Health care-associated infections studies project: An American journal of infection control and national healthcare safety network data quality collaboration case study - Chapter 2 Identifying Healthcare-associated Infections (HAI) for NHSN Surveillance case study vignettes.

This case study is part of a series centered on the Centers for Disease Control and Prevention/National Healthcare Safety Network (NHSN) healthcare-associated infection (HAI) surveillance definitions. This specific case study focuses on the application of three of the surveillance concepts included in the Patient Safety Component, Chapter 2 - Identifying Healthcare-associated Infections (HAI) for NHSN Surveillance. The intent of the case study series is to foster standardized application of the NHSN HAI surveillance definitions and encourage accurate HAI event determination among Infection Preventionists (IPs).

Health care-associated infections studies project: An American Journal of Infection Control and National Healthcare Safety Network Data Quality Collaboration Case Study - Chapter 9 Surgical site infection event (SSI) case study.

This case study is part of a series centered on the Centers for Disease Control and Prevention/National Healthcare Safety Network (NHSN) healthcare-associated infection (HAI) surveillance definitions. This specific case study focuses on the application of common surveillance concepts included in the Patient Safety Component, Chapter 9 - Surgical Site Infection Event (SSI). The intent of the case study series is to foster standardized application of the NHSN HAI surveillance definitions and encourage accurate HAI event determination among Infection Preventionists (IPs).

In adrenal glomerulosa cells, angiotensin II inhibits proliferation by interfering with fibronectin-integrin signaling.

Angiotensin II (Ang II), through the Ang II type 1 receptor subtype, inhibits basal proliferation of adrenal glomerulosa cells by inducing the disruption of actin stress fiber organization. This effect is observed in cells cultured on plastic or on fibronectin. The aim of the present study was to investigate how Ang II may interfere with extracellular matrix/integrin signaling. In cells treated for 3 d with echistatin (EC) (a snake-venom RGD-containing protein that abolishes fibronectin binding to alpha(5)beta(1) or alpha(v)beta(3) integrins), basal proliferation decreased by 38%, whereas Ang II was unable to abolish basal proliferation. In cells grown on fibronectin, Ang II decreased binding of paxillin to focal adhesions and, similarly to EC, induced a rapid dephosphorylation of paxillin (1 min), followed by an increase after 15 min. Fibronectin enhanced RhoA/B and Rac activation induced by Ang II, an effect abolished by EC. Under basal conditions, paxillin was more readily associated with RhoA/B than with Rac. Stimulation with Ang II induced a transient decrease in RhoA/B-associated paxillin (after 5 min), with a return to basal levels after 10 min, while increasing Rac-associated paxillin. Finally, results reveal that glomerulosa cells are able to synthesize and secrete fibronectin, a process by which cells can stimulate their own proliferative activity when cultured on plastic. Together, these results suggest that Ang II acts at the level of integrin-paxillin complexes to disrupt the well- developed microfilament network, a condition necessary for the inhibition of cell proliferation and initiation of steroidogenesis.

From integrative signalling to metabolic disorders.

The adrenal cortex undergoes constant dynamic structural changes, a key element in ensuring integrative functionality of the gland. Studies have shown that the cellular environment can modulate cell functions such as proliferation and steroid secretion. For example, 3-day treatment with angiotensin II promotes protein synthesis with a concomitant decrease in proliferation of glomerulosa cells, when cultured on fibronectin, but not on collagen IV or laminin. These effects involve close interaction between cytoskeleton-associated proteins and activation of p42/p44mapk and p38 MAPK pathways. On the other hand, adrenocorticotropin hormone (ACTH), which is clearly the most potent stimulus of fasciculata cells, induces specific modulation of targeted proteins, when cells are cultured on collagen IV, but not on fibronectin or laminin. In particular, ACTH treatment leads to increased expression of Seladin-1 and induces the relocalization of Seladin-1 from the cytoplasm to the nucleus, both in vivo and in culture conditions, in adult rats and in human fetal adrenal glands. As a whole, these results indicate that Seladin-1, together with collagen IV, is able to modulate ACTH responsiveness. Hence, Seladin-1 may participate in the regulation of steroidogenesis when localized in the cytoplasm, while conversely protecting cells against oxidative stress generated by intense ACTH stimulation when massively localized in the nucleus.

Expression of extracellular matrix proteins and integrins in rat adrenal gland: importance for ACTH-associated functions.

The expression of main extracellular matrix (ECM) and their integrins were studied in the adult rat adrenal gland. Collagen I, IV (CI, CIV), laminin (LN) and fibronectin (FN) expression was observed surrounding each glomerulosa cell and as long fibrils between the cords of fasciculata cells. In the medulla, FN was present around chromaffin cells or bordering blood vessels. Integrin alpha2, alpha3 and alpha5 were present mainly in the cortex, while alpha1 was present in the medulla. In culture, all ECM favoured proliferation of both glomerulosa and fasciculata cells, while protein synthesis was lower on FN and LN in glomerulosa cells. CIV promoted ACTH-induced proliferation whereas FN favoured ACTH-induced protein synthesis in glomerulosa cells. Except for LN, ECM increased expression of 3beta-hydroxysteroid dehydrogenase and enhanced basal aldosterone, although corticosterone secretion was only enhanced by CI and CIV. In fasciculata cells, the potency of ACTH-induced cAMP production was lower on ECM, compared with plastic. Moreover, ACTH, but not ECM, activated mitogenic-activated protein kinase p38 and stress-activated protein kinases. Glomerulosa and fasciculata cells grown on CI and CIV had a polygonal morphology, while cells grown on LN appeared as clusters of small rounded cells. On FN, the glomerulosa cells exhibited polygonal morphology while fasciculata cells appeared as clusters of small rounded cells. Together, these results indicate that ECM modulates basal and ACTH-induced cell functions, with FN, CI and CIV specifically favouring steroid secretion, as opposed to LN which inhibits secretion while promoting proliferation.

Seladin-1 expression in rat adrenal gland: effect of adrenocorticotropic hormone treatment.

Seladin-1 (KIAA0018) gene is the seventh most highlyexpressed gene in the adult adrenal gland, along with genes coding for steroidogenic enzymes. The aim of the present study was to investigate the localization of the Seladin-1 protein in control and ACTH-treated rat adrenal glands and to verify whether Seladin-1 is involved in secretion. Immunofluorescence studies revealed that Seladin-1 was localized principally in the zona fasciculata, cytoplasm, and nucleus. Expression of Seladin-1 was increased by ACTH treatment, in vivo and in culture conditions. Subcellular fractionation offasciculata cells showed that Seladin-1 was mainly present in the nucleus, membrane, and cytoskeleton fractions and, to a lesser extent, in the cytosol. ACTH treatment decreased Seladin-1 expression in the cytosol, with a concomitant increase in the nuclear fraction. In the glomerulosa and fasciculata cells in culture, ACTH induced a relocalization of Seladin-1 into specific nuclear regions. This ACTH-induced relocalization was abrogated by the pre-treatment of cells with 75 nM U18666A (an inhibitor of Seladin-1). In addition, fasciculata cells exhibited an increase in the basal level of steroid secretion when cultured in the presence of U18666A (25 and 75 nM), although ACTH-induced secretion was decreased. In summary, the present study demonstrates that the protein expression of Seladin-1 is more abundant in fasciculata cells than in glomerulosa cells and that the ACTH treatment increases both expression and nuclear localization of the protein. Results also suggest that depending on its cellular localization, the Delta24-reductase activity of Seladin-1 may play a major role in steroid secretion in the adrenal gland.

Role of MAPKs in angiotensin II-induced steroidogenesis in rat glomerulosa cells.

Angiotensin II is one of the most important stimuli of rat adrenal glomerulosa cells, stimulating both steroid secretion and growth. In a previous report, we had shown that Ang II promotes cellular hypertrophy, but not proliferation, in rat adrenal glomerulosa cells maintained in primary culture for 3 days. The inhibition of proliferation and stimulation of hypertrophy induced by Ang II involves both p42/p44(mapk) and p38 MAPK activation. The increase in cell protein content induced by Ang II entails formation of a cortical actin ring and Rac-dependent activation of p42/p44(mapk) and p38 MAPK. The present study summarizes these results and provides evidences that Ang II-induced activation of p42/p44(mapk) and p38 MAPK are implicated in aldosterone secretion by enhancing expression of specific steroidogenic proteins such as StAR and 3beta-HSD.

The growth-promoting effects of angiotensin II in adrenal glomerulosa cells: an interactive tale.

The zona glomerulosa of the adrenal cortex is well-known for its high level of proliferation, compared to the adjacent zona fasciculata, both in in vivo and in vitro conditions. Angiotensin II (Ang II) is a potent growth factor for glomerulosa cells, appearing as a proliferative factor in vivo, under sodium-deficient diet conditions, as well as in vitro, in studies conducted with whole zona glomerulosa. However, in cells maintained in primary culture for 3 days, Ang II rather promotes cellular hypertrophy with a concomitant arrest in basal cell proliferation. The present essay aims at providing experimental arguments supporting such unexpected observations, with particular focus on the modulatory impact of the extracellular environment on Ang II action, namely AT(1) receptor-induced signaling pathways and cell responses.

Differential involvement of cytoskeleton and rho-guanosine 5'-triphosphatases in growth-promoting effects of angiotensin II in rat adrenal glomerulosa cells.

Glomerulosa cells readily proliferate in primary culture. However, 3-d treatment with angiotensin II (Ang II) promotes cellular hypertrophy with a concomitant decrease in proliferation. The aim of the present study was to investigate the manner by which cytoskeleton and Rho-GTPase proteins may be involved in Ang II-induced growth and MAPK activation. Preincubation with Y27632 (an inhibitor of Rho-associated kinase) decreased basal proliferation, as did Ang II, whereas toxin B, which inhibits Rho-GTPases, enhanced the inhibitory effect of Ang II. Conversely, toxin B inhibited protein synthesis induced by Ang II, whereas Y27632 had no effect. Ang II induced a rapid but transient activation of RhoA/B, an effect abolished in Y27632-preincubated cells. Activation of Rac appeared biphasic, with an early activation at 1 min, followed by a more sustained effect at 10 min. Toxin B abolished Rac activation. Immunofluorescence studies revealed that Y27632 and toxin B disrupted the F-actin network similarly to Ang II. Y27632 also abolished the cortical F-actin ring induced by Ang II. Preincubation of cells with toxin B abolished p38 MAPK phosphorylation and early activation of p42(mapk) (ERK2) and decreased p44(mapk) (ERK1) induced by Ang II. In contrast, Y27632, abolished p44(mapk) (ERK1) but had no effect on p42(mapk) (ERK2) or p38. Together these results indicate that, in rat adrenal glomerulosa cells, specific Rho/Rho-associated kinase-dependent activation of p44(mapk) (ERK1) and an intact cytoskeletal organization are necessary in mediating basal cell proliferation, whereas activation of p42/p44(mapk), p38 MAPK, and Rac are essential in mediating Ang II-induced protein synthesis (steroidogenic acute regulatory protein and 3beta-hydroxysteroid dehydrogenase).

Angiotensin II stimulates protein synthesis and inhibits proliferation in primary cultures of rat adrenal glomerulosa cells.

Angiotensin II (Ang II) is one of the most important stimuli of rat adrenal glomerulosa cells. The aim of the present study was to investigate whether Ang II can stimulate cell proliferation and/or hypertrophy and investigate pathways and intracellular targets. A 3-d treatment with Ang II (5-100 nm), through the Ang II type 1 receptor subtype, abolished cell proliferation observed in control cells but increased protein synthesis. Preincubation with PD98059 (a MAPK kinase inhibitor) abolished basal proliferation and had no effect on basal protein synthesis but did reverse the effect of Ang II on protein synthesis. The p38 MAPK inhibitor SB203580 reversed the inhibitory effect on cell proliferation and abolished the increase in protein synthesis, whereas the c-Jun N-terminal kinase inhibitor SP600125 had no effect. Time-course studies revealed that Ang II stimulated phosphorylation of both p42/p44mapk and p38 MAPK but did not activate c-Jun N-terminal kinase. Ang II had no effect on the level of cyclin E expression but increased the expression of the cyclin-dependent kinase, p27Kip1, an effect abolished in cells preincubated with SB203580 and PD98059. In conclusion, in cultured rat glomerulosa cells, a 3-d treatment with Ang II increases protein synthesis, with a concomitant decrease in proliferation. These effects are mediated by both the p42/p44mapk and p38 MAPK pathways, which increase expression of the steroidogenic enzymes, steroidogenic acute regulatory protein and 3beta-hydroxysteroid dehydrogenase and p27Kip1, a protein known to block the cell cycle in G1 phase. Together these results support the key role of Ang II as a stimulus of steroid synthesis rather than a proliferating factor.

Extracellular matrix and hormones modulate DAX-1 localization in the human fetal adrenal gland.

CONTEXT: The orphan nuclear receptor DAX-1 is essential for human adrenal cortex development and functions as a transcriptional repressor of multiple genes implicated in steroidogenic pathways. OBJECTIVE: The aim of this study was to investigate the localization of the DAX-1 protein in human fetal adrenal glands and to assess whether this protein can be modulated by the extracellular matrix and hormones. RESULTS: DAX-1 is localized mainly in the nucleus in the outer definitive zone and in the cytoplasm in the fetal zone, whereas the number of DAX-1 positive cells decreases from the external to the internal portion of the gland. When cultured on a collagen or a fibronectin matrix, DAX-1 is localized in the nucleus of the definitive cells and exhibits a nucleocytoplasmic distribution in the fetal cells. ACTH stimulation induces nuclear localization of DAX-1 in fetal cells cultured on collagen without modifying nucleocytoplasmic localization on fibronectin. In contrast, angiotensin II induces the protein to be localized only in the cytoplasm in fetal cells cultured on either collagen or fibronectin. CONCLUSIONS: The localization of DAX-1 is compatible with the known functional properties of DAX-1 regarding the steroidogenic activity of adrenal cells. Moreover, this study suggests that modulation of DAX-1 localization in the fetal adrenal gland by hormones and components of the extracellular matrix may represent a mechanism for controlling the expression of steroidogenic enzymes in the definitive and fetal zones.

Inhibition of steroidogenic response to adrenocorticotropin by leptin: implications for the adrenal response to maternal separation in neonatal rats.

Previous studies have shown that leptin can regulate the adrenocortical axis. Neonatal rodents exhibit a period of adrenal hyporesponsiveness to stress in the first 2 wk of life, and we determined the role of leptin as a mediator of this process. We examined the direct effects of leptin on neonatal adrenal steroidogenic responses to ACTH under basal conditions and after 24-h maternal separation. In isolated adrenocortical cells from as early as postnatal d 5 (PND5) and throughout the neonatal period, acute (2.5 h) incubation with leptin significantly inhibited ACTH-stimulated corticosterone and aldosterone secretion without affecting cAMP production. In PND10 pups, 24-h maternal separation and the resulting rapid decline in plasma leptin levels increased basal corticosterone and aldosterone secretion in vivo and in isolated cells, but did not modify the ability of leptin to inhibit stimulated steroid production in vitro. Maternal separation in PND10 pups increased adrenal expression of steroidogenic acute regulatory protein (StAR) and peripheral-type benzodiazepine receptor (PBR) proteins as well as all steroidogenic enzymes measured (3beta-hydroxysteroid dehydrogenase, P450C11B1, and P450C11B2). Leptin (1 mg/kg body weight, i.p.) replacement during maternal separation did not affect basal corticosterone output, but reduced corticosterone secretion and StAR and PBR protein expression induced by exogenous ACTH challenge (20 or 80 microg/kg body weight, i.p.). These results indicate that leptin inhibits ACTH-stimulated secretion of corticosterone and aldosterone, at least through a rapid reduction in the expression of StAR and PBR protein in the neonatal adrenal gland. As leptin concentrations in pups are controlled to a large extent by the maternal diet, these results emphasize the key role of leptin to mediate the maternal influence on the adrenocortical axis of the infant.

Connection between integrins and cell activation in rat adrenal glomerulosa cells: a role for Arg-Gly-Asp peptide in the activation of the p42/p44(mapk) pathway and intracellular calcium.

Integrins are responsible for adhesion and activation of several intracellular cascades. The present study was aimed at determining whether the interaction between fibronectin and integrins could generate pathways involved in physiological functions of rat adrenal glomerulosa cells. Immunofluorescence studies and adhesion assays showed that fibronectin was the best matrix in promoting the formation of focal adhesion. Binding of glomerulosa cells to fibronectin, but not to collagen I or poly-L-lysine, involved the integrin-binding sequence Arg-Gly-Asp (RGD). Activation of glomerulosa cells with Arg-Gly-Asp-Ser (RGDS) induced an increase in [Ca(2+)](i), whereas fibronectin triggered a release of Ca(2+) from InsP(3)-sensitive Ca(2+) stores. Aldosterone secretion induced by ACTH, angiotensin II, and RGDS and proliferation were improved on fibronectin, compared with poly-L-lysine. The RGDS peptide induced a transient increase in the activity of the p42/p44(mapk), independent of phosphatidylinositol-3 kinase and protein kinase C. Integrins alpha(5) and alpha(V) as well as their fibronectin receptor partners beta(1) and beta(3), were identified. These results suggest that in rat adrenal glomerulosa cells, binding of the alpha(5)beta(1), alpha(v)beta(1), or alpha(v)beta(3) integrins to fibronectin is involved in the generation of two important signaling events, increase in intracellular calcium, and activation of the p42/p44(mapk) cascade, leading to cell proliferation and aldosterone secretion.

Characterization of blood pressure and morphological traits in cardiovascular-related organs in 13 different inbred mouse strains.

To better understand the contributions of various genetic backgrounds to complex quantitative phenotypes, we have measured several quantitative traits of cardiovascular interest [i.e., systolic blood pressure, weight (corrected by body weight) of several cardiac compartments and adrenals and kidneys, and histological correlates for kidneys and adrenals] in male and female mice from 13 different inbred strains. We selected strains so that each major genealogical group would be represented and to conform to priorities set by the Mouse Phenome Database project. Interstrain comparisons of phenotypes made it possible to identify strains that displayed values that belonged to either the low or the high end of the interstrain variance for quantitative traits, such as systolic blood pressure, body weight, left ventricular weight, and/or adrenocortical structure. For instance, both male and female C3H/HeJ and A/J mice displayed either low systolic blood pressure or low cardiac ventricular mass, respectively, and male C57BL6/J displayed low adrenal weight. Likewise, intersex comparisons made it possible to identify phenotypic values that were sexually dimorphic for some of the same traits. For instance, female AKR/J mice had relatively higher body weight and systolic blood pressure values than their male counterparts, perhaps constituting an animal model of the metabolic X syndrome. These strain- and sex-specific features will be of value both for future genetic and/or developmental studies and for the development of new animal models that will help in the generation of mechanistic hypotheses. All data have been deposited to the Mouse Phenome Database for future integration with the Mouse Genome Database and can be further analyzed and compared with tools available on the site.

Recommendations for the standardisation of oxytocin nasal administration and guidelines for its reporting in human research.

A series of studies have reported on the salubrious effects of oxytocin nasal spray on social cognition and behavior in humans, across physiology (e.g., eye gaze, heart rate variability), social cognition (e.g., attention, memory, and appraisal), and behavior (e.g., trust, generosity). Findings suggest the potential of oxytocin nasal spray as a treatment for various psychopathologies, including autism and schizophrenia. There are, however, increasing reports of variability of response to oxytocin nasal spray between experiments and individuals. In this review, we provide a summary of factors that influence transmucosal nasal drug delivery, deposition, and their impact on bioavailability. These include variations in anatomy and resultant airflow dynamic, vascularisation, status of blood vessels, mode of spray application, gallenic formulation (including presence of uptake enhancers, control release formulation), and amount and method of administration. These key variables are generally poorly described and controlled in scientific reports, in spite of their potential to alter the course of treatment outcome studies. Based on this review, it should be of no surprise that differences emerge across individuals and experiments when nasal drug delivery methods are employed. We present recommendations for researchers to use when developing and administering the spray, and guidelines for reporting on peptide nasal spray studies in humans. We hope that these recommendations assist in establishing a scientific standard that can improve the rigor and subsequent reliability of reported effects of oxytocin nasal spray in humans.

Differential responses to salt supplementation in adult male and female rat adrenal glands following intrauterine growth restriction.

In low sodium-induced intrauterine growth restricted (IUGR) rat, foetal adrenal steroidogenesis as well as the adult renin-angiotensin-aldosterone system (RAAS) is altered. The aim of the present study was to determine the expression of cytochrome P450 aldosterone synthase (P450aldo) and of angiotensin II receptor subtypes 1 (AT(1)R) and 2 (AT(2)R) in adult adrenal glands and whether this expression could be influenced by IUGR and by high-salt intake in a sex-specific manner. After 6 weeks of 0.9% NaCl supplementation, plasma renin activity, P450aldo expression and serum aldosterone levels were decreased in all groups. In males, IUGR induced an increase in AT(1)R, AT(2)R, and P450aldo levels, without changes in morphological appearance of the zona glomerulosa (ZG). By contrast, in females, IUGR had no effect on the expression of AT(1)R, but increased AT(2)R mRNA while decreasing protein expression of AT(2)R and P450aldo. In males, salt intake in IUGR rats reduced both AT(1)R mRNA and protein, while for AT(2)R, mRNA levels decreased whereas protein expression increased. In females, salt intake reduced ZG size in IUGR but had no affect on AT(1)R or AT(2)R expression in either group. These results indicate that, in response to IUGR and subsequently to salt intake, P450aldo, AT(1)R, and AT(2)R levels are differentially expressed in males and females. However, despite these adrenal changes, adult IUGR rats display adequate physiological and adrenal responses to high-salt intake, via RAAS inhibition, thus suggesting that extra-adrenal factors likely compensate for ZG alterations induced by IUGR.

Angiotensin-II acute regulation of rapid response genes in human, bovine, and rat adrenocortical cells.

Angiotensin-II (Ang-II) regulates adrenal steroid production and gene transcription through several signaling pathways. Changes in gene transcription occur within minutes after Ang-II stimulation, causing an increase in aldosterone production and subsequent increase in the overall capacity to produce aldosterone. Our goal was to compare the Ang-II regulation of early gene expression and confirm the up-regulation of selected genes using quantitative real-time RT-PCR (qPCR) across three species, such as, human, bovine, and rat. Microarray analyses were performed using samples from control and Ang-II (10 nM)-treated (1 h) cells from human adrenocortical tumor cell line H295R, and primary adrenal glomerulosa cells from bovine and rat, applied respectively to human, bovine, and rat chips. qPCR was performed to confirm up-regulation of selected genes using mRNA. The microarray comparison revealed 18% similarity among the top 50 up-regulated genes, with human/rat, 20%; human/bovine, 36%; and rat/bovine, 26% similarity. The gene list generated by this comparison included: activating transcription factor 3, B-cell translocation gene (BTG2), Nuclear receptor subfamily 4, group A, member 1 (NR4A1), NR4A2, NR4A3, early growth response 1, v-fos FBJ murine osteosarcoma viral oncogene homolog (c-FOS), FOSB, and Jun family member B (JUNB). Pretreatment of H295R cells with cycloheximide had no effect on Ang-II induction of these genes, suggesting that they are direct targets of Ang-II signaling. The Ang-II gene targets have been defined in three different adrenocortical model systems. Several of the listed genes have previously been described as being key regulators of adrenocortical function. The presence of adrenal cell common genes in such distinct cell models strengthens the hypothesis that these genes are regulators of aldosterone production.

Direct inhibitory effects of leptin on the neonatal adrenal and potential consequences for brain glucocorticoid feedback.

Leptin is most studied for its primary role in the CNS control of energy balance and food intake in humans and rodents, yet it has functions on multiple target sites including the adrenal gland. In adult rodents, leptin has been shown to inhibit adrenal steroidogenesis and we have recently demonstrated that some of the mechanisms responsible for leptin-induced inhibition of adrenal glucocorticoid production, namely a reduction of StAR protein expression are already present in the neonatal adrenal gland. The effect of leptin on the neonatal adrenal gland integrates well with the previously demonstrated effect of this protein to inhibit stress responses, enhance glucocorticoid receptor expression in the CNS and sensitivity to glucocorticoid inhibitory feedback in neonates. The leptin receptor isoform and intracellular mechanisms involved in regulation of the adrenocortical activity at multiple levels might differ between target tissues (CNS vs periphery) and age (neonates vs adult). Neonatal leptin represents an important regulator of adrenocortical function during a critical period of brain development, which is exquisitely sensitive to circulating glucocortcoid concentrations. Since circulating leptin levels in neonates vary according to maternal diet, this protein can be viewed as a critical link between environmental and maternal factors and the developing physiology of the infant.