[Proper use of fluoroquinolones in hospitalized elderly patients: Realisation of a clinical audit in geriatric hospitals of Lyon University Hospital]. / Bon usage des fluoroquinolones chez le sujet âgé hospitalisé : réalisation d'un audit clinique dans les hôpitaux gériatriques des hospices civils de Lyon.

INTRODUCTION: Fluoroquinolones (FQ) are widely used because of their broad spectrum and their ease of use, especially in the elderly. Nevertheless, their misuse is behind the development and the emergence of bacterial resistances. The objective of this study was to evaluate the compliance of FQ prescriptions in the elderly hospitalized, before and after pharmaceutical interventions (PI). METHODS: A prospective clinical audit was conducted for three months in three geriatric hospitals. A pharmacist carried out the evaluation of the prescription's conformity with the help of referent geriatric doctor in infectiology according to the criteria defined by SPILF recommendations (2015). The PIs and their future have been collected and codified. RESULTS: A total of 100 patients were included (mean age: 85.3 years; male female ratio: 1,17). The medical-pharmaceutical collaboration helped to increase the overall compliance rate from 56 to 80%. FQ were used for urinary (72%) or respiratory (20%) infections, first-line (57%), documented (60%) and monotherapy (63%). Our results show that FQ misuse is a not inconsiderable case since in 28% they should not have been prescribed. Thirty-three PI were performed and accepted in 72% cases. CONCLUSION: Actions to promote the FQ proper use have therefore been put in place: communication of the audit results, dissemination of a summary of SPILF recommendations and creation of a tool to assist in the analysis of prescriptions FQ for pharmacists.

[Integrate the surgical hand disinfection as a quality indicator in an operating room of urology]. / Intégrer la désinfection chirurgicale des mains comme indicateur de qualité dans un bloc opératoire d'urologie.

The surgical hand disinfection by friction (SDF) helps to reduce the risk of surgical site infections. For this purpose and in order to promote good compliance to quality care, the urology service of Centre Hospitalier Lyon Sud achieved a continuous internal audit to improve the quality of the SDF. METHODS: An internal audit executed by the medical students of urology was established in 2013. The study population was all operators, instrumentalists and operating aids of urology operating room (OR). Each student realized 5-10 random observations, of all types of professionals. The criteria measured by the audit were criteria for friction. RESULTS: The evolution of indicators was positive. Particularly, the increasing duration of the first and second friction was statistically significant during follow-up (P=0.001). The total duration of friction shows a similar trend for all professionals. CONCLUSION: The surgical hand disinfection by friction in the urology OR of the Centre Hospitalier Lyon Sud has gradually improved over the iterative audits.

Risk factors for urinary tract infections in geriatric hospitals.

BACKGROUND: Urinary tract infection (UTI) is the most frequent nosocomial infection in geriatric units. An understanding of risk factors for infection may help to identify prevention strategies. AIM: Identification of the risk factors for UTI in elderly patients. METHODS: Retrospective analysis of three prospective cohorts. All hospitalized patients present in, or admitted to, a geriatric unit from June 1st to June 28th, for the years 2009, 2012, and 2015 were included and followed until discharge or until June 30th of the year concerned. For each patient, type and dates of stay, type and dates of catheter, risk factors, and nosocomial UTI (NUTI) data were collected. Univariate and multivariate (Cox model) analyses were made using SPSS software. FINDINGS: A total of 4669 patients were included and were followed for a total of 83,068 days. There were 189 NUTIs (4.0% patients). NUTIs were significantly more frequent among female patients, in rehabilitation units, in immunosuppressed patients, among those with acute retention, post-void residual, history of urinary tract infection in the previous six months, and in case of dependency. NUTIs were significantly more frequent among those who had a catheter (Z-test, P < 0.001). NUTIs were more frequent among patients with intermittent, indwelling, or suprapubic catheters. They were also more frequent in acute/subacute care or rehabilitation units, in women, in immunosuppressed patients, and in those with a history of previous UTI; they were less frequent in dementia patients. CONCLUSION: The occurrence of NUTI is an important issue in both catheterized and non-catheterized patients; prevention programmes should be widened to include non-catheterized patients.

The fibrinolytic system: A new target for treatment of depression with psychedelics.

Current understanding of the neurobiology of depression has grown over the past few years beyond the traditional monoamine theory of depression to include chronic stress, inflammation and disrupted synaptic plasticity. Tissue plasminogen activator (tPA) is a key factor that not only promotes fibrinolysis via the activation of plasminogen, but also contributes to regulation of synaptic plasticity and neurogenesis through plasmin-mediated activation of a probrain derived neurotrophic factor (BDNF) to mature BDNF. ProBDNF activation could potentially be supressed by competition with fibrin for plasmin and tPA. High affinity binding of plasmin and tPA to fibrin could result in a decrease of proBDNF activation during brain inflammation leading to fibrosis further perpetuating depressed mood. There is a paucity of data explaining the possible role of the fibrinolytic system or aberrant extravascular fibrin deposition in depression. We propose that within the brain, an imbalance between tPA and urokinase plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) and neuroserpin favors the inhibitors, resulting in changes in neurogenesis, synaptic plasticity, and neuroinflammation that result in depressive behavior. Our hypothesis is that peripheral inflammation mediates neuroinflammation, and that cytokines such as tumor necrosis factor alpha (TNF-α) can inhibit the fibrinolytic system by up- regulating PAI-1 and potentially neuroserpin. We propose that the decrement of the activity of tPA and uPA occurs with downregulation of uPA in part involving the binding and clearance from the surface of neural cells of uPA/PAI-1 complexes by the urokinase receptor uPAR. We infer that current antidepressants and ketamine mitigate depressive symptoms by restoring the balance of the fibrinolytic system with increased activity of tPA and uPA with down-regulated intracerebral expression of their inhibitors. We lastly hypothesize that psychedelic 5-ht2a receptor agonists, such as psilocybin, can improve mood through anti- inflammatory and pro-fibrinolytic effects that include blockade of TNF-α activity leading to decreased PAI-1 activity and increased clearance. The process involves disinhibition of tPA and uPA with subsequent increased cleavage of proBDNF which promotes neurogenesis, decreased neuroinflammation, decreased fibrin deposition, normalized glial-neuronal cross-talk, and optimally functioning neuro-circuits involved in mood. We propose that psilocybin can alleviate deleterious changes in the brain caused by chronic stress leading to restoration of homeostatic brain fibrinolytic capacity leading to euthymia.

Quantitative Susceptibility Mapping in Cerebral Cavernous Malformations: Clinical Correlations.

BACKGROUND AND PURPOSE: Quantitative susceptibility mapping has been shown to assess iron content in cerebral cavernous malformations. In this study, our aim was to correlate lesional iron deposition assessed by quantitative susceptibility mapping with clinical and disease features in patients with cerebral cavernous malformations. MATERIALS AND METHODS: Patients underwent routine clinical scans in addition to quantitative susceptibility mapping on 3T systems. Data from 105 patients met the inclusion criteria. Cerebral cavernous malformation lesions identified on susceptibility maps were cross-verified by T2-weighted images and differentiated on the basis of prior overt hemorrhage. Mean susceptibility per cerebral cavernous malformation lesion (χ̄lesion) was measured to correlate with lesion volume, age at scanning, and hemorrhagic history. Temporal rates of change in χ̄lesion were evaluated in 33 patients. RESULTS: Average χ̄lesion per patient was positively correlated with patient age at scanning (P < .05, 4.1% change with each decade of life). Cerebral cavernous malformation lesions with prior overt hemorrhages exhibited higher χ̄lesion than those without (P < .05). Changes in χ̄lesion during 3- to 15-month follow-up were small in patients without new hemorrhage between the 2 scans (bias = -0.0003; 95% CI, -0.06-0.06). CONCLUSIONS: The study revealed a positive correlation between mean quantitative susceptibility mapping signal and patient age in cerebral cavernous malformation lesions, higher mean quantitative susceptibility mapping signal in hemorrhagic lesions, and minimum longitudinal quantitative susceptibility mapping signal change in clinically stable lesions. Quantitative susceptibility mapping has the potential to be a novel imaging biomarker supplementing conventional imaging in cerebral cavernous malformations. The clinical significance of such measures merits further study.

Ventilator-associated pneumonia in ARDS patients: the impact of prone positioning. A secondary analysis of the PROSEVA trial.

BACKGROUND: The goal of this study was to assess the impact of prone positioning on the incidence of ventilator-associated pneumonia (VAP) and the role of VAP in mortality in a recent multicenter trial performed on patients with severe ARDS. METHODS: An ancillary study of a prospective multicenter randomized controlled trial on early prone positioning in patients with severe ARDS. In suspected cases of VAP the diagnosis was based on positive quantitative cultures of bronchoalveolar lavage fluid or tracheal aspirate at the 10(4) and 10(7) CFU/ml thresholds, respectively. The VAP cases were then subject to central, independent adjudication. The cumulative probabilities of VAP were estimated in each position group using the Aalen-Johansen estimator and compared using Gray's test. A univariate and a multivariate Cox model was performed to assess the impact of VAP, used as a time-dependent covariate for mortality hazard during the ICU stay. RESULTS: In the supine and prone position groups, the incidence rate for VAP was 1.18 (0.86-1.60) and 1.54 (1.15-2.02) per 100 days of invasive mechanical ventilation (p = 0.10), respectively. The cumulative probability of VAP at 90 days was estimated at 46.5 % (27-66) in the prone group and at 33.5 % (23-44) in the supine group. The difference between the two cumulative probability curves was not statistically significant (p = 0.11). In the univariate Cox model, VAP was associated with an increase in the mortality rate during the ICU stay [HR 1.65 (1.05-2.61), p = 0.03]. HR increased to 2.2 (1.39-3.52) (p < 0.001) after adjustment for position group, age, SOFA score, McCabe score, and immunodeficiency. CONCLUSIONS: In severe ARDS patients prone positioning did not reduce the incidence of VAP and VAP was associated with higher mortality.

Controlling urinary tract infections associated with intermittent bladder catheterization in geriatric hospitals.

BACKGROUND: Controlling urinary tract infections (UTIs) associated with intermittent catheterization in geriatric patients. AIM: After a local epidemiological study identified high rates of UTI, a multi-disciplinary working group implemented and evaluated corrective measures. METHODS: In 2009, a one-month prospective study measured the incidence of UTI, controlled for risk factors and exposure, in six geriatric hospitals. In 2010, a self-administered questionnaire on practices was administered to physicians and nurses working in these geriatric units. In 2011, the working group developed a multi-modal programme to: improve understanding of micturition, measurement of bladder volume and indications for catheter drainage; limit available medical devices; and improve prescription and traceability procedures. Detailed training was provided to all personnel on all sites. The epidemiological study was repeated in 2012 to assess the impact of the programme. FINDINGS: Over 1500 patients were included in the 2009 study. The incidence of acquired infection was 4.8%. The infection rate was higher in patients with intermittent catheters than in patients with indwelling catheters (29.7 vs 9.9 UTI per 100 patients, P = 0.1013) which contradicts the literature. In 2010, the 269 responses to the questionnaire showed that staff did not consider catheterization to place patients at risk of infection, staff had poor knowledge of the recommended indications and techniques, and the equipment varied widely between units. Following implementation of the programme, the study was repeated in 2012 with over 1500 patients. The frequency of UTI in patients with intermittent catheters fell to rates in the published literature. CONCLUSION: Multi-modal programmes are an effective means to control UTI.

Lower respiratory events in seated tracheotomized tetraplegic patients.

AIM: Retrospective study on the changing position impact on respiratory events in 14 rehabilitation tracheotomized tetraplegic patients, during 25 months. METHODS: Three positions were compared: permanently supine (16 periods), seated on or=6 days/week (10 periods). The end-point was the incidence of the following respiratory events: pneumonia, atelectasis and plugging of tracheal/bronchial secretions. Patients were considered as their own control but data were pooled for analysis. RESULTS: Pneumonia and plugging incidences were significantly higher in the permanently supine position than in the seated or=6 days position. Atelectasis occurred only in the supine position. CONCLUSION: Plugging prevalence was significantly higher in the permanently supine position (53.3%) than in the seated or=6 days position (14.6%, P=0.001).

Detecting related cases of bloodstream infections using time-interval distribution modelling.

An algorithm was designed to highlight related bloodstream infections using data from a nosocomial infection surveillance system to help local public health authorities direct specific measures towards clusters of cases. The approach was based on a two-step procedure. The first was a test to identify pathogens with an abnormal number of close cases. The second modelled, for the identified pathogens, the distribution of time intervals between successive cases as a mixture of two theoretical distributions in order to determine a threshold below which a specific investigation is required. The algorithm was applied to bloodstream infection surveillance data collected during a 10-year period (1996-2005) in an 878-bed teaching hospital (24 wards) in Lyon, France. The first step identified seven pathogens among the 18 being studied. The modelling succeeded in setting time thresholds to spot clusters of cases requiring further investigation with defined sensitivity and specificity. Setting the sensitivity level at 95%, the threshold values ranged from 24 days (Acinetobacter baumannii) to 294 days (Enterobacter cloacae); the specificity was higher than 70% (up to 97.5% for A. baumannii) except for E. cloacae (52.1%). Setting the specificity level at 95% resulted in a decrease in sensitivity except for A. baumannii (it reached nearly 100%); it fell below 50% for three pathogens: around 40% for Streptococcus pneumoniae and Enterococcus faecalis and 25% for Enterobacter cloacae. The threshold values then ranged from 8 days (S. pneumoniae) to 67 days (Streptococcus pyogenes). The approach proved promising though further refinements are needed before routine use.

[Monoarthritis caused by Streptococcus pyogenes after nasal septoplasty: interest of the nasal decontamination in preoperative care]. / Monoarthrite àStreptococcus pyogenes dans les suites d'une septoplastie: intérêt de la décontamination nasale en préopératoire.

The nasal septoplasty is a very current intervention in otorhinolaryngology surgery. The infectious complications of this intervention are rare and mostly mild. We report here the case of a patient hospitalized in ambulatory surgery within a fracture of the nose with luxation of the septum in the nasal fossa. This patient was operated for a reduction of this fracture with septoplasty. Twelve hours after the intervention the patient presented septic arthritis due to Streptococcus pyogenes. The tracks of prevention are presented.

[Surgical site infections in orthognathic surgery and risk factors associated]. / Les infections du site opératoire en chirurgie orthognathique et leurs facteurs de risque.

INTRODUCTION: Surgical site infections (SSI) in orthognathic surgery are considered infrequent and without any important consequence for the final operative result. A procedure of epidemiological surveillance was implemented to determine the frequency of SSI in orthognathic surgery and to better document their risk factors. MATERIAL AND METHOD: This prospective study included all interventions in our orthognathic surgery department between September 1(st) 2006 and August 31(st) 2007. SSI and their risk factors were documented for up to one year of follow-up. Risk factors were correlated to SSI using monovariate and multivariate analyses. RESULTS: Ten (7%) out of 143 consecutive interventions in orthognathic surgery were complicated by a SSI. All the SSI were secondary to a mandibular ramus sagittal split osteotomy. The two significantly correlated risk factors with the SSI in multivariate analysis were the length of surgery and the type of antibiotic prophylaxis. DISCUSSION: This rate of SSI correlates to published data. To decrease this rate, it would be necessary, in association with the usual precautionary measures, to limit the operating time and to recommend an antibioprophylaxis combining amoxicillin plus clavulanic acid (Augmentin).

[Prediction of successful induction of labor: a comparison between fetal fibronectin assay and the Bishop score]. / Prédiction de la réussite du déclenchement du travail. Comparaison entre le score de Bishop et le dosage de la fibronectine foetale.

OBJECTIVE: The aim of our study is to determine whether the fetal fibronectin is a better predictor of successful induction of labor than the Bishop score. MATERIAL AND METHODS: A prospective observational non-randomized study was conducted in our unit including 234 patients scheduled for induction of labor from October 2000 to June 2004. Fetal fibronectin was assayed by taking sample from the endocervix and the cervical status was evaluated using the Bishop score. Data were analysed by Chi-square test of Mantel-Haenzel and Cox stepwise multiple regression using SPSS version 12 software. RESULTS: The likelihood ratios for predicting that vaginal delivery would occur within 24h of induction for positive fetal fibronectine were 1.34 (95% CI 1.04-1.73, p=0.027) all patients included and 1.51(95% CI 1.00-2.33, p=0.048) for the nulliparas and 1.92 (95% CI 1.51-2.42, p=0.0001) for the Bishop score. On multiple regressions, the only variables independently associated with a successful induction were the Bishop score, the parity and the age of the patient. No significant association was found between the presence of cervical fibronectin and the caesarean section rate: 21.84% for positive fibronectin versus 21.78% for negative fibronectin. CONCLUSION: The fetal fibronectine is probably useless in this context, given the additional cost and no improvement compared with the simple Bishop score.

Field test comparison of two dermal tolerance assessment methods of hand hygiene products.

This study aimed to compare the sensitivity and workload requirement of two dermal tolerance assessment methods of hand hygiene products, in order to select a suitable pilot testing method for field tests. An observer-rating method and a self-assessment method were compared in 12 voluntary hospital departments (autumn/winter of 2005-2006). Three test-periods of three weeks were separated by two-week intervals during which the routine products were reintroduced. The observer rating method scored dryness and irritation on four-point scales. In the self-assessment method, the user rated appearance, intactness, moisture content, and sensation on a visual analogue scale which was converted into a 10-point numerical scale. Eleven products (soaps) were tested (223/250 complete reports for observer rating, 131/251 for self-assessment). Two products were significantly less well tolerated than the routine product according to the observers, four products according to the self-assessments. There was no significant difference between the two methods when products were classified according to tolerance (Fisher's test: P=0.491). For the symptom common to both assessment methods (dryness), there is a good correlation between the two methods (Spearman's Rho: P=0.032). The workload was higher for observer rating method (288 h of observer time plus 122 h of prevention team and pharmacist time compared with 15 h of prevention team and pharmacist time for self-assessment). In conclusion, the self-assessment method was considered more suitable for pilot testing, although further time should be allocated for educational measures as the return rate of complete self-assessment forms was poor.

Tolerance and acceptability of 14 surgical and hygienic alcohol-based hand rubs.

Tests were performed under working practice conditions to measure the tolerance and acceptability of commercially available hand rubs with proven efficacy. The products were compared with those in current use at the Hospices Civils de Lyon for surgical hand disinfection (Sterillium) and hygienic hand disinfection (Purell) to obtain information for public sector purchases. The 12 test products were Alcogel H, Assanis Pro, Clinogel, Dermalcool, Manugel Plus, Manugel Plus NPC, Manurub Liquid, Manurub Gel, Purell 85, Spitacid, Spitagel and Sterillium Gel. They were tested from mid-November to mid-April over four periods of three weeks, separated by two-week intervals during which the customary product was re-introduced. Participation of hospital wards and theatres was voluntary. Skin dryness and irritation were scored before and after each test period. Acceptability and ease of use were assessed by means of a questionnaire. Among the eight surgical hand rubs, only Manurub Liquid, Manurub Gel and Manugel Plus NPC did not cause significantly more dryness and irritation than Sterillium. For the 10 hygienic hand rubs, differences were noted depending upon the test period. Overall, Assanis Pro, Clinogel, Purell 85 and Sterillium Gel did not cause significantly more dryness and irritation than Purell. However, over the (colder) first three test periods, Assanis Pro and Sterillium Gel caused more irritation and Purell 85 caused more dryness than Purell. Responses to the questionnaires on acceptability indicated that users preferred their customary hand rubs (Sterillium and Purell). As these field tests involving many participants did not identify any superior products, previous purchase orders were renewed.

[Screening for bacteriuria in diabetic patients. Is it possible to stop systematic urine cytobacteriological testing?]. / Dépistage des bactériuries à l'admission chez les patients diabétiques: peut-on abandonner les examens cytobactériologiques urinaires systématiques?

OBJECTIVES AND METHOD: The discordance between test by urine dipstick (nitrites and leucocyte-esterase) and analysis in laboratory, with urinary culture on the same sample was studied in diabetic patients, from October 2000 to May 2002, to eventually stop systematic laboratory test. The dipstick result (Clinitek 20 Bayer) was classified as "possibility of bacteriuria" if one of the two tests was positive. Bacteriuria was considered significant if the laboratory test result gave, at least 10(5) bacteria per mL, (one strain), and at least 10(4) leucocytes. The out point was the dipstick negative predictive value (NPV). RESULTS: The study included 683 patients. The dipstick result was "possibility of bacteriuria" in 153 cases (22.4%). Thirty-nine bacteriuria (5.7%) were reported, including 2 dipstick false negatives. The NPV was 99.6% [IC 95% : 99.1-100]. CONCLUSION: The systematic laboratory tests were stopped.

High prevalence of nosocomial infections in rehabilitation units accounted for by urinary tract infections in patients with spinal cord injury.

This study (part of the nationwide French prevalence survey of 2001) was organized to investigate the prevalence and risk factors of nosocomial infections (NIs) and the resistant flora in patients hospitalized in rehabilitation units. Two hundred and eighty-six patients were included from two hospitals in the 'Hospices Civils de Lyon' group. Patients were classified into those with and without a spinal cord injury (SCI). Seventy-eight (27.3%) patients had an SCI. They were younger and more often characterized by a low Activity of Daily Life score, bladder incontinence and chronic respiratory disease. Urinary catheterization and mechanical ventilation were more common in these patients. The NI prevalence rate was higher in the SCI group (21.8% vs 4.3%, P<0.00001), particularly for urinary tract infections (UTIs, 19.2% vs 3.4%, P<0.00001). There was a positive relationship between the number of risk factors and NI acquisition. Multi-variate analysis showed that the only independent risk factor for NI acquisition was indwelling urinary catheterization [odds ratio (OR): 11.64, 95% confidence intervals (CI): 2.53-53.65, P=0.002]. Marginally significant factors were chronic kidney or liver disease (OR: 5.84, 95%CI: 0.80-42.68, P=0.082) and SCI (OR: 2.97, 95%CI: 0.61-14.60, P=0.179). The prevalence of antibiotic-resistant micro-organisms was high (nine cases of resistant organisms for 31 infection sites), but there were no differences between the groups. The high rate of NIs, especially UTIs, in SCI patients was not due to an independent effect of SCI but was probably due to the high number of risk factors. These high-risk patients need targeted NI surveillance.

[Changing medical practices and nosocomial infection rates in French maternity units from 1997 to 2000]. / Surveillance des infections nosocomiales dans les maternités du sud-est de la France entre 1997 et 2000.

OBJECTIVES: In this study we describe the changes in medical practices and nosocomial infection rates in obstetrics observed through a surveillance network in the South East of France. MATERIAL AND METHOD: The maternity units which belong to this network participated in voluntary surveillance using the network's methodology. The criteria for the diagnosis of nosocomial infections were in accordance with the methods described by the Centers for Disease Control and Prevention. RESULTS: 101240 pregnancies including 18503 caesareans (18.3%) were included in the network from 1997 to 2000. During the study period, nosocomial infection rates following caesarean section and vaginal delivery decreased respectively from 7.8% to 4.3% (p < 0.001) and from 2.2% to 0.9% (p < 0.001). This evolution was compared with the evolution of collected risk factors. CONCLUSION: In spite of the methodology limits, general value of surveillance and infection control programs in maternity units has been confirmed by the results of this surveillance network. During the study period, both obstetrics-related risk factors for nosocomial infection and observed hospital-acquired infection rates were dramatically reduced, what prove an improvement of quality of care in maternity units.

Nodal/activin signaling establishes oral-aboral polarity in the early sea urchin embryo.

Components of the Wnt signaling pathway are involved in patterning the sea urchin primary or animal-vegetal (AV) axis, but the molecular cues that pattern the secondary embryonic axis, the aboral/oral (AO) axis, are not known. In an analysis of signaling molecules that influence patterning along the sea urchin embryonic axes, we found that members of the activin subfamily of transforming growth factor-beta (TGF-beta) signaling molecules influence the establishment of AO polarities in the early embryo. Injection of activin mRNAs into fertilized eggs or treatment with exogenously applied recombinant activin altered the allocation of ectodermal fates and ventralized the embryo. The phenotypes observed resemble the ventralized phenotype previously reported for NiCl2, a known disrupter of AO patterning. Sensitivity to exogenous activin occurs between fertilization and the late blastula stage, which is also the time of highest NiCl2 sensitivity. These results argue that specification of fates along the embryonic AO axis involves TGF-beta signaling. To further examine TGF-beta signaling in these embryos, we cloned an endogenous TGF-beta from sea urchin embryos that is a member of the activin subfamily, SpNodal, and show through gain of function analysis that it recapitulates results obtained with exogenous activins and NiCl2. The expression pattern of SpNodal is consistent with a role for nodal signaling in the establishment of fates along the AO axis. Loss of function experiments using SpNodal antisense morpholinos also support a role for SpNodal in the establishment of the AO axis.