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We introduce a method utilizing single laser-generated cavitation bubbles to stimulate cellular mechanotransduction in dermal fibroblasts embedded within 3D hydrogels. We demonstrate that fibroblasts embedded in either amorphous or fibrillar hydrogels engage in Ca 2+ signaling following exposure to an impulsive mechanical stimulus provided by a single 250 µm diameter laser-generated cavitation bubble. We find that the spatial extent of the cellular signaling is larger for cells embedded within a fibrous collagen hydrogel as compared to those embedded within an amorphous polyvinyl alcohol polymer (SLO-PVA) hydrogel. Additionally, for fibroblasts embedded in collagen, we find an increased range of cellular mechanosensitivity for cells that are polarized relative to the radial axis as compared to the circumferential axis. By contrast, fibroblasts embedded within SLO-PVA did not display orientation-dependent mechanosensitivity. Fibroblasts embedded in hydrogels and cultured in calcium-free media did not show cavitation-induced mechanotransduction; implicating calcium signaling based on transmembrane Ca 2+ transport. This study demonstrates the utility of single laser-generated cavitation bubbles to provide local non-invasive impulsive mechanical stimuli within 3D hydrogel tissue models with concurrent imaging using optical microscopy. STATEMENT OF SIGNIFICANCE: Currently, there are limited methods for the non-invasive real-time assessment of cellular sensitivity to mechanical stimuli within 3D tissue scaffolds. We describe an original approach that utilizes a pulsed laser microbeam within a standard laser scanning microscope system to generate single cavitation bubbles to provide impulsive mechanostimulation to cells within 3D fibrillar and amorphous hydrogels. Using this technique, we measure the cellular mechanosensitivity of primary human dermal fibroblasts embedded in amorphous and fibrillar hydrogels, thereby providing a useful method to examine cellular mechanotransduction in 3D biomaterials. Moreover, the implementation of our method within a standard optical microscope makes it suitable for broad adoption by cellular mechanotransduction researchers and opens the possibility of high-throughput evaluation of biomaterials with respect to cellular mechanosignaling., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objectives: Risk indices help quantify the risk of cardiovascular events and death prior to making decisions about prophylactic AAA repair. This paper aims to study the predictive capabilities of 5 validated indices., Design and Methods: A prospective observational multi-centre cohort study from August 2005 to September 2007 in Glasgow recruited 106 consecutive patients undergoing elective open AAA repair. The Glasgow Aneurysm Score (GAS), Vascular physiology only Physiological and Operative Severity Score for enUmeration of Mortality (V(p)-POSSUM), Vascular Biochemical and Haematological Outcome Model (VBHOM), Revised Cardiac Risk Index (RCRI) and Preoperative Risk Score of the Estimation of Physiological Ability and Surgical Stress Score (PRS of E-PASS) were calculated. Indices were compared using receiver operating characteristic (ROC) analysis and area under the curve (AUC) estimates. End points were all-cause mortality, Major Adverse Cardiac Events (MACE) and cardiac death., Results: GAS, VBHOM and RCRI did not predict outcome. V(p)-POSSUM predicted MACE (AUC = 0.681), cardiac death (AUC = 0.762) and all-cause mortality (AUC = 0.780), as did E-PASS (AUC = 0.682, 0.821, 0.703 for MACE, cardiac death and all-cause mortality respectively)., Conclusion: Whilst V(p)-POSSUM and E-PASS predicted outcome, the less complex RCRI and GAS performed poorly which questions the utility of decision making based on these surgical risk indices., (Copyright © 2012 European Society for Vascular Surgery. Published by Elsevier Ltd. All rights reserved.)

Background: The prediction of long-term survival after surgery is complex. Natriuretic peptides can predict short-term postoperative cardiac morbidity and mortality. This study aims to determine the long-term prognostic significance of preoperative B-type natriuretic peptide (BNP) concentration after major non-cardiac surgery., Methods: We conducted a prospective single-centre observational cohort study in a West of Scotland teaching hospital. Three hundred and forty-five patients undergoing major non-cardiac surgery were included. The primary endpoint was long-term all-cause mortality., Results: Overall survival was 67.8% (234/345), with 27 postoperative deaths (within 42 days) and 84 deaths at subsequent follow-up (median follow-up 953 days). A BNP concentration of >87.5 pg ml(-1) best predicted mortality, and the mean survival of patients with an elevated BNP (>87.5 pg ml(-1)) was 731.9 (95% CI 613.6-850.2) days compared with 1284.6 days [(95% CI 1219.3-1350.0), P<0.001] in patients with a BNP<87.5 pg ml(-1). BNP was an independent predictor of survival., Conclusions: BNP is an independent predictor of long-term survival after major non-cardiac surgery. A simple preoperative blood test can provide predictive information on future risk of death, and potentially has a role in preoperative risk assessment.

Introduction: Clinical/microbiological studies have consistently revealed the persistence of some bacteria after conventional root canal debridement. Although this was originally attributed to the complexity of the root canal anatomy and the difficulty of delivering antibacterial agents effectively, it has emerged that the biofilm encasement of bacterial cells may confer a further mechanism of resistance. The purpose of this study was to investigate the relative disruption and bactericidal effects of root canal irrigants on single- and dual-species biofilms of root canal isolates., Methods: Biofilms of Streptococcus sanguinis, Enterococcus faecalis, Fusobacterium nucleatum, and Porphyromonas gingivalis were grown on nitrocellulose membranes for 72 hours and immersed in NaOCl, EDTA, chlorhexidine, and iodine for 1, 5, or 10 minutes. The number of viable and nonviable bacteria disrupted from the biofilm and those remaining adherent were determined by using a viability stain in conjunction with fluorescence microscopy., Results: Gram-negative obligate anaerobe species were more susceptible to cell removal than gram-positive facultative anaerobes. The majority of cells were disrupted after the first minute of exposure; however, the extent varied according to the agent and species. The most effective agent at disrupting biofilms was NaOCl. Iodine was generally effective at bacterial killing but not disruption., Conclusions: Biofilm disruption and cell viability were influenced by the species, their coassociation in dual-species biofilms, the test agent, and the duration of exposure. The effectiveness of NaOCl as an endodontic irrigant was reinforced.

Topically applied retinoic acids have been found to enhance the gene expression for collagen types I and III in the skin of UVB-irradiated hairless mice. Prior damage is required because the effect is not observed in the skin of age-matched, non-irradiated control animals. Immunochemical methods have shown an increase in TGF-beta 1 and, to a lesser extent, of TGF-beta 2 in the epidermis following retinoic acid treatment. There were no changes in mRNA levels for any of the isotypes of TGF-beta induced by retinoic acid treatment. This study suggests that TGF-beta may mediate the effect of retinoic acids on dermal repair through the stimulation of collagen gene expression.

Twenty-one-day-old rats placed on a vitamin D-deficient diet showed no decrease in serum, 1,25-dihydroxy vitamin D levels after 13 days on this diet. Between 13 and 20 days on this D-deficient diet there was a 50% decrease in serum 1,25-dihydroxy vitamin D. After 34 days, the level of 1,25-dihydroxy vitamin D in serum had dropped to near zero. With a vitamin D-deficient diet lacking calcium, there was an apparent stimulation of 1-hydroxylase, resulting in higher 1,25-dihydroxy vitamin D serum levels after 6--20 days on the diet. After 27 days there were no differences in 1,25-dihydroxy-vitamin D levels in animals fed a calcium-replete or calcium-deficient rachitogenic diet. When 1-day-old chickens were maintained on a rachitogenic diet for 1 week, 1,25-dihydroxy vitamin D levels were higher in animals fed the calcium-deficient diet compared with the calcium-replete diet. After 2 weeks on either rachitogenic diet, 1,25-dihydroxy vitamin D levels decreased to near zero. Measurement of 1,25-dihydroxy vitamin D levels has provided a biochemical indicator of vitamin D deficiency in chicks and rats which will complement other established biological criteria for vitamin D deficiency.

This paper describes the use of family therapy with adolescent school refusers. School refusal is assessed and formulated from a family systems perspective, and therapy proceeds with the crisis precipitated by insisting on the adolescent's early return to school. Some of the common problems encountered in this approach are considered along with its results.

The hypercalciuria that occurs when 1,25 (OH)2D3 (calcitriol) is given to humans with normal renal function depends on dietary Ca absorption and may also relate, in part, to enhanced bone resorption. To evaluate the relationship between urinary and dietary Ca during treatment with calcitriol, 12 metabolic balance studies were performed in normal volunteers ingesting a diet containing 350 mg/day of Ca, to which Ca gluconate was added. After 10 days on either 350 mg/day or 1550 mg/day of Ca, calcitriol, 0.5 microgram every 12 hr, was given. Then diet Ca was changed in successive 5-day treatment periods from 350 to 650, 950 and 1550 mg/day (group A) or from 1550 to 950, 650 and 350 mg/day (group B). On the lowest diet Ca, urinary Ca was less than Ca intake during calcitriol treatment (group A, 220 +/- 50 mg/day; group B, 247 +/- 40). As diet Ca was changed during calcitriol treatment, urinary Ca correlated with diet Ca (r = 0.60) until diet Ca reached 950 mg/day. With calcitriol, serum iPTH fell by 18 to 25% (P less than 0.01) and urinary hydroxyproline fell by 11 to 19% (P less than 0.05 to 0.01). Baseline serum levels of 1,25(OH)2D were 47 +/- 8 and 34 +/- 5 pg/ml in group A and B, respectively, and the values increased to 51 +/- 12 and 45 +/- 7.4 pg/ml during treatment with calcitriol. Serum Ca from fasted subjects was not affected by calcitriol, but the mean postabsorptive serum Ca (moon) was increased by 0.35 mg/dl.(ABSTRACT TRUNCATED AT 250 WORDS)

In this study the hypothesis that calcium supplementation during pregnancy can modify blood pressure patterns in a population of normal pregnant women was tested. Thirty-six women with normal single pregnancies, between 20 and 35 years of age, in the second trimester of gestation (15 weeks), were randomly assigned to receive 1 gm of calcium per day (n = 11), 2 gm per day (n = 11), or a placebo (n = 14). No differences were observed at the times of admission into the study (baseline) in demographic and clinical variables or in the calcium intake of each group. Baseline blood pressure measures in several positions also were not different. After the initial blood pressure measures (fifteenth week), five follow-up blood pressure measures were obtained. The supplemented groups had significantly lower diastolic blood pressure than the control subjects between the twentieth and twenty-fourth weeks of gestation. Thereafter, an increase in the control group and the group receiving 1 gm of calcium was observed, but levels were similar at term. On the contrary, patients receiving 2 gm of calcium had blood pressure values that remained significantly lower throughout the third trimester. No differences or clear patterns were observed in the blood levels of calcium, magnesium, phosphorus, and proteins between and within groups during gestation. A possible explanation involving parathyroid hormone is attempted.

The dose response and pharmacokinetics of orally administered calcitriol were investigated in normal humans. In one protocol, six volunteers received calcitriol 0.25 micrograms twice a day, 0.5 micrograms daily, and 0.5 micrograms twice a day, in successive weeks. Peak plasma levels of 1,25(OH)2D occurred 4 to 8 hours after ingestion of a single dose of 0.5 micrograms, with a return to baseline within 24 hours. The 8:00 AM calcitriol plasma levels were raised only when the drug was given twice daily. Urinary calcium excretion (UCa) was significantly increased from 199 +/- 19 mg/24 hr during the control period to similar levels of 302 +/- 26 mg/24 hr after 0.25 microgram twice a day and 284 +/- 31 mg/24 hr after 0.50 microgram daily. With 0.50 microgram twice a day, UCa was 417 +/- 36 mg/24 hr, a value greater than after the lower doses (p less than 0.05). In another protocol, fourteen volunteers received calcitriol 0.25 microgram, 0.5 microgram, and 1.0 microgram twice a day each for 14 days with intervening control periods of 2 weeks. A dose-related response in urinary calcium/creatinine excretion occurred. Thus, UCa (milligrams calcium per milligram creatinine) increased with calcitriol from 0.13 +/- 0.014 mg to 0.15 +/- 0.018 mg with 0.25 microgram twice a day, from 0.13 +/- 0.010 mg to 0.22 +/- 0.022 mg with 0.5 microgram twice a day, and from 0.12 +/- 0.012 mg to 0.23 +/- 0.012 mg with 1 microgram twice a day (p less than 0.05 with 0.25 microgram, p less than 0.01 with 0.5 and 1 microgram twice a day).(ABSTRACT TRUNCATED AT 250 WORDS)

Serum bone Gla protein (BGP) was measured by radioimmunoassay in 166 healthy men and women aged 30-90 years. Serum BGP levels increased with age in both sexes and were higher in women than in men at all ages. The most striking rise occurred in women after age 40-49. BGP was significantly correlated positively with serum alkaline phosphatase and negatively with midshaft and distal bone mass in both sexes. In women only, BGP levels were significantly positively related to levels of immunoreactive parathyroid hormone (iPTH). When age was included in the multiple regression analysis BGP was still correlated with alkaline phosphatase in both sexes and iPTH in women only. Serum BGP levels were significantly higher in 13 osteoporotic patients than in age-matched controls. It is postulated that with increasing age 1,25-dihydroxyvitamin D levels fall, causing a rise in iPTH and thus in bone turnover, which is reflected by a rise in BGP levels.

Chronic irradiation of hairless mice with UVB leads to elastosis as evidenced by both histologic means and an increase in skin desmosine content. Treatment with topical all-trans- or 13-cis-retinoic acid causes dose-dependent increments in the area of the dermal "repair zone"; skin desmosine content increases during irradiation but does not change significantly after irradiation is discontinued and retinoic acid treatment commenced. During the course of the irradiation the animals develop permanent wrinkles on the exposed dorsal surface, which can be recorded in plastic impressions. The extent of wrinkling can be quantitated and it has been demonstrated that topically applied retinoic acids lead to the complete effacement of these surface features and that the process appears to be permanent.