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Neuroinflammation is involved in the pathogenesis of Alzheimer's disease, and the transcription factor NF-kappa B is a player in this event. We found here that the ischemic damage alone or in association with A beta(1-42) activates the NF-kappa B pathway, induces an increase of BACE1 and a parallel inhibition of Uch-L1 and TREM2, both in vitro and in vivo, in Tg 5XFAD and in human brains of sporadic AD. This mechanism creates a synergistic loop that fosters inflammation. We also demonstrated a significant protection exerted by the restoration of Uch-L1 activity. The rescue of the enzyme is able to abolish the decrease of TREM2 and the parameters of neuroinflammation.

Glial fibrillary acidic protein (GFAP), ubiquitin carboxy-terminal hydrolase-L1 (UCH-L1), and matrix metalloproteinase 9 (MMP-9) are potential biomarkers of traumatic brain injury (TBI) but also of secondary insults to the brain. The aim of this study was to describe the cerebral distribution of GFAP, UCH-L1, and MMP-9 in a rat model of diffuse TBI associated with standardized hypoxia-hypotension (HH). Adult male Sprague-Dawley rats were allocated to Sham (n = 10), TBI (n = 10), HH (n = 10), and TBI+HH (n = 10) groups. After 4 hours, brains were rapidly removed and immunostaining of GFAP, UCH-L1, and MMP-9 was performed. Areas of interest that have been described as particularly sensitive to hypoxic insults were analyzed. For GFAP, in the neocortex, immunostaining revealed a significant decrease in strong staining for HH and TBI+HH groups compared with TBI group ( P

BACKGROUND: Glial fibrillary acidic protein (GFAP) and ubiquitin C-terminal hydrolase-L1 (UCH-L1) are promising biomarkers of traumatic brain injury (TBI). OBJECTIVE: We investigated the relation of the GFAP and UCH-L1 levels to the severity of TBI during the first week after injury. METHODS: Plasma UCH-L1 and GFAP were measured from 324 consecutive patients with acute TBI and 81 control subject enrolled in a 2-center prospective study. The baseline measures included initial Glasgow Coma Scale (GCS), head computed tomographic (CT) scan at admission, and blood samples for protein biomarkers that were collected at admission and on days 1, 2, 3, and 7 after injury. RESULTS: Plasma levels of GFAP and UCH-L1 during the first 2 days after the injury strongly correlated with the initial severity of TBI as assessed with GCS. Additionally, levels of UCH-L1 on the seventh day after the injury were significantly related to the admission GCS scores. At admission, both biomarkers were capable of distinguishing mass lesions from diffuse injuries in CT, and the area under the curve of the receiver-operating characteristic curve for prediction of any pathological finding in CT was 0.739 (95% confidence interval, 0.636-0.815) and 0.621 (95% confidence interval, 0.517-0.713) for GFAP and UCH-L1, respectively. CONCLUSION: These results support the prior findings of the potential role of GFAP and UCH-L1 in acute-phase diagnostics of TBI. The novel finding is that levels of GFAP and UCH-L1 correlated with the initial severity of TBI during the first 2 days after the injury, thus enabling a window for TBI diagnostics with latency. ABBREVIATIONS: AUC, area under the curveCI, confidence intervalED, emergency departmentGCS, Glasgow Coma ScaleGRAP, glial fibrillary acidic proteinIMPACT, International Mission for Prognosis and Clinical TrialROC, receiver-operating characteristicTBI, traumatic brain injuryTRACK-TBI, Transforming Research and Clinical Knowledge in Traumatic Brain InjuryUCH-L1, ubiquitin C-terminal hydrolase-L1.

Ubiquitin C-terminal hydrolase L1 (UCH-L1) is a deubiquitinating enzyme that is highly expressed in neurons, and gathering evidence indicates that UCH-L1 may play pathogenic roles in many neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease (PD). Additionally, lipid rafts have attracted interest in neurodegeneration as playing a common role in many neurodegenerative diseases. In the present study, we demonstrated that UCH-L1 associates with lipid rafts as with other PD-associated gene products. In addition, UCH-L1 regulates lipid raft-dependent endocytosis and it is not dependent on the expression and degradation of caveolin-1 or flotillin-1. Finally, UCH-L1 regulates cell-to-cell transmission of α-synuclein. This study provides evidence that many PD-associated gene products share common signaling pathways to explain the pathogenesis of PD., Graphical Abstract

Мета. На основі аналізу результатів клініко-біохімічного і комп’ютерно-томографічного дослідження потерпілих у гострому періоді тяжкої черепно-мозкової травми (ЧМТ) оцінити можливості визначення концентрації у сироватці біомаркера ушкодження нейронів білка UCH-L1 і біомаркера ушкодження астроглії білка S100B у 1-шу добу після травми у визначенні патофізіологічних особливостей ЧМТ, зокрема, в діагностиці вогнищевого і дифузного ушкодження головного мозку (ГМ).Матеріали і методи. Проаналізовані результати діагностичних досліджень і лікувальних маніпуляцій, проведених у 72 потерпілих віком від 16 до 76 років за тяжкої ЧМТ. Оцінювали кореляцію результатів молекулярно-біологічного дослідження (визначення концентрації білків UCH-L1 та S100B у сироватці крові методом твердофазного імуноферментного аналізу — ІФА з використанням наборів виробництва “Sigma-Aldrich”, США у 1-шу добу після тяжкої ЧМТ) з видом ушкодження відповідно до класифікації L.F. Marshall (дифузного або вогнищевого).Результати. За тяжкої ізольованої ЧМТ у 1-шу добу при виключенні супутніх ушкоджень, інтоксикації та інших причин непритомного стану потерпілого, перевищення порогового значення показника відношення концентрації UCH-L1 / S100B у сироватці крові 15,8 дозволяло з високою вірогідністю діагностувати дифузне ушкодження ГМ, при значенні показника, меншого за порогове – діагностувати вогнищеве ушкодження ГМ. Чутливість моделі 77,8%, специфічність — 79,6%.Висновок. Визначення концентрації біомаркера ушкодження нейронів білка UCH-L1 та біомаркера ушкодження астроглії білка S100B у сироватці крові потерпілих у 1-шу добу після травми дозволяє з високою ефективністю діагностувати дифузне й вогнищеве ушкодження ГМ за тяжкої ЧМТ., Purpose: To evaluate the feasibility of determining serum concentration of biomarker of neuronal protein damage UCH-L1 and biomarker of astroglia damage S100 on the 1st day after injury, particularly for the diagnosis of focal and diffuse brain impairments based on clinical and biochemical and computer tomographic study in patients with acute severe brain injury.Methods. We used the results of diagnostic tests and therapeutic manipulations in 72 patients aged 16 to 76 years with severe traumatic brain injury. Correlation of the molecular biological study results (determining the UCH-L1 and S100B serum concentration by solid phase enzyme immunoassay — ELISA using sets of reagents Sigma-Aldrich, USA, on the 1st day after severe TBI) with the injury type (diffuse or focal according to L. F. Marshall classification) was evaluated.Results. In patients with isolated severe TBI, after exclusion of concomitant extracranial injuries, intoxication and other causes for unconsciousness, UCH-L1 / S100B concentration ratio exceeding the cut-off value of 15.8 indicated a high probability of diffuse injury, ratio less than 15.8 was a marker of focal injury. The sensitivity of the model was 77.8%, specificity — 79.6%.Conclusion. It has been shown that the estimation of serum concentration of neuronal damage biomarker UCH-L1 and astroglial damage biomarker S100B in patients with severe TBI on the 1st day after injury allows diagnose diffuse and focal brain damage with high efficiency., Цель. На основе анализа результатов клинико-биохимического и компьютерно-томографического исследования пострадавших в остром периоде тяжелой ЧМТ оценить возможности определения концентрации в сыворотке биомаркера повреждения нейронов белка UCH-L1 и биомаркера повреждения астроглии белка S100B в 1-е сутки после травмы в определении патофизиологических особенностей ЧМТ, в частности, диагностике очагового и диффузного повреждения головного мозга.Материалы и методы. Проанализированы результаты диагностических исследований и лечебных манипуляций, проведенных у 72 пострадавших в возрасте от 16 до 76 лет с тяжелой ЧМТ. Оценивали корреляцию результатов молекулярно-биологического исследования (определение концентрации белков UCH-L1 и S100B в сыворотке крови методом твердофазного иммуноферментного анализа - ИФА с использованием наборов производства «Sigma-Aldrich», США в 1-е сутки после тяжелой ЧМТ) с видом повреждения (диффузного или очагового), определенного в соответствии с классификацией LF Marshall.Результаты. При тяжелой изолированной ЧМТ в 1-е сутки после травмы, при исключении сопутствующих повреждений, интоксикации и других причин нарушения сознания пострадавшего, превышение порогового значения показателя отношение концентрации UCH-L1 / S100B в сыворотке крови» 15,8 позволяло с высокой вероятностью диагностировать диффузное повреждение головного мозга, а при значениях этого показателя ниже порогового 15,8 - диагностировать очаговое повреждение головного мозга. Чувствительность модели 77,8%, специфичность - 79,6%.Вывод. Определение концентрации биомаркера повреждения нейронов белка UCH-L1 и биомаркера повреждения астроглии белка S100B в сыворотке крови пострадавших в 1-е сутки после травмы позволяет с высокой эффективностью диагностировать диффузное и очаговое повреждение головного мозга при тяжелой ЧМТ.

// Hyun Jung Kim 1 , Venkataraman Magesh 1 , Jae-Jin Lee 1 , Sun Kim 1 , Ulla G. Knaus 2 and Kong-Joo Lee 1 1 Graduate School of Pharmaceutical Sciences, College of Pharmacy, Ewha Womans University, Seoul, Korea 2 Conway Institute, University College Dublin, Dublin, Ireland Correspondence to: Kong-Joo Lee, email: // Keywords : UCH-L1, ubiquitination, hydrogen peroxide, NOX4, invasion Received : December 19, 2014 Accepted : March 20, 2015 Published : April 15, 2015 Abstract This study explored the role of ubiquitin C-terminal hydrolase-L1 (UCH-L1) in the production of ROS and tumor invasion. UCH-L1 was found to increase cellular ROS levels and promote cell invasion. Silencing UCH-L1, as well as inhibition of H 2 O 2 generation by catalase or by DPI, a NOX inhibitor, suppressed the migration potential of B16F10 cells, indicating that UCH-L1 promotes cell migration by up-regulating H 2 O 2 generation. Silencing NOX4, which generates H 2 O 2 , with siRNA eliminated the effect of UCH-L1 on cell migration. On the other hand, NOX4 overexpressed in HeLa cells happens to be ubiquitinated, and NOX4 following deubiquitination by UCH-L1, restored H 2 O 2 -generating activity. These in vitro findings are consistent with the results obtained in vivo with catalase (-/-) C57BL/6J mice. When H 2 O 2 and UCH-L1 levels were independently varied in these animals, the former by infecting with H 2 O 2 -scavenging adenovirus-catalase, and the latter by overexpressing or silencing UCH-L1, pulmonary metastasis of B16F10 cells overexpressing UCH-L1 increased significantly in catalase (-/-) mice. In contrast, invasion did not increase when UCH-L1 was silenced in the B16F10 cells. These findings indicate that H 2 O 2 levels regulated by UCH-L1 are necessary for cell invasion to occur and demonstrate that UCH-L1 promotes cell invasion by up-regulating H 2 O 2 via deubiquitination of NOX4.

Diagnosis and monitoring of diseases by measurement of biochemical markers has most commonly been performed on samples of peripheral blood. However, no such markers are available for clinical use in the major diseases of the central nervous system (CNS). In Parkinson's disease circulating biomarkers would find clinical utility in early diagnosis and also monitoring of disease progression. Of particular interest is early diagnosis as this would create .a window of opportunity for treatment with neuroprotective drugs. We have developed a novel strategy for monitoring disease activity in the CNS based on the recognition that tissue injuries incite inflammation and recruitment of phagocytes that engulf debris. We postulated that some of these debris laden phagocytes may return to the peripheral blood and their cargo of CNS proteins could be measured. If CNS antigens can be measured in PBMCs it may be an indicator of active neurodegeneration as the debris engulfed by phagocytes is completely degraded within days. To make this approach more specific to Parkinson's disease we probed PBMC lysates for neuromelanin as a marker of degeneration within the substancia nigra. We performed a proof of principle study in ten subjects with early PD and ten age and sex matched controls. The biomarkers neuromelanin, Tau protein, UCH-L1 and HPCAL-1 were measured in PBMC lysates from these two groups. Neuromelanin and Tau protein mean levels were elevated in PD compared with controls and was extremely statistically significant in both cases. UCH-L1 and HPCAL-1 mean levels were elevated in PD over controls and were not quite significant in both cases. These results suggest that this is a promising new approach for diagnosis and monitoring of PD and potentially other CNS diseases.

WOS: 000412669100014 PubMed ID: 29033641 Introduction: The mechanism of ubiquitination-related abnormalities causing neural development problems is still unclear. We examined the association between autism and serum transactive response DNAbinding protein-43 (TDP-43) and ubiquitin c-terminal hydrolase-L1 (UCH-L1) levels, both of which are members of the ubiquitinproteosome system. Methods: We measured serum levels of TDP-43 and UCH-L1 in 24 children with autism and 24 healthy children. Childhood Autism Rating Scale (CARS) was used to assess symptom severity at admission. Results: The mean serum TDP-43 and UCH-L1 levels in children with autism spectrum disorder (ASD) were found to decrease compared to healthy controls (p< 0.001, 506.21 +/- 780.97 ng/L and 1245.80 +/- 996.76 ng/L, respectively; 3.08 +/- 5.44 ng/mL and 8.64 +/- 6.67 ng/mL, respectively). A positive correlation between serum TDP-43 levels and UCH-L1 levels was found in the ASD group (r= 0.947, n= 24, p< 0.001). The CARS score of children with ASD was 48.91 points (standard deviation [SD]: 5.82). Conclusion: Low serum levels of UCH-L1 and TDP-43 may reflect disturbed ubiquitination in autism. Amaç: Protein ubikitinlenme ile ilgili bozuklukların nöronal gelişim sorunlarına nasıl yol açtığı açıklanamamıştır. Bu çalışmada, ubikuitin proteozom sistemine üye olan ubikuitin c-terminal hidrolaz-L1 (UCH-L1) ve transaktif yanıtlı DNA-bağlayıcı protein (TDP-43) düzylerinin otizm ile ilişkisi araştırılmıştır.Yöntemler: Bu bağlamda 24 otizm tanılı çocuk ve 24 sağlıklı çocukta serum TDP-43 ve UCH-L1 düzeylerine bakılmıştır. Otizm şiddet derecesi Çocukluk Otizm Derecelendirme Ölçeği (ÇODÖ) ile değerlendirilmiştir. Bulgular: Otizm tanılı çocukların başvuru sırasında ortalama ÇODÖ puanları 48.91 (SD: 5.82) idi. Sağlıklı gruba göre otizmli grupta serum TDP-43 ve Serum UCH-L1 düzeyleri anlamlı olarak düşüktü (sırasıyla 506,21±780,97 ng/L ve 1245,80±996,76 ng/L; 3,08±5,44 ng/mL ve 8,64±6,67 ng/mL; p

Glial fibrillary acidic protein (GFAP) and ubiquitin C-terminal hydrolase-L1 (UCH-L1) have been studied as potential biomarkers of mild traumatic brain injury (mTBI). We report the levels of GFAP and UCH-L1 in patients with acute orthopedic injuries without central nervous system involvement, and relate them to the type of extracranial injury, head magnetic resonance imaging (MRI) findings, and levels of GFAP and UCH-L1 in patients with CT-negative mTBI. Serum UCH-L1 and GFAP were longitudinally measured from 73 patients with acute orthopedic injury on arrival and on days 1, 2, 3, 7 after admission, and on the follow-up visit 3–10 months after the injury. The injury types were recorded, and 71% patients underwent also head MRI. The results were compared with those found in patients with CT-negative mTBI ( n = 93). The levels of GFAP were higher in patients with acute orthopedic trauma than in patients with CT-negative mTBI ( p = 0.026) on arrival; however, no differences were found on the following days. The levels of UCH-L1 were not significantly different between these two groups at any measured point of time. Levels of GFAP and UCH-L1 were not able to distinguish patients with CT-negative mTBI from patients with orthopedic trauma. Patients with orthopedic trauma and high levels of UCH-L1 or GFAP values may be falsely diagnosed as having a concomitant mTBI, predisposing them to unwarranted diagnostics and unnecessary brain imaging. This casts a significant doubt on the diagnostic value of GFAP and UCH-L1 in cases with mTBI.

The purpose of the study was to investigate the potential of determining the serum concentration of neuronal damage biomarker ubiquitin C-terminal hydrolase L1 (UCH-L1) for predicting the outcomes of traumatic brain injury (TBI).Methods. We analyzed the results of diagnostic tests and therapeutic interventions in 72 patients aged 16 to 76 years with severe traumatic brain injury of different origin with the assessment of its outcomes. The results of the molecular biological study (determining the UCH-L1 serum concentration by solid phase enzyme immunoassay — ELISA using sets of reagents Sigma-Aldrich, USA, on the 1st day after severe TBI) were compared with the analyzes of 10 healthy donors. The TBI outcomes were assessed by Glasgow Outcome Scale (GOS) in 6 months after injury. A correlation of severe TBI outcomes with UCH-L1 serum levels obtained on the 1st day after injury was evaluated.Results. In patients with isolated severe TBI, after exclusion of concomitant extracranial injuries, intoxication and other causes for unconsciousness, serum levels of UCH-L1 exceeding the cut-off value of 29.5 ng/ml indicated a high probability of patient’s death (sensitivity of the model was 91.2 %, specificity — 94.7 %). The levels of serum UCH-L1 exceeding the cut-off value of 15.2 ng/ml on the 1st day after severe TBI was a marker of a high probability of unfavorable outcomes in 6 months after the trauma, which include death and severe disability (sensitivity of the model was 100 %, specificity — 86.7 %).Conclusion. The estimation of serum concentration of neuronal damage biomarker UCH-L1 in patients with severe TBI on the 1st day after injury demonstrated a high effectiveness for predicting the TBI outcomes in 6 months after injury., Цель исследования — изучение возможностей определения содержания в сыворотке крови биомаркера повреждения нейронов белка убиквитин-С-концевой гидролазы L1 (UCH-L1) в прогнозировании последствий черепно-мозговой травмы (ЧМТ).Материалы и методы. Проанализированы результаты диагностических исследований и лечебных манипуляций у 72 пострадавших в возрасте от 16 до 76 лет с тяжелой ЧМТ различного происхождения с оценкой ее последствий. Результаты молекулярно-биологического исследования (определение концентрации белка UCH-L1 в сыворотке крови методом твердофазного иммуноферментного анализа — ИФА с использованием наборов производства “Sigma-Aldrich”, США в 1-е сутки после тяжелой ЧМТ) сравнивали с соответствующими данными у 10 условно здоровых доноров. Последствия ЧМТ оценивали с помощью шкалы исходов Глазго (ШИГ) через 6 мес после травмы. Установлена корреляция исхода ЧМТ с концентрацией UCH-L1 в сыворотке крови в 1-е сутки после травмы.Результаты. У пострадавших с тяжелой изолированной ЧМТ, при исключении сопутствующих повреждений, интоксикации и других причин потери сознания, превышение уровня UCH-L1 в сыворотке крови порогового значения 29,5 нг/мл позволяло с высокой вероятностью прогнозировать смерть больного (чувствительность модели 91,2%, специфичность — 94,7%). Превышение концентрации белка UCH-L1 в сыворотке крови в 1-е сутки после тяжелой ЧМТ порогового значения 15,2 нг/мл с высокой вероятностью свидетельствовало о возникновении через 6 мес после травмы неблагоприятных последствий, в том числе, смерти и тяжелой инвалидизации (чувствительность модели 100%, специфичность — 86,7%).Вывод. Определение концентрации биомаркера повреждения нейронов белка UCH-L1 в сыворотке крови пострадавших с тяжелой ЧМТ в 1-е сутки после травмы позволяет с высокой вероятностью прогнозировать неблагоприятные последствия травмы, которые возникают через 6 мес после повреждения., Мета дослідження — вивчення можливостей визначення вмісту в сироватці крові біомаркера ушкодження нейронів білка убіквітин-С-кінцевої гідролази L1 (UCH-L1) в прогнозуванні наслідків черепно-мозкової травми (ЧМТ).Матеріали і методи. Проаналізовані результати діагностичних досліджень і лікувальних маніпуляцій у 72 потерпілих віком від 16 до 76 років з тяжкою ЧМТ різного походження з оцінкою її наслідків. Результати молекулярно-біологічного дослідження (визначення концентрації білка UCH-L1 у сироватці крові методом твердофазного імуноферментного аналізу — ІФА з використанням наборів виробництва “Sigma-Aldrich”, USA у 1-шу добу після тяжкої ЧМТ) порівнювали з відповідними даними у 10 умовно здорових донорів. Наслідки ЧМТ оцінювали за шкалою наслідків Глазго (ШНГ) через 6 міс після травми. Встановлено кореляцію наслідків ЧМТ з концентрацією UCH-L1 у сироватці крові у 1-шу добу після травми.Результати. За тяжкої ізольованої ЧМТ, при виключенні супутніх ушкоджень, інтоксикації та інших причин непритомного стану, перевищення рівня UCH-L1 у сироватці крові порогового значення 29,5 нг/мл дозволяло з високою вірогідністю прогнозувати смерть хворого (чутливість моделі 91,2%, специфічність — 94,7%). Перевищення концентрації білка UCH-L1 у сироватці крові у 1-шу добу після тяжкої ЧМТ порогового значення 15,2 нг/мл з високою вірогідністю свідчило про появу через 6 міс після травми несприятливих наслідків, в тому числі смерті та тяжкої інвалідизації (чутливість моделі 100%, специфічність — 86,7%).Висновок. Визначення концентрації біомаркера ушкодження нейронів білка UCH-L1 у сироватці крові потерпілих за тяжкої ЧМТ у 1-шу добу після травми дозволяє з високою вірогідністю прогнозувати несприятливі наслідки травми, що виникають через 6 міс після ушкодження.

Ubiquitin C-terminal Hydrolase L1 (UCH-L1) has oncogenic properties and is highly expressed during malignancies. We recently documented that Epstein-Barr virus (EBV) infection induces uch-l1 expression. Here we show that Kaposi's Sarcoma-associated herpesvirus (KSHV) infection induced UCH-L1 expression, via cooperation of KSHV Latency-Associated Nuclear Antigen (LANA) and RBP-Jκ and activation of the uch-l1 promoter. UCH-L1 expression was also increased in Primary Effusion Lymphoma (PEL) cells co-infected with KSHV and EBV compared with PEL cells infected only with KSHV, suggesting EBV augments the effect of LANA on uch-l1. EBV latent membrane protein 1 (LMP1) is one of the few EBV products expressed in PEL cells. Results showed that LMP1 was sufficient to induce uch-l1 expression, and co-expression of LMP1 and LANA had an additive effect on uch-l1 expression. These results indicate that viral latency products of both human γ-herpesviruses contribute to uch-l1 expression, which may contribute to the progression of lymphoid malignancies.

Objective Biomarkers ubiquitin C-terminal hydrolase-L1 (UCH-L1) and glial fibrillary acidic protein (GFAP) may help detect brain injury, assess its severity, and improve outcome prediction. This study aimed to evaluate the prognostic value of these biomarkers during the first days after brain injury. Methods Serum UCH-L1 and GFAP were measured in 324 patients with traumatic brain injury (TBI) enrolled in a prospective study. The outcome was assessed using the Glasgow Outcome Scale (GOS) or the extended version, Glasgow Outcome Scale–Extended (GOSE). Results Patients with full recovery had lower UCH-L1 concentrations on the second day and patients with favorable outcome had lower UCH-L1 concentrations during the first 2 days compared with patients with incomplete recovery and unfavorable outcome. Patients with full recovery and favorable outcome had significantly lower GFAP concentrations in the first 2 days than patients with incomplete recovery or unfavorable outcome. There was a strong negative correlation between outcome and UCH-L1 in the first 3 days and GFAP levels in the first 2 days. On arrival, both UCH-L1 and GFAP distinguished patients with GOS score 1–3 from patients with GOS score 4–5, but not patients with GOSE score 8 from patients with GOSE score 1–7. For UCH-L1 and GFAP to predict unfavorable outcome (GOS score ≤3), the area under the receiver operating characteristic curve was 0.727, and 0.723, respectively. Neither UCHL-1 nor GFAP was independently able to predict the outcome when age, worst Glasgow Coma Scale score, pupil reactivity, Injury Severity Score, and Marshall score were added into the multivariate logistic regression model. Conclusions GFAP and UCH-L1 are significantly associated with outcome, but they do not add predictive power to commonly used prognostic variables in a population of patients with TBI of varying severities.

Increasing studies identified podocyte injury as a key early risk factor resulting in diabetic nephropathy (DN). The ubiquitin carboxy-terminal hydrolase 1 (UCH-L1) participates in podocyte differentiation and injury, which is elevated in the podocytes of a variety of nephritis. Whether UCH-L1 expression is positively related to podocyte injury of DN remains unclear. In this study, elevated expression of UCH-L1 and its intrinsic mechanism in high glucose (HG)-stimulated murine podocytes were investigated using western blot and real-time quantitative PCR. Kidney biopsies of DN patients and health individuals were stained by immunofluorescence (IF) method. The morphological and functional changes of podocytes were tested by F-actin staining and cell migration assay. Results demonstrated that HG induced upregulation of UCH-L1 and activation of the Wnt/β-catenin signaling pathway in podocytes. However, blocking of the Wnt pathway by dickkopf related protein 1 (DKK1) eliminated the above changes. Furthermore, IF staining confirmed that, compared with healthy individuals, the expression of UCH-L1 and β-catenin were obviously increased in kidney biopsy of DN patients. Overexpression of UCH-L1 remodeled its actin cytoskeleton, increased its cell migration and impacted its important proteins. All the findings manifested that Wnt/β-catenin/UCH-L1 may be a new potential therapy method in the treatment of DN in future.

A rapid and reliable diagnostic test to distinguish ischemic from hemorrhagic stroke in patients presenting with stroke-like symptoms is essential to optimize management and triage for thrombolytic therapy. The present study measured serum concentrations of ubiquitin C-terminal hydrolase (UCH-L1) and glial fibrillary astrocytic protein (GFAP) in acute stroke patients and healthy controls and investigated their relation to stroke severity and patient characteristics. We also assessed the diagnostic performance of these markers for the differentiation of intracerebral hemorrhage (ICH) from ischemic stroke (IS). Both UCH-L1 and GFAP concentrations were significantly greater in ICH patients than in controls (p

Dikkat eksikliği ve hiperaktivite bozukluğunun tam olarak nedeni bilinmemekte ancak, dopaminerjik ve nöradrenerjik aktivitelerin inhibitör etkisi ile serebral korteks içindeki katekolamin metabolizmasındaki düzensizlik DEHB'nin en muhtemel nedenidir. Nörobiyolojikal faktörler DEHB'nin etiyolojisinde rol oynamaktadır. Fakat DEHB'nin prognozunu ve tedavisini etkileyen biyolojik markırlar çok sınırlıdır. Bu çalışmanın Dicle üniversitesi Tıp Fakültesi Çocuk Psikiyatri Polikliniğine başvuran, DEHB tanısı alan ve başka herhangi bir sistemik bozukluğu olmayan 30 çocuk ve 30 sağlıklı çocuk ile yapılması planlandı. Glial fibriler asidik protein (GFAP), Nogo-A, ubikuitin karboksi terminal hidrolaz-L1 (UCH-L1) ve TAR DNA bağlayıcı protein (TDP-43) serum düzeyleri Enzim-bağlı-immunosorbent yöntemi ile belirlendi. Çalışmamızda UCH-L1 ve TDP-43 düzeylerinin kontrol grubu değerlerine göre DEHB grubunda istatistiksel olarak anlamlı düzeyde yüksek olduğu bulunmuştur. Benzer şekilde Nogo-A ve GFAP düzeyleri de DEHB grubunda istatistiksel olarak anlamlı düzeyde yüksek bulunmuştur. Nöroplastik bozuklukların DEHB ile ilişkili olduğunu ve TDP-43 ve UCH-L1 düzeylerinin DEHB bozukluğunun erkenden tanılanmasını büyük oranda kolaylaştırabileceğini öne sürebiliriz. Ancak TDP-43 ve UCH-L1seroprospinal sıvı düzeylerinin DEHB bozukluğu çocuklarında henüz belirsizdir ve bu konu daha fazla araştırma yapılmayı haketmektedir. Dahası, GFAP ve Nogo-A seviyeleri DEHB'de nöroplastisite, microglial ve astroglial değişikliklerinin de bir kanıtı olabilir., Although the reason for attention deficit and hyperactivity disorder is not known exactly, with inhibitor effect of dopaminergic and noradrenergic activities, disorder in catecholamine metabolism in cerebral cortex is the most possible cause of ADHD. Neurobiological factors play an important role in the ethology of ADHD. However, the biological markers effecting the prognosis and treatment of ADHD are very limited. This study was planned to be conducted with the inclusion of 30 children who applied to Dicle University, Medicine Faculty, Psychiatry Polyclinic, and were diagnosed with ADHD yet without any systemic disease; and 30 healthy children. The serum levels of glial fibrillary acidic protein (GFAP), nogo-A, ubiquitin carboxy-terminal hydrolase L1 (UCH-L1) and TAR DNA binding protein (TDP-43) were determined by Enzyme-linked immunosorbent assay (ELISA). In our study, it was found that serum levels of UCH-L1 and TDP-43 levels were statistically significantly higher in ADHD children than those of control group. Similarly, Nogo-A and GFAP levels were also found to be statistically significantly high in ADHD group. It can be suggested that neuroplasticity disorders are associated with ADHD and the levels of TDP-43 and UCH-L1 can ease the early diagnosis of ADHD to a great extent. Nevertheless, the cerebrospinal fluid levels of TDP-43 and UCH-L1 are yet unclear in children with ADHD; therefore, this issue deserves further investigation. In addition, GFAP and Nogo-A levels can be an evidence of neuroplasticity, microglial and astroglial changes in ADHD.

Most of the mammalian genome is transcribed. This generates a vast repertoire of transcripts that includes protein-coding messenger RNAs, long non-coding RNAs (lncRNAs) and repetitive sequences, such as SINEs (short interspersed nuclear elements). A large percentage of ncRNAs are nuclear-enriched with unknown function. Antisense lncRNAs may form sense-antisense pairs by pairing with a protein-coding gene on the opposite strand to regulate epigenetic silencing, transcription and mRNA stability. Here we identify a nuclear-enriched lncRNA antisense to mouse ubiquitin carboxy-terminal hydrolase L1 (Uchl1), a gene involved in brain function and neurodegenerative diseases. Antisense Uchl1 increases UCHL1 protein synthesis at a post-transcriptional level, hereby identifying a new functional class of lncRNAs. Antisense Uchl1 activity depends on the presence of a 5' overlapping sequence and an embedded inverted SINEB2 element. These features are shared by other natural antisense transcripts and can confer regulatory activity to an artificial antisense to green fluorescent protein. Antisense Uchl1 function is under the control of stress signalling pathways, as mTORC1 inhibition by rapamycin causes an increase in UCHL1 protein that is associated to the shuttling of antisense Uchl1 RNA from the nucleus to the cytoplasm. Antisense Uchl1 RNA is then required for the association of the overlapping sense protein-coding mRNA to active polysomes for translation. These data reveal another layer of gene expression control at the post-transcriptional level.

The ubiquitin-proteasome system (UPS) functions as a major degradation pathway for misfolded and damaged proteins with an important neuroprotective role in the CNS against a variety of cellular stresses. Parkin and ubiquitin C-terminal hydrolase L1 (UCH-L1) are two relevant components of the UPS associated with a number of neurodegenerative disorders. We here address the expression profile of parkin and UCH-L1 in the mammalian retina, with special emphasis on primates. We describe for the first time the presence of parkin in the retina of mammals, including humans. Parkin and UCH-L1 genes were expressed at the mRNA and protein levels in the retina of all species examined. The immunolocalization pattern of parkin was quite widespread, being expressed by most retinal neuronal types, including photoreceptors. UCH-L1 was localized to horizontal cells and specific subtypes of bipolar and amacrine cells, as well as to ganglion cells and their axons forming the nerve fiber layer. In rodents no UCH-L1 immunoreactivity was found in cone or rod photoreceptors, whereas this protein was present along the whole length of cones in all other mammals. Remarkably, UCH-L1 was expressed by dopaminergic amacrine cells of primates. The ample distribution of parkin and UCH-L1 in the mammalian retina, together with the crucial role played by the UPS in normal neuronal physiology in the brain, points to a participation of these two proteins in the ubiquitin-proteasomal pathway of protein degradation in most retinal cell types, where they could exert a protective function against neuronal stress. This research was supported by grants from the Spanish Instituto de Salud Carlos III (ISCIII; PI09/1623) to J.M.-N., Ministerio de Ciencia e Innovación (MICINN; SAF2007-62262), Fundación Séneca (FS/05662/PI/07) and Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED) to M.-T.H.; and MICINN (BFU2009-07793/BFI), ISCIII (RETICS RD07/0062/0012), Generalitat Valenciana (ACOMP/2009/139), Organización Nacional de Ciegos de España (ONCE) and Fundación Lucha contra la Ceguera (FUNDALUCE) to N.C. J.E.-R. and L.C. are recipients of predoctoral contracts from the Universidad de Alicante.

Bu tez çalışması Batman Üniversitesi Bilimsel Araştırma Projeleri birimi tarafından BTÜBAP_2015_Yüksek Lisans_1 nolu proje ile desteklenmiştir., Çalışmamızda otizmde diagnostik marker olarak kullanılma potansiyeli olduğu düşünülen nöroplastisit, nörodejeneratif ve nöroprotektif molekül düzeylerinin incelenmesi ile periferik kan düzeyinde teşhis sağlayabilecek, uygulanabilirliği kolay biyomarkırların belirlenmesi ve otistik bozukluğun patofizyolojik mekanizmasın anlaşılması için farklı bir bakış açısının oluşturulması amaçlanmıştır Bu çalışmanın İstanbul Üniversitesi, Cerrahpaşa Tıp Fakültesi Çocuk Psikiyatri Polikliniğine başvuran, otizm tanısı alan ve başka herhangi bir sistemik bozukluğu olmayan 30 çocuk ve 30 sağlıklı çocuk ile yapılması planlandı. Glial fibriler asidik protein (GFAP), Nogo-A, ubikuitin karboksi terminal hidrolaz-L1 (UCH-L1) ve TAR DNA bağlayıcı protein (TDP-43) serum düzeyleri Enzim-bağlı-immunosorbent tahlili ELISA yöntemi ile belirlendi. Çalışmamızda kontrol grubu ile otistik grubuna ait GFAP, Nogo-A, UCH-L1 ve TDP-43 düzeyleri karşılaştırıldığında; otizm grubuna ait serum GFAP, UCH-L1 ve TDP-43 düzeylerinin kontrol grubu değerlerine göre istatistiksel olarak anlamlı düzeyde düşük olduğu bulunmuş, Nogo-A değerleri açısından ise iki grup arasında istatistiksel olarak anlamlı bir farklılık bulunmamıştır. Otistik çocuklarda serum GFAP düzeylerinin düşük bulunması, otizmde mikroglial ve astroglial aktivasyonun intrasellüler alanda birikmesinin bir işareti olarak düşünülebilir. UCH-L1 ve TDP-43 moleküllerinin otistik çocuklardaki düşük düzeyleri ve birbirleri ile olan ilişkisi; bu moleküllerin otistik çocuklarda meydana gelen ubikuitinasyonun bozulmasında rol üstlendiği şeklinde yorumlanabilir., We aimed to determine the applicable easily biomarkers in peripheral blood level and create a different perspective for understanding the pathophysiologic mechanisms of autistic disorder by examination of neuroplasticity, neurodegenerative and neuroprotective molecules levels which it is thought to be used as a diagnostic marker for potential. This study planned to be done with 30 children who were diagnosed with autism and without any systemic disease admitted to child and adolescent mental health outpatient clinic of Cerrahpaşa Medicine Faculty, İstanbul University, and 30 healthy children were included in the study as a control group. The serum levels of glial fibrillary acidic protein (GFAP), nogo-A, ubiquitin carboxy-terminal hydrolase L1 (UCH-L1) and TAR DNA binding protein (TDP-43) were determined by Enzyme-linked immunosorbent assay (ELISA). In our study, the control and autistic children were compared in terms of neuroplasticity, neurodegeneration and neuroprotection molecular levels; it was found that serum levels of GFAP, UCH-L1 and TDP-43 levels was statistically significantly lower in autistic children than those of control group and there was no statistically significant difference between two groups in terms of Nogo-A. The presence of low serum levels of GFAP, UCH-L1 and TDP-43, and correlations with each other in children with autism may be suggests that occurring changes the microglial and astroglial activation and the existence of the deterioration in the ubiquitination process in children autism.

Objective - We examined two potential biomarkers of brain damage in HIE neonates: glial fibrillary acidic protein (GFAP; a marker of gliosis) and ubiquitin C-terminal hydrolase L1 (UCH-L1; a marker of neuronal injury). We hypothesized the biomarkers would be measurable in cord blood of healthy neonates and could serve as a normative reference for brain injury in HIE infants. Further, we hypothesized that serum samples of HIE neonates would have higher levels and would correlate with brain damage on MRI and later developmental outcomes.Study Design - Serum UCH - L1 and GFAP concentrations from HIE neonates(n = 16) were compared with controls(n = 11).Pearson correlation coefficients and a mixed model design examined the relationship between biomarker concentrations of HIE neonates and brain damage(MRI) and developmental outcomes(Bayley - III).Result– Both biomarkers were detected in cord blood from control subjects.UCH - L1 concentrations were higher in HIE neonates(p < 0.001) and associated with cortical injury(p < 0.055) and later motor and cognitive developmental outcomes(p < 0.05).The temporal change in GFAP concentrations from birth to 96 hours of age predicted motor developmental outcomes(p < 0.05) and injury to the basal ganglia and white matter.Conclusion– UCH - L1 concentrations correlated with cortical injury and developmental delays and GFAP concentrations correlated with basal ganglia and white matter injury and motor delay in HIE affected patients.Researchers should continue to explore UCH - L1 and GFAP as promising serum biomarkers of brain damage and predictors of neurodevelopmental outcomes in neonates with HIE.

Objective α-synuclein, Nogo-A and Ubiquitin C-terminal hydrolase L1 (UCH-L1) have neuromodulatory roles for human brain. Therefore, abnormalities of these molecules are associated with neuropsychiatric disorders. Although some serum studies in the other disorders have been made, serum study of α-synuclein, Nogo-A and UCH-L1 is not present in patients with schizophrenia and healthy controls. Therefore, our aim was to compare serum levels of α-synuclein, Nogo-A and UCH-L1 of the patients with schizophrenia and healthy controls. Methods Forty-four patients with schizophrenia who is followed by psychotic disorders unit, and 40 healthy control were included in this study. Socio-demographic form and Positive and Negative Syndrome Scale (PANSS) was applied to patients, and sociodemographic form was applied to control group. Fasting bloods were collected and the serum levels of α-synuclein, Nogo-A and UCH-L1 were measured by ELISA method. Results Serum α-synuclein [patient: 12.73 (5.18–31.84) ng/mL; control: 41.77 (15.12–66.98) ng/mL], Nogo-A [patient: 33.58 (3.09–77.26) ng/mL; control: 286.05 (136.56–346.82) ng/mL] and UCH-L1 [patient: 5.26 (1.64–10.87) ng/mL; control: 20.48 (11.01–20.81) ng/mL] levels of the patients with schizophrenia were significianly lower than healthy controls (p

The ubiquitin-proteasome system (UPS) plays a fundamental role in regulating various biological activities. Ubiquitin C-terminal hydrolase L1 (UCH-L1) is a deubiquitinating enzyme, belonging to the UPS. To date, it has been reported that UCH-L1 is highly and restrictedly expressed in neural and reproductive tissues and plays significant roles in these organs. Although the expression of UCH-L1 in the anterior pituitary gland has been reported, the detailed localization and the role of UCH-L1 remain obscure. In the present study, we detected UCH-L1 protein exclusively in hormone-producing cells, but not non-hormone producing folliculostellate cells in the anterior pituitary lobe. In addition, the cytoplasmic expression of UCH-L1 varied and was limited to gonadotropes and mammotropes. To investigate the role of UCH-L1 in anterior pituitary cells, we performed a comparative analysis using genetically UCH-L1-deficient gad mice. Significant decreases in the numbers of gonadotropes and mammotropes were observed in gad mice, suggesting a close involvement of UCH-L1 in these cells. Moreover, we also determined the expression of UCH-L1 in cultured gonadotropes. Taken together, this is the first report to definitely demonstrate the presence of UCH-L1 in mouse anterior pituitary gland, and our results might provide a novel insight for better understanding the role of UCH-L1 in the hypothalamic-pituitary-gonadal axis and in the reproduction.

Introduction: There is an unmet need for biomarkers for Parkinson's disease (PD) and atypical parkinsonian disorders (APD). α-Synuclein, linked to the pathogenesis of PD, is a promising biomarker candidate in need of further investigation. The ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), a pivotal component of the ubiquitin proteasome system which seems to be disturbed in PD, may also be involved in the pathogenesis of this disorder. Methods: We investigated cerebrospinal fluid (CSF) α-synuclein and UCH-L1 levels from 22 healthy controls, 52 patients with PD, 34 with multiple system atrophy (MSA), 32 with progressive supranuclear palsy, and 12 with corticobasal degeneration. Results: α-Synuclein levels were significantly decreased in PD and in MSA compared with controls, and in synucleinopathies compared with tauopathies. UCH-L1 levels were significantly decreased in PD, MSA as well as PSP compared with controls, and in PD compared with APD (p

The use of biomarkers of brain injury in pediatric neurocritical care has been explored for at least 15 years. Two general lines of research on biomarkers in pediatric brain injury have been pursued, 1) studies of “bio-mediators” in cerebrospinal fluid (CSF) of children after traumatic brain injury (TBI) to explore the components of the secondary injury cascades in an attempt to identify potential therapeutic targets and 2) studies of the release of structural proteins into the CSF, serum, or urine in order to diagnose, monitor, and/or prognosticate in patients with TBI or other pediatric neurocritical care conditions. Unique age-related differences in brain biology, disease processes, and clinical applications mandate the development and testing of brain injury bio-mediators and biomarkers specifically in pediatric neurocritical care applications. Finally, although much of the early work on biomarkers of brain injury in pediatrics has focused on TBI, new applications are emerging across a wide range of applications specifically for pediatric neurocritical care including abusive head trauma, cardiopulmonary arrest, septic shock, extracorporeal membrane oxygenation, hydrocephalus, and cardiopulmonary bypass. The potential scope of the utility of biomarkers in pediatric neurocritical care is thus also discussed.

El marcaje de proteínas con ubiquitina, conocido como ubiquitinación, cumple diferentes funciones que incluyen la regulación de varios procesos celulares, tales como: la degradación de proteínas por medio del proteosoma, la reparación del ADN, la señalización mediada por receptores de membrana, y la endocitosis, entre otras (1). Las moléculas de ubiquitina pueden ser removidas de sus sustratos gracias a la acción de un gran grupo de proteasas, llamadas enzimas deubiquitinizantes (DUBs) (2). Las DUBs son esenciales para la manutención de la homeostasis de la ubiquitina y para la regulación del estado de ubiquitinación de diferentes sustratos. El gran número y la diversidad de DUBs descritas refleja tanto su especificidad como su utilización para regular un amplio espectro de sustratos y vías celulares. Aunque muchas DUBs han sido estudiadas a profundidad, actualmente se desconocen los sustratos y las funciones biológicas de la mayoría de ellas. En este trabajo se investigaron las funciones de las DUBs: USP19, USP4 y UCH-L1. Utilizando varias técnicas de biología molecular y celular se encontró que: i) USP19 es regulada por las ubiquitin ligasas SIAH1 y SIAH2 ii) USP19 es importante para regular HIF-1α, un factor de transcripción clave en la respuesta celular a hipoxia, iii) USP4 interactúa con el proteosoma, iv) La quimera mCherry-UCH-L1 reproduce parcialmente los fenotipos que nuestro grupo ha descrito previamente al usar otros constructos de la misma enzima, y v) UCH-L1 promueve la internalización de la bacteria Yersinia pseudotuberculosis. The conjugation of ubiquitin to proteins, known as ubiquitination, has different cellular functions; they include targeting proteins for degradation by the proteasome, regulation of DNA damage repair signaling, membrane receptor signaling and endocytosis (1). The ubiquitin moieties can be de-conjugated from their substrates or other ubiquitin moieties by a large group of proteases named deubiquitinating enzymes (DUBs) (2). DUBs are essential for the maintenance of the ubiquitin homeostasis in the cell and regulation of the ubiquitination status of the different substrates. The diversity of these proteases hints on their specificity for certain targets and participation in particular cellular pathways. Although several DUBs have been thoroughly studied, at present the targets and physiological roles of most of them remain unknown. Here, we studied the functional roles of the ubiquitin specific protease 19 (USP19), USP4 and the ubiquitin C-terminal hydrolase (UCH-L1), using several cellular and molecular techniques. We found that, i) USP19 can be regulated by SIAH ubiquitin ligases, ii) USP19 is important for controlling the key regulator of response to hypoxia, HIF-1α, iii) USP4 is a proteasome-interacting DUB, iv) an mCherry-UCH-L1 chimera reproduces only partially previous phenotypes described for UCH-L1, and v) UCH-L1 promotes Yersinia pseudotuberculosis internalization. ERACOL Universidad del Rosario Karolinska Institutet

Glutamate excitotoxicity plays a key role in the etiology of a variety of neurological, psychiatric, and neurodegenerative disorders. The goal of this study was to investigate spatiotemporal distribution in the brain and cerebrospinal fluid (CSF) concentrations of ubiquitin C-terminal hydrolase-1 (UCH-L1), glial fibrillary acidic protein (GFAP), αII-spectrin breakdown products (SBDP150, SBDP145, and SBDP120), and their relationship to neuropathology in an animal model of kainic acid (KA) excitotoxicity. Triple fluorescent labeling and Fluoro-Jade C staining revealed a reactive gliosis in brain and specific localization of degenerating neurons in hippocampus and entorhinal cortex of KA-treated rats. Immunohistochemistry showed upregulation of GFAP expression in hippocampus and cortex beginning 24h post KA injection and peaking at 48h. At these time points concurrent with extensive neurodegeneration all SBDPs were observed throughout the brain. At 24h post KA injection, a loss of structural integrity was observed in cellular distribution of UCH-L1 that correlated with an increase in immunopositive material in the extracellular matrix. CSF levels of UCH-L1, GFAP, and SBDPs were significantly increased in KA-treated animals compared with controls. The temporal increase in CSF biomarkers correlated with brain tissue distribution and neurodegeneration. This study provided evidence supporting the use of CSF levels of glial and neuronal protein biomarkers to assess neurotoxic damage in preclinical animal models that could prove potentially translational to the clinic. The molecular nature of these biomarkers can provide critical information on the underlying mechanisms of neurotoxicity that might facilitate the development of novel drugs and allow physicians to monitor drug safety.

The success of proteasome inhibition in multiple myeloma highlights the critical role for the ubiquitin-proteasome system (UPS) in this disease. However, there has been little progress in finding more specific targets within the UPS involved in myeloma pathogenesis. In an unbiased screen of de-ubiquitinase activity, we previously found the ubiquitin hydrolase UCH-L1 to be frequently over-expressed in B-cell malignancies, including myeloma. Using a transgenic mouse model, we also found that aberrant expression of UCH-L1 is sufficient to drive the development of spontaneous B-cell lymphomas and plasmacytomas, demonstrating its oncogenic activity. Whether and how UCH-L1 affects the clinical behavior of myeloma, however, is not known. Here we show that UCH-L1 is a potent negative prognostic factor that is essential for the dissemination and progression of myeloma. We found high levels of UCHL1 to predict early progression in newly diagnosed patients; a finding reversed in studies including bortezomib. We also found high UCHL1 levels to be a critical factor in the superiority of bortezomib over high-dose dexamethasone in relapsed patients. High UCHL1 partially overlaps with, but is distinct from, known genetic risks including 4p16 rearrangement and 1q21 amplification. Using an orthotopic mouse model, we found UCH-L1 depletion delays myeloma dissemination and causes regression of established disease. We conclude that UCH-L1 is a biomarker of aggressive myeloma that may be an important marker of bortezomib response, and may itself be an effective target in disseminated disease. Disclosures Galardy: Mission Therapeutics: Research Funding.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.