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PubMed2005

Phylogeography of Mabuya maculilabris (Reptilia) from São Tomé Island (Gulf of Guinea) inferred from mtDNA sequences.

The pattern of genetic variation of the lizard Mabuya maculilabris from São Tomé Island (Gulf of Guinea) was investigated using a combination of three mitochondrial DNA gene fragments. Forty-eight haplotypes were recovered among 66 individuals covering the whole island. The genealogy inferred from the most parsimonious network of haplotypes allows us to detect two main and long branches departing from the putative group of oldest haplotypes. The tips of these branches exhibit star-like phylogenies, which may indicate of recently expanded populations, most probably from a small number of founders. A nested clade analysis suggests a complex pattern of past events that gave rise to the extant geographical pattern found in the haplotype distribution: past and allopatric fragmentation, range expansion, restricted gene flow and long-distance dispersal. These results are consistent with the complex geological history of the island where important volcanic activity with extensive lava flows has occurred during several periods. Mismatch-distribution analysis and AMOVA also support these conclusions. Substantial genetic structuring among these lizards was detected as well as high levels of differentiation between the southern edge populations (particularly those from the Rolas Islet) and the remaining ones. However, variation is low relative to the geological age of the island. Our results indicate that patterns of variation observed in reptiles in other oceanic islands are not indicative of those observed in the islands of the Gulf of Guinea.

Animals↗
PubMed2003

Large-scale variation among human and great ape genomes determined by array comparative genomic hybridization.

Large-scale genomic rearrangements are a major force of evolutionary change and the ascertainment of such events between the human and great ape genomes is fundamental to a complete understanding of the genetic history and evolution of our species. Here, we present the results of an evolutionary analysis utilizing array comparative genomic hybridization (array CGH), measuring copy-number gains and losses among these species. Using an array of 2460 human bacterial artificial chromosomes (BACs) (12% of the genome), we identified a total of 63 sites of putative DNA copy-number variation between humans and the great apes (chimpanzee, bonobo, gorilla, and orangutan). Detailed molecular characterization of a subset of these sites confirmed rearrangements ranging from 40 to at least 175 kb in size. Surprisingly, the majority of variant sites differentiating great ape and human genomes were found within interstitial euchromatin. These data suggest that such large-scale events are not restricted solely to subtelomeric or pericentromeric regions, but also occur within genic regions. In addition, 5/9 of the verified variant sites localized to areas of intrachromosomal segmental duplication within the human genome. On the basis of the frequency of duplication in humans, this represents a 14-fold positional bias. In contrast to previous cytogenetic and comparative mapping studies, these results indicate extensive local repatterning of hominoid chromosomes in euchromatic regions through a duplication-driven mechanism of genome evolution.

Animals↗
PubMed2003

Cell proliferation, apoptosis, and expression of Bcl-2 and Bax in non-lactating human breast epithelium in relation to the menstrual cycle and reproductive history.

Cell proliferation, apoptosis, and the expression of Bcl-2 and Bax were investigated in breast tissue of healthy premenopausal women in order to study the effect of the menstrual cycle and reproductive history on the cell turnover in the non-lactating mammary gland epithelium. Immunohistochemistry was used to detect the proliferation-associated antigen Ki-67, as well as Bcl-2 and Bax. Apoptotic cells were identified by enzymatic labelling of fragmentized DNA (TUNEL-technique) and morphologic analysis. Consistent with published data, the proliferative activity and the frequency of apoptotic events as detected by morphologic analysis was higher in the luteal than in the follicular phase of the menstrual cycle. Parity, lactation, and age correlated with lower proliferative activity, whereas the frequency of apoptosis was not significantly influenced by the reproductive history. Staining patterns for Bax and Bcl-2 showed characteristic changes due to the menstrual cycle with a maximum of immunoreactivity for Bcl-2 in the follicular phase and for Bax in the luteal phase. However, there was no statistically significant association between Bcl-2/Bax immunoreactivity and menstrual cycle or reproductive parameters. We conclude that other molecular pathways than the Bax/Bcl-2 antagonism may additionally be involved in the regulation of apoptotic cell death in the breast epithelium. Knowledge of the entire complexity of apoptosis regulation is necessary to understand the observed effects of parity and lactation on mammary epithelial biology, and possibly to be able to influence pathological processes caused by an imbalance between cell renewal and elimination.

Adolescent↗
PubMed2004

A socio-ecological autopsy of the E. coli O157:H7 outbreak in Walkerton, Ontario, Canada.

The socio-political context of modern environmental health disasters tends to be defined as being outside the scope of official public health and epidemiological investigations into the causes of such disasters. On the other hand, popular accounts of these disasters tend to focus exclusively on the role of particular individuals and/or political actors, while minimizing the role of ecological factors. It is argued that an exclusive focus on either set of causal factors gives an incomplete or distorted picture of the origins of an environmental health disaster. In this paper, a socio-ecological analysis is developed to demonstrate how the largest outbreak of waterborne E. coli O157:H7 in Canadian history was the emergent product of a complex interplay and intertwining of social and ecological processes. The socio-ecological autopsy approach that is developed here traces the social and ecological chain of events that ultimately led to the outbreak and demonstrates, in particular, the need for investigative analysis to focus on the socio-ecological "incubation" of an environmental health disaster. Drawing from both the social sciences (particularly, the sociology of disasters and organizational sociology), and from the ecological sciences (particularly disease ecology), the analysis developed here responds to the call for the application of a more transdisciplinary approach to the study of contemporary environmental health problems.

Agriculture↗
PubMed1999

Predictors of decreased renal function in patients with heart failure during angiotensin-converting enzyme inhibitor therapy: results from the studies of left ventricular dysfunction (SOLVD)

BACKGROUND: Although angiotensin-converting enzyme inhibitor therapy reduces mortality rates in patients with congestive heart failure (CHF), it may also cause decreased renal function. Little information is available to predict which patients are at highest risk for this complication. OBJECTIVE: To quantify specific clinical predictors of reduction in renal function in patients with CHF who are prescribed angiotensin-converting enzyme inhibitor therapy. METHOD: We analyzed data from the Studies of Left Ventricular Dysfunction (SOLVD), a randomized, double-blind, placebo-controlled trial of enalapril for the treatment of CHF. There were 3379 patients randomly assigned to enalapril with a median follow-up of 974 days and 3379 patients randomly assigned to placebo with a mean follow-up of 967 days. Decreased renal function was defined as a rise in serum creatinine >/=0.5 mg/dL (44 micromol/L) from baseline. We used time-to-event analysis to identify potential predictors of decrease in renal function including age, baseline ejection fraction, baseline creatinine, low systolic blood pressure (<100 mm Hg), history of hypertension, diabetes, and use of antiplatelet, diuretic, and beta-blocker therapy. RESULTS: Patients randomly assigned to enalapril had a 33% greater likelihood of decreased renal function than controls (P =.003). By multivariate analysis, in both the placebo and enalapril groups older age, diuretic therapy, and diabetes were associated with decreased renal function, whereas beta-blocker therapy and higher ejection fraction were renoprotective. Older age was associated with a greater risk of developing decreased renal function in both groups, but significantly more so in the enalapril group (enalapril: risk ratio [RR] 1.42 per 10 years, 95% confidence interval [CI] 1.32-1.52 with enalapril; placebo: RR 1.18, 95% CI 1.12-1.25). Diuretic therapy was likewise associated with a greater risk of decreased renal function in the enalapril group (RR 1.89, 95% CI 1.70-2.08) than in the placebo group (RR 1.35, 95% CI 1.09-1.66). Conversely, enalapril had a relative renoprotective effect (RR 1.33, 95% CI 1.13-1.53) compared with placebo (RR 1.96, 95% CI 1.57-2.44) in patients with diabetes. A lower risk of renal impairment was seen in both groups with beta-blocker therapy (RR 0.70, 95% CI 0.57-0.85) and higher baseline ejection fraction (RR 0.93 per 5% increment, 95% CI 0.91-0. 96). CONCLUSIONS: Enalapril use caused a 33% increase in the risk of decreased renal function in patients with CHF. Diuretic use and advanced age increased this risk. Diabetes was associated with an increased risk of renal impairment in all patients with CHF, but this risk was reduced in the enalapril group compared with the placebo group. beta-Blocker therapy and higher ejection fraction were renoprotective in all patients regardless of therapy.

Adrenergic beta-Antagonists↗
PubMed1987

Polymorphism and gene conversion in mouse alpha-globin haplotypes.

We have cloned and characterized three distinct alpha-globin haplotypes obtained from inbred strains of the mouse, Mus domesticus. We report here the complete nucleotide sequence of the six alpha-globin genes that the haplotypes contain. Our analysis of these genes and those from one other previously described haplotype indicates that recurrent gene conversion events have played a major role in their history. The pattern of nucleotide substitutions suggests that conversions have occurred both within and between haplotypes. Limited segments of coding and noncoding DNA have been involved in these gene conversion events. In two of the haplotypes, the nonallelic genes of each maintain DNA sequence identity over discrete intervals and encode the same alpha-globin polypeptide. On the other hand, the coding regions of some genes have accumulated replacement changes that result in distinct alpha-globins. In one instance, these changes appear to reflect positive selection of advantageous mutations.

Amino Acid Sequence↗
PubMed2001

Fetal antecedents of male factor sub-fertility: how important is birthweight?

BACKGROUND: Severely reduced fetal growth has been linked to gonadal dysfunction. The aim of this study was to explore the link between birthweight and subsequent male infertility using the Aberdeen Maternity and Neonatal Databank (AMND). METHODS AND RESULTS: Assuming 80% power at the 5% significance level, a minimum of 126 men (63 in each group) was required to show a mean difference of 0.5 (SD = 1) in the standardized birthweight scores between cases and controls. Men born locally with unexplained male factor infertility (n = 79) were identified from the infertility clinic database. Men with normal semen parameters, born in Aberdeen and attending the infertility clinic with their partners (n = 104), served as controls. The pregnancy records of these men's mothers were obtained from the AMND and a standard case-control analysis performed. In addition to standardized birthweight scores, the following variables were studied: crude birthweight, social class, maternal age, parity, obstetric history, antepartum and peripartum events. CONCLUSIONS: No differences were found between the two groups, suggesting that low birthweight is unlikely to affect male fertility in later life.

Birth Weight↗
PubMed2004

A general rule for the dependence of developmental rate on temperature in ectothermic animals.

In animals that do not regulate their body temperature by the production of heat, the proportion of the total developmental time spent in a particular developmental stage does not change with temperature. In the quasi-linear region of the relationship between developmental rate and temperature, all of the developmental stages appear to have the same species-specific lower developmental threshold. This trait, which is called developmental isomorphy, constrains developmental adaptations of ectotherms to their environments and facilitates the precise timing of life-history events.

Amphibians↗
PubMed1989

Erythrocyte sodium-lithium countertransport in primary and renal hypertension: relation to family history.

Sodium-lithium countertransport (Na-Li CT) has been reported to be increased in essential hypertension (EHT) but the nature and degree of distinction from normal controls in unclear. Of 44 unselected patients with EHT in the hospital hypertension clinic 36% had Na-Li CT greater than the normal control range and 70% of these had a family history of hypertension. Almost all the patients with normal Na-Li CT had no family history of hypertension. Analysis of variance showed that raised Na-Li CT was related to both a family history of hypertension and a family history of a cardiovascular event. Of 23 patients with hypertension secondary to renal disease, 43% had Na-Li CT greater than the normal control range and raised Na-Li CT was related to both a family history of hypertension and a family history of cardiovascular event in the same way as EHT. Raised Na-Li CT was not characteristic of EHT but identified a subgroup of patients with EHT and a family history of hypertension, some of whom also had renal disease.

Adult↗
PubMed2025

The auxin gatekeepers: Evolution and diversification of the YUCCA family.

The critically important YUCCA (YUC) gene family is highly conserved and specific to the plant kingdom, primarily responsible for the final and rate-limiting step for indole-3-acetic acid (IAA) biosynthesis. IAA is an essential phytohormone, involved in virtually all aspects of plant growth and development. In addition, IAA is involved in fine-tuning plant responses to biotic and abiotic interactions and stresses. While the YUC gene family has significantly expanded throughout the plant kingdom, a detailed analysis of the evolutionary patterns driving this diversification has not been performed. Here, we present a comprehensive phylogenetic analysis of the YUC family, combining YUCs from species representing key evolutionary plant lineages. The evolutionary history of YUCs is complex and suggests multiple recruitment events via horizontal gene transfer from bacteria. We identify and hierarchically classify the YUC family into an early diverging grade, five distinct classes and 41 subclasses. Angiosperm YUC diversity and expansion are explained in the context of protein sequence conservation, as well as spatial and gene expression patterns. The presented YUC gene landscape offers new perspectives on the distribution and evolutionary trends of this crucial family, which facilitates further YUC characterization within plant development and response to environmental change.

Indoleacetic Acids↗
PubMedDate not supplied

Relationship of family history scores for stroke and hypertension to quantitative measures of white-matter hyperintensities and stroke volume in elderly males.

White-matter hyperintensities (WMHI) are frequently associated with cerebrovascular risk factors in the elderly, particularly hypertension, and have been interpreted as a subclinical form of ischemic brain damage. WMHI, clinical stroke and blood pressures show significant genetic influences. The objective of this study was to determine whether a relationship exists between family history of stroke and/or hypertension in first degree relatives and WMHI in the elderly. WMHI and stroke (CVA) volumes were quantified from brain MRI performed on 414 white, male twins born between 1917 and 1927 (average age 72.3 +/- 2.9 years). WMHI, adjusted for age and head size, was significantly correlated with the family history score (r = 0.21, p < 0.001). Dividing the family history scores into quintiles revealed significant differences in WMHI by quintile mean (p < 0.05). Subjects in the highest quintile of family history score had the highest mean WMHI. Recalculation of the family history score, by only counting relatives reported to have had a clinical stroke as a positive event, revealed a nonsignificant correlation with WMHI, but the correlation of the family history score with MRI CVA volume was significant (p < 0.05). Stepwise multivariate analysis including ApoE status, current smoking status, smoking packyear history, Doppler ankle/arm blood pressure ratios, current and long term hypertensive status and current systolic and diastolic pressures indicated that the stroke/hypertension family history score was the single best predictor (p < 0.01) of WMHI volumes. Family history was not an independent predictor of CVA volume.

Aged↗
PubMed2006

Immunizations for patients with metabolic disorders.

Individuals with underlying metabolic disorders are a potential high-risk group for vaccine-preventable diseases. Newborn metabolic screening has provided a means of early identification and treatment for many of these disorders, whereas childhood immunization is one of the most effective means of decreasing the morbidity and mortality resulting from communicable diseases worldwide. There are very few contraindications to the routine administration of vaccines to the healthy, immunocompetent individual. In certain high-risk groups, such as immunocompromised patients, gravid females, and those with a history of previous anaphylactic reaction to a vaccine or its components, selective withholding of immunizations must be considered to decrease potential adverse events. A detailed analysis of the medical literature revealed few specific recommendations regarding appropriate immunization techniques for patients with metabolic disorders. In this review we detail the major metabolic disorder subtypes, elaborate on the available literature on immunizations for patients with these disorders, and provide suggested vaccine recommendations.

Adolescent↗
PubMed1995

Bleeding and thromboembolism during anticoagulant therapy: a population-based study in Rochester, Minnesota.

OBJECTIVE: To estimate the incidence of and identify risk factors for hemorrhage and thromboembolism during long-term anticoagulant therapy. DESIGN: We conducted a population-based retrospective cohort study of all residents of Rochester, Minnesota, in whom a course of warfarin therapy intended to last for more than 4 weeks was initiated between Sept. 1, 1987, and Dec. 31, 1989. METHODS: Medical records were reviewed, and pertinent data were compiled. All bleeding complications were classified as minor or major on the basis of the bleeding severity index, and thromboembolic events were classified as major if they were fatal or life-threatening. Cumulative incidences of adverse events were analyzed statistically. RESULTS: During the study period, 261 patients had incident courses of anticoagulation (52% were male, 61% were 65 years of age or older, and 31% were 75 years of age or older), with 221 patient-years of warfarin exposure. The primary indications for anticoagulation were venous thromboembolism (39%); stroke or transient ischemic attack (21%); atrial fibrillation (11%); and coronary artery disease, procedures for coronary artery disease, or cardiomyopathy (7%). The cumulative incidence of major hemorrhage at 1, 3, 12, and 24 months was 1.6%, 3.3%, 5.3%, and 10.6%, respectively, and of major or minor thromboembolic events was 2.3%, 5.0%, 7.4%, and 13.1%, respectively. In multivariate analysis, (1) a malignant condition was significantly associated with major hemorrhage; (2) malignant disease and history of peptic ulcer were significantly associated with the combined outcome of major or minor hemorrhage; and (3) malignant disease was significantly associated with any thromboembolism. Age, sex, atrial fibrillation, history of gastrointestinal hemorrhage, history of peptic ulcer, alcohol abuse, hypertension, stroke, and the Charlson comorbidity index were not significantly associated with major hemorrhage. CONCLUSION: In this population-based study, including a high proportion of elderly patients, malignant disease at initiation of warfarin anticoagulation was significantly associated with both major hemorrhage and any thromboembolism. Advanced age is not a contraindication to anticoagulant therapy.

Age Factors↗
PubMed1996

Meta-analysis of intravenous dipyridamole-thallium-201 imaging (1985 to 1994) and dobutamine echocardiography (1991 to 1994) for risk stratification before vascular surgery.

OBJECTIVES: This study evaluated the prognostic value of abnormal test results with pharmacologic stress with regard to perioperative and long-term outcomes in a large population of candidates for vascular surgery. BACKGROUND: Although numerous studies have demonstrated the prognostic value of dipyridamole-thallium-201 myocardial perfusion and dobutamine echocardiography in vascular surgery candidates, a synopsis of predictive estimates is difficult because of individual study variability in pretest clinical risk, sample size and study design. METHODS: A systematic review of published reports on preoperative pharmacologic stress risk stratification from the MEDLINE data base (1985 to 1994) identified 10 reports on dipyridamole-thallium-201 myocardial perfusion (1,994 patients) and 5 on dobutamine stress echocardiography (446 patients). Random effects models were used to calculate summary odds ratios and 95% confidence intervals. RESULTS: Summary odds ratios for death or myocardial infarction and secondary cardiac end points were greater for dobutamine echocardiographic dyssynergy (14- to 27-fold) than for dipyridamole-thallium-201 redistribution (4-fold); wider confidence intervals were noted with dobutamine echocardiography. Pretest coronary disease probability was correlated with the positive predictive value of a reversible thallium-201 defect (r=0.70), increasing sixfold from low to high risk patient subsets. Cardiac event rates were low in patients without a history of coronary artery disease (1% in 176 patients) compared with patients with coronary disease and a normal or fixed-defect pattern (4.8% in 83 patients) and one or more thallium-201 redistribution abnormality (18.6% in 97 patients, p=0.0001). CONCLUSIONS: Meta-analysis of 15 studies demonstrated that the prognostic value of noninvasive stress imaging abnormalities for perioperative ischemic events is comparable between available techniques but that the accuracy varies with coronary artery disease prevalence.

Adrenergic beta-Agonists↗
PubMed2002

Prediction of the long-term survival in breast cancer patients according to the present oncological status.

After a breast cancer diagnosis, single or multiple events can occur during follow-up (recurrence, metastasis, and death). An analysis of long-term survival should take into account not only the initial characteristics of the patient, but also her oncological status (that is, her history) after surgery. For this purpose, we used a technique proposed by Klein, Keiding and Copelan (1994), to predict the probability of a patient being alive 20 years after surgery for a breast cancer, based on data concerning her oncological status at time t. The first step of the model was to estimate the hazard function for each event of interest (recurrence, metastasis, and death) in a Cox model including initial patient characteristics (age, tumour size, number of involved axillary lymph nodes and the Scarff, Bloom and Richardson (SBR) histo-prognostic grade) and time-dependent covariates representing the occurrence of intermediate events (recurrence and metastasis). The second step was to use these estimations to calculate the conditional probability of being alive 20-t years later for a patient, given her oncological status at time t (t<10 years). In this second step, the method presented by Klein, Keiding and Copelan was extended to include non-proportional hazards. This model has been applied to a population of 3180 patients operated on for a breast cancer at the Institut Gustave Roussy between 1 January 1954 and 31 December 1983. At the time of surgery, the probability of survival at 20 years is 0.78 for all patients. Ten years after surgery, if no recurrence or metastasis are observed, the probability of survival at 20 years will rise to 0.89. If only a recurrence is observed, the probability of a patient being alive at 20 years will drop to 0.72. If a metastasis and no recurrence is observed, the probability of survival at 20 years will be only 0.18. If both recurrence and metastasis are observed the probability of survival at 20 years will be equal to 0.09. In conclusion, the model used dynamically appraises the prognosis and represents a new approach for studying the outcome of breast cancer patients having undergone surgery.

Adult↗
PubMed1984

A predictor analysis of patients' assessment of outcome after operation for duodenal ulcer. A one-year prospective study.

During a period of one year data were obtained concerning life events, non-specific psychological symptoms, individual social history and ulcer history in consecutive cases of patients about to undergo elective surgical treatment for duodenal ulcer. At a one-year follow-up, a blind clinical evaluation was performed, and information concerning the patients' assessment of outcome was obtained. Those patients who at the one-year follow-up stated no improvement due to the operation could be predicted to some extent from postoperative complications, partly from a long ulcer history. The patients who stated that the result did not come up to their expectations were predicted from older age, and from certain symptoms, especially severe headache. It is suggested that it is relevant to apply the patients' assessment of outcome for the purpose of evaluation, supplementing the clinical assessment of the more biomedical aspects of outcome.

Adult↗
PubMed2001

A system of analyzing medical errors to improve GME curricula and programs.

The report of the Institute of Medicine (IOM) To Err is Human recommended that both mandatory and voluntary event-reporting systems be established to identify and learn from errors. Because of the tight coupling of graduate medical education (GME) programs and the delivery of care, any event-reporting system used in a teaching hospital should be able to document the types of errors that are being made by graduate medical trainees (GTs). The authors performed an analysis of the root causes of events involving GTs that were recorded in hospital-based near-miss reporting systems. The root causes were classified using the Eindhoven Classification Model, medical version. Case histories of three separate events, one from an accident and emergency department in the United Kingdom, and two from a large teaching hospital in the United States, are used to illustrate the method. In all three cases, lack of knowledge on the part of the trainee contributed to the incident. Inadequate educational preparation had the potential for causing significant harm to the patient. Organizational causes were also present in each case, which illustrates the need to examine not only educational issues but also procedural and management issues related to GME. In each case, the analysis revealed in striking clarity deficiencies of educational content and problems of program structure. The authors conclude that doing a root-cause analysis in conjunction with a near-miss event-reporting system in a teaching hospital can be a valuable source of documented information to guide needed educational and system changes to GME programs.

Curriculum↗
PubMed2002

Antegrade selective cerebral perfusion during operations on the thoracic aorta: factors influencing survival and neurologic outcome in 413 patients.

OBJECTIVE: We retrospectively analyzed hospital mortality and neurologic outcome after operations on the thoracic aorta with the aid of antegrade selective cerebral perfusion to determine a predictive risk model. METHODS: Between October 1995 and May 2001, 413 patients (mean age, 63.0 +/- 11.5 years) underwent operations on the thoracic aorta with antegrade selective cerebral perfusion. Indications for surgical intervention were acute type A dissection in 116 (28.1%) patients, degenerative aneurysm in 227 (55.0%) patients, and postdissection aneurysm in 70 (16.9%) patients. One hundred twenty-five (30.3%) patients were operated on urgently; concomitant procedures were performed in 171 (41.4%) patients. Mean cerebral perfusion time was 63.0 +/- 38.7 minutes (range, 16-220 minutes). Preoperative and intraoperative factors were evaluated by means of univariate and multivariate analysis to identify predictors of hospital mortality and neurologic outcome. RESULTS: The hospital mortality was 9.4%. Stepwise logistic regression revealed urgency status (P =.000; odds ratio, 19.9) and recent history of a recent central neurologic event (P =.004; odds ratio, 8.0) to be independent determinants for hospital mortality. Temporary neurologic dysfunction occurred in 20 (5.1%) patients. Urgency status (P =.005; odds ratio, 7.5), history of a central neurologic event (P =.003; odds ratio, 8.6), and coronary artery bypass grafting (P =.019; odds ratio, 6.0) were independent determinants of temporary neurologic dysfunction. Urgency status (P =.003; odds ratio, 8.6) was the only independent determinant for permanent neurologic dysfunction, and it occurred in 15 (3.7%) patients. CONCLUSION: Antegrade selective cerebral perfusion is an effective method of brain protection. Cerebral perfusion times of longer than 90 minutes were not associated with an increased risk of hospital mortality or poorer neurologic outcome. Urgency status and recent history of central neurologic events were retained as important risk factors for hospital mortality and neurologic outcome.

Aged↗