Ampa Receptor Dysregulation And Therapeutic Interventions In A Mouse Model Of Cdkl5 Deficiency Disorder

CDKL5 Deficiency Disorder (CDD) is a rare disease that presents as a set of neurological deficits including early-life epilepsy, intellectual disability, and autistic-like behaviors. It results from pathogenic mutations in the gene for cyclin-dependent kinase-like 5 (CDKL5), a protein that is highly expressed in brain. There is no cure for CDD and seizures in this disorder are typically resistant to traditional anti-epileptic drugs, although some patients respond well to cannabidiol. However, underlying mechanisms of what causes hyperexcitability and neurological deficits in CDD is poorly understood. We investigated the novel Cdkl5R59X mouse (R59X), and observed that mutant mice have social interaction and memory deficits, and decreased latency to seizure after administration of pentylenetetrazol. Given the observed behavioral alterations and hyperexcitability in R59X mice, we hypothesized that mutant mice would exhibit underlying molecular and functional alterations in proteins involved in regulating the E:I balance. Indeed, we observed a specific decrease in membrane-bound AMPA receptor (AMPAR) subunit GluA2 as well as decreased GluA2:GluA1 in the hippocampus of R59X mice, suggesting an increase in hippocampal GluA2-lacking AMPARs, which are calcium permeable and have significant roles in regulating neuronal plasticity and excitability. Indeed, decreased hippocampal GluA2 was accompanied by inward rectification of AMPAR currents in whole-cell patch recordings from hippocampal CA1 neurons, indicative of an increased population of functional GluA2-lacking AMPARs, and elevated early-phase long-term potentiation at Schaffer collateral-CA1 synapses. Finally, we evaluated the therapeutic potential of GluA2-lacking AMPAR blocker IEM-1460 as well as cannabidiol to observe 1) whether blocking increased hippocampal GluA2-lacking AMPARs would attenuate behavioral alterations and hyperexcitability in R59X mice and 2) whether cannabidiol shows therapeutic efficacy in R59X mice similar to human CDD patients, thus validating R59X mice as a relevant model to test potential therapeutics in CDD. Indeed, IEM-1460 significantly rescued deficits in social behavior, short-term memory, and latency to seizure while cannabidiol significantly rescued deficits in short and long-term memory and latency to seizure. These results verify that blocking increased GluA2-lacking AMPARs may be a successful therapeutic strategy in CDD, and that both IEM-1460 and cannabidiol may attenuate hyperexcitability as well as autistic-like behaviors and memory deficits in CDD

CLINICAL EVALUATION OF SHATAPUSHPA SHATAVARI CHURNA AND O.C. PILLS IN THE MANAGEMENT OF ARTAVA KSHAYA W.S.R. TO CERTAIN MENSTRUAL DISORDERS

A randomised control clinical trial was carried out on 60 Artava Kshaya (irregular, scanty and painful menses) patients aged between 18 and 40 years having complaints of irregular, scanty and painful menstruations. The patients were registered from OPD of CSMSS Ayurved Rugnalaya & Research Centre, Aurangabad. Divided into 2 groups. They were administrated Shatapuspa and Shatawari churna (group A) and OC pill (group B) for three months in a dose of 5 g daily with cow Ghrita and Guda; OC pills (Mala N) once in a day as per standard schedule. The specific investigations were done in order to exclude congenital anomalies, endometritis, endocrine disorders, diabetes and heart disease. The clinical assessment was carried out in 3 intervals 30 days apart. It is inferred that the study shows the effect of both treatments were significant in clinical study. In group A, none of the patient showed excellent improvement in Artava Kshaya, while moderate improvement was observed in one (3.33%) patient, mild improvement was observed in 25 (83.25%) patients and 4 (13.32%) of the patients showed ineffective improvement after treatment. In group B, none of the patient showed excellent improvement in Artava Kshaya, while moderate improvement was observed in 2 (6.66%) patients, mild improvement was observed in 25 (83.25%) patients and 3 (9.99%) of the patients showed ineffective improvement after treatment. The Shatapushpa Shatavari Churna is more effective on Yoni Vedana, Artava strava Pramana and Artava strava kalvavadhi.-Both Shatapushpa Shatavari Churna and O.C. pill are effective on and duration between two Artava Darshana but O.C. pill is comparatively more effective

Crystal structure of N-[(2S,5R)-4-oxo-2,3-diphenyl-1,3-thiazinan-5-yl]acetamide 0.375-hydrate

Acknowledgements We acknowledge NSF funding (CHEM-0131112) for the X-ray diffractometer. We also express gratitude to Euticals for the gift of T3P in 2-methyl­tetra­hydro­furan, and to Oakwood Products for the gift of N-acetyl-L-cysteine.Peer reviewedPublisher PD

Redetermination of {5-[(7-chloro­quinolinium-4-yl)amino]-2-hy­droxy­benz­yl}diethyl­ammonium dichloride dihydrate

The structure of the title compound (common name: amodiaquinium dichloride dihydrate), C20H24ClN3O2 +·2Cl−·2H2O, was previously determined from powder diffraction data [Llinàs et al. (2006 ▶). Acta Cryst. E62, o4196-o4199]. It has now been refined from diffractometer data to a significantly higher precision. The dihedral angle between the quinoline and benzene rings is 54.57 (6)°. The central amino N atom inter­acts more strongly with the quinoline ring than with the benzene ring, as indicated by the shorter C—N bond length [1.341 (2) Å compared to 1.431 (2) Å]. In the crystal, mol­ecules are packed into a three-dimensional network/supra­molecular structure through hydrogen bonds between the amodiaquinium cations, chloride anions and water mol­ecules

2-[(2,3,6,7-Tetra­hydro-1H,5H-benzo[ij]quinolizin-9-yl)methyl­ene]propane­dinitrile

The π system of the title compound, known as julolidinemalononitrile, C16H15N3, is nearly planar, with a 3.5 (1)° twist between the aromatic and dicyano­vinyl groups. The bond lengths indicate significant zwitterionic character in the ground state

Conformational ensemble of the poliovirus 3CD precursor observed by MD simulations and confirmed by SAXS: A strategy to expand the viral proteome?

The genomes of RNA viruses are relatively small. To overcome the small-size limitation, RNA viruses assign distinct functions to the processed viral proteins and their precursors. This is exemplified by poliovirus 3CD protein. 3C protein is a protease and RNA-binding protein. 3D protein is an RNA-dependent RNA polymerase (RdRp). 3CD exhibits unique protease and RNA-binding activities relative to 3C and is devoid of RdRp activity. The origin of these differences is unclear, since crystal structure of 3CD revealed “beads-on-a-string” structure with no significant structural differences compared to the fully processed proteins. We performed molecular dynamics (MD) simulations on 3CD to investigate its conformational dynamics. A compact conformation of 3CD was observed that was substantially different from that shown crystallographically. This new conformation explained the unique properties of 3CD relative to the individual proteins. Interestingly, simulations of mutant 3CD showed altered interface. Additionally, accelerated MD simulations uncovered a conformational ensemble of 3CD. When we elucidated the 3CD conformations in solution using small-angle X-ray scattering (SAXS) experiments a range of conformations from extended to compact was revealed, validating the MD simulations. The existence of conformational ensemble of 3CD could be viewed as a way to expand the poliovirus proteome, an observation that may extend to other viruses