Journal:

Article Title: Cathepsin-L, a Key Molecule in the Pathogenesis of Drug-Induced and I-Cell Disease-Mediated Gingival Overgrowth

doi:

Figure Lengend Snippet: Latent and active cathepsin activity of cells cultured with or without nifedipine. Enzyme activities of active forms of cathepsin-(B+L) (A), active form of cathepsin-B (B), total forms of cathepsin-(B+L) (C), and total form of cathepsin-B (D) were measured in gingival fibroblasts cultured with or without 100 ng/ml of nifedipine. Gingival fibroblasts cultured with or without nifedipine were subjected to measurement of cathepsin activities as described in Materials and Methods. Values are representative of the results obtained from gingival fibroblasts isolated from normal gingival tissues and nifedipine-induced gingival overgrowth, since gingival fibroblasts obtained from nifedipine-induced gingival overgrowth and from normal tissues were similarly affected by nifedipine. Values are expressed as the % activity against that of controls (day 0). Both active and total forms of cathepsin-(B+L) activity were significantly suppressed by nifedipine, while that of cathepsin-B was not influenced.

Article Snippet: Reagents The blocker of L-type Ca (2+) channels, nifedipine, was from Bayer AG Pharma Research Center (Wuppertal, Germany).

Techniques: Activity Assay, Cell Culture, Isolation

Journal:

Article Title: Cathepsin-L, a Key Molecule in the Pathogenesis of Drug-Induced and I-Cell Disease-Mediated Gingival Overgrowth

doi:

Figure Lengend Snippet: mRNA expression of various lysosomal enzymes cultured with or without nifedipine in gingival fibroblasts. The expressions of mRNA encoding various lysosomal enzymes were examined in gingival fibroblasts cultured with (nifedipine) or without (−) 100 ng/ml of nifedipine for the indicated time periods. Total RNA was isolated and was subjected to RT-PCR analysis as described in Materials and Methods. Values are representative of the results obtained from several gingival fibroblasts from different donors, since all cells tested showed similar results. Expression of cathepsin-L mRNA was markedly suppressed in gingival fibroblasts after 1 day of incubation with nifedipine, while that of other lysosomal enzymes was not apparently influenced. HeLa cells served as the positive control.

Article Snippet: Reagents The blocker of L-type Ca (2+) channels, nifedipine, was from Bayer AG Pharma Research Center (Wuppertal, Germany).

Techniques: Expressing, Cell Culture, Isolation, Reverse Transcription Polymerase Chain Reaction, Incubation, Positive Control

Journal:

Article Title: Cathepsin-L, a Key Molecule in the Pathogenesis of Drug-Induced and I-Cell Disease-Mediated Gingival Overgrowth

doi:

Figure Lengend Snippet: The effects of TG on the activity of cathepsins. Gingival fibroblasts were cultured with or without 100 ng/ml of nifedipine in the presence or absence of the indicated concentrations of Ca mobilizer TG. Twenty-four hours later, cellular proteins were extracted and were subjected to enzyme assay. TG suppressed both cathepsin-(B+L) and -B activity, while nifedipine alone specifically suppressed cathepsin-(B+L) activity. Values are expressed as the % activity against that of controls (day 0).

Article Snippet: Reagents The blocker of L-type Ca (2+) channels, nifedipine, was from Bayer AG Pharma Research Center (Wuppertal, Germany).

Techniques: Activity Assay, Cell Culture, Enzymatic Assay

Journal: Frontiers in Physiology

Article Title: Abnormal Calcium Handling in Duchenne Muscular Dystrophy: Mechanisms and Potential Therapies

doi: 10.3389/fphys.2021.647010

Figure Lengend Snippet: Schematic representation of intracellular Ca 2+ cycling in a normal muscle cell. Dystrophin stabilizes muscle cells by linking the cytoskeleton (F-actin, intermediate filaments, microtubule) to the extracellular matrix via the dystrophin-associated glycoprotein complex (DAGC). Ca 2+ -induced Ca 2+ release (CICR) occurs through activation of voltage-gated, L-type Ca 2+ channels (Ca v ) and the release of Ca 2+ from the sarcoplasmic/endoplasmic reticulum (SR) via ryanodine receptor (RyR). CICR activates muscle contraction. Resequestration of Ca 2+ back into the SR by SR Ca 2+ ATPase (SERCA) initiates muscle relaxation. The physiological intracellular Ca 2+ level regulates mitochondrial Ca 2+ content and function and maintains cellular energetics. DG, dystroglycan; NCX, sodium–calcium exchanger; NHE, sodium–proton exchanger; PMCA, plasma membrane Ca 2+ ATPase; SAC, stretch-activated channels; SOCC, store-operated Ca 2+ channel; TRPC1, transient receptor potential channel 1.

Article Snippet: Although these observations signify the therapeutic benefits of L-type Ca 2+ -channel blockers in animal models, several clinical trials have revealed no clinical benefit in DMD patients (Spinazzola and Kunkel, ).

Techniques: Activation Assay, Clinical Proteomics, Membrane

Journal: Frontiers in Physiology

Article Title: Abnormal Calcium Handling in Duchenne Muscular Dystrophy: Mechanisms and Potential Therapies

doi: 10.3389/fphys.2021.647010

Figure Lengend Snippet: Schematic representation of mechanisms causing Ca i 2 + overload in a dystrophin-deficient muscle cell. Loss of dystrophin causes destabilization of dystrophin-associated glycoprotein complex (DAGC), membrane tear, and activation of store-operated calcium channels (SOCCs), resulting in abnormal Ca 2+ entry. High concentrations of extracellular ATP can activate abnormal Ca 2+ influx via P2X7. RyR dysfunction causes Ca 2+ leak from the SR and SERCA dysfunction compromises Ca 2+ resequestration. NOX activation can also increase RyR Ca 2+ leak. These changes result in abnormal and chronic elevation of the cytoplasmic Ca 2+ levels. Supraphysiological level Ca 2+ activates Ca 2+ -dependent proteases and phospholipase and causes muscle necrosis and replacement of muscle by fatty and fibrotic tissues. Sustained elevation of cytoplasmic Ca 2+ levels also affects mitochondrial function and increases reactive oxygen species (ROS). Collectively, these changes lead to muscle wasting and contractile dysfunction. Bold and broken arrows indicate the enhanced and decreased function of the Ca 2+ channels, respectively. Cx43, connexin 43; NCX, sodium–calcium exchanger; NHE, sodium–proton exchanger; NOX, NADPH oxidases; PMCA, plasma membrane Ca 2+ ATPase; P2X7, P2X purinoceptor 7; SAC, stretch-activated channels; SOCC, store-operated Ca 2+ channel; TRPC1, transient receptor potential channel 1.

Article Snippet: Although these observations signify the therapeutic benefits of L-type Ca 2+ -channel blockers in animal models, several clinical trials have revealed no clinical benefit in DMD patients (Spinazzola and Kunkel, ).

Techniques: Membrane, Activation Assay, Clinical Proteomics

Journal: Frontiers in Physiology

Article Title: Abnormal Calcium Handling in Duchenne Muscular Dystrophy: Mechanisms and Potential Therapies

doi: 10.3389/fphys.2021.647010

Figure Lengend Snippet: Mitochondrial dysfunction in DMD. Schematic representation of mitochondrial structural and functional alterations in dystrophin-deficient cardiac/skeletal muscle cells. An abnormal elevation of Ca 2+ in the cytoplasm and mitochondria-associated membrane (MAM) region resulted in increased mitochondrial Ca 2+ uptake and enhanced activation of mitochondrial permeability transition pore (mPTP) opening. These changes impair mitochondrial function and mitochondrial dynamics and contributing to the metabolic crisis. Currently, there is no experimental evidence for the role of the MAM region and mitochondrial Ca 2+ uniporter (MCU) in the mitochondrial Ca 2+ overload in DMD. Bold and broken arrows indicate the enhanced and decreased function of the Ca 2+ channels, respectively. ER, endoplasmic reticulum; ETC, electron transport chain; GRP75, glucose-regulated protein 75; IP3R, inositol trisphosphate receptor; MFN, mitofusin; NCLX, Na + -Ca 2+ -Li + exchanger; PLN, phospholamban; ROS, reactive oxygen species; RyR, ryanodine receptor; SERCA, sarcoplasmic/endoplasmic reticulum Ca 2+ ATPase; SLN, sarcolipin; SR, sarcoplasmic reticulum; VDAC, voltage-dependent anion channel.

Article Snippet: Although these observations signify the therapeutic benefits of L-type Ca 2+ -channel blockers in animal models, several clinical trials have revealed no clinical benefit in DMD patients (Spinazzola and Kunkel, ).

Techniques: Functional Assay, Membrane, Activation Assay, Permeability

Journal: Marine Drugs

Article Title: Biomedical Potential of the Neglected Molluscivorous and Vermivorous Conus Species

doi: 10.3390/md20020105

Figure Lengend Snippet: Drug development with conotoxins.

Article Snippet: 2 , ω-CVID , AM336 , Neuropathic pain , Ca 2+ channel (CaV2.2) N-type calcium channels/blocker , Phase IIa * , Relevare Pharmaceuticals LTD., Australia , Conus catus (p) , [ ] .

Techniques: