Potential of dietary polyphenols for protection from age-related decline and neurodegeneration: a role for gut microbiota?

Many epidemiological studies have shown the beneficial effects of a largely plant-based diet, and the strong association between the consumption of a Mediterranean-type diet with healthy aging including a lower risk of cognitive decline. The Mediterranean diet is characterized by a high intake of olive oil, fruits and vegetables and is rich in dietary fiber and polyphenols - both of which have been postulated to act as important mediators of these benefits. Polyphenols are large molecules produced by plants to protect them from environmental threats and injury. When ingested by humans, as little as 5% of these molecules are absorbed in the small intestine with the majority metabolized by the gut microbiota into absorbable simple phenolic compounds. Flavan-3-ols, a type of flavonoid, contained in grapes, berries, pome fruits, tea, and cocoa have been associated with many beneficial effects on several risk factors for cardiovascular disease, cognitive function and brain regions involved in memory formation. Both preclinical and clinical studies suggest that these brain and heart benefits can be attributed to endothelial vascular effects and anti-inflammatory properties among others. More recently the gut microbiota has emerged as a potential modulator of the aging brain and intriguingly polyphenols have been shown to alter microbiota composition and be metabolized by different microbial species. However, there is a need for well controlled studies in large populations to identify predictors of response, particularly given the vast inter-individual variation of human gut microbiota.

Neuroprotection after cardiac arrest with 2-iminobiotin: a single center phase IIa study on safety, tolerability, and pharmacokinetics.

Background: Brain injury is a serious problem in patients who survive out-of-hospital cardiac arrest (OHCA). Neuroprotective drugs could reduce hypoxic-ischemic reperfusion injury. The aim of this study was to investigate the safety, tolerability, and pharmacokinetics (PK) of 2-iminobiotin (2-IB), a selective inhibitor of neuronal nitric oxide synthase. Methods: Single-center, open-label dose-escalation study in adult OHCA patients, investigating three 2-IB dosing schedules (targeting an AUC0-24h of 600-1,200 ng*h/m in cohort A, of 2,100-3,300 ng*h/mL in cohort B, and 7,200-8,400 of ng*h/mL in cohort C). Safety was investigated by monitoring vital signs until 15 min after study drug administration and adverse events up to 30 days after admission. Blood sampling for PK analysis was performed. Brain biomarkers and patient outcomes were collected 30 days after OHCA. Results: A total of 21 patients was included, eight in cohort A and B and five in cohort C. No changes in vital signs were observed, and no adverse events related to 2-IB were reported. A two-compartment PK model described data the best. Exposure in group A (dosed on bodyweight) was three times higher than targeted (median AUC0-24h 2,398 ng*h/mL). Renal function was an important covariate; therefore, in cohort B, dosing was performed on eGFR on admission. In cohort B and C, the targeted exposure was met (median AUC0-24h 2,917 and 7,323 ng*h/mL, respectively). Conclusion: The administration of 2-IB to adults after OHCA is feasible and safe. PK can be well predicted with correction for renal function on admission. Efficacy studies with 2-IB after OHCA are needed.

Heifers don't care: no evidence of negative impact on animal welfare of growing heifers when using virtual fences compared to physical fences for grazing.

Virtual fencing (VF) represents a way to simplify traditional pasture management with its high labour and cost requirements for fencing and to make better use of the 'beneficial' agronomic and ecological effects of livestock grazing. In this study, the VF technology (® Nofence, AS, Batnfjordsøra Norway) was used with Fleckvieh heifers to investigate possible welfare impacts on the animals compared to conventionally fenced animals when they were trained to respond correctly to the system. The Nofence® collars (attached to the neck of the heifers) send acoustic signals as a warning when the animals approach the VF line, which was set up by GPS coordinates within the Nofence®-App, followed by an electric pulse when they do not stop or return. The heifers had no experience with VF prior to the study. Two treatments (VF versus physical fencing (PF)) were applied to six groups of four heifers each (three groups per treatment) over three 12-day time replicates. One VF line separated the pasture of the VF group into an accessible or non-accessible area. The control group had a PF line. Both groups were equipped with Nofence® collars (deactivated for the PF group). The trial took place on two adjacent paddocks of 1 000 m2 each following a 12-day schedule which was divided into three sections: visual support of the VF line by a physical barrier (first 2 days), only virtual border without visual support, moving the VF line (on day 8). Each time replicate followed the next successively on different paddocks with two new groups of heifers, which were grazed 5 h daily. During the whole experiment, the behaviour of each of the four animals per group was continuously observed; 2 h a.m., 2 h p.m. Exclusion by the VF line was effective in our trial. None of the heifers crossed the virtual boundary, i.e. the time spent in exclusion zone was zero. The heifers received 2.70 ± 2.63 acoustic signals and 0.30 ± 0.36 electric pulses (mean ± SD) per heifer and hour during all time replicates. Main cattle behaviour on pasture was not affected by the fencing system. Live weight gain, herbage consumption and faecal cortisol metabolites also revealed no significant differences. The duration until the heifers restarted grazing after an electric pulse from the Nofence® collar was significantly shorter than after an electric pulse from the physical fence. We can summarise that in our study, cattle well-being on pasture was not negatively affected by VF compared to PF.

[Functioning of parents and adolescents after family intervention Parenting with Love and Limits]. / Functioneren van ouders en jongeren na gezinsinterventie Ouderschap met Liefde en Grenzen.

BACKGROUND: Parenting with Love and Limits (PLL) is a family intervention aimed at reducing behavioral problems in adolescents up to the age of 18 by, among other things, improving communication between family members and the parenting skills of the parents. AIM: This prospective study examined whether parental communication and parenting skills and youth externalizing problem behavior were positively changed after PLL. METHOD: The study group consisted of 48 adolescents who had completed the PLL-intervention with their primary caregivers. Data from two measurements completed by the primary caregivers (44 mothers and 4 fathers) were used: prior to PLL (T0) and after PLL (T1). RESULTS: Results of the paired t-test indicated strong positive changes in communication, parenting and aggressive behavior after nearly eight months of PLL. A statistically significant (reliable change) decrease in aggressive behavior was found in one third of the adolescents, but not in deviant behavior. CONCLUSION: PLL improved functioning in some of the families, but further research is needed into the effective elements and for whom the intervention is effective under what circumstances.

Role of diet and its effects on the gut microbiome in the pathophysiology of mental disorders.

There is emerging evidence that diet has a major modulatory influence on brain-gut-microbiome (BGM) interactions with important implications for brain health, and for several brain disorders. The BGM system is made up of neuroendocrine, neural, and immune communication channels which establish a network of bidirectional interactions between the brain, the gut and its microbiome. Diet not only plays a crucial role in shaping the gut microbiome, but it can modulate structure and function of the brain through these communication channels. In this review, we summarize the evidence available from preclinical and clinical studies on the influence of dietary habits and interventions on a selected group of psychiatric and neurologic disorders including depression, cognitive decline, Parkinson's disease, autism spectrum disorder and epilepsy. We will particularly address the role of diet-induced microbiome changes which have been implicated in these effects, and some of which are shared between different brain disorders. While the majority of these findings have been demonstrated in preclinical and in cross-sectional, epidemiological studies, to date there is insufficient evidence from mechanistic human studies to make conclusions about causality between a specific diet and microbially mediated brain function. Many of the dietary benefits on microbiome and brain health have been attributed to anti-inflammatory effects mediated by the microbial metabolites of dietary fiber and polyphenols. The new attention given to dietary factors in brain disorders has the potential to improve treatment outcomes with currently available pharmacological and non-pharmacological therapies.

Protective factors as uni- or bipolar factors and their incremental validity and accuracy in predicting general recidivism.

This study examined whether protective factors are unique or the opposite of risk factors and whether they have incremental validity in the prediction of general recidivism. Using a sample of 3306 Dutch forensic outpatients, this study was the first large-sample study ever performed on this topic. Results from exploratory factor analyses demonstrated a relatively stable factor structure of 14 factors, consisting of 32 of the initially included 68 risk factors and 11 of the initially included 17 protective items. The protective factors were found to be either bipolar (i.e., mirror images of risk factors) or responsivity characteristics (i.e., motivation for treatment, cognitive disability). Incremental validity for the recidivism prediction was found in one factor with internal protective items (e.g., empathy, financial management, life goals). This factor decreased the recidivism risk by 6%. However, weak predictive accuracy was found for this factor. Implications for clinical forensic practice are discussed with special focus on the risk-need-responsivity model.

Practice of tracheostomy in patients with acute respiratory failure related to COVID-19 - Insights from the PRoVENT-COVID study.

OBJECTIVE: Invasively ventilated patients with acute respiratory failure related to coronavirus disease 2019 (COVID-19) potentially benefit from tracheostomy. The aim of this study was to determine the practice of tracheostomy during the first wave of the pandemic in 2020 in the Netherlands, to ascertain whether timing of tracheostomy had an association with outcome, and to identify factors that had an association with timing. METHODS: Secondary analysis of the 'PRactice of VENTilation in COVID-19' (PRoVENT-COVID) study, a multicenter observational study, conducted from March 1, 2020 through June 1, 2020 in 22 Dutch intensive care units (ICU) in the Netherlands. The primary endpoint was the proportion of patients receiving tracheostomy; secondary endpoints were timing of tracheostomy, duration of ventilation, length of stay in ICU and hospital, mortality, and factors associated with timing. RESULTS: Of 1023 patients, 189 patients (18.5%) received a tracheostomy at median 21 [17 to 28] days from start of ventilation. Timing was similar before and after online publication of an amendment to the Dutch national guidelines on tracheostomy focusing on COVID-19 patients (21 [17-28] vs. 21 [17-26] days). Tracheostomy performed ≤ 21 days was independently associated with shorter duration of ventilation (median 26 [21 to 32] vs. 40 [34 to 47] days) and higher mortality in ICU (22.1% vs. 10.2%), hospital (26.1% vs. 11.9%) and at day 90 (27.6% vs. 14.6%). There were no patient demographics or ventilation characteristics that had an association with timing of tracheostomy. CONCLUSIONS: Tracheostomy was performed late in COVID-19 patients during the first wave of the pandemic in the Netherlands and timing of tracheostomy possibly had an association with outcome. However, prospective studies are needed to further explore these associations. It remains unknown which factors influenced timing of tracheostomy in COVID-19 patients.

Quantitative analysis of EEG reactivity for neurological prognostication after cardiac arrest.

OBJECTIVE: To test whether 1) quantitative analysis of EEG reactivity (EEG-R) using machine learning (ML) is superior to visual analysis, and 2) combining quantitative analyses of EEG-R and EEG background pattern increases prognostic value for prediction of poor outcome after cardiac arrest (CA). METHODS: Several types of ML models were trained with twelve quantitative features derived from EEG-R and EEG background data of 134 adult CA patients. Poor outcome was a Cerebral Performance Category score of 3-5 within 6 months. RESULTS: The Random Forest (RF) trained on EEG-R showed the highest AUC of 83% (95-CI 80-86) of tested ML classifiers, predicting poor outcome with 46% sensitivity (95%-CI 40-51) and 89% specificity (95%-CI 86-92). Visual analysis of EEG-R had 80% sensitivity and 65% specificity. The RF was also the best classifier for EEG background (AUC 85%, 95%-CI 83-88) at 24 h after CA, with 62% sensitivity (95%-CI 57-67) and 84% specificity (95%-CI 79-88). Combining EEG-R and EEG background RF classifiers reduced the number of false positives. CONCLUSIONS: Quantitative EEG-R using ML predicts poor outcome with higher specificity, but lower sensitivity compared to visual analysis of EEG-R, and is of some additional value to ML on EEG background data. SIGNIFICANCE: Quantitative EEG-R using ML is a promising alternative to visual analysis and of some added value to ML on EEG background data.

Effectiveness, safety and efficacy of INTELLiVENT-adaptive support ventilation, a closed-loop ventilation mode for use in ICU patients - a systematic review.

Introduction: INTELLiVENT-Adaptive Support Ventilation (INTELLiVENT-ASV), an advanced closed-loop ventilation mode for use in intensive care unit (ICU) patients, is equipped with algorithms that automatically adjust settings on the basis of physiologic signals and patient's activity. Here we describe its effectiveness, safety, and efficacy in various types of ICU patients.Areas covered: A systematic search conducted in MEDLINE, EMBASE, the Cochrane Central register of Controlled Trials (CENTRAL), and in Google Scholar identified 10 randomized clinical trials.Expert opinion: Studies suggest INTELLiVENT-ASV to be an effective automated mode with regard to the titrations of tidal volume, airway pressure, and oxygen. INTELLiVENT-ASV is as safe as conventional modes. However, thus far studies have not shown INTELLiVENT-ASV to be superior to conventional modes with regard to duration of ventilation and other patient-centered outcomes. Future studies are needed to test its efficacy.

Early or late pregnancy loss and development of clinical cardiovascular disease risk factors: a prospective cohort study.

OBJECTIVE: To assess the association between the outcome of a woman's first pregnancy and risk of clinical cardiovascular disease risk factors. DESIGN: Prospective cohort study. SETTING AND POPULATION: Nurses' Health Study II. METHODS: Multivariable-adjusted Cox proportional hazards models were used to compute hazard ratios (HRs) and 95% confidence intervals (CIs) for the associations between first pregnancy outcome and hypertension, type 2 diabetes, and hypercholesterolemia. MAIN OUTCOME MEASURES: Hypertension, type 2 diabetes, and hypercholesterolemia. RESULTS: Compared to women who reported a singleton live first birth, women with early spontaneous abortion (<12 weeks) had a greater rate of type 2 diabetes (HR: 1.20; 95% CI: 1.07-1.34) and hypercholesterolemia (HR: 1.06; 95% CI: 1.02-1.10), and a marginally increased rate of hypertension (HR: 1.05, 95% CI: 1.00-1.11). Late spontaneous abortion (12-19 weeks) was associated with an increased rate of type 2 diabetes (HR: 1.38; 95% CI: 1.14-1.65), hypercholesterolemia (HR: 1.11; 95% CI: 1.03-1.19), and hypertension (HR: 1.15; 95% CI: 1.05-1.25). The rates of type 2 diabetes (HR: 1.45; 95% CI: 1.13-1.87) and hypertension (HR: 1.15; 95% CI: 1.01-1.30) were higher in women who delivered stillbirth. In contrast, women whose first pregnancy ended in an induced abortion had lower rates of hypertension (HR: 0.87; 95% CI: 0.84-0.91) and type 2 diabetes (HR: 0.89; 95% CI: 0.79-0.99) than women with a singleton live birth. CONCLUSIONS: Several types of pregnancy loss were associated with an increased rate of hypertension, type 2 diabetes, and hypercholesterolemia, which may provide novel insight into the pathways through which pregnancy outcomes and CVD are linked. TWEETABLE ABSTRACT: Pregnancy loss is associated with later maternal risk of hypertension, type 2 diabetes, and hypercholesterolemia.

Co-assemblies of polyoxometalate {Mo72Fe30}/double-tailed magnetic-surfactant for magnetic-driven anchorage and enrichment of protein.

Covalent grafting and electrostatic-driven assembly have been two strategies in constructing well-defined polyoxometalate (POM) assemblies to produce specific morphologies and desirable properties. The modification of anionic counter-ions of amphiphilic compounds in POM-surfactant hybrid systems is still unexploited. Herein, we report the co-assembly of a synthetic double-tailed magnetic surfactant (MagSurf), (C18)2C2N+[FeCl4]-, and POM, {Mo72Fe30}. The magnetic aggregate (POM/MagSurf) results from the building up hierarchical structures at a time-dependent interface. In this construct, both the MagSurfs and {Mo72Fe30} POMs contribute to and mutually strengthen the magnetization of the designed magnetic assembles. Interestingly, the POM/MagSurf aggregates are compatible with aqueous mixtures and successfully employed to serve as magnetic transporting vehicles to anchor and deliver a protein molecule, myoglobin (Mb). Upon applying a magnetic field (0.3 T), the magnetic aggregates induced a directional migration and enrichment of the Mb protein (71-90%). During this process, the protein/POM/MagSurf complexes exhibited strong interactions facilitating stable anchoring and efficient enrichment of the Mb.

[Forensic (F)ACT: a study exploring indication, risk assessment and recidivism]. / Forensische (F)ACT: een onderzoek naar indicatiestelling, risicotaxatie en recidive.

BACKGROUND: Cutbacks in clinical beds in regular and forensic psychiatry increase the burden on outpatient forensic care in The Netherlands.
AIM: Since 2007, Dutch forensic (flexible) assertive community treatment (For(F)ACT) teams offer outpatient, intensive treatment to forensic clients with complex mental health issues. As the need for this form of intensive treatment increases, so does the need for unambiguous indication criteria to facilitate adequate care and accompanied reduction in criminal behavior.
METHOD: The present study investigated the correlation between the clinical indication criteria and risk factors for criminal behavior in 76 For(F)ACT-clients, reviewing which criteria best predicted recidivism.
RESULTS: A weak correlation was found between the indication criteria and risk factors. Further receiver operating characteristic (ROC) analysis showed that a combination of clinical indication criteria best predicted recidivism.
CONCLUSION: The influential risk factors for For(F)ACT-clients are different compared to those for other groups of delinquents. Important treatment factors are breaking patterns, increasing safety and offering social and financial support. The clinical indication criteria should not be replaced by the START risk factors.

International consensus on EEG reactivity testing after cardiac arrest: Towards standardization.

BACKGROUND: In patients after cardiac arrest (CA), EEG reactivity (EEG-R) is proposed as a prognostic marker. However, no clear guidelines exist on how to test EEG-R and definitions are unspecific. Therefore, we aimed at forming international consensus regarding a stimulus protocol for EEG-R testing and the interpretation of EEG-R in daily clinical care. METHODS: We invited 30 international experts on EEG in patients after CA for participation in a two round Delphi study. Consensus was defined as ≥75% agreement, 66-75% agreement was included as recommendation. RESULTS: In the first round 24 experts participated (80% response rate) of whom 22 finished the second round (8% drop-out). Consensus was reached on several parts of the stimulus protocol: Clapping, calling out the patient's name and nail bed pressure should be executed and each stimulus at least three times with recommended duration of at least 5 s. The patient should not be stimulated before EEG-R testing and information on sedation/analgesics should be provided. The consensus definition of EEG-R is "A reproducible change in the EEG in response to stimulation" and appearance of muscle-, movement- and eye blink artefacts, spinal movements and electrographic seizure induction do not qualify as reactive. Almost all respondents agreed that this consensus protocol should also be used in comatose patients with other etiologies. CONCLUSION: This international consensus statement on EEG-R in patients after CA can be regarded as starting point. At the moment evidence is limited and our study can provide best-practice guidance in patients after CA as well as other comatose patients.

Highly malignant routine EEG predicts poor prognosis after cardiac arrest in the Target Temperature Management trial.

INTRODUCTION: Routine EEG is widely used and accessible for post arrest neuroprognostication. Recent studies, using standardised EEG terminology, have proposed highly malignant EEG patterns with promising predictive ability. OBJECTIVES: To validate the performance of standardised routine EEG patterns to predict neurological outcome after cardiac arrest. METHODS: In the prospective multicenter Target Temperature Management trial, comatose cardiac arrest patients were randomised to different temperature levels (950 patients, 36 sites). According to the prospective protocol a routine EEG was performed in patients who remained comatose after the 36 h temperature control intervention. EEGs were retrospectively reviewed blinded to outcome using the standardised American Clinical Neurophysiology Society terminology. Highly malignant, malignant and benign EEG patterns were correlated to poor and good outcome, defined by best achieved Cerebral Performance Category up to 180 days. RESULTS: At 20 sites 207 patients had a routine EEG performed at median 76 h after cardiac arrest. Highly malignant patterns (suppression or burst-suppression with or without discharges) had a high specificity for poor outcome (98%, CI 92-100), but with limited sensitivity (31%, CI 24-39). Our false positive patient had a burst-suppression pattern during ongoing sedation. A benign EEG, i.e. continuous normal-voltage background without malignant features, identified patients with good outcome with 77% (CI 66-86) sensitivity and 80% (CI 73-86) specificity. CONCLUSION: Highly malignant routine EEG after targeted temperature management is a strong predictor of poor outcome. A benign EEG is an important indicator of a good outcome for patients remaining in coma.

Time to epileptiform activity and EEG background recovery are independent predictors after cardiac arrest.

OBJECTIVE: Investigate the temporal development of EEG and prognosis. METHODS: Prospective observational substudy of the Target Temperature Management trial. Six sites performed simplified continuous EEG-monitoring (cEEG) on comatose patients after cardiac arrest, blinded to treating physicians. We determined time-points of recovery of a normal-voltage continuous background activity and the appearance of an epileptiform EEG, defined as abundant epileptiform discharges, periodic/rhythmic discharges or electrographic seizure activity. RESULTS: 134 patients were included, 65 had a good outcome. Early recovery of continuous background activity (within 24 h) occurred in 72 patients and predicted good outcome since 55 (76%) had good outcome, increasing the odds for a good outcome seven times compared to a late background recovery. Early appearance of an epileptiform EEG occurred in 38 patients and 34 (89%) had a poor outcome, increasing the odds for a poor outcome six times compared to a late debut. The time to background recovery and the time to epileptiform activity were highly associated with outcome and levels of neuron-specific enolase. Multiple regression analysis showed that both variables were independent predictors. CONCLUSIONS: Time to epileptiform activity and background recovery are independent prognostic indicators. SIGNIFICANCE: Patients with early background recovery combined with late appearance of epileptiform activity may have a good outcome.

Punch biopsies shorten time to clearance of high-risk human papillomavirus infections of the uterine cervix.

BACKGROUND: The primary objective was to determine human papilloma virus (HPV) clearance rate after cervical biopsy among women with persistent high-risk HPV infection compared with spontaneous HPV clearance rate in the absence of biopsy. METHODS: We collected data from a dedicated screening program of women aged 30-70 years old. Inclusion criteria for the baseline non-interventional cohort were a positive HPV test (hybrid capture 2, HC2) and normal cytology. In the baseline cohort women were followed with approximately yearly HPV-tests and cytology until HPV regressed (one negative HPV test) or interventions in the form of diagnostic biopsies or therapy. Women who had a diagnostic biopsy were included in the biopsy cohort and followed until HPV regression or therapy. Observed HPV regression rates and time to HPV regression were compared between baseline and biopsy cohorts. For the comparison, we used Fisher's exact test for the HPV regression rates and interval-censored, accelerated failure time model for time to HPV regression. RESULTS: Among the 1079 women included in the baseline cohort, 499 (46.3%) had HPV clearance and 475 were referred for colposcopy with biopsy. The biopsy cohort comprised all women who were not treated and had at least one HC2 test after biopsy (201/475; 42.3%). Of those, 138 (68.7%) experienced HPV regression. In the biopsy cohort, time to clearance of HPV infection was approximately halved (0.46, 95% CI 0.38-0.56) compared with the baseline cohort. This result was robust in a wide range of sensitivity analyses. CONCLUSIONS: A higher proportion of women cleared their HPV infection, and time to HPV clearance was shorter in the biopsy cohort than in the baseline cohort. It is reassuring for clinicians to know that conservative management of patients with HPV persistency is successful when colposcopy with biopsies excludes high-grade disease.

Full and Short Versions of the Coping Inventory for Stressful Situations and Social Problem-Solving Inventory-Revised: Examining Their Factor Structure in a Dutch Outpatient Sex Offender Sample.

Inadequate problem solving is associated with recidivism in sex offenders. However, the psychometric properties of two of the most widely used coping self-report questionnaires, the Social Problem-Solving Inventory-Revised and Coping Inventory for Stressful Situations, have not been established for outpatient sex offenders. Using structural equation modeling, the present study aims to contribute to an empirically based decision as to which version of the Coping Inventory for Stressful Situations or Social Problem-Solving Inventory-Revised is best suited to use with forensic outpatient sex offenders. The sample consisted of 530 Dutch sex offenders treated in an outpatient forensic facility. Results support the use of the Coping Inventory for Stressful Situations-Short Form 20 for identifying problem-solving deficits among outpatient sex offenders, consequently facilitating the assessment of treatment needs.

Shrinking Daughters: Rlm1-Dependent G1/S Checkpoint Maintains Saccharomyces cerevisiae Daughter Cell Size and Viability.

The Rlm1 transcription factor is a target of the cell wall integrity pathway. We report that an rlm1Δ mutant grown on a nonfermentable carbon source at low osmolarity forms cell groups in which a mother cell is surrounded by smaller "satellite-daughter" cells. Mother cells in these groups progressed through repeated rounds of cell division with normal rates of bud growth and genetic stability; however, these cells underwent precocious START relative to wild-type mothers. Thus, once activated, Rlm1 delays the transition from G1 to S, a mechanism we term the cell wall/START (CW/START) checkpoint. The rlm1Δ satellite-cell phenotype is suppressed by deletion of either SLT2, which encodes the kinase that activates Rlm1, or SWI4, which is also activated by Slt2; suggesting that Slt2 can have opposing roles in regulating the START transition. Consistent with an Rlm1-dependent CW/START checkpoint, rlm1Δ satellite daughters were unable to grow or divide further even after transfer to rich medium, but UV irradiation in G1 could partially rescue rlm1Δ satellite daughters in the next division. Indeed, after cytokinesis, these satellite daughters shrank rapidly, displayed amorphous actin staining, and became more permeable. As a working hypothesis, we propose that duplication of an "actin-organizing center" in late G1 may be required both to progress through START and to reestablish the actin cytoskeleton in daughter cells.