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Актуальність. Як відомо, тяжка черепно-мозкова травма супроводжується розвитком гіпоксії з подальшою загибеллю нервових клітин. Важливу роль у патологічному процесі відіграють автоімунні реакції клітинного і гуморального типу. Мета роботи: вивчити вміст у крові автоантитіл до нейроантигенів у хворих на тяжку черепно-мозкову травму. Матеріали та методи. Обстежено 40 хворих з ізольованою тяжкою черепно-мозковою травмою, які перебували на лікуванні у відділенні інтенсивної терапії в післяопераційному періоді. Контрольну групу становили 20 здорових добровольців. Вміст автоантитіл до мозкових антигенів (основного білка мієліну (MBP), кальційзв’язуючого білка (S-100), нейроспецифічної енолази (NSE) та загального мозкового антигенa) визначали методом імуноферментного аналізу на 1-шу, 3-тю, 5-ту ,7-му, 14-ту добу після оперативного втручання. Результати. Було виявлено, що рівень автоантитіл до основного білка мієліну на 1-шу добу, а також упродовж усього дослідження вірогідно не відрізнявся від рівня здорових добровольців. Концентрація автоантитіл до кальційзв’язуючого білка на 1-шу добу вірогідно не відрізнялась від рівня здорових добровольців, але на 3-тю добу вона збільшилась, різниця між показниками пацієнтів і здорових добровольців була статистично вірогідною. На 5-ту добу відбувалося зниження показників зі збереженням вірогідної різниці. На 7-му та 14-ту добу рівні автоантитіл пацієнтів поступово знижувалися (р > 0,05). Рівень автоантитіл до нейроспецифічної енолази на 1-шу добу не перевищував показники здорових добровольців (р > 0,05). На 3-тю добу рівень збільшився, досягши піка на 5-ту добу, коли різниця порівняно з показниками здорових добровольців була статистично вірогідною. В подальшому відбувалося зниження показників зі збереженням статистично вірогідної різниці. Рівні автоантитіл до загального мозкового антигена на 1-шу добу були підвищені, але вірогідно не відрізнялися від показників здорових добровольців. В подальшому показники зростали, досягши піка на 5-ту добу, коли вони статистично різнилися з показниками здорових добровольців. Надалі спостерігалося зниження показників на 7-му та 14-ту добу зі збереженням статистично вірогідної різниці. Висновки. Результати нашого дослідження показали, що тяжка черепно-мозкова травма викликає підвищення рівнів специфічних автоантитіл до мозкових антигенів у сироватці крові хворих. Зафіксований високий рівень автоантитіл до нейроспецифічної енолази, білка S-100, загального мозкового антигена і низький — до основного білка мієліну, що дозволяє зробити висновок про те, що саме нейрони і астроглія схильні насамперед до змін та пошкодження, оскільки до їх антигенів активується автоімунний процес.

Background Immunoassays used to measure total and free thyroxine (T4 and FT4) and total and free triiodothyronine (T3 and FT3) can provide inaccurate results due to interference from endogenous autoantibodies. Case Report: A 74-year-old female treated for hypothyroidism with levothyroxine replacement had elevated thyroid stimulating hormone (TSH), FT4, and FT3. Due to concern for hyperthyroidism, levothyroxine was discontinued and further workup was initiated. A pituitary MRI revealed a microadenoma but the alpha-subunit was normal. She was given octreotide for suspected TSH secreting pituitary adenoma without improvement in her TSH, FT4, or FT3 levels. She was referred to our clinic, where inaccurate lab values for FT4 and FT3 were suspected. Results Testing via equilibrium dialysis liquid chromatography tandem mass spectrometry (LC-MS/MS) method revealed lower levels of FT4 and FT3. Subsequent testing included heterophile blocking tube treatment, polyethylene glycol (PEG) precipitation, and anti-T3/T4 autoantibody levels. The tests revealed thyroid hormone autoantibodies (THAAs) as the cause of immunoassay interference. Conclusions When thyroid hormones are elevated and TSH is not suppressed, confirmatory testing with another method such as equilibrium dialysis LC-MS/MS, which is not susceptible to interference from autoantibodies, should be considered.

Objective To report a case series of thyroid cancer patients in whom false positive results in immunometric assays for thyroglobulin (TgIMA) were caused by heterophilic antibody interference, describe the clinical scenario in which this interference should be suspected, and recommend methods to demonstrate the interference. Methods Three patients with unexpectedly elevated thyroglobulin results (range, 1.6-75 ng/mL) were studied. In the first patient, thyroglobulin was elevated despite the presence of Tg antibody. In the second patient, suppressed thyroglobulin was higher than a recent stimulated thyroglobulin. In the third patient, thyroglobulin became detectable years after treatment and did not change after thyroid-stimulating hormone stimulation. TgIMA concentration determination was compared to determination by a mass spectrometry method (TgMS). Thyroglobulin was also remeasured after preabsorption with heterophile antibody blocking reagents and after serial dilutions. Results In all cases, thyroglobulin was undetectable by TgMS. In 2 of 3 patients, dilutions provided nonlinear thyroglobulin results. After blocking agent preabsorption, thyroglobulin dropped by 35%, 45%, and 91% in the 3 samples. Conclusion False positive thyroglobulin concentrations from heterophilic antibody interference have significant impact on the management of thyroid cancer. Here we show that TgMS assays can be used to rule out heterophilic antibody interference. This interference should be suspected when a detectable thyroglobulin by TgIMA does not respond to thyroid-stimulating hormone or is discordant from the clinical assessment.

Parathyroid hormone (PTH) measurement using immunoassays is inherently vulnerable to interferences due to the presence of different proteins such as heterophile antibodies, human anti-animal antibodies, auto-analyte antibodies, the rheumatoid factor, and others. The frequency of immunoassay interference can be as high as 6%. We report the case of a patient showing persistent high levels of PTH without impact on calcium and bone metabolism. The patient was a 59-year-old asymptomatic woman who consistently showed elevated PTH levels (385-482 pg/ml) using the Roche Elecsys (Cobas e-411) and ADVIA Centaur assays, with normal calcium, phosphorus, vitamin D, and renal function parameters. She had no history of fractures, nephrolithiasis, gastrointestinal complaints, renal insufficiency, or autoimmune diseases. Her physical examination revealed no abnormalities. Biomarkers of bone metabolism were within the reference range. To rule out falsely elevated PTH levels, we initially performed serial dilutions using both assays, which revealed nonlinearity. After a polyethylene glycol precipitation test, less than 10% of PTH was recovered from the supernatant. These results suggested the presence of heterophile antibodies as the cause of the falsely elevated PTH levels. Physicians should be aware of this issue in order to avoid unnecessary clinical investigations and inappropriate treatments.

Objective: The study was done to detect possible causes of arthritis in turkey flock with emphasis on molecular detection of bacterial pathogens direct from samples and estimation of their pathology pattern. Procedure: A total of 100 arthritic legs from 125 French turkey birds were collected from different farms in Sharkia Governorate. Specimens from joint capsule, tendon, and synovial fluid of leg joints were subjected to bacteriological and direct PCR examinations. Moreover, skin, cartilage of articular surface and epiphysis were undergone pathological examination. Blood samples were collected before scarification for both hematological and biochemical examinations. Postmortem examinations revealed joints swelling, hyperkeratosis, erosions, and/or ulcerations. Results: Forty percent (40%) bacterial incidence rate including Staphylococcus spp., E. coli, Salmonella spp., Klebsiella spp., and Pseudomonas aeruginosa were recorded. Molecular detection of these pathogens revealed rapid and more accurate results reach 46%.A high prevalence of multidrug resistance was detected and all isolates were susceptible to amikacin and florfenicol. Genotyping of isolates revealed the presence of strong virulence markers. Further investigations on positive infected samples revealed a significant decrease in RBCs, Hb, PCV %, lymphocytes, total protein, and albumin in addition to a significant increase in the leucocytic count and heterophile, CRP, ALT, AST, uric acid, creatinine, and total globulin. Epidermal leucocytic cells infiltrations, tendonitis, synovium fibrosis, erosions, and/or ulcerations of articular surface cartilage were recorded. Conclusion: Multiplex real-time PCR assay offered an effective alternative to traditional typing methods for the identification and simultaneous detection of pathogens involved in arthritis in turkey.

Pseudomonas aeruginosa is known to be multidrug-resistant organisms so its infection is challenging. This study aimed to use the sponge-like reduced protocol in preparation of P. aeruginosa ghosts (PAGs) as a vaccine alternative and evaluate its competency in stimulating the immune response in Nile tilapia. Characterization of PAGs was done using the light, fluorescent and transmission electron microscope. 240 fish have been divided into four groups (60 fish each; three groups were injected with a high, medium, or low dose of PAGs, and a control group. Total and differential leukocyte count, phagocytic activity, lysozyme activity, and gene expression of some immune-related genes were done after the first and booster ghost dose injection. All fish groups were challenged intraperitoneally with P. aeruginosa (1 × 107 CFU ml−1); the mean percent mortality was calculated. Significant immunization potency of the large dose of PAGs was prominent. It is associated with a significant increased number of WBCs, heterophile, lymphocytes, and monocytes. Also the phagocytic activity and indexes were significantly increased by the high and moderate dose of PAGs. Additionally, significant upregulation of immune-related gene (MHC-II) and inflammatory-related gene (IL8) and maintained expression level of TLR7 were recorded. We conclude that chemically prepared P. aeruginosa ghosts maintained both the 3D structure and surface molecules of live bacteria and also are capable of inducing specific immune responses in Nile Tilapia, and reduced mortalities in challenged fish. Our results indicate the advantage of this procedure for production of bacterial ghost, which can be used in wide a range of hosts.

Detectable serum levels of beta subunit of human chorionic gonadotropin (βhCG) in nonpregnant, perimenopausal women bring confusion in both clinician and patient and could lead to unnecessary diagnostic and therapeutic procedures. A 45-year-old woman with the continuous elevation of βhCG underwent hysteroscopy, two laparoscopic surgeries, two explorative uterine cavity curettages and three cycles of cytostatic therapy. No ultrasonographic, macroscopic or histological signs of pregnancy, both uterine and ectopic or gestational trophoblastic disease were found. Both radiographic and computed tomography reports ruled out the presence of a nongynecological neoplasm. All recommended steps were taken for confirmation or ruling out the possibility of heterophile antibodies interference, but the results were not absolute. Finally, the referent laboratory confirmed the presence of the antibodies, but their exact type remains unknown. This case underlines the importance of the universally accepted protocol in the cases of persistent βhCG elevation in nonpregnant, perimenopausal women.

Black Soldier Fly larvae are natural antibiotics. The use of methanol extract in BSF larvae has a dual function, namely high protein content and high antibiotic content to kill Gram-negative bacteria. The biological structure of BSF larvae with antimicrobial peptide (AMP) acts as an inhibitor of pathogenic microorganisms, has a high lauric acid function as a natural antimicrobial and chitin, the polysaccharide plays a role in enhancing the immune response of the animal. This study aims to gain the best type of BSF protein from feed treatments containing live BSF, dry BSF, and BSF extract to increase blood productivity and hematology. The results of this study show that the best P3 treatment for the productivity of laying hens at the age of 18 to 26 weeks with the use of BSF extract since it contains the highest protein compared to other treatments. Productivity variables show the best P3 treatment, each egg weight 57.17 g bird -1 , daily egg production 90.88%, egg mass 2408.16 g bird -1 and ration conversion 2.0. The mean hematology of the blood showed the highest P3 treatment, each erythrocyte 2.91±0.13 x 10 6 mm -3 , leucocytes 17.69±3.68 x 10 3 mm -3 , hematocrit 27.8±1.14%, hemoglobin 9.96±1.12%, MCV 95.77±4.41 fl, MCHC 35.99±5.21%, MCH 34.38±4.53 pg, lymphocytes 61.2 ± 4.86%, heterophile 31.2±2%, monocytes 5±0.62% and ratio H/L 0.51±0.04.

We present the case of a female patient who developed persistently elevated levels of cardiac troponin (cTn) after a previous episode of clinically presumed myocarditis. Extensive investigation concluded that the presence of heterophile antibodies was causing false positive cTn elevation. (Level of Difficulty: Intermediate.), Graphical abstract, We present the case of a female patient who developed persistently elevated levels of cardiac troponin (cTn) after a previous episode of clinically…

Leukocyte profiles, such as the heterophile/lymphocyte (H/L) ratio, are influenced by several ecological and ecophysiological factors and are generally considered to be independent and robust indicators of stress levels in birds. We tested the hypotheses that Semipalmated Sandpiper Calidris pusilla presents a positive association between physiological stress (H/L ratio) and molt, and that there should be a negative association between physiological stress (H/L ratio) and body condition (mass). During its annual cycle, Semipalmated Sandpiper performs long-distance migrations and often arrives in South America in September/October when it can undergo molting before its migration to the north. In May, while preparing to return to its reproductive sites, Semipalmated Sandpiper increases its body mass. The research was carried out on Restinga Island of the Rio Paraiba estuary in northeast Brazil. Data collection took place during the wintering of Semipalmated Sandpiper in October 2017 and February and May 2018. We found a positive association between the H/L ratio and the presence of molt in October and February, but a negative association between the H/L ratio and body mass in May. Our results support the hypotheses that the presence of molt should be positively associated with the H/L ratio due to the high energy and nutritional costs that molting requires and that the H/L ratio and body mass should be negatively associated due to the favorable nutritional status of Semipalmated Sandpiper during its preparation to migrate to its reproductive areas.

Epstein-Barr virus (EBV) primary infection usually presents with classic symptoms of infectious mononucleosis (IM) like fever, lymphadenopathies and tonsillopharyngitis. Liver damage is frequently mild and self-limited and there are only a few cases of severe EBV-induced cholestatic hepatitis and jaundice reported in the literature. The authors present the case of a 22-year-old woman who was admitted with acute fever and jaundice. Physical examination revealed posterior cervical lymphadenopathies and painful hepatosplenomegaly. Laboratorial findings suggested an obstructive cause for jaundice but ultrasound and magnetic resonance cholangiopancreatography excluded biliary duct pathology. Heterophile antibodies were negative but EBV-specific antibodies revealed isolated positive viral capsid antigen (VCA) immunoglobin (Ig) M suggesting the diagnosis of early phase of EBV primary infection. The diagnosis of EBV-induced cholestatic hepatitis was confirmed after identification of EBV deoxyribonucleic acid (DNA) in blood and by liver biopsy. Supportive management was provided and, despite an initial clinical deterioration, the patient had a favorable outcome. EBV is a virus with a high prevalence worldwide, mainly subclinical, and jaundice is a rare manifestation of the infection. Although the majority of the patients recover without sequelae, progression to liver failure has been described and a careful assessment for complications is mandatory. Therefore, EBV infection should be included in the comprehensive differential diagnosis of jaundice in all age groups.

In this report we evaluated a diagnostic algorithm, proposed by the Belgian Superior Health Council, to detect acute and past Epstein-Barr virus (EBV) infections by means of serology in donors of human body material for transplantation. The available EBV serology parameters were tested on eighty serum samples on three random access analysers: Architect i2000 SR, Liasion XL and BioPlex 2200. The EBV sero-status was determined according to the proposed algorithm and results were compared between the different analysers. Seventy one % of the samples gave concordant interpretations on the three analysers. Most of the discordant results were attributable to early antigen (EA) IgG. The knowledge of the EA IgG and heterophile antibodies (HA) IgM status provided only limited added value and was only useful to distinguish between a very early acute infection and false positivity of viral capsid antigen IgM. The diagnostic algorithm proposed by the Belgian Superior Health Council is merely directive and each individual lab remains responsible for the interpretation and implementation of test combinations for the detection of EBV infections. Our study shows the limited added value of testing for EA IgG and HA IgM, based both on clinical and technical performance.

Introduction: The aim of this study is to assess epidemiological, clinical and laboratory characteristics of primary infection by Epstein–Barr virus (EBV) in children without previous diagnosis of any immune disease and its relationship with clinical presentation. Patients and methods: A retrospective study was conducted on all children from 0 to 15 years with IgM against viral capsid of EBV positive or indeterminate during a 22 month period. Epidemiological, clinical and laboratory data were analysed and compared between typical (mononucleosis syndrome) and non-typical clinical symptoms. Results: The study included a total of 103 children, with a median age of 7 years (3–12.5 years). Almost two-thirds (63%) of patients had typical clinical signs, with a mononucleosis syndrome, and 37% had a non-typical presentation. The non-typical clinical group had a lower age (P = .03) and took less antibiotic than the typical clinical group (P = .015). From laboratory studies, there were no differences between the groups, except in RCP, which was higher in typical clinical group (P = .04). Heterophile antibodies were positive in 33% of patients. An indeterminate IgM against viral capsid was present in 20% of the patients, and most of them had an oligosymptomatic or atypical presentation. An IgM positive for other viruses was found in 21%, and 3 of them were suspicious of false positive for EBV. Conclusions: In the studied population, a primary infection due to EBV is common in younger ages, and they have usually an oligosymptomatic clinical presentation. A very low percentage of positive heterophile antibodies were found. Cases with indeterminate IgM against viral capsid are more frequent in the non-typical clinical group. Co-infection with other viruses is common. Resumen: Introducción: El objetivo de este estudio es conocer las características epidemiológicas, clínicas y analíticas de la primoinfección por el virus de Epstein-Barr (VEB) en niños sin diagnóstico previo de ninguna enfermedad inmune y su relación con la forma de presentación clínica. Pacientes y métodos: Estudio retrospectivo de pacientes entre 0 y 15 años con IgM sérica frente a la cápside viral del VEB positiva o indeterminada, durante un período de 22 meses. Se analizaron datos epidemiológicos, clínicos y de laboratorio y se compararon según tuvieran una clínica típica (síndrome mononucleósico) o no típica. Resultados: Se incluyeron 103 niños. La mediana de la edad fue de 7 años (3-12.5 años). El 63% de los pacientes presentaron clínica típica o síndrome mononucleósico y el 37% una clínica no típica. La edad fue significativamente menor en el grupo de clínica no típica (p = 0,03) y recibieron menos tratamiento antibiótico (p = 0,015). En los parámetros analíticos no hubo diferencias estadísticamente significativas excepto en la PCR, discretamente más elevada en el grupo de clínica típica (p = 0,04). El 33% de los pacientes tuvieron anticuerpos heterófilos positivos. El 20% tuvieron una IgM frente a la cápside viral indeterminada, la mayoría con clínica oligosintomática o atípica. El 21% tuvieron IgM positivas para otros virus y 3 de ellos fueron posibles falsos positivos para el VEB. Conclusiones: En nuestra población, la primoinfección por VEB es frecuente en niños de menor edad, y en ellos predominan las formas oligosintomáticas. El porcentaje de anticuerpos heterófilos positivos ha sido muy bajo en nuestra muestra. Los casos con IgM frente a la cápside viral indeterminada son más frecuentes en el grupo de clínica no típica. Es común detectar coinfección con otros virus.

Background Heterophile antibody (HAb) interferences in immunoassays can cause falsely elevated hCG concentrations leading to incorrect diagnosis and treatments options. When results are not consistent with the clinical findings, hCG HAb interference investigation may be requested by the physician. A retrospective evaluation of the frequency of HAb interference was performed among cases of physician-requested investigations and the effectiveness of commercially available blocking reagents to detect HAb interference in two immunoassay systems was evaluated. Methods One hundred and thirteen physician requests for hCG HAb investigation from 2008 to 2017 were reviewed. The primary method used to measure hCG was the Beckman Coulter Access Total βhCG (2008–2010) and the Roche Elecsys HCG+β (2014–2017). HAb investigation included measurement by two immunoassays before and after treatment of samples with heterophile blocking reagents and serial dilution studies. Results Five cases of HAb and HAb-like interference were identified. The interference frequency was 6.7% for the Beckman assay and 2.9% for the Roche assay. The presence of HAb was detected using heterophile blocking reagents and an alternative method in three cases. The other two cases were detected due to discrepant results with an alternative method and non-linear serial dilutions (HAb-like). Conclusions HAb interference was observed in the Beckman and the Roche assays. The heterophile blocking reagents failed to detect 40% of interference cases. Blocking reagents should not solely be used for these investigations. Multiple strategies including the use of serial dilutions and using an alternative platform are critical when troubleshooting interferences in hCG immunoassays.

The proper clinical evaluation of pituitary and adrenal disorders depends on the accurate measurement of plasma ACTH. The modern two-site sandwich ACTH immunoassay is a great improvement compared with older methods but still has the potential for interferences such as heterophile antibodies and pro-opiomelanocortin (POMC) and ACTH fragments. We report the cases of five patients in whom the diagnosis or differential diagnosis of Cushing syndrome was confounded by erroneously elevated results from the Siemens ACTH Immulite assay [ACTH(Immulite)] that were resolved using the Roche Cobas or Tosoh AIA [ACTH(Cobas) and ACTH(AIA), respectively]. In one case, falsely elevated ACTH(Immulite) results owing to interfering antibodies resulted in several invasive differential diagnostic procedures (including inferior petrosal sinus sampling), MRI, and unnecessary pituitary surgery. ACTH(Cobas) measurements were normal, and further studies excluded the diagnosis of Cushing syndrome. In three cases, either Cushing disease or occult ectopic ACTH were suspected owing to elevated ACTH(Immulite) results. However, adrenal (ACTH-independent) Cushing syndrome was established using ACTH(AIA) or ACTH(Cobas) and proved surgically. In one case, ectopic ACTH was suspected owing to elevated ACTH(Immulite) results; however, the ACTH(Cobas) findings led to the diagnosis of alcohol-induced hypercortisolism that resolved with abstinence. We have concluded that ACTH(Immulite) results can be falsely increased and alternate ACTH assays should be used in the diagnosis or differential diagnosis of clinical disorders of the hypothalamic–pituitary–adrenal axis.

A plethora of studies have described heterosis or hybrid vigor; however, a global understanding of its regulation and the transmission of transcriptional levels between parents and hybrid has yet to be attained. To improve our understanding the molecular mechanisms controlling maize heterosis, we used an incomplete diallel cross design consisting of four elite maize inbred lines and six of their hybrids to measure the degree of variation in gene expression between the parents and their hybrids. We found that differentially expressed genes (DEGs) drove diversity of tissue specific heterosis and that heterophile expression was a generally complementary mechanism of gene expression in hybrids. However, the full expression of heterosis was due to the proportion of super dominant gene expression patterns that aggregate the regulatory network of dominant genes in response to adversity, and thus promotes heterosis in hybrids. Our results provide a new understanding and perspective into the regulatory mechanisms that control heterosis and represent an important step towards a more comprehensive explanation of heterosis in maize.

We present the case of an 83-year-old man with metastatic prostate cancer who had testosterone levels reading above castration range despite appropriate medical and surgical castration. Mass spectrometry was performed to confirm presence of testosterone, but no testosterone was detected. The elevated testosterone as measured by standard immunoassay was postulated to be secondary to heterophile antibodies in the patient's serum. This report highlights the need for a high index of suspicion for interference in testosterone immunoassays when levels remain mildly elevated. Mass spectrometry may provide a more reliable method by which to detect testosterone concentration prior to escalation of care.

Immunoassays are powerful qualitative and quantitative analytical techniques. Since the first description of an immunoassay method in 1959, advances have been made in assay designs and analytical characteristics, opening the door for their widespread implementation in clinical laboratories. Clinical endocrinology is closely linked to laboratory medicine because hormone quantification is important for the diagnosis, treatment, and prognosis of endocrine disorders. Several interferences in immunoassays have been identified through the years; although some are no longer encountered in daily practice, cross-reaction, heterophile antibodies, biotin, and anti-analyte antibodies still cause problems. Newer interferences are also emerging with the development of new therapies. The interfering substance may be exogenous (e.g., a drug or substance absorbed by the patient) or endogenous (e.g., antibodies produced by the patient), and the bias caused by interference can be positive or negative. The consequences of interference can be deleterious when clinicians consider erroneous results to establish a diagnosis, leading to unnecessary explorations or inappropriate treatments. Clinical laboratories and manufacturers continue to investigate methods for the detection, elimination, and prevention of interferences. However, no system is completely devoid of such incidents. In this review, we focus on the analytical interferences encountered in daily practice and possible solutions for their detection or elimination.

Background Heterophile antibodies are one of the most common causes of false-positive troponin. Case summary We report a case of a 53-year-old woman with false-positive troponin elevation and a clinical presentation understood and treated as non-ST-elevation acute coronary syndrome. Because of chronic basal elevation of troponin (at a ‘plateau’ level) and chest pain, the patient underwent several invasive coronary angiograms until false-positive increase of troponin due to heterophile antibodies was suspected. Borderline stenosis of a left circumflex coronary artery found on first coronary angiogram was a coincidental finding and heterophile antibodies in the patient’s serum were confirmed. Discussion This interesting case report aims to remind the clinicians about the possibility of false-positive troponin level due to laboratory analytical interference caused by heterophile antibodies. In this case, it is important to suspect false-positive troponin elevation, even when coronary artery disease is found. This rare and less mentioned and/or recognized cause of troponin elevation may lead to unnecessary invasive diagnostics and aggressive treatment of patients.

Hyperprolactinemia can have different causes: physiological, pharmacological, and pathological. When investigating the etiology of hyperprolactinemia, clinicians need to be aware of several conditions leading to misdiagnosis. The most popular pitfalls are: acute physical and psychological stress, macroprolactin, hook effect, even though antibodies interferences and biotine use have to be considered. A 52-year-old woman was referred to Endocrinology clinic for oligomenorrhoea and headache. She worked as a butcher. Hormonal evaluation showed very high PRL (305 ng/ml, reference interval

The adoptive transfer of ex vivo-expanded natural killer (NK) cells has recently been employed as an alternative cancer treatment in certain institutions. However, the safety profiles of this strategy remain uncharacterized. We evaluated three patients who exhibited elevated serum parathyroid hormone (PTH) levels without the relevant clinical manifestations and had a history of autologous NK cell therapy. The serum PTH concentration was measured using a second-generation PTH assay, and the serum thyroglobulin concentration was measured using a second-generation thyroglobulin assay. Subsequently, the PTH or thyroglobulin concentration obtained using heterophile-blocking tube (HBT) for a secondary confirmation assay was measured and compared with the result of the initial assay. The three patients had falsely elevated serum PTH and thyroglobulin levels owing to heterophile antibody interference associated with NK cell therapy that persisted for at least up to 12 months after the treatment and was confirmed by normalization of hormone levels after HBT treatment. We propose that certain types of mouse monoclonal antibodies used to stimulate NK cells can induce heterophile antibodies. Abnormal laboratory test results in individuals administered NK cell therapy without the relevant clinical manifestations must be examined in the context of heterophile antibody interference to avoid misdiagnosis and unnecessary testing.

A discordance between sex hormone-binding globulin (SHBG) measurements by 2-site ELISAs was investigated using pairings of various "in house" SHBG antibodies together with a concordant control. The 2-site monoclonal ELISAs used the same base coat (11F11) and discordance was observed with one top coat monoclonal antibody (7H9) and also when a polyclonal SHBG antibody was paired with the basecoat antibody (11F11). Sialidase treatment of the discordant sample and purified SHBG revealed increased levels using 7H9 whereas there was no change in SHBG in the concordant sample. Conversely, following sialidase treatment, the discordant sample showed no change in SHBG measured using the other monoclonal antibody pairings whereas the SHBG levels in the concordant sample declined following sialidase using the same monoclonal antibody pairings. This implicated glycosylation as a factor in antibody recognition and synthetic peptides spanning the two N-linked and one O-linked glycosylation regions showed that SHBG recognition by monoclonal antibody 7H9 could be disrupted by a peptide spanning the O-linked glycosylation site. Hence rather than immunoassay discordance being attributed to heterophile antibodies or other circulating antibodies here it can be likely attributed to glycosylation affecting antibody recognition and hence the measurement of SHBG.

Introduction: Interpretation of thyroid function tests (TFTs) in background of non-specific symptoms concerning for hypothyroidism, has become challenging with assay interference. Heterophile antibody interferences are both test and laboratory platform dependent. We present a case of a toxic nodule erroneously diagnosed as isolated central hypothyroidism due to heterophile antibody interference. Case: A 38 year old lady was referred to Endocrinology for possible central hypothyroidism with symptoms of progressive fatigue, 40lb unintentional weight loss, poor appetite, nausea, intermittent diarrhea/constipation, occasional cold intolerance and intermittent lightheadedness. TFTs suggested low TSH 0.175 mcIU/ml (N 0.35-3.8 mcIU/ml) and low FT4 0.74 ng/dl (N 0.8-1.8 ng/dl). Pituitary hormone evaluation revealed normal ACTH, AM cortisol, IGF-1, prolactin and FSH. A pituitary MRI was normal. Two months later, she had worsening fatigue, anxiety with palpitations and tremors, and 14lb weight gain. TFTs again confirmed low TSH (0.575 mcIU/ml) and lower FT4 (0.70 ng/dl). Empirical weight-based levothyroxine (LT4) supplementation was initiated for central hypothyroidism. Six weeks later, TFTs showed hyperthyroidism (TSH Due to persistent discordance in symptoms and biochemical tests, TFTs were evaluated for heterophile antibody interference. While HAMA-treated TSH remained low (0.04 mIU/l), FT4 by equilibrium dialysis (FT4 ED) was normal (1.1 ng/dl, regular assay 0.74 ng/dl), suggesting subclinical hyperthyroidism. A thyroid uptake scan confirmed an autonomous toxic right thyroid nodule with suppressed remaining gland. Patient ultimately underwent 15 mCi RAI ablation of the nodule. TFTs done 1 month post-RAI suggested normal TSH (1.2 mcIU/ml) and normal FT4 ED (1 ng/dl). Previous symptoms have since resolved. Conclusion: Current methods of TFTs are generally reliable in diagnosing and monitoring thyroid disease. Rarely, medications, supplements, and endogenous antibodies can bind to TSH, T4 or lab reagent, resulting in inaccurate values. This has become increasingly common with use of high dose biotin. TSH with HAMA/heterophile antibodies and FT4 ED are more accurate forms of diagnostic testing. When approaching patients with symptoms not consistent with typical hypo- or hyperthyroidism and are not responding as expected to therapy, it is important to consider more accurate testing to rule out assay errors. References: 1. Soh SB, Aw TC. Laboratory Testing in Thyroid Conditions - Pitfalls and Clinical Utility. Ann Lab Med. 2019;39(1):3-14. doi:10.3343/alm.2019.39.1.3 2. Spencer C., PhD. Assay of Thyroid Hormones and Related Substances

Background: Epstein–Barr virus (EBV) infection is associated with significant morbidity and mortality in renal transplant (RT) recipients. The spectrum of illness ranges from infectious mononucleosis (IM) to post-transplant lymphoproliferative disorder (PTLD). In association with clinical signs and symptoms, virus-specific serology and heterophile antibody tests are widely used in confirming the diagnosis of IM in the general population. However, these tests may have a limited role in immunosuppressed RT recipients from seropositive donor, especially in children who were EBV-seronegative prior to the transplant. The aim of this study is to evaluate the utility of these tests in the early diagnosis of IM in this subset of patients. Methods: This is a case study with a review of literature. Results: Here, we present a 14-year-old male with hemophilia B who presented with fever, fatigue, sore throat, palatal petechial rash, exudative tonsillitis and cervical lymphadenopathy 3 months post-RT. He was EBV seronegative prior to RT and received a deceased donor kidney transplant from a seropositive donor. Induction was done with Thymoglobulin and maintenance immunosuppression consisted of tacrolimus and mycophenolate. Initial heterophile antibody test (monospot) was negative, but became positive at 5 months and remained positive at 9 months follow-up post-RT. EBV viral capsid antigens (VCA) IgM and IgG, early antigen (EA) and nuclear antigen (EBNA) were all negative at the time of presentation. VCA IgM and IgG both became positive at 5 months and peaked at 9 months follow-up, however the EA and EBNA remained negative. EBV viral load as measured by polymerase chain reaction (PCR) was negative for the first 3 months post-RT but became positive at presentation, peaked at 6 months and started declining thereafter. Peripheral blood smear examination showed no absolute and atypical lymphocytosis. Cytomegalovirus PCR in the blood and throat culture for streptococcus were negative. There was no splenomegaly. He was managed conservatively with intravenous fluids, bed rest, antipyretics and reduction of immunosuppression. Conclusions: EBV serological markers have a limited role in the early diagnosis of EBV-IM following RT in prior seronegative children. Initial heterophile antibody test may also be negative, and hence a repeat test may be necessary. Once becoming positive, the VCA IgM may remain persistently elevated for prolonged duration. In addition to the suppressed cellular immunity secondary to immunosuppression, humoral response to viral infections is also delayed in transplant recipients, especially in the early transplant period. Hence, routine monitoring with PCR is superior to serology in diagnosing IM early and monitoring the EBV infection post-RT for timely evaluation and management.

Broiler transportation is an important factor in poultry industry. Transportation duration can affect oxidative stress, physiological, and broiler's performance. Transportation process from loading to unloading is a critical point. This study aims to evaluate effects of different transportation durations on physiological responses and performance of broilers. There were two durations treatment in this study, namely short (30 mins) and long (180 mins). Traits measured in this study were physiological responses heterophile lymphocytes (HL) ratio and weight loss percentage. The result showed that transportation duration significantly affects on weight loss. Long transportation duration resulted in higher weight loss (0.065±0.034 gram). It can be concluded long-duration transportation decreased HL ratio and affected performance indicators.

Poultry producers are challenged to improve production while using fewer antibiotics due to increased restriction on antimicrobial usage so prebiotics and probiotics are being tested under different experimental conditions to study the pathways used by these substances to assist in the prevention of carcass contamination and in the elimination of pathogens present in the bird's organisms. The current study aimed to investigate the effect of spray and orally administered Kanne-Brottrunk® which contain Lactobacillus and Bifidobacterium at which lactic acids bacteria concentration is 5x106 cfu/ml in addition to vitamins, minerals, essential amino acids and trace elements on the mucosal trachea IgA, IL-6 and INF-γ as well as healthiness of intestinal and tracheal mucosa using electron microscopy. Six hundred one day old Cobb broilers chickens were reared on litter in three separated groups each one contain 200 chicks, 1st group "Drinking Water Administrated", 2ndgroup "Spray Administrated" and "control non Kanne-Brottrunk® medicated group". Serum, trachea and intestine collected for evaluation of immune status and epithelium healthiness at 14, 21, 28 and 35 day old. Immunological Parameters measured were IL-6, INF-γ and Mucosal IgA concentration using EISA kits accompanied by tracheal and intestinal electron microscopy histomorphology. The results proved that Kanne-Brottrunk® administrated in water stimulate immune response by increased level of IL-6, INF-γ and Mucosal IgA and also increase immune cells infiltration as plasma cell, lymphoid cells, macrophages, and heterophile in trachea and intestine with protection of their tracheal cilia and intestinal villi more than aerosol administration.

In this case report, we describe a young athletic male with a family history of early sudden cardiac death who presented with atypical chest pain and was found to have a positive serum troponin. Although his symptoms resolved without intervention, workup revealed hypertension, hyperlipidemia, mild left ventricular hypertrophy, non-obstructive coronary artery disease, and the presence of serum heterophile antibodies. Ultimately, it was concluded that his rigorous exercise regimen as well as the presence of heterophile antibodies may have contributed to his positive serum troponin. This case serves as a reminder of the nonspecific diagnostic value of modern troponin assays, and that the results of these tests should always be incorporated into the clinical context.

Aim: This study aimed to evaluate the effects of Meniran extract (Phyllanthus niruri L.) administration on leukocyte profile of broiler chickens infected with Mycoplasma gallisepticum. Materials and Methods: Thirty broiler chickens, 21 days old were divided into five treatment groups. P0 (-): Chickens without any treatment; P0 (+), P1, P2, and P3: Chickens were infected with M. gallisepticum 108 cells/ml/animal orally, then given no treatment, Meniran extract 60%, 62.5%, and 65% orally at a dose of 1 ml/kg body weight, respectively. The treatment of Meniran extract was given for 7 days. Results: Leukocyte count with the lowest number showed in Group P0 (-) and Group P3 (p>0.05). Increased number of basophils was found in Group P0 (+), Group P1, and Group P2. The highest number of heterophils was found in Group P0 (+) and was significantly different from Group P0 to P3 (p0.05). Discussion: Increased the number of leukocytes is often observed in inflammation due to general infections, trauma, or toxicity. Shifting in the number of heterophile or lymphocytes, an increase in the number of monocytes, basophils, and eosinophils may also be associated with various infectious or inflammatory conditions. Heterophils play a role as an antibacterial defense through several effective mechanisms. When infections and inflammation occur, the heterophils will increase to phagocytosis microbe. Conclusion: It can be concluded that Meniran extract (P. niruri L.) at a dose of 65% can decrease the total number of leukocytes in broilers infected with M. gallisepticum.

The post-operative management of patients with differentiated thyroid carcinoma (DTC) relies on serial measurements of serum thyroglobulin. Current methodologies for thyroglobulin quantitation vary in their analytical and clinical performance. For years, thyroglobulin radioimmunoassays (RIA) and immunometric assays (IMA) have been used despite analytical interferences from anti-thyroglobulin autoantibodies (TgAb) as well as heterophile antibodies (HAb). TgAb interference limits Tg utility as a tumor marker in ∼30% of TgAb-positive patients. Consequently, additional studies are necessary to rule out persistent or recurrent disease in these patients. Recently, thyroglobulin mass spectrometry assays have been introduced as a solution to the interference problems observed in immunoassays. However, their analytical sensitivity is inferior to the high sensitivity immunoassays. The aims of this review are to: (i) review current thyroglobulin assays; (ii) discuss technical limitations of each assay; and (iii) discuss the clinical uses of thyroglobulin in serum and fine-needle aspirate biopsy washouts for the management of DTC patients. An understanding of the technical advantages and disadvantages of Tg assays is critical for clinicians and laboratorians to effectively use and interpret this test in the management of DTC patients.

Infectious mononucleosis (IM) is often an uncomplicated self-limited illness resulting from Epstein-Barr virus (EBV) in 90% cases. This is a case report of 21-year-old female whose initial clinical and laboratory presentation suggested Heterophile antibody negative Epstein–Barr Viral capsid Antigen (VCA) IgM positive infectious Mononucleosis. Our case was complicated by biliary stasis, cold autoimmune hemolytic anemia with acrocyanosis, thrombocytopenia and some of the features of hemophagocytic lymphohistiocytosis (HLH). Following symptomatic management patient recovered. Physicians should routinely counsel their patients with IM for these complications and should avoid overzealous treatment