi , Lord Martin , Frejd Fredrik Y. , Ståhl Stefan , Löfblom John
Published: March 2014

Emerging strategies in cancer biotherapy include the generation and application of bispecific antibodies, targeting two tumor‐associated antigens for improved tumor selectivity and potency. Here, an alternative format for bispecific molecules was designed and investigated, in which two Affibody molecules were linked by an albumin‐binding domain (ABD). Affibody molecules are small (6 kDa) affinity proteins and this new format allows for engineering of molecules with similar function as full‐length bispecific antibodies, but in a dramatically smaller size (around eight‐fold smaller). The ABD was intended to function both as a tag for affinity purification as well as for in vivo half‐life extension in future preclinical and clinical investigations. Affinity‐purified bispecific Affibody molecules, targeting HER2 and HER3, showed simultaneous binding to the three target proteins (HER2, HER3, and albumin) when investigated in biosensor assays. Moreover, simultaneous interactions with the receptors and albumin were demonstrated using flow cytometry on cancer cells. The bispecific Affibody molecules were also able to block ligand‐induced phosphorylation of the HER receptors, indicating an anti‐proliferative effect. We believe that this compact and flexible format has great potential for developing new potent bispecific affinity proteins in the future, as it combines the benefits of a small size (e.g. improved tissue penetration and reduced cost of goods) with a long circulatory half‐life.
Simultaneous transapical aortic and mitral valve‐in‐valve implantation for double prostheses dysfunction: Case report and technical insights
Authors: D'Onofrio Augusto , Zucchetta Fabio , Gerosa Gino
Published: March 2014

Transcatheter “Valve‐in‐Valve” implantation (ViV) has shown promising results in high‐risk patients suffering from structural valve deterioration (SVD) of a previously implanted heart valve bioprosthesis. We present a case of a 68‐year‐old woman with a history of three previous cardiac operations on the aortic and mitral valve. At the time of admission she was severely symptomatic due to a simultaneous SVD of a 23 mm aortic and of a 29 mm mitral St. Jude Biocor bioprosthesis. Because of the history of several cardiac operations and to her comorbidities, the patient was considered with an extremely high surgical risk profile and was therefore scheduled for double concomitant mitral and aortic ViV. Through a trans‐apical approach, the patient underwent 23 and 29 mm Edwards Sapien XT implantation in the aortic and mitral bioprosthesis, respectively. The procedure was uneventful as well as the following hospital stay. At 6‐months follow‐up the patient is in NYHA class I. Echocardiography shows that the aortic bioprosthesis has no leak and the mean gradient is 20 mm Hg while the mitral valve has mild leak and maximum and mean gradients are 21 and 10 mm Hg, respectively. The three main technical aspects that should be carefully considered in double concomitant ViV are: sequence of valve deployment (whether to implant the mitral or the aortic valve first), choice of access and valve sizing. In conclusion, double simultaneous trans‐apical mitral and aortic ViV is technically feasible. © 2014 Wiley Periodicals, Inc.
Organic–Inorganic Interface‐Induced Multi‐Fluorescence of MgO Nanocrystal Clusters and Their Applications in Cellular Imaging
Authors: Xie Shuifen , Bao Shixiong , Ouyang Junjie , Zhou Xi , Kuang Qin , Xie Zhaoxiong , Zheng Lansun
Published: March 2014

Surface functionalization of inorganic nanomaterials through chemical binding of organic ligands on the surface unsaturated atoms, forming unique organic–inorganic interfaces, is a powerful approach for creating special functions for inorganic nanomaterials. Herein, we report the synthesis of hierarchical MgO nanocrystal clusters (NCs) with an organic–inorganic interface induced multi‐fluorescence and their application as new alternative labels for cellular imaging. The synthetic method was established by a dissolution and regrowth process with the assistance of carboxylic acid, in which the as‐prepared MgO NCs were modified with carboxylic groups at the coordinatively unsaturated atoms of the surface. By introducing acetic acid to partially replace oleic acid in the reaction, the optical absorption of the produced MgO NCs was progressively engineered from the UV to the visible region. Importantly, with wider and continuous absorption profile, those MgO NCs presented bright and tunable multicolor emissions from blue‐violet to green and yellow, with the highest absolute quantum yield up to (33±1) %. The overlap for the energy levels of the inorganic–organic interface and low‐coordinated states stimulated a unique fluorescence resonance energy transfer phenomenon. Considering the potential application in cellular imaging, such multi‐fluorescent MgO NCs were further encapsulated with a silica shell to improve the water solubility and stability. As expected, the as‐formed MgO@SiO2 NCs possessed great biocompatibility and high performance in cellular imaging.
On the Mechanism of Bifunctional Squaramide‐Catalyzed Organocatalytic Michael Addition: A Protonated Catalyst as an Oxyanion Hole
Authors: Kótai Bianka , Kardos György , Hamza Andrea , Farkas Viktor , Pápai Imre , Soós Tibor
Published: March 2014

A joint experimental–theoretical study of a bifunctional squaramide‐amine‐catalyzed Michael addition reaction between 1,3‐dioxo nucleophiles and nitrostyrene has been undertaken to gain insight into the nature of bifunctional organocatalytic activation. For this highly stereoselective reaction, three previously proposed mechanistic scenarios for the critical CC bond‐formation step were examined. Accordingly, the formation of the major stereoisomeric products is most plausible by one of the bifunctional pathways that involve electrophile activation by the protonated amine group of the catalyst. However, some of the minor product isomers are also accessible through alternative reaction routes. Structural analysis of transition states points to the structural invariance of certain fragments of the transition state, such as the protonated catalyst and the anionic fragment of approaching reactants. Our topological analysis provides deeper insight and a more general understanding of bifunctional noncovalent organocatalysis.
Metal‐Free CC Coupling Reactions with Tetraguanidino‐Functionalized Pyridines and Light
Authors: Stang Simone , Kaifer Elisabeth , Himmel Hans‐Jörg
Published: March 2014

Herein we report on metal‐free CC coupling reactions mediated by the pyridine derivative 2,3,6,7‐tetrakis(tetramethylguanidino)pyridine under the action of visible light. The rate‐determining step is the homolytic NC bond cleavage of the initially formed N‐alkyl pyridinium ion upon excitation with visible light. The released alkyl radicals subsequently dimerize to the CC coupling product. 2,3,6,7‐Tetrakis(tetramethylguanidino)pyridine, which is a strong electron donor (E1/2(CH2Cl2)=−0.76 V vs. ferrocene) is oxidized to the dication. For alkyl=benzyl and allyl, relatively high first‐order rate constants of 0.23±0.03 and 0.13±0.03 s−1 were determined. Regeneration of neutral 2,3,6,7‐tetrakis(tetramethylguanidino)‐pyridine by reduction allows to drive the process in a cycle.
One‐Pot Synthesis of Highly Luminescent Carbon Quantum Dots and Their Nontoxic Ingestion by Zebrafish for In Vivo Imaging
Authors: Huang Yi‐Fan , Zhou Xin , Zhou Rong , Zhang Hong , Kang Kai‐Bin , Zhao Min , Peng Yong , Wang Qiang , Zhang Hao‐Li , Qiu Wen‐Yuan
Published: March 2014

Photoluminescent carbon and/or silicon‐based nanodots have attracted ever increasing interest. Accordingly, a myriad of synthetic methodologies have been developed to fabricate them, which unfortunately, however, frequently involve relatively tedious steps, such as initial surface passivation and subsequent functionalization. Herein, we describe a green and sustainable synthetic strategy to combine these procedures into one step and to produce highly luminescent carbon quantum dots (CQDs), which can also be easily fabricated into flexible thin films with intense luminescence for future roll‐to‐roll manufacturing of optoelectronic devices. The as‐synthesized CQDs exhibited enhanced cellular permeability and low or even noncytotoxicity for cellular applications, as corroborated by confocal fluorescence imaging of HeLa cells as well as cell viability measurements. Most strikingly, zebrafish were directly fed with CQDs for in vivo imaging, and mortality and morphologic analysis indicated ingestion of the CQDs posed no harm to the living organisms. Hence, the multifunctional CQDs potentially provide a rich pool of tools for optoelectronic and biomedical applications.
Ruthenium(II)‐Catalyzed Synthesis of Isochromenes by CH Activation with Weakly Coordinating Aliphatic Hydroxyl Groups
Authors: Nakanowatari Sachiyo , Ackermann Lutz
Published: March 2014

Cationic ruthenium(II) complexes have been employed for the highly effective oxidative annulation of alkynes with benzyl alcohols to deliver diversely decorated isochromenes. The hydroxyl‐directed CH/OH functionalization process proceeded efficiently under an atmosphere of air. Detailed mechanistic studies were indicative of a kinetically relevant CH metalation.
1‐Alkenylcalcium Iodide: Synthesis and Stability
Authors: Köhler Mathias , Görls Helmar , Langer Jens , Westerhausen Matthias
Published: March 2014

To enhance the scope of heavy calcium‐based Grignard reagents, 1,2‐dihydro‐4‐iodonaphthalene (1) was reduced with calcium in THF giving tetrakis(thf) (1,2‐dihydronaphth‐4‐yl)calcium iodide (2). This derivative represents a 1‐alkenylcalcium complex based on X‐ray structure determination and NMR data. The stability of this compound is significantly reduced compared with the aromatic naphthylcalcium iodide.
Conformational Changes in Calcium‐Sensor Proteins under Molecular Crowding Conditions
Authors: Sulmann Stefan , Dell'Orco Daniele , Marino Valerio , Behnen Petra , Koch Karl‐Wilhelm
Published: March 2014

Fundamental components of signaling pathways are switch modes in key proteins that control start, duration, and ending of diverse signal transduction events. A large group of switch proteins are Ca2+ sensors, which undergo conformational changes in response to oscillating intracellular Ca2+ concentrations. Here we use dynamic light scattering and a recently developed approach based on surface plasmon resonance to compare the protein dynamics of a diverse set of prototypical Ca2+‐binding proteins including calmodulin, troponin C, recoverin, and guanylate cyclase‐activating protein. Surface plasmon resonance biosensor technology allows monitoring conformational changes under molecular crowding conditions, yielding for each Ca2+‐sensor protein a fingerprint profile that reflects different hydrodynamic properties under changing Ca2+ conditions and is extremely sensitive to even fine alterations induced by point mutations. We see, for example, a correlation between surface plasmon resonance, dynamic light scattering, and size‐exclusion chromatography data. Thus, changes in protein conformation correlate not only with the hydrodynamic size, but also with a rearrangement of the protein hydration shell and a change of the dielectric constant of water or of the protein–water interface. Our study provides insight into how rather small signaling proteins that have very similar three‐dimensional folding patterns differ in their Ca2+‐occupied functional state under crowding conditions.
ChemInform Abstract: Identification of the Chromophore in the Apatite Pigment [Sr10(PO4)6 (CuOH1‐x‐y)2]: Linear OCuO‐ Featuring a Resonance Raman Effect, an Extreme Magnetic Anisotropy, and Slow Spin Relaxation.
Authors: Kazin Pavel E. , Zykin Mikhail A. , Zubavichus Yan V. , Magdysyuk Oxana V. , Dinnebier Robert E. , Jansen Martin
Published: March 2014

The copper‐doped apatite pigments [Sr10(PO4)6 (CuOH1‐x‐y)2] (x = 0.1, 0.3; y = 0.01—0.42) are characterized by powder XRD, XANES, EXAFS, magnetic measurements, Raman, and UV/VIS diffuse reflectance spectroscopy.
ChemInform Abstract: Enantio‐ and Regioselective CuH‐Catalyzed Hydroamination of Alkenes.
Authors: Zhu Shaolin , Niljianskul Nootaree , Buchwald Stephen L.
Published: March 2014

A mild method for the synthesis of chiral tertiary amines using an asymmetric copper‐catalyzed hydroamination is reported.
ChemInform Abstract: Photocatalysis by 3,6‐Disubstituted‐s‐tetrazine: Visible‐Light Driven Metal‐Free Green Synthesis of 2‐Substituted Benzimidazole and Benzothiazole.
Authors: Samanta Suvendu , Das Sudipto , Biswas Papu
Published: March 2014

3,6‐Di(pyridin‐2‐yl)‐1,2,4,5‐tetrazine exhibits effective visible light absorption and reversible one‐electron reduction behavior.
ChemInform Abstract: Formation of Tertiary Amides and Dihydrogen by Dehydrogenative Coupling of Primary Alcohols with Secondary Amines Catalyzed by Ruthenium Bipyridine‐Based Pincer Complexes.
Authors: Srimani Dipankar , Balaraman Ekambaram , Hu Peng , Ben‐David Yehoshoa , Milstein David
Published: March 2014

The direct amidation of primary alcohols with challenging secondary amines is achieved in the presence of small amounts of Ru‐catalysts.
ChemInform Abstract: A Concise Cross‐Metathesis Route to Enantiopure 1‐Azaspirocycles.
Authors: Wang Zhen James , Spiccia Nicolas Daniel , Gartshore Christopher James , Illesinghe Jayamini , Jackson William Roy , Robinson Andrea Jane
Published: March 2014

The title reaction leading to synthetically valuable spirocyclic products in high yields is described.
ChemInform Abstract: Palladium(II)‐Catalyzed ortho‐Arylation of Benzylic Phosphonic Monoesters Using Potassium Aryltrifluoroborates.
Authors: Meng Xiangjian , Kim Sunggak
Published: March 2014

The methylene monophosphonic acid is found to be an effective directing group which allows clean and direct ortho‐arylation of several arenes.
ChemInform Abstract: Asymmetric Epoxidation with H2O2 by Manipulating the Electronic Properties of Non‐Heme Iron Catalysts.
Authors: Cusso Olaf , Garcia‐Bosch Isaac , Ribas Xavi , Lloret‐Fillol Julio , Costas Miquel
Published: March 2014

Electronic effects in the iron catalyst are used to control the activation of hydrogen peroxide and oxygen transfer in a non‐porphyrinic catalyst.
ChemInform Abstract: Silver‐Catalyzed C—C Bond Formation with Carbon Dioxide: Significant Synthesis of Dihydroisobenzofurans.
Authors: Sekine Kohei , Takayanagi Ayano , Kikuchi Satoshi , Yamada Tohru
Published: March 2014

The C—C bond forming reaction of o‐alkynylacetophenone derivatives and CO2 to afford dihydroisobenzofurans such as (IV), (VI), and (VIII)/(IX) via selective 5‐exo‐dig cyclization is reported.
ChemInform Abstract: Metal‐Free Synthesis of 3,3‐Disubstituted Oxindoles via 1,2‐Alkylarylation of Activated Alkenes with Alcohols.
Authors: Zhou Zhao‐Zhao , Hua Hui‐Liang , Luo Jian‐Yi , Chen Zi‐Sheng , Zhou Ping‐Xin , Liu Xue‐Yuan , Liang Yong‐Min
Published: March 2014

A great variety of title compounds are economically and efficiently synthesized by a metal‐free strategy through an oxidative radical cyclization.
ChemInform Abstract: Asymmetric Mannich Reactions of Imidazo[2,1‐b]thiazole‐derived Nucleophiles with (Ss)‐N‐tert‐Butanesulfinyl (3,3,3)‐Trifluoroacetaldimine.
Authors: Mei Haibo , Xie Chen , Wu Lingmin , Soloshonok Vadim A. , Han Jianlin , Pan Yi
Published: March 2014

In the title reaction, the N‐tert‐butylsulfinyl group serves not only as an efficient chiral auxiliary, but also as an amine protecting group.
ChemInform Abstract: Synthesis and Evaluation of α‐Ketotriazoles and α,β‐Diketotriazoles as Inhibitors of Mycobacterium tuberculosis.
Authors: Lherbet Christian , et al. et al.
Published: March 2014

New α,β‐diketotriazoles (IIIb,c) exhibit interesting antituberculosis activities and no apparent cytotoxicity towards colorectal cancer HCT116 and normal fibroblast GM637H human cell lines.
ChemInform Abstract: Preparation of 1,4,5‐Trisubstituted 5‐Acyl‐1,2,3‐triazoles by Selective Acylation Between Copper(I)‐Carbon(sp) and Copper(I)‐Carbon(sp2) Bonds with Acyl Chlorides.
Authors: Wang Bo , Liu Nan , Shao Changwei , Zhang Qun , Wang Xinyan , Hu Yuefei
Published: March 2014

An efficient and operationally simple one‐pot procedure is developed for the synthesis of a wide range of triazoles through cycloaddition of copper(I)‐alkynes with azides and subsequent acylation.
ChemInform Abstract: Synthesis and anti‐Parasitic Activity of a Novel Quinolinone—Chalcone Series.
Authors: Roussaki Marina , Hall Belinda , Costa Lima Sofia , Cordeiro da Silva Anabela , Wilkinson Shane , Detsi Anastasia
Published: March 2014

A series of novel quinolinone‐chalcone hybrids (III) (14 examples) and pyrazole analogues such as (V) are designed, synthesized and evaluated for their antiparasitic activity.
ChemInform Abstract: Dynamic Kinetic Cross‐Coupling Strategy for the Asymmetric Synthesis of Axially Chiral Heterobiaryls.
Authors: Ros Abel , Estepa Beatriz , Ramirez‐Lopez Pedro , Alvarez Eleuterio , Fernandez Rosario , Lassaletta Jose M.
Published: March 2014

The deracemization of configurationally stable 2‐arylpyridines (I) and analogues with aryl boroxines (II) is achieved by applying a dynamic kinetic asymmetric cross‐coupling strategy.
ChemInform Abstract: New Di(hetero)arylethers and Di(hetero)arylamines in the Thieno[3,2‐b]pyridine Series: Synthesis, Growth Inhibitory Activity on Human Tumor Cell Lines and Non‐tumor Cells, Effects on Cell Cycle and on Programmed Cell Death.
Authors: Queiroz Maria‐Joao R. P. , et al. et al.
Published: March 2014

New compounds (IIIc), (Vb), and (Vc) show the strongest growth inhibitory activities on human tumor cell lines and cause pronounced alterations in the cell cycle profile of NCI‐H460 cells.
ChemInform Abstract: Phosphine‐Catalyzed Formal Vinylogous Aldol Reaction of γ‐Methyl Allenoates with Aldehydes: Easy Access to 1,3‐Dioxanes and Dienols.
Authors: Qin Zifeng , Ma Renqin , Xu Silong , He Zhengjie
Published: March 2014

The reaction of γ‐methyl allenoates is investigated in the presence of different triarylphosphines as catalysts delivering chemo‐ and stereoselective formation of 1,3‐dioxanes or 6‐hydroxy‐2,4‐dienoates.
ChemInform Abstract: Microwave Assisted Synthesis of Dihydrobenzo[4,5]imidazo[1,2‐a]pyrimidin‐4‐ones; Synthesis, in vitro Antimicrobial and Anticancer Activities of Novel Coumarin Substituted Dihydrobenzo[4,5]imidazo[1,2‐a]pyrimidin‐4‐ones.
Authors: Puttaraju Kallimeledoddi B. , Shivashankar Kalegowda , Chandra Chandra , Mahendra M. , Rasal Vijaykumar P. , Vivek Ponnuru N. Venkata , Rai Khushboo , Chanu Maibam Beebina
Published: March 2014

Most of the synthesized title compounds (V) (8 examples) exhibit good antibacterial, antifungal, and cytotoxic activities.
ChemInform Abstract: Stereoselective Palladium‐Catalyzed Alkenylation and Alkynylation of Thioglycosides.
Authors: Brachet Etienne , Brion Jean‐Daniel , Alami Mouad , Messaoudi Samir
Published: March 2014

A method to prepare thioglycoside derivatives is developed using Pd‐catalyzed glycosylation of thioglycosides with alkenyl and alkynyl halides.
ChemInform Abstract: The Synthetic Preparation of Naturally‐Occurring Aromatase Inhibitors, Morachalcone A (Ia), Isogemichalcone B (Ib), and Isogemichalcone C (Ic).
Authors: Brandt Drew R. , Pannone Kristina M. , Romano Joseph J. , Casillas Eduard G.
Published: March 2014

Title compounds (I) are synthesized in up to 10 steps in 4 to 8% overall yields.
ChemInform Abstract: Palladium‐Catalyzed Cross‐Dehydrogenative Functionalization of C(sp2)—H Bonds
Authors: Wu Yinuo , Wang Jun , Mao Fei , Kwong Fuk Yee
Published: March 2014

Review: recent advances with a focus on the period from 2011 to early 2013; 96 refs.
ChemInform Abstract: Landau Theory Description of Polar Smectic Structures
Authors: Dolganov P. V. , Kats E. I.
Published: March 2014

Review: 91 refs.