Investigating Inflammatory Mediators
Order ID 53563633773 Type Essay Writer Level Masters Style APA Sources/References 4 Perfect Number of Pages to Order 5-10 Pages
Investigating Inflammatory Mediators
Specific findings from prior clinical work specifically investigating inflammatory mediators have demonstrated differential biomarker patterns in umbilical cord serum from infants stratified by preterm and CP status . Of the
potential inflammation markers that differed between cases and controls, the markers were lower (based on medians) in the preterm CP cases relative to controls but were higher, relative to controls, in the term CP cases.
Sampling from a different compartment (plasma) and using a different approach, a study by Lin et al. (2010) comparing school-age children who were former preterm births reported higher cytokine responses (increased tumor
necrosis factor-alpha [TNF-α] plasma levels and greater mRNA levels of toll- like receptor four [TLR4]) among those children with CP who were preterm relative to controls (children born preterm with normal development) .
Another study of blood, using a serial approach (i.e., repeated measures) in over 900 preterm infants, documented elevated concentration values of myriad inflammatory- relevant mediators which were associated with different
risk profiles depending on the sampling day . Using CSF from preterm infants with brain injury, Douglas- Escobar and Weiss (2012) documented combinations of biomarker concentration values that could be used to
inform clinical decision making . Using blood from a sample of children with and without CP, Zareen (2020)  found significantly increased levels of erythropoie- tin at baseline in children with CP compared with chil- dren in
the comparison group. In response to challenge (lipopolysaccharide), both groups had appropriate and comparable response profiles for interleukin-8, vascular endothelial growth factor (VEGF), TNF-α, and granu- locyte–
macrophage colony-stimulating factor (GM- CSF) levels. The children with CP showed a statistically significant lipopolysaccharide hypo-responsiveness profile for interleukin-1a, interleukin-1b, interleukin-2, and interleukin-6
levels. Collectively, the work to date consistently shows immune and inflammatory differ- ences in children with CP.
There remain gaps in our knowledge about the spe- cific linkages among various immune and related mechanisms driving hypothesized persistent inflamma- tory states in children with CP, and the relation among the various
biomarkers, risk factors, and specific out- comes. The overall generality of the findings to date for the CP population is limited by two kinds of problems, namely the relative difficulties in establishing valid pre- clinical models for
this purpose  and the extreme paucity of clinically-relevant biomarker research within this high-need vulnerable patient group. For example, the relation among the CSF biomarkers investigated by Douglas-Escobar and Weiss
(2012) and CP as an outcome was not clear . More biomarker data of a comparable kind from the same and different compart- ments across the same and different age groups are needed. To the best of our knowledge, no
comparable work has investigated inflammatory-relevant molecu- lar biomarkers in CSF from children, adolescents, and young adults with CP.
The purpose of this preliminary investigation was exploratory. The design was cross-sectional using a single time point for specimen collection from a clini- cal sample. There were two specific aims. The first aim was to
document levels of inflammatory and related molecules in CSF in a sample primarily of school-age children with CP. To do so, for detectable analytes, participants were arrayed along each analyte’s con- centration gradient. The
second aim was to examine clinically relevant grouping variables (e.g., CP sever- ity, term/preterm birth) to identify any potentially rel- evant correlational patterns among the molecules. Our intent was to extend the work initiated
by Kaukola and Lin (described above) using a clinical sample in which standard-of-care surgical interventions were leveraged to gain access to CSF for future hypothesis-generating research purposes.
Page 3 of 13Goracke‑Postle et al. BMC Neurol (2021) 21:384
Methods Protocol approval This study was approved by the Institutional Review Board (IRB, #0809M46301) of the University of Minne- sota. Written informed consent was obtained from each participant or legal representative
Participants This study utilized a single-time point cross-sectional design. Twenty-eight individuals (82% male) with CP participated (mean age = 9.74 years, SD = 4.36; range = 4–23). Specific CP diagnoses included: quad-
riplegia (n = 17), diplegia (n = 5), and triplegia (n = 5). Participants were included in the study if they (a) had cerebral palsy, (b) were between 3–25 years of age, and (c) were scheduled for initial intrathecal baclofen (ITB) pump
implant. Individuals were excluded if (a) they had an existing cerebral shunt; or (b) they had compounded dosing (i.e., opioid adjunctive to baclofen) through their pump. The participants were already characterized clini- cally
using the Gross Motor Functional Classification System for Cerebral Palsy (GMFCS) to categorize gross motor function. The GMFCS is a 5-level classification system based on self-initiated movement with emphasis on truncal
control and walking. The GMFCS is widely used clinically and in classifying individuals with CP for research studies .
For the subgroup comparisons, the breakdown of par- ticipant demographics was as follows: males (n = 23) and females (n = 5); non-quadriplegia (n = 11) and quad- riplegia (n = 17); Caucasian (n = 26) and other (n = 2); spastic
CP (n = 13) and mixed tone CP (n = 12); term birth, defined as 37 weeks or later (n = 6), preterm birth, defined as 28–37 weeks (n = 12) and extremely preterm birth, defined as less than 28 weeks (n = 9); seizure (n = 6) and no
seizure (n = 16) (Table 1).
CSF collection Patients were consented in accordance with an approved IRB protocol. If consent was given, CSF was collected during a standard-of-care surgical procedure (ITB pump implant). In all cases, the surgery proceeded
as usual until the spinal catheter had been placed. Then, the neurosur- geon collected 10–20 ml of CSF from the spinal catheter placed well above the spinal puncture site. This method avoided contaminating the collected CSF
Immediately following collection, the CSF was placed on wet ice (+ 4 °C) and transported to a cold room for processing, centrifuged at 3000 rpm × 5 min, pipetted into 100 and 250 µL aliquots, flash frozen in liquid nitro- gen
and archived at -80 °C. After specimen collection,
the patient was monitored closely following routine operative and post-operative procedures. There were no adverse events.
CSF analyte analysis CSF was analyzed using conventional biochemical meth- ods based on commercially available enzyme-linked immunosorbent assay (ELISA) kits and expression lev- els of each marker were quantified.
Specifically, samples were tested by the Cytokine Reference Laboratory (CRL, University of Minnesota). This is a CLIA’88-licensed facility (license #24D0931212). Samples were analyzed for adrenocorticotropic hormone (ACTH),
agouti- related peptide (AgRP), brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), follicle-stimulating hormone (FSH), growth hormone (GH), luteinizing hormone (LH), prolactin (PRL), and thyroid
stimulating hormone (TSH) using the “Human Brain-Derived Protein Panel” on the Luminex platform and done as a multi-plex (Luminex instrument—Bioplex 100 [Bio-Rad, 1000 Alfred Nobel Drive, Hercules, CA, 94547], Software:
bio-plex Manager 4.0). The polysty- rene bead set (cat. # HPT-66 K-09) with kit lot number 1757143 was used. Kits/reagents were purchased from EMD Millipore Corporation, Billerica, MA. Interferon α2 (IFNα2), interleukin-1α (IL-
1α), interleukin-1ra (IL-1ra), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), interleukin-12p40 (IL-12p40), interleukin-12p70 (IL-12p70), interferon gamma-induced protein 10 (IP- 10), monocyte chemotactic protein
(MCP-1), mac- rophage inflammatory protein 1β (MIP1β), regulated on activation normal T expressed and secreted (RANTES), and tumor necrosis factorα (TNFα) were analyzed using the “Cytokine/Chemokine Panel 1” on the
Luminex plat- form and done as a multi-plex (Luminex instrument— Bioplex 100 [Bio-Rad, 1000 Alfred Nobel Drive, Hercules, CA, 94547], Software: bio-plex Manager 4.0). The poly- styrene bead set (cat. # MPXHCYTO-60 K-14)
with kit lot number 1757142 was used. Kits/reagents were pur- chased from EMD Millipore Corporation, Billerica, MA. Dynorphin A, neuropeptide Y, somatostatin, β endor- phin, cortisol, neurotensin, orexin A, substance P, mela-
tonin, oxytocin, and melanocyte-stimulating hormone (α-MSH) were analyzed using the “Human Neuropeptide Panel” on the Luminex platform and done as a multi-plex (Luminex instrument—Bioplex 100 Bio-Rad, 1000 Alfred
Nobel Drive, Hercules, CA, 94547], Software: bio-plex Manager 4.0). The polystyrene bead set (cat. # HNP- 35 K-08) with kit lot number 1823005 was used. Kits/rea- gents were purchased from EMD Millipore Corporation, Billerica,
Samples were assayed according to manufacturer’s instructions. ELISA employ the quantitative sandwich
Page 4 of 13Goracke‑Postle et al. BMC Neurol (2021) 21:384
enzyme immunoassay technique. The absorbance is measured on the microtiter plate reader (Bio-Rad model 550). The intensity of the color formed is proportional to the concentration of the sample. Fluorescent color- coded
beads coated with a specific capture antibody were added to each sample. After incubation, and washing, biotinylated detection antibody was added, followed by phycoerythrin-conjugated streptavidin. The beads were read on a
Luminex instrument (Bioplex 100) which is a dual-laser fluidics-based instrument. One laser deter- mines the analyte being detected via the color coding; the other measures the magnitude of the PE signal from the
detection antibody which is proportional to the amount of analyte bound to the bead. Samples were tested in duplicate and values were interpolated from 5 parameter- fitted (5PL) standard curves.
Statistical analyses Data analysis was exploratory and relied on visual analy- sis, descriptive statistics, and correlational analyses. First, visual analysis of each analyte was conducted to under- stand its distributional form and to
identify potential outliers. Further, measures of central tendency (means,
Table 1 Participant health information; M ± SD or n (%)
Note: gestational age was not available for one participant; Term birth = born 37 weeks gestation or later; preterm birth = born at 28–37 weeks gestation; extremely preterm birth = born at less than 28 weeks gestation; CP
Cerebral Palsy, GMFCS Gross Motor Function Classification System, level I ambulant without assistance, level II ambulant without assistive devices, limitations outside the home, level III ambulant with assistive devices,
wheelchair required outside the home, level IV non‑ ambulatory, self‑mobile in wheelchair with limitations, level V non‑ambulatory, self‑mobility very limited
Complete sample (n = 28)
Term birth (n = 6)
Preterm birth (n = 12)
Extremely preterm birth (n = 9)
Male 23 (82.1) 3 (50.0) 11 (91.7) 9 (100)
Caucasian 26 (92.9) 6 (100.0) 12 (100.0) 8 (88.9)
African American 1 (3.6) 0 (0) 0 (0) 1 (11.1)
Other (not specified) 1 (3.6) 0 (0) 0 (0) 0 (0)
None 16 (57.1) 3 (50.0) 7 (58.3) 6 (66.7)
History of seizure/diagnosis of epilepsy 6 (21.4) 1 (16.7) 4 (33.3) 1 (11.1)
Questionable seizure activity 3 (10.7) 1 (16.7) 0 (0) 1 (11.1)
Missing 3 (10.7) 1 (16.7) 1 (8.3) 1 (11.1)
Hemiplegia 1 (3.6) 0 (0) 1 (8.3) 0 (0)
Diplegia 5 (17.9) 2 (33.3) 3 (25.0) 0 (0)
Triplegia 5 (17.9) 1 (16.7) 0 (0) 4 (44.4)
Quadriplegia 17 (60.7) 3 (50.0) 8 (66.7) 5 (55.6)
Level I 3 (10.7) 0 (0) 3 (25.0) 0 (0)
Level II 3 (10.7) 1 (16.7) 0 (0) 2 (22.2)
Level III 7 (25.0) 3 (50.0) 2 (16.7) 2 (22.2)
Level IV 8 (28.6) 1 (16.7) 4 (33.3) 3 (33.3)
Level V 6 (21.4) 1 (16.7) 2 (16.7) 2 (22.2)
Missing 1 (3.6) 0 (0) 1 (8.3)
Spastic 13 (46.4) 4 (66.7) 5 (41.7) 4 (44.4)
Mixed tone 12 (42.9) 2 (33.3) 5 (41.7) 4 (44.4)
Missing 3 (10.7) 0 (0) 2 (16.7) 1 (11.1)
Current feeding tube
Yes 8 (28.6) 2 (33.3) 2 (16.7) 4 (44.4)
No 19 (67.9) 4 (66.7) 10 (83.3) 5 (55.5)
Missing 1 (3.6) 0(0) 0 (0) 0 (0)
Days in NICU at birth Range of NICU days
76.25 ± 67.04 (0–300)
3.50 ± 7.00 (0–14)
52.73 ± 29.72 (10–120)
137.33 ± 64.76 (90–300)
Page 5 of 13Goracke‑Postle et al. BMC Neurol (2021) 21:384
medians) and variation (standard deviations, coefficients of variation) were calculated for each analyte.
Second, to understand the associations between ana- lytes a series of pairwise scatterplots and Pearson prod- uct-moment correlations were computed between each possible pair of analytes for the entire sample. The corre-
lations were tested for statistical significance against the null hypothesis of r = 0. Parallel analyses and plots were generated for the data set with missing data imputed with the lower limit / sensitivity number. Given the large
number of correlations tested (528), Type 1 errors were controlled for by using the false discovery rate correction discussed by Benjamini & Hochberg (1995). Correlations were considered against an alpha = 0.05, after the false
discovery rate correction was applied.
Third, to understand how the associations between analytes varied by subgroups, the correlational analyses described above were repeated for each of the gestational term subgroups. Given the large number of compari- sons
involved in these analyses, an alpha = 0.001 was set
for each test after the false discovery correction rate was applied.
Finally, differences in correlations between subgroups were also tested. To be included in between-group analysis, each correlation first had to be statistically significant within the subgroups. Thirty-two pairs of correlations were
statistically significant across all sub- groups. Then the identified correlations were tested against one another to check whether they were sig- nificantly different by birth status. To do this, Fisher’s r to z transformation was used
to calculate the differ- ence in correlations that met the criteria for inclusion, and tested for statistical significance of that difference; p ≤ 0.05.
Results CSF analyte expression in CP subjects Detectable CSF analytes were broken down, broadly, by the following categories: hormonal/endocrine brain-derived peptides or proteins: ACTH, AgRP,
Fig. 1 Visual representation of the direction and strength of the Pearson’s correlation coefficients between all 33 analytes assayed. Positive (blue), negative (red), strong (dark shading), and weak (light shading) correlations are
Investigating Inflammatory Mediators
NO RESPONSE POOR / UNSATISFACTORY SATISFACTORY GOOD EXCELLENT
50% of the total points)
Zero points: Student failed to submit the final paper. 20 points out of 50: The essay illustrates poor understanding of the relevant material by failing to address or incorrectly addressing the relevant content; failing to identify or inaccurately explaining/defining key concepts/ideas; ignoring or incorrectly explaining key points/claims and the reasoning behind them; and/or incorrectly or inappropriately using terminology; and elements of the response are lacking. 30 points out of 50: The essay illustrates a rudimentary understanding of the relevant material by mentioning but not full explaining the relevant content; identifying some of the key concepts/ideas though failing to fully or accurately explain many of them; using terminology, though sometimes inaccurately or inappropriately; and/or incorporating some key claims/points but failing to explain the reasoning behind them or doing so inaccurately. Elements of the required response may also be lacking. 40 points out of 50: The essay illustrates solid understanding of the relevant material by correctly addressing most of the relevant content; identifying and explaining most of the key concepts/ideas; using correct terminology; explaining the reasoning behind most of the key points/claims; and/or where necessary or useful, substantiating some points with accurate examples. The answer is complete. 50 points: The essay illustrates exemplary understanding of the relevant material by thoroughly and correctly addressing the relevant content; identifying and explaining all of the key concepts/ideas; using correct terminology explaining the reasoning behind key points/claims and substantiating, as necessary/useful, points with several accurate and illuminating examples. No aspects of the required answer are missing. Use of Sources (worth a maximum of 20% of the total points). Zero points: Student failed to include citations and/or references. Or the student failed to submit a final paper. 5 out 20 points: Sources are seldom cited to support statements and/or format of citations are not recognizable as APA 6th Edition format. There are major errors in the formation of the references and citations. And/or there is a major reliance on highly questionable. The Student fails to provide an adequate synthesis of research collected for the paper. 10 out 20 points: References to scholarly sources are occasionally given; many statements seem unsubstantiated. Frequent errors in APA 6th Edition format, leaving the reader confused about the source of the information. There are significant errors of the formation in the references and citations. And/or there is a significant use of highly questionable sources. 15 out 20 points: Credible Scholarly sources are used effectively support claims and are, for the most part, clear and fairly represented. APA 6th Edition is used with only a few minor errors. There are minor errors in reference and/or citations. And/or there is some use of questionable sources. 20 points: Credible scholarly sources are used to give compelling evidence to support claims and are clearly and fairly represented. APA 6th Edition format is used accurately and consistently. The student uses above the maximum required references in the development of the assignment. Grammar (worth maximum of 20% of total points) Zero points: Student failed to submit the final paper. 5 points out of 20: The paper does not communicate ideas/points clearly due to inappropriate use of terminology and vague language; thoughts and sentences are disjointed or incomprehensible; organization lacking; and/or numerous grammatical, spelling/punctuation errors 10 points out 20: The paper is often unclear and difficult to follow due to some inappropriate terminology and/or vague language; ideas may be fragmented, wandering and/or repetitive; poor organization; and/or some grammatical, spelling, punctuation errors 15 points out of 20: The paper is mostly clear as a result of appropriate use of terminology and minimal vagueness; no tangents and no repetition; fairly good organization; almost perfect grammar, spelling, punctuation, and word usage. 20 points: The paper is clear, concise, and a pleasure to read as a result of appropriate and precise use of terminology; total coherence of thoughts and presentation and logical organization; and the essay is error free. Structure of the Paper (worth 10% of total points) Zero points: Student failed to submit the final paper. 3 points out of 10: Student needs to develop better formatting skills. The paper omits significant structural elements required for and APA 6th edition paper. Formatting of the paper has major flaws. The paper does not conform to APA 6th edition requirements whatsoever. 5 points out of 10: Appearance of final paper demonstrates the student’s limited ability to format the paper. There are significant errors in formatting and/or the total omission of major components of an APA 6th edition paper. They can include the omission of the cover page, abstract, and page numbers. Additionally the page has major formatting issues with spacing or paragraph formation. Font size might not conform to size requirements. The student also significantly writes too large or too short of and paper 7 points out of 10: Research paper presents an above-average use of formatting skills. The paper has slight errors within the paper. This can include small errors or omissions with the cover page, abstract, page number, and headers. There could be also slight formatting issues with the document spacing or the font Additionally the paper might slightly exceed or undershoot the specific number of required written pages for the assignment. 10 points: Student provides a high-caliber, formatted paper. This includes an APA 6th edition cover page, abstract, page number, headers and is double spaced in 12’ Times Roman Font. Additionally, the paper conforms to the specific number of required written pages and neither goes over or under the specified length of the paper.
GET THIS PROJECT NOW BY CLICKING ON THIS LINK TO PLACE THE ORDER
CLICK ON THE LINK HERE: https://phdwriters.us/orders/ordernow
Also, you can place the order at www.collegepaper.us/orders/ordernow / www.phdwriters.us/orders/ordernow
Do You Have Any Other Essay/Assignment/Class Project/Homework Related to this? Click Here Now [CLICK ME] and Have It Done by Our PhD Qualified Writers!!
Investigating Inflammatory Mediators