Neuropathologic Role of Inositol (1,4,5)-Trisphosphate Receptor Proteolysis

Ischemic brain injury represents a major cause of death and disability. Developing targeted therapies for neuroprotection requires increased understanding of the molecular mechanisms of neuronal injury following ischemia. Both disruption of Ca2+ homeostasis and pathological activation of proteases are believed to play causal roles in delayed neuronal death after ischemia-reperfusion. Presented here are data supporting a novel role for proteolysis of an intracellular Ca2+ release channel, inositol 1,4,5-trisphosphate receptor (InsP3R), in ischemic brain injury. We identified a unique calpain cleavage site in the type 1 InsP3R (InsP3R1) and utilized a recombinant truncated form of the channel (capn-InsP3R1) to investigate the functional consequences of InsP3R1 proteolysis. Using a combination of single-channel electrophysiology and single-cell Ca2+ imaging, we determined that capn-InsP3R1 has InsP3-independent gating and constitutive channel activity. This constitutive channel activity decreased Ca2+ content of intracellular stores in Neuro-2A cells by increasing endoplasmic reticulum (ER) Ca2+ leak. Additionally, capn-InsP3R1 compromised ER Ca2+ buffering capacity in primary cortical cultures, leading to decreased neuronal viability and enhanced sensitivity to excitotoxic injury. Using stereotaxic intracerebral injection of viral vectors, we transduced neurons in vivo with capn-InsP3R1 and observed spontaneous degeneration of subpopulations of neurons in the hippocampus. Together, these results reveal a previously unknown role of calpain-cleaved InsP3R1 in disruption of intracellular Ca2+ homeostasis and neuronal death. Importantly, we also provide evidence of calpain-mediated proteolysis of InsP3R1 in neurons in the cerebellum after ischemic brain injury. The findings presented here provide the first functional studies of calpain-cleaved InsP3R1, and advance our understanding of the pathological role of the cleaved channel in neurodegeneration. Inhibiting Ca2+ release through calpain-cleaved InsP3R1 may emerge as a novel therapeutic strategy for intervention in ischemic brain injury and other neurodegenerative diseases associated with disruption of Ca2+ homeostasis

Architecture education: rubrics in Google Classroom as a tool of improving the assessment and learning

Blended learning has emerged as a promising approach in architecture education, combining face-to-face instruction with online tools such as Google Classroom, etc. for teaching and learning as well as assessment. The rubrics is a helpful grading tool that the instructors can use to rate the students’ work in a more standardized, transparent, and easy way. The well-designed rubrics can enable the students in recognising their areas of strength and weakness and it will also help them to improve the quality of their work. Although the rubrics have some limitations, several studies show that they can help teachers communicate their expectations more effectively. In this paper, the researchers discussed the effect of using the rubrics in the Google Classroom in architecture education. The rubrics were developed in Google Classroom to evaluate the main task of the subject i.e., assignments for the students of Bachelor of Architecture. The data was collected from the students through a questionnaire that was developed using the web-based Google Forms platform. The marks obtained in the assignments and the data collected through the questionnaire were presented graphically for ease of understanding. Results indicate that rubrics using Google Classroom facilitate a clearer understanding of expectations, promote consistency in grading and feedback, increased transparency, and support student self-assessment and reflection. The integration of rubrics within Google Classroom enhances students’ engagement, motivation, and overall learning experience

The Atypical Calpains: Evolutionary Analyses and Roles in Caenorhabditis elegans Cellular Degeneration

The calpains are physiologically important Ca2+-activated regulatory proteases, which are divided into typical or atypical sub-families based on constituent domains. Both sub-families are present in mammals, but our understanding of calpain function is based primarily on typical sub-family members. Here, we take advantage of the model organism Caenorhabditis elegans, which expresses only atypical calpains, to extend our knowledge of the phylogenetic evolution and function of calpains. We provide evidence that a typical human calpain protein with a penta EF hand, detected using custom profile hidden Markov models, is conserved in ancient metazoans and a divergent clade. These analyses also provide evidence for the lineage-specific loss of typical calpain genes in C. elegans and Ciona, and they reveal that many calpain-like genes lack an intact catalytic triad. Given the association between the dysregulation of typical calpains and human degenerative pathologies, we explored the phenotypes, expression profiles, and consequences of inappropriate reduction or activation of C. elegans atypical calpains. These studies show that the atypical calpain gene, clp-1, contributes to muscle degeneration and reveal that clp-1 activity is sensitive to genetic manipulation of [Ca2+]i. We show that CLP-1 localizes to sarcomeric sub-structures, but is excluded from dense bodies (Z-disks). We find that the muscle degeneration observed in a C. elegans model of dystrophin-based muscular dystrophy can be suppressed by clp-1 inactivation and that nemadipine-A inhibition of the EGL-19 calcium channel reveals that Ca2+ dysfunction underlies the C. elegans MyoD model of myopathy. Taken together, our analyses highlight the roles of calcium dysregulation and CLP-1 in muscle myopathies and suggest that the atypical calpains could retain conserved roles in myofilament turnover

Immunohistochemical Method and Histopathology Judging for the Systemic Synuclein Sampling Study (S4)

Immunohistochemical (IHC) α-synuclein (Asyn) pathology in peripheral biopsies may be a biomarker of Parkinson disease (PD). The multi-center Systemic Synuclein Sampling Study (S4) is evaluating IHC Asyn pathology within skin, colon and submandibular gland biopsies from 60 PD and 20 control subjects. Asyn pathology is being evaluated by a blinded panel of specially trained neuropathologists. Preliminary work assessed 2 candidate immunoperoxidase methods using a set of PD and control autopsy-derived sections from formalin-fixed, paraffin-embedded blocks of the 3 tissues. Both methods had 100% specificity; one, utilizing the 5C12 monoclonal antibody, was more sensitive in skin (67% vs 33%), and was chosen for further use in S4. Four trainee neuropathologists were trained to perform S4 histopathology readings; in subsequent testing, their scoring was compared to that of the trainer neuropathologist on both glass slides and digital images. Specificity and sensitivity were both close to 100% with all readers in all tissue types on both glass slides and digital images except for skin, where sensitivity averaged 75% with digital images and 83.5% with glass slides. Semiquantitative (0-3) density score agreement between trainees and trainer averaged 67% for glass slides and 62% for digital images

The genetic architecture of the human cerebral cortex

The cerebral cortex underlies our complex cognitive capabilities, yet little is known about the specific genetic loci that influence human cortical structure. To identify genetic variants that affect cortical structure, we conducted a genome-wide association meta-analysis of brain magnetic resonance imaging data from 51,665 individuals. We analyzed the surface area and average thickness of the whole cortex and 34 regions with known functional specializations. We identified 199 significant loci and found significant enrichment for loci influencing total surface area within regulatory elements that are active during prenatal cortical development, supporting the radial unit hypothesis. Loci that affect regional surface area cluster near genes in Wnt signaling pathways, which influence progenitor expansion and areal identity. Variation in cortical structure is genetically correlated with cognitive function, Parkinson's disease, insomnia, depression, neuroticism, and attention deficit hyperactivity disorder

Longitudinal clinical and biomarker characteristics of non-manifesting LRRK2 G2019S carriers in the PPMI cohort

We examined 2-year longitudinal change in clinical features and biomarkers in LRRK2 non-manifesting carriers (NMCs) versus healthy controls (HCs) enrolled in the Parkinson’s Progression Markers Initiative (PPMI). We analyzed 2-year longitudinal data from 176 LRRK2 G2019S NMCs and 185 HCs. All participants were assessed annually with comprehensive motor and non-motor scales, dopamine transporter (DAT) imaging, and biofluid biomarkers. The latter included cerebrospinal fluid (CSF) Abeta, total tau and phospho-tau; serum urate and neurofilament light chain (NfL); and urine bis(monoacylglycerol) phosphate (BMP). At baseline, LRRK2 G2019S NMCs had a mean (SD) age of 62 (7.7) years and were 56% female. 13% had DAT deficit (defined as <65% of age/sex-expected lowest putamen SBR) and 11% had hyposmia (defined as ≤15th percentile for age and sex). Only 5 of 176 LRRK2 NMCs developed PD during follow-up. Although NMCs scored significantly worse on numerous clinical scales at baseline than HCs, there was no longitudinal change in any clinical measures over 2 years or in DAT binding. There were no longitudinal differences in CSF and serum biomarkers between NMCs and HCs. Urinary BMP was significantly elevated in NMCs at all time points but did not change longitudinally. Neither baseline biofluid biomarkers nor the presence of DAT deficit correlated with 2-year change in clinical outcomes. We observed no significant 2-year longitudinal change in clinical or biomarker measures in LRRK2 G2019S NMCs in this large, well-characterized cohort even in the participants with baseline DAT deficit. These findings highlight the essential need for further enrichment biomarker discovery in addition to DAT deficit and longer follow-up to enable the selection of NMCs at the highest risk for conversion to enable future prevention clinical trials

ВИРОБНИЧИЙ ТРАВМАТИЗМ В УКРАЇНІ: ПРИЧИНИ ТА ШЛЯХИ ЗАПОБІГАННЯ

The basic causes of occupational injuries in the workplace. The main factors that cause high rates of occupational injuries in Ukraine. The state of occupational injuries 2011-2014 years by industries and regions of Ukraine. The groups of occupations for which it was registered the most cases of fatal injuries. Designed prevention of occupational injuries in the workplace. It proposes steps of prevention of occupational injuries in the enterprise.Сформульовано основні причини виробничого травматизму на підприємствах. Визначено основні фактори, які зумовлюють високі показники виробничого травматизму в Україні. Досліджено стан виробничого травматизму за 2011-2014 роки по галузях економіки та по регіонах України. Виявлено групи професій, за якими було зареєстровано найбільше випадків смертельного травматизму. Розроблено заходи профілактики виробничого травматизму на підприємствах. Запропоновано етапи профілактики виробничого травматизму на підприємстві