Retinoic acid (RA) induces neural differentiation of SH-SY5Y neuroblastoma cells. We show that the mRNA levels of the differentiation-inhibiting basic helix-loop-helix transcription factors ID1,ID2, and ID3 are down-regulated during RA-induced differentiation of SH-SY5Y cells. The levels of ID proteins decreased in parallel to the observed transcriptional repression. The expression of other basic helix-loop-helix genes changed during RA-induced differentiation: expression of neuroblast-specificASCL1 (HASH-1) gene was promptly reduced after RA treatment, whereas expression of differentiation-promoting genes NEUROD6 (NEX-1, HATH-2) andNEUROD1 was increased. Treatments with 12-O-tetradecanoylphorbol-13-acetate, another inducer of neuroblastoma cell differentiation, also resulted in coordinated down-regulation of ID gene expression, underscoring the role of ID genes in differentiation. Down-regulation ofID gene expression by RA involves a complex mechanism because full transcriptional repression required newly synthesized proteins and signaling by phosphatidylinositol 3-kinase (PI3K). RA treatment activates the PI3K/Akt signaling pathway, resulting in increased PI3K activity in extracts from RA-treated cells and a rapid increase in phosphorylation of Akt in Ser-473. Inhibition of PI3K by LY294002 impaired RA-induced differentiation, as assessed by morphological and biochemical criteria. We propose that RA, by activating the PI3K/Akt signaling pathway, plays an important role in the regulation of neuronal cell survival.

The increase in life expectancy has also been accompanied by an increase in the use of medication to treat chronic diseases. Polypharmacy is associated with medication-related problems such as the increase in the anticholinergic burden. Older people are more susceptible to anticholinergic effects on the central nervous system and this, in turn, may be related to cognitive impairment. In this paper, we develop an updated anticholinergic burden scale, the CRIDECO Anticholinergic Load Scale (CALS) via a systematic review of the literature and compare it with the currently most used Anticholinergic Burden Scale (ACB). Our new scale includes 217 different drugs with anticholinergic properties, 129 more than the ACB. Given the effect that anticholinergic medications have on cognitive performance, we then used both scales to investigate the relationship between anticholinergic burden and cognitive impairment in adult Spanish subjects with subjective memory complaint. In our population, we observed an association between cognitive impairment and the anticholinergic burden when measured by the new CALS, but not when the ACB was applied. The use of a more comprehensive and upgraded scale will allow better discrimination of the risk associated with the use of anticholinergic medications on cognitive impairment. CALS can help raise awareness among clinicians of the problems associated with the use of medications, or combinations of them, with large anticholinergic effect, and promote a better personalized pharmacological approach for each patient.

The mRNA levels for the three α₁-adrenoceptor subtypes, α_1A, α_1B, and α_1D, were quantified by real-time RT-PCR in arteries from Wistar rats. The α_1D-adrenoceptor was prominent in both aorta (79.0%) and mesenteric artery (68.7%), α_1A predominated in tail (61.7%) and small mesenteric artery (73.3%), and both α_1A- and α_1D-subtypes were expressed at similar levels in iliac artery. The mRNA levels of the α_1B-subtype were a minority in all vessels (1.7–11.1%). Concentration-response curves of contraction in response to phenylephrine or relaxation in response to α₁-adrenoceptor antagonists on maximal sustained contraction induced by phenylephrine were constructed from control vessels and vessels pretreated with 100 μmol/l chloroethylclonidine (CEC) for 30 min. The significant decrease in the phenylephrine potency observed after CEC treatment together with the inhibitory potency displayed by 8-{2-[4-(2-methoxyphenyl)-1-piperazinyl]-8-azaspiro ( 4 , 5 ) decane-7-dionedihydrochloride} (BMY-7378, an α_1D-adrenoceptor antagonist) confirm the relevant role of α_1D-adrenoceptors in aorta and iliac and proximal mesenteric arteries. The potency of 5-methylurapidil (an α_1A-adrenoceptor antagonist) and the changes in the potency of both BMY-7378 and 5-methylurapidil after CEC treatment provided evidence of a mixed population of α_1A- and α_1D-adrenoceptors in iliac and distal mesenteric arteries. The low potency of prazosin (pIC₅₀ < 9) as well as the high 5-methylurapidil potency in tail and small mesenteric arteries suggest the main role of α_1A/α_1L-adrenoceptors with minor participation of the α_1D-subtype. The mRNA levels and CEC treatment corroborated this pattern and confirmed that the α_1L-adrenoceptor could be a functional isoform of the α_1A-subtype.

Increasing technological advances have generated a digital dependency in the population, resulting in a group of digitally excluded vulnerable people that lack basic digital skills. The aim of this study was to assess the digital divide in patients in relation to the healthcare environment. We explored the extent and effects of the digital health divide by undertaking a systematic review of the academic literature and comparing our findings with the results of a cross-sectional in-person survey answered by 881 people at four community pharmacies. In terms of the sociodemographic profile of the patients, we collected data regarding their gender, age, education level, and location (periphery or urban). The parameters evaluated were use of the internet to search for health information, use of telemedicine, use of different medical/healthcare applications, understanding explanations given by physicians regarding health, and asking pharmacists for help about newly prescribed treatments. Moreover, 168 pharmacists answered an online survey about how often they helped patients to make health center appointments or to download their COVID-19 vaccination certificate. Gender did not influence these results, but age, education level, and population location did. Those with the lowest levels of education required more help to request a health center appointment. People with high education levels and those living in an urban environment more often searched the internet for information about treatments that were new to them. Finally, people living in periphery areas received more help from their pharmacists, 60% of which said they had helped patients to download their COVID-19 vaccination certificate, with 24% of them saying they helped patients with this on a daily basis.

In human and animal hypertension models, increased activity of G-protein-coupled receptor kinase (GRK) 2 determines a generalized decrease of β-adrenergic vasodilatation. We analyzed the possibility of differential changes in the expression and functionality of α_1A, α_1B, α_1D, β₁, β₂, and β₃-ARs also being involved in the process. We combined the quantification of mRNA levels with immunoblotting and functional studies in aortas of young and adult spontaneously hypertensive rats (SHRs) and their controls (Wistar Kyoto). We found the expression and function of β₁-adrenoceptors in young prehypertensive SHRs to be higher, whereas a generalized increase in the expression of the six adrenoceptors and GRK2 was observed in aortas of adult hypertensive SHRs. α_1D- and β₃-Adrenoceptors, the subtypes that are more resistant to GRK2-mediated internalization and mostly expressed in rat aorta, exhibited an increased functional role in hypertensive animals, showing two hemodynamic consequences: 1) an increased sensitivity to the vasoconstrictor stimulus accompanied by a decreased sensitivity to the vasodilator stimulus (α_1D-ARs are the most sensitive to agonists, and β₃-ARs are the least sensitive to agonists); and 2) a slower recovery of the basal tone after adrenergic stimulus removal because of the kinetic characteristic of the α_1D subtype. These functional changes might be involved in the greater sympathetic vasoconstrictor tone observed in hypertension.