Effects of caffeine on cerebral blood flow.

OBJECTIVE: The objective of the present study is to evaluate whether, after caffeine ingestion, there are variations in blood velocity of the middle cerebral arteries in clinically healthy young people as well as to evaluate whether this variation is dependent on the administered dose. METHODS: We used transcranial Doppler ultrasonography to record blood velocities of the middle cerebral arteries in three groups of 15 clinically healthy young adults each: no caffeine, a45 mg, and 120 mg of caffeine groups. Transcranial Doppler ultrasonography provided simultaneous bilateral velocity of the middle cerebral arteries measurements while participants performed functional tests (hyperventilation and hypoventilation orders) and three cognitive activities (test 1, short-term memory; test 2, solving a vocabulary problem; and test 3, solving a math problem) each in 31-s tests with 1-min rests between them. Participants were assessed before and 30 min after caffeine ingestion. RESULTS: There was a significant decrease in mean velocity, peak systolic velocity, end-diastolic velocity, and heart rate after high caffeine intake, except in hyperventilation, which was only observed in peak systolic velocity. With the intake of a lower dose, significant decreases were seen with hypoventilation and with test 1. In hyperventilation, there was only a significant decrease in end-diastolic velocity and heart rate; in test 2, it was found in mean velocity and peak systolic velocity; and in test 3, only in heart rate. CONCLUSION: With this study, we conclude that caffeine influences the cardiovascular system acutely, interfering with the velocity of the middle cerebral arteries, causing its decrease. We also conclude that this acute effect causes vasodilation of the cerebral arteries, more accentuated with higher doses of caffeine.

Xport-A functions as a chaperone by stabilizing the first five transmembrane domains of rhodopsin-1.

Rhodopsin-1 (Rh1), the main photosensitive protein of Drosophila, is a seven-transmembrane domain protein, which is inserted co-translationally in the endoplasmic reticulum (ER) membrane. Biogenesis of Rh1 occurs in the ER, where various chaperones interact with Rh1 to aid in its folding and subsequent transport from the ER to the rhabdomere, the light-sensing organelle of the photoreceptors. Xport-A has been proposed as a chaperone/transport factor for Rh1, but the exact molecular mechanism for Xport-A activity upon Rh1 is unknown. Here, we propose a model where Xport-A functions as a chaperone during the biogenesis of Rh1 in the ER by stabilizing the first five transmembrane domains (TMDs) of Rh1.

Normal percentile reference curves for skin ultrasound thickness and stiffness at Rodnan sites.

OBJECTIVES: Our primary objective was to establish preliminary normal reference curves for ultrasound-dermal thickness and skin stiffness in the 17 Rodnan skin sites, considering the effect of gender and age on these measures. As an exploratory objective, we investigated the effect of body mass index and the menopause on skin ultrasound measures. METHODS: A cross-sectional study was conducted involving 140 healthy volunteers, aged 20-79 years. Recruitment was stratified by gender and age (10-year categories). Ultrasound-dermal thickness and skin stiffness were assessed by high-frequency ultrasound and shear-wave elastography, respectively, at the 17 Rodnan skin sites. Outcomes were evaluated through a mixed linear model, univariate and multivariate regressions. Normal reference curves were derived for both ultrasound measures in each skin site. An online calculator of the percentiles of skin ultrasound measures was developed. RESULTS: Ultrasound-dermal thickness and stiffness measures were higher in men than women in all Rodnan skin sites (except in chest for ultrasound-dermal thickness). Age had also a significant impact in both ultrasound measures, but only in some skin sites. Gender and age percentile curves (97.5th, 95th, 75th, 50th, 25th, 5th, 2.5th) were plotted for each of the measures in each skin site. CONCLUSIONS: Gender and age are strongly associated with skin ultrasound parameters, imposing the need for gender-specific and age-specific reference values. Normal reference percentile curves are provided as a basis for future cooperative work to strengthen its evidence basis, representativeness and refinement regarding potentially influential factors.

EMC is required for biogenesis of Xport-A, an essential chaperone of Rhodopsin-1 and the TRP channel.

The ER membrane protein complex (EMC) is required for the biogenesis of a subset of tail anchored (TA) and polytopic membrane proteins, including Rhodopsin-1 (Rh1) and the TRP channel. To understand the physiological implications of EMC-dependent membrane protein biogenesis, we perform a bioinformatic identification of Drosophila TA proteins. From 254 predicted TA proteins, screening in larval eye discs identified two proteins that require EMC for their biogenesis: fan and Xport-A. Fan is required for male fertility in Drosophila and we show that EMC is also required for this process. Xport-A is essential for the biogenesis of both Rh1 and TRP, raising the possibility that disruption of Rh1 and TRP biogenesis in EMC mutants is secondary to the Xport-A defect. We show that EMC is required for Xport-A TMD membrane insertion and that EMC-independent Xport-A mutants rescue Rh1 and TRP biogenesis in EMC mutants. Finally, our work also reveals a role for Xport-A in a glycosylation-dependent triage mechanism during Rh1 biogenesis in the endoplasmic reticulum.

Phosphorylation of iRhom2 Controls Stimulated Proteolytic Shedding by the Metalloprotease ADAM17/TACE.

Cell surface metalloproteases coordinate signaling during development, tissue homeostasis, and disease. TACE (TNF-α-converting enzyme), is responsible for cleavage ("shedding") of membrane-tethered signaling molecules, including the cytokine TNF, and activating ligands of the EGFR. The trafficking of TACE within the secretory pathway requires its binding to iRhom2, which mediates the exit of TACE from the endoplasmic reticulum. An important, but mechanistically unclear, feature of TACE biology is its ability to be stimulated rapidly on the cell surface by numerous inflammatory and growth-promoting agents. Here, we report a role for iRhom2 in TACE stimulation on the cell surface. TACE shedding stimuli trigger MAP kinase-dependent phosphorylation of iRhom2 N-terminal cytoplasmic tail. This recruits 14-3-3 proteins, enforcing the dissociation of TACE from complexes with iRhom2, promoting the cleavage of TACE substrates. Our data reveal that iRhom2 controls multiple aspects of TACE biology, including stimulated shedding on the cell surface.

Ire1 mediated mRNA splicing in a C-terminus deletion mutant of Drosophila Xbp1.

The Unfolded Protein Response is a homeostatic mechanism that permits eukaryotic cells to cope with Endoplasmic Reticulum (ER) stress caused by excessive accumulation of misfolded proteins in the ER lumen. The more conserved branch of the UPR relies on an ER transmembrane enzyme, Ire1, which, upon ER stress, promotes the unconventional splicing of a small intron from the mRNA encoding the transcription factor Xbp1. In mammals, two specific regions (the hydrophobic region 2--HR2--and the C-terminal translational pausing site) present in the Xbp1unspliced protein mediate the recruitment of the Xbp1 mRNA-ribosome-nascent chain complex to the ER membrane, so that Xbp1 mRNA can be spliced by Ire1. Here, we generated a Drosophila Xbp1 deletion mutant (Excision101) lacking both HR2 and C-terminal region, but not the Ire1 splicing site. We show that Ire1-dependent splicing of Xbp1 mRNA is reduced, but not abolished in Excision101. Our results suggest the existence of additional mechanisms for ER membrane targeting of Xbp1 mRNA that are independent of the C-terminal domain of Drosophila Xbp1unspliced.