Two New C19-Diterpenoid Alkaloids from Delphinium shawurense

Shawurenine C ( 1a ) and D ( 1b ), a new pair of regioisomeric C₁₉-diterpenoid alkaloids, and five known C₁₉-diterpenoid alkaloids ( 2 – 6 ) were isolated from the aerial part of Delphinium shawurense W. T. Wang. The chemical structures of new compounds were established based on spectroscopic analyses: HR-ESI-MS, and 1D, 2D NMR spectroscopic data. The anti-inflammatory and cytotoxic activities of these diterpenoid alkaloids were also evaluated.     

Design of Tripeptides as a Competitive Inhibitor for HMG-CoA Reductase

International Journal of Peptide Research and Therapeutics - This study presents a simple approach in design of tripeptides as a competitive inhibitor for 3-hydroxy-3-methylglutaryl CoA reductase...     

Correlated Spontaneous Activity Persists in Adult Retina and Is Suppressed by Inhibitory Inputs

Spontaneous rhythmic activity is a hallmark feature of the developing retina, where propagating retinal waves instruct axonal targeting and synapse formation. Retinal waves cease around the time of eye-opening; however, the fate of the underlying synaptic circuitry is unknown. Whether retinal waves are unique to the developing retina or if they can be induced in adulthood is not known. Combining patch-clamp techniques with calcium imaging, we demonstrate that propagative events persist in adult mouse retina when it is deprived of inhibitory input. This activity originates in bipolar cells, resembling glutamatergic stage III retinal waves. We find that, as it develops, the network interactions progressively curtail this activity. Together, this provides evidence that the correlated propagative neuronal activity can be induced in adult retina following the blockade of inhibitory interactions.     

Presynaptic inhibition modulates spillover, creating distinct dynamic response ranges of sensory output

Sensory information is thought to be modulated by presynaptic inhibition. Although this form of inhibition is a well-studied phenomenon, it is still unclear what role it plays in shaping sensory signals in intact circuits. By visually stimulating the retinas of transgenic mice lacking GABAc receptor-mediated presynaptic inhibition, we found that this inhibition regulated the dynamic range of ganglion cell (GC) output to the brain. Presynaptic inhibition acted differentially upon two major retinal pathways; its elimination affected GC responses to increments, but not decrements, in light intensity across the visual scene. The GC dynamic response ranges were different because presynaptic inhibition limited glutamate release from ON, but not OFF, bipolar cells, which modulate the extent of glutamate spillover and activation of perisynaptic NMDA receptors at ON GCs. Our results establish a role for presynaptic inhibitory control of spillover in determining sensory output in the CNS.     

Failure to maintain eye-specific segregation in nob, a mutant with abnormally patterned retinal activity

Axon terminals from the two eyes initially overlap in the dorsal-lateral geniculate nucleus (dLGN) but subsequently refine to occupy nonoverlapping territories. Retinal activity is required to establish and maintain this segregation. We show that despite the presence of retinal activity, segregated projections desegregate when the structure of activity is altered. Early in development, spontaneous retinal activity in the no b-wave (nob) mouse is indistinguishable from that of wild-type mice, and eye-specific segregation proceeds normally. But, around eye-opening, spontaneous and visually evoked activity in nob retinas become abnormal, coincident with a failure to preserve precise eye-specific territories. Dark-rearing studies suggest that altered visual experience is not responsible. Transgenic rescue of the mutated protein (nyctalopin) within nob retinal interneurons, without rescuing expression in either retinal projection neurons or their postsynaptic targets in the dLGN, restores spontaneous retinal activity patterns and prevents desegregation. Thus, normally structured spontaneous retinal activity stabilizes newly refined retinogeniculate circuitry.     

Olfactory CNG channel desensitization by Ca2+/CaM via the B1b subunit affects response termination but not sensitivity to recurring stimulation

Ca2+/calmodulin-mediated negative feedback is a prototypical regulatory mechanism for Ca2+-permeable ion channels. In olfactory sensory neurons (OSNs), such regulation on the cyclic nucleotide-gated (CNG) channel is considered a major mechanism of OSN adaptation. To determine the role of Ca2+/calmodulin desensitization of the olfactory CNG channel, we introduced a mutation in the channel subunit CNGB1b in mice that rendered the channel resistant to fast desensitization by Ca2+/calmodulin. Contrary to expectations, mutant OSNs showed normal receptor current adaptation to repeated stimulation. Rather, they displayed slower response termination and, consequently, reduced ability to transmit olfactory information to the olfactory bulb. They also displayed reduced response decline during sustained odorant exposure. These results suggest that Ca2+/calmodulin-mediated CNG channel fast desensitization is less important in regulating the sensitivity to recurring stimulation than previously thought and instead functions primarily to terminate OSN responses.     

Optimized Protocol for Retinal Wholemount Preparation for Imaging and Immunohistochemistry

Working with delicate tissue can be a complicating factor when performing immunohistochemical assessment. Here, we present a method that utilizes a ring-supported hydrophilized PTFE membrane to provide structural support to both living and fixed tissue during immunohistochemical processing, which allows for the use of a variety of protocols that would otherwise cause damage to the tissue. First, this is demonstrated with bolus loading of fluorescent markers into living retinal tissue. This method allows for quick visualization of targeted structures, while the membrane support maintains tissue integrity during the injection and allows for easy transfer of the preparation for further imaging or processing.Second, a procedure for antibody staining in tissue fixed with carbodiimide is described. Though paraformaldehyde fixation is more common, carbodiimide fixation provides a superior substrate for the visualization of synaptic proteins. A limitation of carbodiimide is that the resulting fixed tissue is relatively fragile; however, this is overcome with the use of the supporting membrane. Retinal tissue is used to demonstrate these techniques, but they may be applied to any fragile tissue.     

Various structural aspects of the functional transformation of the kidneys during adaptation to fasting

Electron microscopic and morphometric investigations of the rat kidneys in 3-6 days after a complete fasting have revealed certain ultrastructural signs of an increased proteolysis in the proximal parts of the nephron canaliculi and structural manifestation in the glomerular blood stream changes, that represents an important condition for passing proteins the glomerular membrane. In this connection, it is supposed that at adaptation to fasting the kidney ensures proteolysis of endogenic-proteins--the first stage of their reconstruction. This function is ensured by structural-functional reconstruction of the juxtaglomerular complex, vessels of the filtrative apparatus and by the proteolytic system in the proximal part of the nephron canaliculus.     

Cellular mechanisms of compensatory processes of the kidney in relation to the mass resected

As a result of comparative investigation of compensatory processes in the kidneys at various degree of their resection, the most general regularities in these processes have been revealed: an initial functional strain of renal cells and their ultrastructures, the secondary increase in hypertrophy and hyperplasia of the renal cells and their organelles. The degree of manifestation of these regular reactions directly depends on the mass of the resected part of the kidney. Subtotal resection of the kidney (80%) is an operation that gets out of the limits of the organ's compensatory possibilities. A sharp decrease, up to complete disappearance of its cellular division in 2-3 weeks after the operation is of great importance. Mechanism for development of renal insufficiency is evidently connected with absence of cellular renewal against the background of their hyperfunction and their quick worn out, that is, evidently, resulted in deficiency of renal epithelium and functional defectiveness connected with it in the remained renal mass.