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1.
J Hand Surg Glob Online ; 6(3): 355-362, 2024 May.
Article in English | MEDLINE | ID: mdl-38817753

ABSTRACT

Purpose: Repair of perilunate dislocations requires stabilization of the carpal bones, although there is limited research on the preferred method(s) of stabilization. We compared the biomechanical characteristics of K-wire versus staple fixation of scapholunate (SL) and lunotriquetral (LT) intervals in the wrist. Methods: After creating a grade IV instability model, 20 cadaver wrists were randomized into 2 stabilization model groups using K-wires or surgical staples across the SL and LT intervals. For each ligament, two K-wires or one surgical staple was used to stabilize the proximal carpal row. Each wrist was loaded with 15 N of force and cycled 5,000 times at an angular velocity of 10 rpm. Displacement between bones was measured at different cycles (10, 100, 500, 1,000, 2,000, 3,000, 4,000, and 5,000). A t test was used for comparisons. Results: For the LT K-wire group, the average displacement at 10 cycles was 0.08 mm, and at 5,000 cycles, 0.163 mm. For the SL K-wire group, the average displacement at 10 cycles was 0.025 mm, and at 5,000 cycles, 0.129 mm. For the LT and SL staple fixation models, the average shifts at 10 cycles were 0.029 and 0.063 mm, and at 5,000 cycles, 0.098 and 0.176 mm, respectively. No significant difference between the fixation models was found at any interval. Notably, in each group, a similar number of small outliers with higher amounts of displacement was observed. Conclusions: Either K-wire or surgical staple stabilization is suitable for perilunate dislocations. The surgical method should be selected based on patient-specific circumstances and the provider's judgment. Type of study/level of evidence: Therapeutic III.

2.
Sci Data ; 10(1): 491, 2023 07 27.
Article in English | MEDLINE | ID: mdl-37500627

ABSTRACT

As the COVID-19 pandemic unfolded in the spring of 2020, governments around the world began to implement policies to mitigate and manage the outbreak. Significant research efforts were deployed to track and analyse these policies in real-time to better inform the response. While much of the policy analysis focused narrowly on social distancing measures designed to slow the spread of disease, here, we present a dataset focused on capturing the breadth of policy types implemented by jurisdictions globally across the whole-of-government. COVID Analysis and Mapping of Policies (COVID AMP) includes nearly 50,000 policy measures from 150 countries, 124 intermediate areas, and 235 local areas between January 2020 and June 2022. With up to 40 structured and unstructured characteristics encoded per policy, as well as the original source and policy text, this dataset provides a uniquely broad capture of the governance strategies for pandemic response, serving as a critical data source for future work in legal epidemiology and political science.


Subject(s)
COVID-19 , Humans , COVID-19/epidemiology , COVID-19/prevention & control , Pandemics , Physical Distancing , Policy Making , SARS-CoV-2
3.
Am J Case Rep ; 23: e936654, 2022 Aug 08.
Article in English | MEDLINE | ID: mdl-35934869

ABSTRACT

BACKGROUND Intramuscular myxomas are rare and benign soft-tissue tumors of uncertain differentiation. Predisposing or precipitating factors have not yet been reported. Activating mutation in GNAS (exons 8 and 9) is detected in >90% of sporadic cases. The role of chronic myopathy, tendinopathy, or trauma to muscles in the etiology of these neoplasms is not known. We report an unusual case of a deltoid mass found following longstanding rotator cuff tendinopathy and a recent fall, later confirmed to be an intramuscular myxoma on biopsy. CASE REPORT A 73-year-old man with a 5-year history of left shoulder pain and rotator cuff tear presented with intractable pain in his left shoulder after a recent fall at home. Physical examination was suggestive of a rotator cuff injury and magnetic resonance imaging (MRI) of the left shoulder revealed a 2.7×2.5×3.7cm T1 hypo- and T2 hyperintense oblong mass-like signal abnormality with heterogeneous, predominantly peripheral enhancement within the deltoid muscle concerning for a malignant mass. Surgical resection was carried out along with left reverse total shoulder replacement, and histopathology revealed findings consistent with an intramuscular myxoma. CONCLUSIONS Intramuscular myxomas are rare, benign tumors. This case report presents one such myxoma incidentally found in a patient with longstanding rotator cuff tendinopathy and a recent fall. Although this co-occurrence is likely incidental, further research and case series review of similar presentations may influence postulations of the pathophysiology of myxomas.


Subject(s)
Muscular Diseases , Myxoma , Rotator Cuff Injuries , Tendinopathy , Aged , Humans , Male , Myxoma/diagnosis , Myxoma/surgery , Shoulder Pain
4.
J Hand Surg Am ; 46(10): 908-916, 2021 10.
Article in English | MEDLINE | ID: mdl-34376294

ABSTRACT

A persistent challenge that has limited access and delivery of digit replantation surgery is timing, as ischemia time has traditionally been considered an important determinant of success. However, reports that the viability of amputated digits decreases after 6 hours of warm ischemia and 12 hours of cold ischemia are largely anecdotal. This review evaluates the quality and generalizability of available evidence regarding ischemia times after digit amputation and reported outcomes of "delayed" replantation. We identify substantial limitations in the literature supporting ischemia time cutoffs and recent evidence supporting the feasibility of delayed digit replantation. The current treatment approach for amputation injuries often necessitates transfers or overnight emergency procedures that increase costs and limit availability of digit replantation nationwide. Evidence-based changes to digit replantation protocols could lead to broader availability of this service, as well as improved care quality.


Subject(s)
Amputation, Traumatic , Finger Injuries , Amputation, Surgical , Amputation, Traumatic/surgery , Finger Injuries/surgery , Fingers/surgery , Humans , Replantation
5.
Cureus ; 13(3): e13881, 2021 Mar 14.
Article in English | MEDLINE | ID: mdl-33868845

ABSTRACT

Bell's palsy is acute peripheral facial nerve palsy; its cause is often unknown but it can be triggered by acute viral infection. Coronavirus disease 2019 (COVID-19) infection commonly presents with respiratory symptoms, but neurologic complications have been reported. A few studies have reported the occurrence of facial nerve palsy during the COVID-19 pandemic. We present a case of Bell's palsy in a 36-year-old man with COVID-19 infection and a past medical history of nephrolithiasis. He presented to the emergency room with a day history of sudden right facial weakness and difficulty closing his right eye four weeks following a diagnosis of COVID-19 infection. Physical examination revealed right lower motor neuron facial nerve palsy (House-Brackmann grade IV). Serologic screen for Lyme disease, human immunodeficiency virus (HIV), and herpes simplex virus (HSV) 1 and 2 were negative for acute infection; however, neuroimaging with MRI confirmed Bell's palsy. He made remarkable improvement following treatment with a course of valacyclovir and methylprednisolone. This case adds to the growing body of literature on neurological complications that should be considered when managing patients with COVID-19 infection.

6.
J Community Hosp Intern Med Perspect ; 11(2): 224-227, 2021 Mar 23.
Article in English | MEDLINE | ID: mdl-33889325

ABSTRACT

Sarcoidosis is a systemic inflammatory condition causing increased immune system activity and manifesting as noncaseating granulomatous disease with the ability to affect multiple organ systems. Neurosarcoidosis is an uncommon presentation, with just 5-10% of patients with sarcoidosis experiencing intracranial disease. The diagnosis of neurosarcoidosis can be difficult, especially given the overlap of imaging findings with more common intracranial lesions. This case presents trigeminal neuralgia as the initial symptom of neurosarcoidosis and emphasizes the importance of a high clinical index of suspicion for neurosarcoidosis in patients with otherwise unexplained neurological symptoms.

7.
Clin Orthop Surg ; 13(1): 76-82, 2021 Mar.
Article in English | MEDLINE | ID: mdl-33747382

ABSTRACT

BACKGROUD: Many U.S. health care institutions have adopted compensation models based on work relative value units (wRVUs) to standardize payments and incentivize providers. A major determinant of payment and wRVU assignments is operative time. We sought to determine whether differences in estimated operative times between the Centers for Medicare & Medicaid Services (CMS) and the National Surgical Quality Improvement Program (NSQIP) contribute to payment and wRVU misvaluation for the most common shoulder/elbow procedures. METHODS: We collected data on wRVUs, payments, and operative times from CMS for 29 types of isolated arthroscopic and open shoulder/elbow procedures. Using regression analysis, we compared relationships between these variables, in addition to median operative times reported by NSQIP (2013-2016). We then determined the relative valuation of each procedure based on operative time. RESULTS: Seventy-nine percent of CMS operative time were longer than NSQIP time (R2 = 0.58), including, but not limited to, shoulder arthroplasty and arthroscopic shoulder surgery. The correlation between payments and operative times was stronger between CMS data (R2 = 0.61) than NSQIP data (R2 = 0.43). Similarly, the correlation between wRVUs and operative times was stronger when using CMS data (R2 = 0.87) than NSQIP data (R2 = 0.69). Nearly all arthroscopic shoulder procedures (aside from synovectomy, debridement, and decompression) were highly valued according to both datasets. Per NSQIP, compensation for revision total shoulder arthroplasty ($10.14/min; 0.26 wRVU/min) was higher than that for primary cases ($9.85, 0.23 wRVU/min) and nearly twice the CMS rate for revision cases ($5.84/min; 0.13 wRVU/min). CONCLUSIONS: CMS may overestimate operative times compared to actual operative times as recorded by NSQIP. Shorter operative times may render certain procedures more highly valued than others. Case examples show that this can potentially affect patient care and incentivize higher compensating procedures per operative time when less-involved, shorter operations have similar patient-reported outcomes.


Subject(s)
Elbow Joint/surgery , Operative Time , Orthopedic Procedures/economics , Orthopedic Procedures/methods , Shoulder Joint/surgery , Aged , Humans , Medicare , United States
8.
Iowa Orthop J ; 40(1): 173-183, 2020.
Article in English | MEDLINE | ID: mdl-32742227

ABSTRACT

Background: Many US health care institutions have adopted compensation models based on work relative value units (wRVUs) to standardize payments and incentivize providers. Among other factors, a major determinant of payment and wRVU assignments is operative time. Our objective was to determine whether differences in estimated operative times between the Centers for Medicare & Medicaid Services (CMS) and the National Surgical Quality Improvement Program (NSQIP) contribute to payment and wRVU misvaluation for the most common hospital-based hand and upper extremity procedures. Methods: Data on wRVUs, surgeon payment, and estimated operative times were collected from CMS for 53 procedures. We used regression models to compare relationships between these variables, in addition to actual median operative times as reported in the NSQIP database, from 2011 to 2016. We then determined the relative valuation of each procedure based on operative time. Results: There was a wide discrepancy between CMS and NSQIP operative times (R2=0.49), with 60% of CMS times being longer than NSQIP times. Payment correlated more strongly with CMS operative times (R2=0.55) than with NSQIP operative times (R2=0.24). Similarly, wRVUs more strongly correlated with CMS operative times (R2=0.84) than with NSQIP operative times (R2=0.51). In general, for trauma-related procedures, any distal radius open reduction internal fixation (ORIF) had the highest valuation while any ORIF proximal to the distal radius had lower valuation in analysis of both databases. While 61% of trauma procedures were highly valued, 70% of elective procedures had a low valuation, including nearly all elective tendon procedures. Notable compensation differences were found between trapeziectomy versus ligament reconstruction and tendon interposition, epicondyle debridement with tendon repair versus denervation, proximal row carpectomy versus four corner fusion, and distal radius open versus percutaneous fixation. Conclusions: CMS may misvalue payment and wRVU rates of hospital-based hand procedures due to inaccurate operative time estimates. By identifying which procedures are misvalued in terms of payment and wRVU per operative time, providers and payors may be able to address these imbalances and maximize appropriate care delivery incentives.Level of Evidence: III.


Subject(s)
Health Care Costs , Hospitals , Medicare/economics , Operative Time , Quality Improvement/economics , Upper Extremity/surgery , Humans , United States
9.
Exp Brain Res ; 238(3): 551-563, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31980847

ABSTRACT

Prenatal exposure to the antiepileptic valproic acid (VPA) is associated with an increased risk of autism spectrum disorder (ASD) in humans. Accordingly, in utero exposure to VPA is a validated and biologically relevant animal model of ASD. The majority of individuals with ASD exhibit some degree of auditory dysfunction, ranging from deafness to hypersensitivity. Animals exposed to VPA in utero have abnormal tonotopic maps and responses in the cerebral cortex and hyperactivation, hypoplasia, abnormal neuronal morphology and reduced calcium binding protein expression throughout the auditory brainstem nuclei. Further, our previous work suggests that GABAergic neuronal populations may be more severely impacted by in utero VPA exposure. However, the axonal projection patterns of brainstem nuclei to the inferior colliculus (IC) have not been investigated in VPA-exposed animals. Herein, we use stereotaxic injections of the retrograde tracer Fast Blue into the central nucleus of the IC (CNIC) and examine the proportions of retrogradely labeled neurons in the nuclei of the lateral lemniscus, superior olivary complex and cochlear nuclei. Our results indicate that not only are there fewer neurons in the auditory brainstem after VPA exposure, but also that fewer neurons are retrogradely labeled from the CNIC. Together, our results indicate that in utero VPA exposure may result in altered patterns of input to the auditory midbrain.


Subject(s)
Auditory Pathways/drug effects , Brain Stem/drug effects , Inferior Colliculi/metabolism , Valproic Acid/pharmacology , Animals , Auditory Pathways/physiology , Autism Spectrum Disorder/drug therapy , Brain Stem/metabolism , Disease Models, Animal , Female , Inferior Colliculi/drug effects , Mesencephalon/metabolism , Neurons/drug effects , Neurons/metabolism , Pregnancy , Prenatal Exposure Delayed Effects/drug therapy , Valproic Acid/metabolism
10.
Arch Med Sci ; 15(1): 1-11, 2019 Jan.
Article in English | MEDLINE | ID: mdl-30697249

ABSTRACT

INTRODUCTION: Competency-based educational models recommend trainee exposure to research, but the best methods for Graduate Medical Education (GME) programs to accomplish this have not been clarified. The objective of this study was to quantify published interventions to generate resident research and compare effectiveness among those interventions. MATERIAL AND METHODS: A systematic review of English-language articles of studies of GME programs was performed, describing resident research interventions and quantifying the number of publications as an outcome. RESULTS: The search produced 13,688 potentially relevant articles, and included 47 articles in the final synthesis. Publication effectiveness was calculated as publications per year. The top ten programs for publication effectiveness were compared to others for interventions chosen. Interventions were characterized as research director, protected time, research requirement, research mentor, curricula, research assistant, biostatistician, information technology support, research fund, pay-for-performance plans, and celebration of accomplishments. Total number of different interventions was not significantly associated with primary outcome (r = 0.20, p = 0.18). When comparing the top ten programs to the others, appointment of a research director was statistically more prevalent in those programs (70% vs. 30%, p = 0.02), while presence of a defined curriculum was more common (90% vs. 57%, p = 0.052) but not statistically significantly. CONCLUSIONS: Leadership interventions (directors, curricula) are associated with successful GME research efforts.

11.
Article in English | MEDLINE | ID: mdl-30559945

ABSTRACT

Acute Generalized Exanthematous Pustulosis (AGEP) is a rare drug reaction manifesting as pustular lesions with surrounding erythema following exposure. The disease is often self-limited and treatment is supportive. It may present in an atypical variant with vesicles that desquamate into erosions, which classifies the disease as an AGEP/SJS Overlap. This overlap syndrome can carry a substantial mortality rate and necessitate elevation in the level of care. Hydroxychloroquine has been implicated in cases of AGEP, and we present a case of AGEP/SJS overlap attributed to this common medication. Given the prevalence of drug eruptions, it is critical for the physicians to recognize and not overlook this rare and potentially fatal dermatological emergency.

12.
Neuroscience ; 377: 53-68, 2018 05 01.
Article in English | MEDLINE | ID: mdl-29510209

ABSTRACT

Auditory dysfunction is a common occurrence in individuals with autism spectrum disorder (ASD). While most cases of ASD are of unknown etiology, in utero exposure to the antiepileptic valproic acid (VPA) significantly increases risk. We have previously identified significant dysmorphology and hypoplasia in the auditory brainstem of humans with ASD and rodents exposed to VPA in utero. Further, we have identified abnormal c-Fos immunolabeling patterns after exposure to pure tone stimuli in VPA-exposed animals. Herein, we describe the impact of repeated exposure to VPA on key components of the auditory hindbrain, the ventral cochlear nucleus (VCN) and superior olivary complex (SOC). Specifically, we examined neuronal number, neuronal morphology, immunolabeling for the calcium binding proteins calbindin (CB) and calretinin (CR), dopaminergic innervation and the structure of calyx terminals in the medial nucleus of the trapezoid body (MNTB). VPA-exposed animals had significantly fewer neurons in both the VCN and SOC. VPA had a differential impact on the size of neurons in the VCN and SOC. VPA-exposed animals have reduced CB and CR immunolabeling and a lower density of dopaminergic terminals. Finally, we saw no difference in the surface area or volume of calyx terminals in the MNTB, although there was a relative increase in the surface area and volume of calyces in VPA-exposed animals. These results indicate hypotrophy of the auditory brainstem, abnormal calcium regulation and reduced dopaminergic input. Together, such alterations suggest abnormal brainstem circuitry and significant auditory dysfunction in VPA-exposed animals.


Subject(s)
Anticonvulsants/adverse effects , Auditory Pathways/drug effects , Brain Stem/drug effects , Calcium-Binding Proteins/metabolism , Prenatal Exposure Delayed Effects , Valproic Acid/adverse effects , Animals , Auditory Pathways/metabolism , Auditory Pathways/pathology , Autism Spectrum Disorder/metabolism , Autism Spectrum Disorder/pathology , Brain Stem/growth & development , Brain Stem/metabolism , Brain Stem/pathology , Disease Models, Animal , Female , Male , Neurons/drug effects , Neurons/metabolism , Neurons/pathology , Pregnancy , Rats, Sprague-Dawley
13.
J Hand Surg Am ; 43(1): 24-32.e1, 2018 01.
Article in English | MEDLINE | ID: mdl-29103849

ABSTRACT

PURPOSE: Timely identification of tissue ischemia is critical, both in the traumatized limb and following free tissue transfer. The purpose of this study was to determine if skin pigmentation affects the ability to detect limb ischemia. METHODS: We conducted a study of healthy controls exposed to limb ischemia. The subjects were classified based on skin pigmentation using a defined skin type assessment tool, a visual color scale, and self-description of race. Participants were randomized by limb and tourniquet status; surgeons were blinded to both. Ischemia was induced by tourniquet insufflations, and board-certified orthopedic and plastic surgeons who had completed an accredited hand surgery fellowship conducted physical examinations. The surgeons monitored the forearms at 2, 6, and 10 minutes based on appearance of ischemia, capillary refill, and color in 3 locations on the limbs (posterior interosseous artery flap skin territory, radial forearm flap skin territory, and the digits). RESULTS: We found a significant decrease in the ability to detect ischemia in participants with increased skin pigmentation, as documented by all metrics, when evaluating the posterior interosseous artery and radial forearm flap skin territories at all time points. For example, when monitoring the posterior interosseous artery flap with the tourniquet insufflated at time 10 minutes, 92.9% of Caucasians were correctly identified as being ischemic whereas only 23.3% of African Americans were correctly identified. CONCLUSIONS: Skin pigmentation significantly affects the identification of an ischemic limb/skin flaps on physical examination. Whereas the standard treatment for monitoring of free tissue transfer is clinical examination, that may not be sufficient for patients with increased skin pigmentation. Surgeons should exercise particular vigilance during physical examination of a potentially ischemic limb/skin flaps with greater skin pigmentation. TYPE OF STUDY/LEVEL OF EVIDENCE: Diagnostic II.


Subject(s)
Forearm/blood supply , Ischemia/diagnosis , Physical Examination , Skin Pigmentation/physiology , Adult , Free Tissue Flaps , Healthy Volunteers , Humans , Middle Aged , Racial Groups , Tourniquets , Young Adult
14.
JBJS Rev ; 4(6)2016 06 07.
Article in English | MEDLINE | ID: mdl-27486724

ABSTRACT

Carpal tunnel syndrome is the most common peripheral nerve compression syndrome. Treatment options include wrist-neutral bracing, corticosteroid injections, operative release of the transverse carpal ligament, and symptom-relief options. Endoscopic carpal tunnel release may give patients a faster recovery compared with traditional open release, but there are no ultimate differences in outcome among the various surgical options.


Subject(s)
Carpal Tunnel Syndrome/surgery , Endoscopy , Humans , Ligaments, Articular , Nerve Compression Syndromes , Treatment Outcome
15.
JBJS Rev ; 4(12)2016 12 20.
Article in English | MEDLINE | ID: mdl-28060785

ABSTRACT

Arthritis and instability represent 2 of the most common pathological processes affecting the distal radioulnar joint (DRUJ). These conditions can present in isolation or as components of a multifactorial process. Nonoperative treatment is indicated for most acute injuries to the DRUJ. The joint should be immobilized in a position of stability to allow for ligament healing. Likewise, early arthritis responds favorably to rest, immobilization, corticosteroids, and nonsteroidal anti-inflammatory drugs (NSAIDs). When DRUJ instability is refractory to nonoperative measures, native ligament repair is the preferable method of treatment. When this method is not possible, anatomical reconstruction of the distal radioulnar ligaments should be performed. For advanced DRUJ arthritis Darrach resection should be reserved for the elderly, low-demand patient. The Sauve-Kapandji procedure allows for arthrodesis of the DRUJ while maintaining forearm rotation and a stable base for the ulnar carpus. DRUJ hemiarthroplasty procedures have been associated with favorable preliminary results. These implants attempt to reproduce native biomechanics and may be used in lieu of or as a salvage procedure after resection arthroplasty. DRUJ arthroplasty should be used as a salvage procedure.


Subject(s)
Arthritis , Arthroplasty , Joint Instability , Wrist Joint/surgery , Arthrodesis , Forearm , Humans
16.
J Appl Psychol ; 101(4): 498-519, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26595754

ABSTRACT

Psychological individual differences, such as personality, affectivity, and general mental ability, have been shown to predict numerous work-related behaviors. Although there is substantial research demonstrating relationships between psychological individual differences and withdrawal behaviors (i.e., lateness, absenteeism, and turnover), there is no integrative framework providing scholars and practitioners a guide for conceptualizing how, why, and under what circumstances we observe such relationships. In this integrative conceptual review we: (a) utilize the Cognitive-Affective Processing System framework (Mischel & Shoda, 1995) to provide an overarching theoretical basis for how psychological individual differences affect withdrawal behaviors; (b) create a theoretical model of the situated person that summarizes the existing empirical literature examining the effect of psychological differences on withdrawal behavior; and (c) identify future research opportunities based on our review and integrative framework.


Subject(s)
Employment/psychology , Individuality , Models, Psychological , Personnel Turnover/statistics & numerical data , Absenteeism , Affect , Humans , Job Satisfaction , Personality
17.
JBJS Case Connect ; 6(4): e90, 2016.
Article in English | MEDLINE | ID: mdl-29252744

ABSTRACT

CASE: A 30-year-old woman underwent arthroscopy for wrist pain 3.5 years following a medial femoral trochlea (MFT) osteocartilaginous free flap for Kienböck disease, which provided the opportunity to examine the transferred cartilage in vivo. Arthroscopy revealed no evidence of chondrolysis, and histologic examination revealed uniformly viable chondrocytes within a matrix consistent with fibrocartilage. CONCLUSION: To the best of our knowledge, the long-term viability of chondrocytes following an MFT free flap has not been previously documented. This case provides early evidence that free tissue transfer based on the descending genicular artery can provide a durable solution for osteocartilaginous defects.


Subject(s)
Cartilage/physiology , Chondrocytes/physiology , Lunate Bone/transplantation , Osteonecrosis/surgery , Adult , Arthroscopy , Bone Transplantation , Cell Survival , Female , Free Tissue Flaps , Humans , Wrist Joint/surgery
18.
J Hand Surg Am ; 40(10): 2116, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26408382
20.
Cells ; 4(1): 84-95, 2015 Feb 11.
Article in English | MEDLINE | ID: mdl-25679284

ABSTRACT

ELISPOT, ELISA and flow cytometry techniques are often used to study the function of immune system cells. It is tempting to speculate that these assays can be used interchangeably, providing similar information about the cytokine secreting activity of cells: the higher the number of cytokine-positive cells measured by flow cytometry, the higher the number of cytokine-secreting cells expected to be detected by ELISPOT and the larger the amount of secreted cytokine expected to be measured by ELISA. We have analyzed the expression level and secretion capacity of IFNγ from peripheral blood mononuclear cells isolated from five healthy donors and stimulated by calcium ionomycin mixed with phorbol 12-myristate 13-acetate in a non-specific manner in side-by-side testing using ELISPOT, ELISA and flow cytometry assays. In our study, we observed a general correlation in donors' ranking between ELISPOT and flow cytometry; ELISA values did not correlate with either ELISPOT or flow cytometry. However, a detailed donor-to-donor comparison between ELISPOT and flow cytometry revealed significant discrepancies: donors who have similar numbers of IFNγ-positive cells measured by flow cytometry show 2-3-fold differences in the number of spot-forming cells (SFCs) measured by ELISPOT; and donors who have the same number of SFCs measured by ELISPOT show 30% differences in the number of IFNγ-positive cells measured by flow cytometry. Significant discrepancies between donors were also found when comparing ELISA and ELISPOT techniques: donors who secreted the same amount of IFNγ measured by ELISA show six-fold differences in the number of SFCs measured by ELISPOT; and donors who have 5-7-times less secreted IFNγ measured by ELISA show a two-fold increase in the number of SFCs measured by ELISPOT compared to donors who show a more profound secretion of IFNγ measured by ELISA. The results of our study suggest that there can be a lack of correlation between IFNγ values measured by ELISPOT, ELISA and flow cytometry. The higher number of cytokine-positive cells determined by flow cytometry is not necessarily indicative of a higher number of cytokine-secreting cells when they are analyzed by either ELISPOT or ELISA. Our ELISPOT vs. ELISA comparison demonstrates that the higher number of SFCs observed in ELISPOT does not guarantee that these cells secrete larger amounts of cytokines compared to donors with lower SFC numbers. In addition, our data indicate that ELISPOT, ELISA and flow cytometry should be performed as complementary, rather than stand-alone assays: running these assays in parallel on samples from the same donors may help to better understand the mechanisms underlying the physiology of cytokine-secreting cells.

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