Inducible heat shock protein A1A (HSPA1A) is markedly expressed in rat myometrium by labour and secreted via myometrial cell-derived extracellular vesicles.

The myometrium goes through physiological, cellular and molecular alterations during gestation that necessitate effective cellular proteostasis. Inducible heat shock protein A1A (HSPA1A) is a member of the 70-kDa heat shock protein A (HSPA) family, which acts as a chaperone to regulate proteostasis; however, HSPA1A also participates as a cytokine in inflammatory regulation, leading to its designation as a chaperokine. This study examined the spatiotemporal expression of HSPA1A protein in the rat myometrium throughout gestation and assessed whether it is secreted as cargo of myometrial cell-derived extracellular vesicles (EVs). Immunoblot analysis demonstrated that HSPA1A expression was markedly elevated during late pregnancy and labour and increased by uterine distension. Myometrial HSPA1A expression insitu increased in myocytes of longitudinal and circular muscle layers from Day 19 through to postpartum, specifically in the cytoplasm and nuclei of myocytes from both muscle layers, but frequently detectable just outside myocyte membranes. Scanning electron microscopy examination of samples isolated from hTERT-HM cell-conditioned culture medium, using EV isolation spin columns, confirmed the presence of EVs. EV lysates contained HSPA8, HSPA1A and the EV markers apoptosis-linked gene 2-interacting protein X (Alix), the tetraspanin cluster of differentiation 63 (CD63), tumour susceptibility gene 101 (TSG101) and HSP90, but not the endoplasmic reticulum protein calnexin. These results indicate that HSPA1A may act as a chaperokine in the myometrium during pregnancy.

Root canal obturation by ultrasonic condensation of gutta-percha. Part II: an in vitro investigation of the quality of obturation.

AIM: To compare the quality of root canal obturation using ultrasonic or cold condensation of gutta-percha and to determine the effect of power setting and activation time on the quality of obturation using the former technique. METHODOLOGY: An extracted human maxillary canine was used in an in vitro split tooth model to allow repeated obturation of the same root canal system using an ultrasonic device to thermocompact gutta-percha without sealer. After each obturation, the root filling was removed from the tooth to allow evaluation of its quality and for the tooth to be re-obturated. The influence of combinations (n = 10 per combination) of power setting (1, 3, 5) and activation times (4, 10, 15 s) was tested on the quality of root filling, assessed by measuring the voids within the body of the root filling as well as at the surface. Image analysis was used to quantify the voids within the body of the root filling. Cold lateral condensation of gutta-percha served as a control. RESULTS: Both surface and cross-sectional analyses revealed that different power setting and activation time combinations produced significantly fewer voids than cold lateral condensation (P < 0.05) at the apical, mid-root and coronal levels. CONCLUSIONS: Taking surface and cross-sectional analysis together only power setting 5 and activation times of 10 and 15 s consistently produced ultrasonically thermocompacted root canal fillings with fewer voids than cold lateral condensation without sealer.

Ultrasonic condensation of gutta-percha: the effect of power setting and activation time on temperature rise at the root surface - an in vitro study.

AIM: To determine the effect of power setting and duration of activation on the temperature rise at the root surface during root canal obturation by ultrasonic condensation of gutta-percha. METHODOLOGY: A human maxillary canine was used in an in vitro split tooth model to allow repeated obturation of the root canal system using an ultrasonic device to thermocompact gutta-percha. Combinations of power settings (1, 3 and 5) and durations of activation (4, 10 and 15 s) were used to test their effect on temperature rise at the root surface using eight K-type thermocouples at the mid-root and apical levels. At the end of each obturation, the tooth was disassembled to remove the gutta-percha in preparation for the next obturation (n = 10 for each combination). Multiple linear regression models were used to investigate the effects of power setting, duration of activation and thermocouple location on the maximum temperature rise recorded. RESULTS: Only one combination of power setting (5) and duration of activation (15 s) resulted in temperature rise in excess of 10 degrees C. The maximum temperature rise at the mid-root level was significantly (P < 0.001) greater than that recorded apically. It is also significantly affected by the combination of power setting and duration of activation. CONCLUSIONS: Temperature rises at the root surface during ultrasonic condensation of gutta-percha in excess of 10 degrees C were evident in only one combination of power (5) and time (15 s) settings at the mid-root level.