Parental control is characterized by excessive monitoring of their children’s activities, discouragement of independent thinking and high levels of intrusion

this method holds the potential to become a diagnostic tool for individuals with Long COVID, enabling comprehensive evaluation of microclot presence, concentration, and size

erall, our novel, rapid, cell-free flow cytometric detection method, utilizing imaging flow cytometry, reveals the presence of amyloid fibrin(ogen) microclots in platelet poor plasma. These microclots represent a potential breakthrough in clinical diagnostics, offering valuable insights into clotting pathologies.

standardized

addition, these parameters may be useful in monitoring the effectiveness of treatment by tracking the changes in them

By measuring these variables, clinicians can potentially guide treatment decisions by tailoring therapies to address the specific clotting abnormalities observed in a patient.

we can anticipate the ability of non-imaging flow cytometers to prove effective in making similar disseminations for the purposes of high-throughput diagnostics.

These variables have the potential to guide clinical decisions about diagnosis and treatment

re we have demonstrated significant differences in the concentration, mean area, and count in various area ranges between Long COVID and controls.

ow cytometry has been utilized in pathology laboratories for a considerable amount of time, making it a more appropriate and feasible means of identifying microclots in people with Long COVID and other conditions with clotting pathologies

results demonstrate that even at a 20× objective, there are notable variations in size and fluorescence between Long COVID and controls

we determined that some of this background activity could be attributed to cellular debris from damaged endothelial cells and fibrinogen strands. These findings align with our previous publications, which suggest the presence of endothelial damage in Long COVID patients

which indicates larger sized amyloid

provide additional evidence of significant variations in the density and distribution of microclots between the control group and individuals with Long COVID

in Long COVID not only is the number and concentration of microclots significantly higher, but their size is significantly larger when compared to controls

indicate that there are significant differences in the microclot area distribution between controls and Long COVI

our flow cytometry analysis confirmed that amyloid microclot objects/mL (*p < 0.05) and microclot mean area (*p < 0.05) were significantly increased in Long COVID compared to controls.

Furthermore, conventional flow cytometers are incapable of providing spatially resolved information, which is frequently vital for quantifying intricate cellular phenotypes

ontrast to the purely numerical quantitative measurements provided by conventional flow cytometry, analysing cell images can also help eliminate erroneous results, such as distinguishing between cells and debris

Due to its quantitative and multiparametric characteristics, as well as the analytical capacity to process up to 50,000 cells per second, flow cytometry is widely recognized as the gold standard technique for counting and characterizing cells in complex samples

Flow cytometry is a widely used analytical approach for counting, analysing, and segregating cells that are suspended in a fluid stream

imaging flow cytometer that allowed us to combine cell imaging as well as the high-event-rate nature of a conventional flow cytometer.

Previously, we have used fluorescence microscopy and a semi-quantitative grading system to evaluate the presence of significant microclotting in Long COVID compared to healthy individuals

detection of fibrin(ogen) amyloid microclots using imaging flow cytometry in Long COVID patients.

Representative micrographs of microclots in (A) controls and (B) Long COVID

amyloid in nature

it is evident that controls exhibit smaller microclots with a lower fluorescence intensity. Conversely, Long COVID patients tend to have larger microclots

we realized that whilst bigger microclots are obviously problematic due to the potential for vessel blockage, an increase in smaller microclots has potential clinical significance as well. These smaller microclots may not cause a significant blood vessel blockage but may contribute to platelet activation and endothelial damage.

Thereafter, each area range increased by 10 μm

Long COVID patients showed a significant increase

The first area range was between 0 and 100 μm2 (10 μm), which is the approximate equivalent to a capillary diameter.

microclot area distribution between controls and Long COVID patients. Here a clear dissimilarity is observed in the microclot area distribution between the control and Long COVID patients.

results showed that the Long COVID patients had significantly higher concentration of microclots (*p < 0.05) as well as a higher microclot mean area (*p < 0.05) compared to controls.

there is currently no clinical pathology test available to identify patients [

2.2.4. Data analysis All flow cytometry data analysis was performed using the IDEAS 6.2 software developed for the Amnis® FlowSight® Imaging Flow Cytometer. For analysis, only ThT positive events were collected during acquisition. A gate was also established to eliminate events with weak fluorescence signal (see supplementary material). For analysis, we included the following microclot parameters: objects/mL, mean area, and area distribution. GraphPad Prism software (version 9.5.0) was used to perform statistical analysis on all data. To determine whether data were normally distributed, the Shapiro-Wilk normality test was applied. Parametric data were analyzed using unpaired, one-tailed t-tests with Welch's correction to test for statistical significance. For non-parametric data, an unpaired, one-tailed Mann-Whitney test was utilized to test for statistical significance. Statistical significance was taken to be at p < 0.05 (and was also marked with asterisks with * = p < 0.05; ** = p < 0.01). Data are represented as either mean ± standard deviation (parametric data) or as median ± [Q1-Q3] (non-parametric data).

460

217

100

31

415

289

228

185

2686282

88644

52934

21026

20 controls and 40 Long COVID patients.

2.2.3. Flow cytometry For data acquisition, we used an Amnis® FlowSight® Imaging Flow Cytometer from Luminex. Imaging flow cytometry combines the advantages of microscopy's single-cell image acquisition with the high-event-rate capacity, quantification, and statistical reliability of traditional flow cytometry. For acquisition, a template was created where the 405 nm and 488 nm lasers were turned on and a gate was established to collect all ThT positive (ThT +) events by using a negative control (water and ThT), a second negative control (PPP without ThT), and a standard PPP control as reference (see supplementary material). To reduce background noise and file size, we only collected events within the ThT-positive gate for analysis. The brightfield images were viewed in Channel 1 and ThT-positive events were viewed in Channel 7 using the 405 laser. All samples were acquired using the same acquisition template for 5 min each. Given that the number of microclots per mL ranged from 21,000–52,000 (Table 2) per sample, this typically involved the detection of 400–3000 microclots per sample. The time required for IDEAS to analyse a sample using the analysis template was approximately 2 min, which could vary based on the number of events in the sample. Overall, the total time required to process a single sample from acquisition to analysis was approximately 7 min.

2.2.2. Blood sample collection and sample preparation Either a qualified phlebotomist or a medical practitioner drew blood samples. To obtain PPP, whole blood (WB) from sodium citrate tubes was centrifuged at 3000×g for 15 min at room temperature, and the supernatant PPP samples were collected in 1.5 mL Eppendorf tubes and stored at −80 °C. On the day of analysis, the stored PPP was brought to room temperature and exposed to thioflavin T (ThT), a fluorescent dye that binds to amyloid protein, at a final concentration of 0.03 mM (obtained from Sigma-Aldrich, St. Louis, MO, USA). The exposure lasted 30 min, during which time the plasma was protected from light

evidence that supports the use of imaging flow cytometry

we used an imaging flow cytometer, the results indicate that these signals will be readily detectable on a non-imaging instrument.

flow cytometric methods are widely available, and since the 1980s have been capable of detecting micron-sized objects [

fluorescence microscopy can identify microclots and platelet hyperactivation

We also previously confirmed entrapped inflammatory molecules within these samples using proteomics analysis

Previously

flow cytometric method for detecting fibrin(ogen) amyloid microclots using long COVID

we provided fluorescence microscopic evidence that there is a significant insoluble fibrin amyloid microclot load along with platelet hyperactivation in both acute and Long COVID

the most rapid gain in immunologic experience

SUPREME COURT OF COLORADOOFFICE OF THE CHIEF JUSTICECOURT APPOINTMENTS THROUGHTHE OFFICE OF RESPONDENT PARENTS’ COUNSEL

The mental and physical health of all individuals involved

The interaction and interrelationship of the child with his or her parents, his or her siblings, and any other person who may significantly affect the child's best interests

wishes of the child's parents as to parenting time

Rule 38 - Right to Trial by Jury(a) Exercise of Right. Upon the filing of a demand and the simultaneous payment of the requisite jury fee by any party in actions wherein a trial by jury is provided by constitution or by statute, including actions for the recovery of specific real or personal property, with or without damages, or for money claimed as due on contract, or as damages for breach of contract, or for injuries to person or property, all issues of fact shall be tried by a jury. The jury fee is not refundable; however, a demanding party may waive that party's demand for trial by jury pursuant to section (e) of this rule.

II. Legal Standards Under C.R.C.P. 56.Summary judgment “shall be rendered forthwith, if the pleadings, depositions, answers tointerrogatories, and admissions on file, together with the affidavits, if any, show that there is nogenuine issue as to any material fact and the moving party is entitled to a judgment as a matter oflaw.” C.R.C.P. Rule 56(c). A material fact is one whose resolution will affect the outcome ofthe case. Peterson v. Halsted, 829 P.2d 373, 375 (Colo. 1992). “A genuine issue of material factcannot be established simply by allegations in pleadings or argument; rather, the opposing partymust set forth specific facts by affidavit or otherwise showing that there is a genuine issue fortrial.” People ex rel A.C., 170 P.3d 844, 846 (Colo. App. 2007). “The purpose of summaryjudgment is to permit the parties to pierce the formal allegations of the pleadings and save thetime and expense connected with trial when, as a matter of law, based on undisputed facts, oneparty could not prevail.” Id. An uncontradicted showing of facts probative of right to judgmentleaves a trial court with no alternative but to conclude that no genuine issues of material factexist. Civil Service Comm’n v. Pinder, 812 P.2d 645, 649 (Colo. 1991); Terrell v. Walter E.Heller & Co., 439 P.2d 989, 991 (Colo. 1968).

(d) If it appears from the evidence that the child may have a mental health disorder

he hearing should be held with-in 60-90days of the date of service of the petitio

demand that the case then be heard at trial by a jury of six people, by a judge, or by ajuvenile magistrate

Dependency&Neglect

SFAs have been shown to act as ligands for toll-like receptors (TLRs), which are usually reserved for pathogen recognition and thereby promote pro-inflammatory gene expression in BV2 and primary mouse microglia

when there is such variation in the connection in which the words are used as reasonably to warrant the conclusion that they were employed in different parts of the act with different intent."95 Context and statutory history can override the presumption.

A motion can be written or spoken, as the relevant rules require.

Summary Judgment

motion for summary judgment shall be filed no later than 91 days (13 weeks) prior to tria

prima facie evidence that such child is neglected or dependent,

on the motion of any interested party

but in no instance shall such hearing be held later than ninety days

Requirements The health center must operate in a manner such that no patient shall be denied service due to an individual’s inability to pay.1

discounts are adjusted on the basis of the patient's ability to pay.

Provide services regardless of the patient’s ability to pay

oral health

access to

What is a Health Center?

Early recognition is typically an important factor in delivering effective therapy.

in the vast majority of these conditions defined by and/or accompanied by microvascular thrombosis, the diagnosis is typically made clinically and sometimes confirmed histologically if possible.

The resolution of a computed tomography angiogram or magnetic resonance angiogram typically does not provide information about the structure of these microscopic vessels. Even a conventional angiogram of the brain does not resolve vessels smaller than 500 µm

correctly identifying that there is an issue in that physiologic compartment.

Clinical overview of microvascular thrombosis

Imaging of the microvasculature

key obstacle

Neutrophils play a major role in tissue damage incurred during IRI. Activated neutrophils are a major source of ROS,

An anti IL-8 antibody prevented pulmonary neutrophil infiltration and tissue injury in a rabbit model of lung IR

The introduction of humanised antibodies including etanercept and infliximab, has provided encouraging results in the treatment of other TNF-α-mediated inflammatory diseases, including a number of forms of arthritis and inflammatory bowel disease (reviewed in18)

Hypoxia and IRI both induce the expression of numerous cytokines, including tumour necrosis factor-alpha (TNF- α,) interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8) and platelet activating factor (PAF), in association with elevations in activity of the transcription factor, NF-kB (reviewed in 16) these cytokines are released systemically and are thus important in the development of systemic inflammatory response syndrome and ultimately multi-system organ failure.

Endothelin-1 inhibitors, including bosentan and tezosentan, inhibit neutrophil infiltration, increase functional capillary density, microvascular perfusion and hence tissue viability and function Following IR

systemic distortion of NO kinetics by administering NOS inhibitors would be likely to induce wide-ranging physiological disturbances.

inhibition of NOS activity using a non-specific NOS inhibitor greatly reduced the severity of muscle damage

considerable evidence for a principal role for nitric oxide in the aetiology of IRI. Myocardial IRI has been well studied, with paradoxical results, where low doses of no were found to be protective and high doses harmful. The influence of NO in skeletal muscle IRI has been less well characterized, with some studies suggesting that NO may potentiate cytotoxicity and others suggesting a beneficial role for NO in extremity IR

The reduction in ENOS levels that occurs in IRI may therefore predispose to vasoconstriction, a common response seen in IRI

activated neutrophils, leads to further pulmonary neutrophil accumulation.

The lung produces leukotrienes following remote IRI. The direct effects of leukotrienes on pulmonary micro vessels lead to increased permeability, transient pulmonary hypertension and the activation of the endothelium to produce thromboxane, resulting in additional vaso-constriction.

leukotrienes C4, D4, and E4 modify the endothelial cytoskeleton, leading to increased vascular permeability and also enhance smooth muscle contraction, resulting in vasoconstriction

Leukotrienes

administration of thromboxane A2 antagonists may offer therapeutic potential

thromboxane synthase inhibitors and synthetic thromboxane A2 receptor antagonists prevented pulmonary leukosequestration, thereby increasing blood flow to reperfused tissues and preserving tissue viability and function

rapid rise in pulmonary artery pressure and a subsequent increase in pulmonary microvascular permeability

promoting vasoconstriction and platelet aggregation

thromboxane A2

demonstrated better post-operative myocardial oxygen consumption after coronary artery bypass surgery8 and improved muscle blood flow following skeletal muscle IRI.9

administration of prostacyclin was shown to improve postoperative graft function.

Prostaglandins, synthesised from arachidonic acid via the cyclo-oxygenase pathway, have a protective vasodilatory effect in IRI. However, since prostaglandins are short-lived molecules, their rapid depletion subsequently leads to uninhibited vasoconstriction, reduced local blood flow and exacerbation of ischaemia

ROS initiate lipid peroxidation of cellular membranes, releasing arachidonic acid, the main substrate for the production of prostaglandins, thromboxanes and leukotrienes

A systematic review6 provided evidence that allopurinol was effective in some studies in reducing the severity of post-operative cardiac dysfunction and arrhythmias after coronary artery bypass grafting,

administration of allopurinol prior to ischaemia, reduces the production of superoxide and hence reduces the severity of reperfusion injury

superoxide in inducing damage to distant organs during skeletal muscle reperfusion injury

During IRI, ROS also activate endothelial cells, elevating the activity of the transcription factor, NF-κB. Once activated, the endothelial cell produces E-selectin, vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), endothelial-leukocyte adhesion molecule (EMLMl Am -1) plasminogen activator inhibitor-1 (PAi-1), tissue factor and interleukin-8 (il-8). These adhesion molecules contribute to important interactions between the neutrophil and the endothelium and will be discussed in more detail later.

The major consequence of hydroxyl radical production is peroxidation of the lipid structures of cell membranes resulting in the production and systemic release of proinflammatory eicosanoids, disruption of cell permeability and ultimately cell death

Once the ischaemic tissue is reperfused, an influx of molecular oxygen catalyses xanthine oxidase to degrade hypoxanthine to uric acid and thereby liberating the highly reactive superoxide anion (O2-) (Figure 18.2, lower panel).

COX-2 may promote the inflammatory response through the rapid and exaggerated production of nitric oxide and prostaglandins, contributing to organ damage.4

Hypoxia itself also activates a number of genes, particularly transcription factors, including activating protein-1 (AP-1), hypoxia-inducible factor-1 (HIF-1) and nuclear factor-kappab (NF-kb). HIF-1 then activates transcription of other genes such as vascular endothelial growth factor (VEGF), erythropoietin and glucose transporter-1, which all play an important role in the cells’ adaptive responses to hypoxia (reviewed in3)

repressed genes were largely involved in energy production, including mitochondrial respiration and fatty acid oxidation

activated genes were largely clustered into cytokine genes and mediators of inflammation and immune cell infiltration

found that expression of 962 genes was induced and 327 genes were repressed.

multi-factorial process resulting in extensive tissue destruction

IRI occurs in a wide range of organs including the heart, lung, kidney, gut, skeletal muscle and brain and may involve not only the ischaemic organ itself but may also induce systemic damage to distant organs, potentially leading to multi-system organ failure

Serum creatine phosphokinase (CK) levels >4000 U/L, chloride (Cl) levels >104 mg/dL, and blood urea nitrogen (BUN) levels <10 mg/dL were found to have 100% association with the diagnosis of acute compartment syndrome in hospitalized trauma patients

Serum creatinine kinase (CK) levels >4000 U/L, chloride (Cl) levels >104 mg/dL, and blood urea nitrogen (BUN) levels <10 mg/dL were found to have 100% association with the diagnosis of acute compartment syndrome.

Previous studies show that physicians have a low sensitivity for detecting elevated compartment pressures when relying on physical examination findings alone

public assistance for needy individuals and families who are residents of the state and whose income and property are insufficient to meet the costs of necessary maintenance and services as determined by the state department and to assist such individuals and families to attain or retain their capabilities for independence, self-care, and self-support, as contemplated by article XXIV of the state constitution and the provisions of the social security act and the food stamp act. The state of Colorado and its various departments, agencies, and political subdivisions are authorized to promote and achieve these ends by any appropriate lawful means through cooperation with and utilization of available resources of the federal government and private individuals and organizations.

Nothing in this article shall be construed to prevent the state department from complying with federal requirements for public assistance programs expressly provided by law

(2) There is created in the state department the Colorado teen parent driver's license program to provide financial assistance for the cost of driver's education school training for eligible individuals and the cost to obtain a driver's license or permit.

Section 26-2-710 - Administrative review

If a participant does not agree with

(12) "Individual responsibility contract" or "IRC" means the contract entered into by the participant and the county department pursuant to section 26-2-708.

(15) "Participant" means an individual who receives any assistance or who participates in a specific component of the Colorado works program.

IRC is a contract

2. A client shall be given notice of any action by the county department, or any person or agency acting on its behalf, which adversely affects the client's eligibility for, or right to grant payments authorized under the Colorado Works program. Failure to give notice of an adverse action shall be grounds for setting aside the action on appeal. The notice must meet the following standards:

3. Failure of the client to request a county conference within ninety (90) calendar days from the date timely notice of the proposed action was mailed, absent the client requesting a postponement within that same ninety (90) days, shall constitute abandonment of the right to a conference. The client does not lose the right to appeal directly to the OAC pursuant to section 3.609.7.

Present new information or documentation to support reversal or modification of the proposed adverse action.

confidential informants, privileged communications between the county department and its attorney,

A. The county department, prior to taking action to deny, terminate, recover, initiate vendor payments, or modify financial assistance provided under the Colorado Works program to a client, shall,

C. Clients shall receive written timely notice, giving at least eleven (11) calendar days advance notice before any adverse action taken during the certification period, becomes effective, except as specified in section 3.609.7.C

A. Each client of Colorado Works shall receive prior written notice of any agency action affecting his or her eligibility for or receipt of grant payments.

3.602.1Applications

B. Administrative Review All Colorado Works clients whose benefits have been denied,

"Verified upon receipt" means information that is provided directly from the primary source and is not questionable and no additional verification is required.

"Adverse action" means a county action to reduce grant payments or to deny an application. A reduction may be the result of a sanction, a demonstrable evidence closure, or ineligibility based on income or household changes.

Not be receiving financial assistance from other financial assistance programs administered by the state of Colorado;

shall be eligible for emergency assistance

Section 26-5.3-104 - Emergency assistance for families with children at imminent risk of being placed out of the home

substantial limiting factor in permitting them

no State agency shall impose any other standards of eligibility as a condition for participating in the program.

shall be

Eligible households

other financial resources

incomes

except

Notwithstanding any other provisions of this chapter

parents and their children 21 years of age or younger who live together

(2)

individual or group of individuals constitute a household if they reside in an institution or boarding house,

(m) (1) “Household” means—

Nothing in this article shall be construed to prevent the state department from complying with federal requirements for the food stamp program expressly provided by federal statute

No later than October 1, 2010, the state department shall create a program or policy that, in compliance with federal law, establishes broad-based categorical eligibility for federal food assistance benefits pursuant to the supplemental nutrition assistance program.

provide phased-in services aimed at reunifying families where a child has been placed out of the home, where appropriate, by tapping into other available federal funds or through moneys realized from cost avoidance in prevention of placement;

Out-of-home placement is often the most expensive and disruptive method of providing services to troubled families

The general assembly finds and declares that: (a) Maintaining a family structure to the greatest degree possible is one of the fundamental goals that all state agencies must observe

Family preservation services

(1) "At-risk family" means a family unit with a child who meets out-of-home placement criteria as established by the state board or who, without intervention, risks continued involvement with the child welfare system as established by the state board.

Upon application made to any court of competent jurisdiction by a party to any agency proceeding or by a person adversely affected by agency action and a showing to the court that there has been undue delay in connection with such proceeding or action, the court may direct the agency to decide the matter promptly.

Every agency shall proceed with reasonable dispatch to conclude any matter presented to it with due regard for the convenience of the parties or their representatives, giving precedence to rehearing proceedings after remand by court order

respondent, the guardian ad litem for the child, or a child who is twelve years of age or older may demand a trial by jury of six persons at the adjudicatory hearing pursuant to section 19-3-505 , or the court, on its own motion, may order such a jury to try any case at the adjudicatory hearing pursuant to section 19-3-505 .

the court shall advise the respondent of the minimum and maximum time frames for the dependency and neglect process, including the minimum and maximum time frames for adjudication, disposition, and termination of parental rights for

will permit low-income households to obtain a more nutritious diet through normal channels of trade by increasing food purchasing power for all eligible households who apply for participation

Congress further finds that increased utilization of food in establishing and maintaining adequate national levels of nutrition will promote the distribution in a beneficial manner of the Nation's agricultural abundance and will strengthen the Nation's agricultural economy, as well as result in more orderly marketing and distribution of foods

shall be eligible to participate in the supplemental nutrition assistance program. Assistance under this program shall be furnished to all eligible households who make application for such participation.

households in which each member receives benefits under a State program funded under part A of title IV of the Social Security Act (42 U.S.C. 601 et seq.),

Notwithstanding any other provisions

section 2012(m)(4) of this title

except subsections (b), (d)(2), (g), and (r) of section 2015

If a participant does not agree with or fails to participate in a program or service identified in the IRC, the participant shall continue to receive the basic cash assistance grant that the participant received at the time the appeal is requested during the pendency of any appeal process.

(3) If a participant does not agree with or fails to participate in a program or service identified in the IRC, the participant shall continue to receive the basic cash assistance grant that the participant received at the time the appeal is requested during the pendency of any appeal process.

exercises responsibility

(II) Legal custody is held by an agency that does not have physical possession of the child; or

of the following occurs:

or the care of the child

as long as the parent

who lives in the home

The parents of a dependent child

those persons or families who may receive assistance under the Colorado works program include:

Subject to the provisions of this section

26-2-706. Target populations.

even though one

benefits shall not be reduced or terminated prior to the receipt of the final agency decision unless:

Freetext search

Kendra Dunn Strengthening Families Director State - OEC

Colorado Family First Implementation Team was the Services Continuum workgroup

The Court Improvement Program (CIP) is working collaboratively with CDHS, the Colorado County Attorney Association, the Office of Respondent Parents Counsel, and the Office of the Child’s Representative to maintain alignment and consistent messaging with Family First requirements

Megan KearsleyFamily UnitCourt Improvement Program CoordinatorState Court Administrator’s Officemegan.kearsley@judicial.state.co.usCristina Ritchie Cooper, Esq.Senior AttorneyABA Center on Children and the Lawcristina.cooper@americanbar.orgJennifer Mullenbach, Esq.Deputy County AttorneyCounty Attorney's Officejmullenb@co.jefferson.co.usAshley Chase, Esq.Staff Attorney & Legislative LiaisonOffice of the Child’s Representativeashleychase@coloradochildrep.orgAlison Butler, Esq.Carrie Ann Lucas Disability Advocacy DirectorOffice of Respondent Parents Counselabutler@coloradoorpc.org

Administrative Review Division (ARD)