Regulation and dysregulation of intracellular calcium (Ca 2+ ) signaling via the inositol 1,4,5-trisphosphate receptor (InsP 3 R) has been linked to many cellular processes and pathological conditions. In the present study, addition of neuronal calcium sensor-1 (NCS-1), a high-affinity, low-capacity, calcium-binding protein, to purified InsP 3 R type 1 (InsP 3 R1) increased the channel activity in both a calcium-dependent and -independent manner. In intact cells, enhanced expression of NCS-1 resulted in increased intracellular calcium release upon stimulation of the phosphoinositide signaling pathway. To determine whether InsP 3 R1/NCS-1 interaction could be functionally relevant in bipolar disorders, conditions in which NCS-1 is highly expressed, we tested the effect of lithium, a salt widely used for treatment of bipolar disorders. Lithium inhibited the enhancing effect of NCS-1 on InsP 3 R1 function, suggesting that InsP 3 R1/NCS-1 interaction is an essential component of the pathomechanism of bipolar disorder.

Testosterone plays a crucial role in neuronal function, but elevated concentrations can have deleterious effects. Here we show that supraphysiological levels of testosterone (micromolar range) initiate the apoptotic cascade. We used three criteria, annexin V labeling, caspase activity, and DNA fragmentation, to determine that apoptotic pathways were activated by testosterone. Micromolar, but not nanomolar, testosterone concentrations increased the response in all three assays of apoptosis. signals lead to apoptotic cell death. These effects of testosterone on neurons will have long term effects on brain function.Neurosteroids have been implicated as components essential for the normal function of the central nervous system (1-4). The gonadal steroid hormones are required for reproductive function, but androgens also affect areas of the brain that are not primarily involved in reproduction such as the hippocampus (5), preoptic area, amygdala, and medial hypothalamic area (6). At physiological levels, androgens are involved in neuronal differentiation, neuroprotection, neuronal survival and development (7-9). These responses occur slowly (over hours) and are mediated through the intracellular androgen receptor. In the developing brain, androgens are capable of changing the ultrastructural characteristics of the neuronal plasma membrane with a relatively fast pace (10, 11). Recently, we have shown that nanomolar levels of testosterone induce rapid intracellular Ca 2ϩ increases in neuroblastoma cells (within seconds), which begin as Ca 2ϩ transients in the cytosol, propagate as waves of Ca 2ϩ in the cytoplasm and nucleus, and develop into an oscillatory pattern (12). These Ca 2ϩ signals depend on an interplay between Ca 2ϩ efflux from the endoplasmic reticulum through inositol 1,4,5-trisphosphate-sensitive Ca 2ϩ release channels (InsP 3 Rs) 4 and Ca 2ϩ reuptake into the endoplasmic reticulum by Ca 2ϩ pumps. This new testosterone-induced pathway in neuronal cells leads to neurite outgrowth (12), an essential event in neuronal differentiation (13). These results suggest an important physiological mechanism for the action of testosterone in neurons at physiological concentrations. However, it is unknown how these cells respond to high plasma levels of this neurosteroid, generally administered exogenously to achieve an increase in muscle mass (14, 15) or for replacement therapy (16). In vivo administration of large doses of androgens has been correlated with neurobehavioral changes like hyperexcitability, supra-aggressive nature, and suicidal tendencies (17, 18). These behavioral changes could be the outward manifestation of neuronal damage resulting from exposure to high concentrations of testosterone.In this investigation, we evaluated the hypothesis that high concentrations of testosterone can induce deleterious effects in neurons. We show that high levels of testosterone initiate an apoptotic program in neuroblastoma cells. Apoptosis is the normal and controlled process of cell death (19). Precise control of apoptos...

Background Specific treatment for COVID-19 is still an unmet need. Outcomes of clinical trials on repurposed drugs have not been yielding success. Therefore, it is necessary to include complementary approaches of medicine against COVID-19. Purpose This study was designed to evaluate the impact of traditional Indian Ayurvedic treatment regime on asymptomatic patients with COVID-19 infection. Study design It is a placebo controlled randomized double-blind pilot clinical trial. Methods The study was registered with Clinical Trial Registry-India (vide Registration No. CTRI/2020/05/025273) and conducted at the Department of Medicine in National Institute of Medical Sciences and Research, Jaipur, India. 1 g of Giloy Ghanvati ( Tinospora cordifolia ) and 2 g of Swasari Ras (traditional herbo-mineral formulation) and 0.5 g each of Ashwagandha ( Withania somnifera ) and Tulsi Ghanvati ( Ocimum sanctum ) were given orally to the patients in treatment group twice per day for 7 days. Medicines were given in the form of tablets and each tablet weighed 500 mg. While, Swasari Ras was administered in powdered form, 30 min before breakfasts and dinners, rest were scheduled for 30 min post-meals. Patients in the treatment group also received 4 drops of Anu taila (traditional nasal drop) in each nostril every day 1 h before breakfast. Patients in the placebo group received identical-looking tablets and drops, post randomization and double blinded assortments. RT-qPCR test was used for the detection of viral load in the nasopharyngeal and oropharyngeal swab samples of study participants during the study. Chemiluminescent immunometric assay was used to quantify serum levels of interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α) and high sensitivity C-reactive protein (hs-CRP) on day 1 and day 7 of the study. Results By day 3, 71.1 % and 50.0 % patients recovered in the treatment and placebo groups, respectively. Treatment group witnessed 100% recovery by day 7, while it was 60.0 % in the placebo group. Average fold changes in serum levels of hs-CRP, IL-6 and TNF-α in treatment group were respectively, 12.4, 2.5 and 20 times lesser than those in the placebo group at day 7. There was 40% absolute reduction in the risk of delayed recovery from infection in the treatment group. Conclusions Ayurvedic treatment can expedite virological clearance, help in faster recovery and concomitantly reduce the risk of viral dissemination. Reduced inflammation markers suggested less severity of SARS-CoV-2 infection in the treatment group. Moreover, there was no adverse effect observed to be associated with this treatment.

Background Newly emerged COVID-19 has been shown to engage the host cell ACE2 through its spike protein receptor binding domain (RBD). Here we show that natural phytochemical from a medicinal herb, Withania somnifera, have distinct effects on viral RBD and host ACE2 receptor complex. Methods We employed molecular docking to screen thousands of phytochemicals against the ACE2-RBD complex, performed molecular dynamics (MD) simulation, and estimated the electrostatic component of binding free energy, along with the computation of salt bridge electrostatics. Results We report that W. somnifera compound, Withanone, docked very well in the binding interface of AEC2-RBD complex, and was found to move slightly towards the interface centre on simulation. Withanone significantly decreased electrostatic component of binding free energies of ACE2-RBD complex. Two salt bridges were also identified at the interface; incorporation of Withanone destabilized these salt bridges and decreased their occupancies. We postulate, such an interruption of electrostatic interactions between the RBD and ACE2 would block or weaken COVID-19 entry and its subsequent infectivity. Conclusion Our data, for the first time, show that natural phytochemicals could well be the viable options for controlling COVID-19 entry into host cells, and W. somnifera may be the first choice of herbs in these directions to curb the COVID-19 infectivity.

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