Growth factors and growth associated protein (GAP-43) in the peripheral nervous system

The factors required to stimulate short-term Schwann cell DNA synthesis in serum-free medium (containing insulin) are thoroughly analysed and compared with the growth factor requirements of long-term ''autocrine" Schwann cells. Evidence is presented to demonstrate the importance of cAMP in Schwann cell DNA synthesis. Measurements of intracellular cAMP levels by radioimmunoassay in both Schwann cell types is related to the differing mitogen requirements of these cells. Further tissue culture studies show that insulin-like growth factor-I (IGF-I) plays a role in Schwann cell DNA synthesis, and that cultured Schwann cells are IGF immunoreactive. Binding studies and Scatchard analysis using 125I-IGF-I provide evidence for a type 1 IGF receptor on cultured Schwann cells. An immunohistochemical analysis on the distribution of IGF in vivo in the rat sciatic nerve is reported. Dried cell preparations and and 2hr cultures are used to document the presence of IGF in the Schwann cells from embryonic to adult rats. Teased nerve preparations from postnatal rats are used to study the distribution of IGF in neurones. Results obtained suggest that IGF-I may be acting as an autocrine/paracrine Schwann cell mitogen in vivo. An immunohistochemical investigation reveals that the growth associated protein GAP-43, hitherto considered to be associated with neurones and certain CNS glia, is present in non-myelin-forming Schwann cells. Nerve sections and dried cell preparations are used to document the developmental distribution of GAP-43 in Schwann cells in vivo. Further analyses on the developmental regulation of GAP-43 expression in Schwann cells has been carried out in tissue culture. Gel electrophoresis and western blotting techniques are used to confirm the identity of Schwann cell GAP-43. Evidence that GAP-43 is widely distributed in the neurones of the adult peripheral nervous system (PNS) is provided. Immunohistochemical studies and western blotting demonstrate that GAP-43 is present in high amounts in all three sub divisions of the autonomic nervous system (ANS) of the adult rat. Taken together with known distribution of GAP-43 in areas of the CNS associated with plasticity, these findings suggest a role for GAP-43 in the plasticity of the ANS

Carbon turnover in the water-soluble protein of the adult human lens.

PurposeHuman eye lenses contain cells that persist from embryonic development. These unique, highly specialized fiber cells located at the core (nucleus) of the lens undergo pseudo-apoptosis to become devoid of cell nuclei and most organelles. Ostensibly lacking in protein transcriptional capabilities, it is currently believed that these nuclear fiber cells owe their extreme longevity to the perseverance of highly stable and densely packed crystallin proteins. Maintaining the structural and functional integrity of lenticular proteins is necessary to sustain cellular transparency and proper vision, yet the means by which the lens actually copes with a lifetime of oxidative stress, seemingly without any capacity for protein turnover and repair, is not completely understood. Although many years of research have been predicated upon the assumption that there is no protein turnover or renewal in nuclear fiber cells, we investigated whether or not different protein fractions possess protein of different ages by using the (14)C bomb pulse.MethodsAdult human lenses were concentrically dissected by gently removing the cell layers in water or shaving to the nucleus with a curved micrometer-controlled blade. The cells were lysed, and the proteins were separated into water-soluble and water-insoluble fractions. The small molecules were removed using 3 kDa spin filters. The (14)C/C was measured in paired protein fractions by accelerator mass spectrometry, and an average age for the material within the sample was assigned using the (14)C bomb pulse.ResultsThe water-insoluble fractions possessed (14)C/C ratios consistent with the age of the cells. In all cases, the water-soluble fractions contained carbon that was younger than the paired water-insoluble fraction.ConclusionsAs the first direct evidence of carbon turnover in protein from adult human nuclear fiber cells, this discovery supports the emerging view of the lens nucleus as a dynamic system capable of maintaining homeostasis in part due to intricate protein transport mechanisms and possibly protein repair. This finding implies that the lens plays an active role in the aversion of age-related nuclear (ARN) cataract

Baited technique improves censuses of cryptic fish in complex habitats

Many fish species are cryptic by nature or show a negative response to divers. This may make traditional censusing techniques difficult to perform and the results obtained using these methods may be inaccurate. This problem will be exacerbated in complex habitats, such as those found on coral reefs, because the cryptic species present will be even more difficult to locate and identify. Predation studies on coral reefs have been especially hampered by this problem because many predatory species are cryptic and so it has been difficult to obtain reliable abundance estimates. This study tested a visual census technique that used bait to bring cryptic predatory fish into view. Results from this census were then compared to those from a traditional survey using belt transects, and to a patch reef tagging study where all individuals of 3 key species were located. The baited technique produced significantly higher density estimates for 3 of the 4 most abundant cryptic species. The patch reef experiment demonstrated that the baited technique accounted for 85 to 96% of fish present. Censuses without bait observed only 40 to 61% of fish present. For mobile species, on the other hand, the baited census appeared to overestimate abundance, due to movement of fish into the census area. We therefore recommend combining baited censuses of sedentary cryptic species with traditional censuses of mobile species to gain an accurate picture of piscivorous reef fish communities. Using this approach at Lizard Island on the Great Barrier Reef, the proportion of cryptic piscivores in the community was found to be almost double that which would have otherwise been observed. We also found considerable spatial variation in the abundance and distribution of piscivorous fish. These patterns would have been quite different had the survey been based on belt transects alone. Previous studies using belt transects may therefore have underestimated the importance of cryptic piscivorous species in communities of coral reef fish

Regenerative function of immune system: Modulation of muscle stem cells

Ageing is characterised by progressive deterioration of physiological systems and the loss of skeletal muscle mass is one of the most recognisable, leading to muscle weakness and mobility impairments. This review highlights interactions between the immune system and skeletal muscle stem cells (widely termed satellite cells or myoblasts) to influence satellite cell behaviour during muscle regeneration after injury, and outlines deficits associated with ageing. Resident neutrophils and macrophages in skeletal muscle become activated when muscle fibres are damaged via stimuli (e.g. contusions, strains, avulsions, hyperextensions, ruptures) and release high concentrations of cytokines, chemokines and growth factors into the microenvironment. These localised responses serve to attract additional immune cells which can reach in excess of 1 × 105 immune cell/mm3 of skeletal muscle in order to orchestrate the repair process. T-cells have a delayed response, reaching peak activation roughly 4 days after the initial damage. The cytokines and growth factors released by activated T-cells play a key role in muscle satellite cell proliferation and migration, although the precise mechanisms of these interactions remain unclear. T-cells in older people display limited ability to activate satellite cell proliferation and migration which is likely to contribute to insufficient muscle repair and, consequently, muscle wasting and weakness. If the factors released by T-cells to activate satellite cells can be identified, it may be possible to develop therapeutic agents to enhance muscle regeneration and reduce the impact of muscle wasting during ageing and disease

Small molecule additive for low-power accumulation mode organic electrochemical transistors

A small molecule additive, dodecylbenzenesulfonate (DBSA), is added to the electrolyte in OECTs to improve the device performance.ERC IMBIBE EPSRC CDT Plastic Electronic

The lymphocyte secretome from young adults enhances skeletal muscle proliferation and migration, but effects are attenuated in the secretome of older adults.

Older people experience skeletal muscle wasting, in part due to impaired proliferative capacity of quiescent skeletal muscle satellite cells which can be reversed by exposure to young blood. To investigate the role of immune cells in muscle regeneration, we isolated lymphocytes from whole blood of young and older healthy volunteers and cultured them with, or without, anti-CD3/CD28 activators to induce release of cytokines, interleukins, and growth factors into the media. The secreted proteins were collected to prepare a conditioned media, which was subsequently used to culture C2C12 myoblasts. The conditioned media from the activated young lymphocytes increased the rate of proliferation of myoblasts by around threefold (P < 0.005) and caused an approximate fourfold (P < 0.005) increase in migration compared with nonactivated lymphocyte control media. These responses were characterized by minimal myotube formation (2%), low fusion index (5%), low myosin heavy chain content, and substantial migration. In contrast, myoblasts treated with conditioned media from activated old lymphocytes exhibited a high degree of differentiation, and multi-nucleated myotube formation that was comparable to control conditions, thus showing no effect on proliferation or migration of myoblasts. These results indicate that secreted proteins from lymphocytes of young people enhance the muscle cell proliferation and migration, whereas secreted proteins from lymphocytes of older people may contribute to the attenuated skeletal muscle satellite cell proliferation and migration

A comprehensive individual patient data meta‐analysis of the effects of cardiac contractility modulation on functional capacity and heart failure‐related quality of life

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Decitabine impact on the endocytosis regulator RhoA, the folate carriers RFC1 and FOLR1, and the glucose transporter GLUT4 in human tumors.

BackgroundIn 31 solid tumor patients treated with the demethylating agent decitabine, we performed tumor biopsies before and after the first cycle of decitabine and used immunohistochemistry (IHC) to assess whether decitabine increased expression of various membrane transporters. Resistance to chemotherapy may arise due to promoter methylation/downregulation of expression of transporters required for drug uptake, and decitabine can reverse resistance in vitro. The endocytosis regulator RhoA, the folate carriers FOLR1 and RFC1, and the glucose transporter GLUT4 were assessed.ResultsPre-decitabine RhoA was higher in patients who had received their last therapy &gt;3&nbsp;months previously than in patients with more recent prior therapy (P = 0.02), and varied inversely with global DNA methylation as assessed by LINE1 methylation (r = -0.58, P = 0.006). Tumor RhoA scores increased with decitabine (P = 0.03), and RFC1 also increased in patients with pre-decitabine scores ≤150 (P = 0.004). Change in LINE1 methylation with decitabine did not correlate significantly with change in IHC scores for any transporter assessed. We also assessed methylation of the RFC1 gene (alias SLC19A1). SLC19A1 methylation correlated with tumor LINE1 methylation (r = 0.45, P = 0.02). There was a small (statistically insignificant) decrease in SLC19A1 methylation with decitabine, and there was a trend towards change in SLC19A1 methylation with decitabine correlating with change in LINE1 methylation (r = 0.47, P &lt;0.15). While SLC19A1 methylation did not correlate with RFC1 scores, there was a trend towards an inverse correlation between change in SLC19A1 methylation and change in RFC1 expression (r = -0.45, P = 0.19).ConclusionsIn conclusion, after decitabine administration, there was increased expression of some (but not other) transporters that may play a role in chemotherapy uptake. Larger patient numbers will be needed to define the extent to which this increased expression is associated with changes in DNA methylation

Paediatric obsessive-compulsive disorder and depressive symptoms: clinical correlates and CBT treatment outcomes.

Depression frequently co-occurs with paediatric obsessive-compulsive disorder (OCD), yet the clinical correlates and impact of depression on CBT outcomes remain unclear. The prevalence and clinical correlates of depression were examined in a paediatric specialist OCD-clinic sample (N = 295; Mean = 15 [7 - 18] years, 42 % female), using both dimensional (Beck Depression Inventory-youth; n = 261) and diagnostic (Development and Wellbeing Assessment; n = 127) measures of depression. The impact of depressive symptoms and suspected disorders on post-treatment OCD severity was examined in a sub-sample who received CBT, with or without SSRI medication (N = 100). Fifty-one per-cent of patients reported moderately or extremely elevated depressive symptoms and 26 % (95 % CI: 18 - 34) met criteria for a suspected depressive disorder. Depressive symptoms and depressive disorders were associated with worse OCD symptom severity and global functioning prior to CBT. Individuals with depression were more likely to be female, have had a psychiatric inpatient admission and less likely to be attending school (ps < 0.01). OCD and depressive symptom severity significantly decreased after CBT. Depressive symptoms and depressive disorders predicted worse post-treatment OCD severity (βs = 0.19 and 0.26, ps < 0.05) but became non-significant when controlling for pre-treatment OCD severity (βs = 0.05 and 0.13, ns). Depression is common in paediatric OCD and is associated with more severe OCD and poorer functioning. However, depression severity decreases over the course of CBT for OCD and is not independently associated with worse outcomes, supporting the recommendation for treatment as usual in the presence of depressive symptoms