The caregiver burden in lupus: findings from UNVEIL, a national online lupus survey in the United States.

OBJECTIVES: Lupus imposes a substantial burden on patients; however, little is known about its impact on those caring for patients with the disease. In this study, we examined the impact 'caring for patients with lupus' has on caregivers from their own perspective. METHODS: UNVEIL was a one-time online national cross-sectional survey developed in partnership with the Lupus Foundation of America and fielded targeting the US Lupus Foundation of America constituents in 2014. Eligible caregivers were adults who self-identified as unpaid caregivers of patients with lupus. Eligible caregivers had to complete a series of sociodemographic questions as well as a series of well established outcome measures, such as the Short Form 12v2 Health Survey, the Work Productivity and Activity Index, the Caregiver Burden Inventory, and the Perceived Benefits of Caregiving Scale. RESULTS: A total of 253 caregivers completed the survey. The majority of caregivers (90.1%) were aged 60 years or younger, more than half (54.2%) were men, and more than half (59.7%) identified themselves as either a spouse or a partner to the patient with lupus they were caring for. Overall health-related quality of life was close to the norm mean of the general US population. Caregivers who were employed missed an average of 12.8% of paid work time due to caregiving responsibilities and reported a 33.5% reduction in on-the-job effectiveness. Nearly half of the caregivers surveyed (49.4%) indicated that their caregiving responsibilities impacted their ability to socialize with friends, and almost all caregivers (97.6%) reported experiencing increased anxiety and stress in relation to their caregiving role. CONCLUSIONS: Caregiving for patients with lupus has a substantial impact on the work productivity and the social and emotional functioning of caregivers. Healthcare professionals and policymakers should continually assess the impact of healthcare decisions on the well-being of those caring for patients with lupus.

Life-history plasticity in female threespine stickleback.

The postglacial adaptive radiation of the threespine stickleback fish (Gasterosteus aculeatus) has been widely used to investigate the roles of both adaptive evolution and plasticity in behavioral and morphological divergence from the ancestral condition represented by present-day oceanic stickleback. These phenotypes tend to exhibit high levels of ecotypic differentiation. Population divergence in life history has also been well studied, but in contrast to behavior and morphology, the extent and importance of plasticity has been much less well studied. In this review, we summarize what is known about life-history plasticity in female threespine stickleback, considering four traits intimately associated with reproductive output: age/size at maturation, level of reproductive effort, egg size and clutch size. We envision life-history plasticity in an iterative, ontogenetic framework, in which females may express plasticity repeatedly across each of several time frames. We contrast the results of laboratory and field studies because, for most traits, these approaches give somewhat different answers. We provide ideas on what the cues might be for observed plasticity in each trait and, when possible, we inquire about the relative costs and benefits to expressed plasticity. We end with an example of how we think plasticity may play out in stickleback life history given what we know of plasticity in the ancestor.

Iterative development and the scope for plasticity: contrasts among trait categories in an adaptive radiation.

Phenotypic plasticity can influence evolutionary change in a lineage, ranging from facilitation of population persistence in a novel environment to directing the patterns of evolutionary change. As the specific nature of plasticity can impact evolutionary consequences, it is essential to consider how plasticity is manifested if we are to understand the contribution of plasticity to phenotypic evolution. Most morphological traits are developmentally plastic, irreversible, and generally considered to be costly, at least when the resultant phenotype is mis-matched to the environment. At the other extreme, behavioral phenotypes are typically activational (modifiable on very short time scales), and not immediately costly as they are produced by constitutive neural networks. Although patterns of morphological and behavioral plasticity are often compared, patterns of plasticity of life history phenotypes are rarely considered. Here we review patterns of plasticity in these trait categories within and among populations, comprising the adaptive radiation of the threespine stickleback fish Gasterosteus aculeatus. We immediately found it necessary to consider the possibility of iterated development, the concept that behavioral and life history trajectories can be repeatedly reset on activational (usually behavior) or developmental (usually life history) time frames, offering fine tuning of the response to environmental context. Morphology in stickleback is primarily reset only in that developmental trajectories can be altered as environments change over the course of development. As anticipated, the boundaries between the trait categories are not clear and are likely to be linked by shared, underlying physiological and genetic systems.

Reproductive success in a natural population of male three-spined stickleback Gasterosteus aculeatus: effects of nuptial colour, parasites and body size.

The effects of nuptial colour, parasites and body size on reproductive success were examined in a natural population of three-spined stickleback Gasterosteus aculeatus. Reproductive males were collected, with the contents of their nests, during the embryo-guarding stage from Lynne Lake (Cook Inlet, Alaska, U.S.A.), and nuptial colour, infection status and body size were recorded. Regression analysis revealed that male body size was the only predictor, of those measured, of reproductive success in nature.

The impact of teriparatide adherence and persistence on fracture outcomes.

UNLABELLED: The study investigated the real-world relationship between teriparatide adherence and persistence and fracture outcomes in a US claims database. Fracture risk was estimated to decrease as adherence and persistence increased for any clinical, vertebral, and non-vertebral fractures. Greater emphasis on programs to increase patient adherence may improve clinical outcomes. INTRODUCTION: Adherence to osteoporosis treatment is essential for achieving optimal therapeutic outcomes. Previous findings from clinical trials and observational studies demonstrate that longer teriparatide (TPTD) exposure is associated with fewer fractures. The study aim was to investigate real-world relationships between TPTD adherence and persistence and fracture outcomes. METHODS: The Thomson Reuters MarketScan® database, 2004-2008, was used to identify TPTD users with continuous enrollment 12 months pre- and 24 months post-TPTD initiation. Post-index fractures included vertebral and non-vertebral. Adherence (medication possession ratio, MPR) groups were defined as high (MPR ≥ 0.80), medium (0.5 ≤ MPR < 0.8), and low (MPR < 0.5). Persistence groups were defined by periods 1-6, 7-12, 13-18, and 19-24 months. Logistic regressions modeled fracture risk for any clinical, hip, vertebral, and non-vertebral fractures, controlling for patient characteristics, insurance and healthcare provider types, Charlson comorbidity index, bone mineral density screening, medication use, and fracture history. RESULTS: Among 3,587 TPTD patients (mean age 68.9 years; 91% female), fracture risk was lowest in high MPR patients in all models except hip (OR = 1.17; p = 0.64). Medium versus high MPR was a significant risk factor for any fracture (OR = 1.49; p = 0.004) and non-vertebral fracture (OR = 1.45; p = 0.014); low-MPR was a significant risk factor for any fracture (OR = 1.64; p < 0.01), vertebral fracture (OR = 2.56; p = 0.001), and non-vertebral fracture (OR = 1.44; p = 0.013). Persistence of 1-6 months versus 19-24 months was associated with higher risk for any clinical (OR = 1.88, p < 0.001), vertebral (OR = 3.69; p < 0.001), and non-vertebral fracture (OR = 1.51; p = 0.011), but not hip (OR = 1.93; p = 0.08). CONCLUSIONS: Fracture risk decreased as TPTD adherence and persistence increased for any clinical, vertebral, and non-vertebral fractures.

Rapid shifts in multiple life history traits in a population of threespine stickleback.

Measurement of the rate of phenotypic or genetic change provides data bearing on many questions of fundamental interest to biologists, including how fast changes can proceed, whether shifts occur gradually or in bursts and how long high rates of change can be sustained. Because traits exist in functionally and genetically correlated suites, studies tracking many traits are likely to be the most informative. We quantify very rapid phenotypic changes in egg size (now smaller), clutch size (larger) and the age/size of both breeding females and males (younger, smaller) in an Alaskan population, with these traits shifting at rates from 0.13 to 0.30 haldanes over a 10-year period. In contrast, female reproductive effort and the allometric relationship of clutch size to body size changed little. These shifts appear to be caused by an altered selective landscape, with the presumed selective agent being increasing lake productivity. Some of the traits undoubtedly have at heritable component and thus represent genetic evolution as well as phenotypic.

Adherence and persistence with teriparatide among patients with commercial, Medicare, and Medicaid insurance.

UNLABELLED: Adherence to, and persistence with, treatments for osteoporosis are low. Adherence with teriparatide decreases over time. Higher copayments in the commercial/Medicare population were associated with worse persistence. Understanding factors such as prior screening, prior treatment history, and out of pocket costs that influence persistence with teriparatide may help clinicians make informed decisions. INTRODUCTION: The purpose of this study was to evaluate adherence and persistence with teriparatide. METHODS: Beneficiaries with at least one claim for teriparatide in 2003 or 2004 and continuous enrollment in the previous 12 months and subsequent 6 months were identified in a national commercial/Medicare and Medicaid administrative claims database (MarketScan®). Adherence was assessed through calculation of the medication possession ratio (MPR). Persistence was measured by time until discontinuation and time until first 60-day gap in treatment. Factors associated with persistence were assessed using Cox proportional hazards models. RESULTS: The average MPR at 6 months was 0.74 (N=2,218) and at 12 months, was 0.66 (N=1,303). At 6 months, 64.6% of patients remained on therapy and at 12 months, 56.7% remained. Bone mineral density screening and use of antiresorptive therapy within the 12 months pre-period, and lower patient copayments were associated with increased persistence. CONCLUSION: Patients appear to have good adherence with teriparatide over the first 6 months which declines over time. Prior screening and treatment of osteoporosis and out of pocket costs appear to impact persistence. To optimize patient outcomes, clinicians should consider clinical factors that impact persistence, while healthcare decision makers should consider the negative effect of higher patient copayments on persistence.

Characteristics of patients initiating teriparatide for the treatment of osteoporosis.

UNLABELLED: The demographic and clinical characteristics of patients initiating teriparatide were compared with those of patients initiating bisphosphonates for the treatment of osteoporosis. In these samples of commercially insured, Medicare, and Medicaid patients, patients initiating teriparatide were older, in poorer health, and appeared to have more severe osteoporosis than patients initiating bisphosphonates. INTRODUCTION: The demographic and clinical characteristics of patients initiating teriparatide are compared with those of patients initiating bisphosphonates. METHODS: Beneficiaries (45 years and older) with at least one claim for teriparatide or a bisphosphonate from 2003 to 2005 and continuous enrollment in the previous 12 months and subsequent 6 months were identified from commercial, Medicare, and Medicaid administrative claims databases. Patients initiating teriparatide (commercial/Medicare (N = 2,218); Medicaid (N = 824)) were compared to patients initiating bisphosphonates (commercial/Medicare (N = 97,570); Medicaid (N = 77,526)) in terms of age, provider specialty, comorbidities, prior use of osteoporosis medications, fractures, BMD screening, health status, and resource utilization. RESULTS: Teriparatide patients were older and in poorer health than bisphosphonate patients. Approximately 38% of teriparatide patients in both groups had fractured in the pre-period compared to 16% of commercial/Medicare and 15% of Medicaid bisphosphonate patients. Teriparatide patients were more likely to have used osteoporosis medications in the pre-period (79.9% versus 32.1% (commercial/Medicare); 82.2% versus 19.6% (Medicaid)). CONCLUSIONS: In these samples of patients, those initiating teriparatide differed from those initiating bisphosphonates. Teriparatide patients were older, in poorer health, and appeared to have more severe osteoporosis than bisphosphonate patients. Comparisons of treatment outcomes should take these differences in patient characteristics into consideration.

MicroCT quantification of in vitro bone resorption of neonatal murine calvaria exposed to IL-1 or PTH.

This study investigated how effectively a laboratory microCT (X-ray micro-computed tomography) system can quantify bone resorption in an in vitro calvarial model and how well this measure correlates with a conventional assay for calcium release (fluorometric titration). In vitro bone resorption in neonatal murine calvaria was quantified for 0.3 or 1.0 nM interleukin-1 (IL-1) or for 1.0 or 10.0 nM parathyroid hormone (PTH) treatment. Compared to control calvaria, a significantly greater fraction F of the calvarial "shell" (computed from the volumetric microCT data) was resorbed in treated calvaria of 5- to 7-day-old pups from the same litter. Excellent correlation (R2 = 0.8234) was observed between F and calcium release, and, unlike the calcium assay, the 3-D maps revealed where bone was resorbed. Mineral was preferentially lost near the sutures, and areas away from the suture were left relatively intact. MicroCT of calvaria before and after 96 h culture demonstrated that this X-irradiation neither increased control resorption nor prevented responses in the treated calvaria. Observations on calvaria from intact mice aged 1, 3, 5, 8, and 11 days showed uniformly distributed mineral (not a pronounced patchwork of "high" and "low" mineral regions) and increasing levels of mineral with age; this suggested that the spatial patterns of resorption were not related to inhomogeneities in the starting mineral distribution.

Responsiveness of the Smoking Cessation Quality of Life (SCQoL) questionnaire.

BACKGROUND: The Smoking Cessation Quality of Life (SCQoL) questionnaire was developed to quantify changes in self-reported functioning and well-being associated with the smoking cessation process and to facilitate comparisons among smokers, former smokers, and nonsmokers. The SCQoL includes 5 cessation-targeted scales and the 8 multi-item scales of the Medical Outcomes Study 36-Item Short-Form Health Survey. OBJECTIVE: This study was conducted to assess the responsiveness of the SCQoL by analyzing associations between SCQoL scale scores and duration of smoking abstinence. METHODS: The SCQoL was administered at a screening visit and 2 to 6 weeks after screening as part of a longitudinal study. Study participants included smokers who intended to quit smoking. Subjects were required to purchase nicotine inhalers and were prompted to quit smoking before follow-up. Based on self-reported duration of abstinence at follow-up, subjects were categorized as recidivists (0 days smoke free), short-term abstainers (1-7 days smoke free), or longer-term abstainers (>7 days smoke free). Kruskal-Wallis tests were used to compare changes in scale scores from screening to follow-up among the 3 groups. RESULTS: The internal consistency reliability of the 13 SCQoL scales ranged from 0.67 to 0.92. Subjects who maintained abstinence for longer periods experienced smaller declines in health-related quality of life between the screening and follow-up assessments. Differences among the 3 groups were driven primarily by differences between recidivists and longer-term abstainers. CONCLUSIONS: The results are consistent with research indicating that recidivists report a greater number of (and more severe) cessation-related symptoms than abstainers. The findings of this investigation provide evidence for the responsiveness of the SCQoL.

Field data do not support a textbook example of convergent character displacement.

Experimental evidence supporting convergent character displacement is rare; only one example exists and it is in the form of orientation and territory competition experiments performed in the laboratory. However, outcomes of laboratory experiments involving behaviour or competition can be artefacts of unnatural conditions and, therefore, the results of the previous experiments supporting convergent character displacement are equivocal. In this study, we re-examine the evolution of melanic nuptial coloration in male three-spined stickleback (Gasterosteus aculeatus) inhabiting the Chehalis River drainage in Washington State. This novel nuptial coloration has been thought to have evolved in response to competition for nesting territories with the co-distributed Olympic mudminnow (Norzumbra hubbsi), which is also melanic and breeds at the same time. I found that melanic stickleback males did not have an advantage over their red counterparts from typical populations when competing for nesting territories with Olympic mudminnows. Additionally competitive interactions between sticklebacks and mudminnows were rare in both cage experiments and naturally breeding sticklebacks. Finally, melanic coloration in the Chehalis populations did not develop until males were parental, well after the hypothesized territory establishment period. These results refute the only experimental support for convergent character displacement and emphasize the importance of conducting behavioural experiments and observations under natural conditions.

Progressive region-specific de novo methylation of the p16 CpG island in primary human mammary epithelial cell strains during escape from M(0) growth arrest.

CpG island methylation plays an important role in normal cellular processes, such as genomic imprinting and X-chromosome inactivation, as well as in abnormal processes, such as neoplasia. However, the dynamics of de novo CpG island methylation, during which a CpG island is converted from an unmethylated, active state to a densely methylated, inactive state, are largely unknown. It is unclear whether the development of de novo CpG island methylation is a progressive process, in which a subset of CpG sites are initially methylated with a subsequent increase in methylation density, or a single event, in which the initial methylation event encompasses the entire CpG island. The tumor suppressor gene p16/CDKN2a/INK4a (p16) is inactivated by CpG island methylation during neoplastic progression in a variety of human cancers. We investigated the development of methylation in the p16 CpG island in primary human mammary epithelial cell strains during escape from mortality stage 0 (M(0)) growth arrest. The methylation status of 47 CpG sites in the p16 CpG island on individual DNA molecules was determined by sequencing PCR clones of bisulfite-treated genomic DNA. The p16 CpG island was initially methylated at a subset of sites in three discrete regions in association with p16 transcriptional repression and escape from M(0) growth arrest. With continued passage, methylation gradually increased in density and methylation expanded to sites in adjacent regions. Thus, de novo methylation in the p16 CpG island is a progressive process that is neither site specific nor completely random but instead is region specific. Our results suggest that early detection of methylation in the CpG island of the p16 gene will require methylation analysis of the three regions and that the identification of region-specific methylation patterns in other genes may be essential for an accurate assessment of methylation-mediated transcriptional silencing.

Home blood glucose monitoring.

OBJECTIVE: To provide a review of self-monitoring blood glucose including home blood glucose meters and patient education. DATA SOURCES: A MEDLINE search (January 1966-January 1998) was conducted to identify original and review articles. Search terms included self-monitoring blood glucose and blood glucose monitoring. Owner's manuals and package inserts were reviewed to determine specific characteristics for each glucose meter. DATA EXTRACTION: All current original and review articles about self-monitoring blood glucose and home blood glucose meters were included if they contained information about benefits of self-monitoring blood glucose, technology and performance of blood glucose meters, quality control, selection characteristics of blood glucose meters, and patient education. DATA SYNTHESIS: Self-monitoring of blood glucose has become an increasingly vital component of the care of the diabetic patient. Many glucose monitors are available with various features that may be confusing to pharmacists. Pharmacists need to be able to aid patients in the selection of an appropriate glucose meter and provide the education necessary for proper use and follow-up. Patient education is a key component in optimizing the potential benefits of self-monitoring. CONCLUSIONS: Self-monitoring of blood glucose, if used properly, can have a positive effect by increasing patient involvement in overall diabetes care. Pharmacists are accessible and can teach patients necessary skills that will enhance their ability to self-manage blood glucose.

Development of the Smoking Cessation Quality of Life questionnaire.

This paper describes the development of the Smoking Cessation Quality of Life (SCQoL) questionnaire, a self-reported measure designed to quantify the impact of smoking cessation on perceived functioning and well-being in adults. In addition to incorporating the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) as a generic core, the SCQoL contains 5 multi-item cessation-targeted scales: social interactions, self-control, sleep, cognitive functioning, and anxiety. The draft SCQoL was developed through a series of focus groups and was pilot-tested in a sample of 101 adults. Respondents were predominantly male (59.2%), with a mean (SD) age of (48.6) (12.7) years and a mean (SD) smoking history of 29.3 (14.7) years. Of the respondents, 76.5% identified themselves as current smokers and 23.5% indicated that they were former smokers. The majority of former smokers (82.6%) reported being abstinent for > or =2 weeks. Multivariate analysis of variance was used to compare scale scores between smokers and former smokers who had been abstinent for > or =2 weeks. Former smokers reported significantly higher scores than did current smokers on 3 of 8 SF-36 scales and 3 of 5 cessation-targeted scales (P < 0.05). In no case did current smokers report significantly higher scale scores than did former smokers. The internal-consistency reliability of the SCQoL scales ranged from 0.68 to 0.96, exceeding 0.70 on 12 of 13 scales. These findings provide preliminary evidence for the reliability and construct validity of the SCQoL.

Both Rb/p16INK4a inactivation and telomerase activity are required to immortalize human epithelial cells.

Normal human cells undergo a limited number of divisions in culture and enter a non-dividing state called replicative senescence. Senescence is accompanied by several changes, including an increase in inhibitors of cyclin-dependent kinases and telomere shortening. The mechanisms by which viral oncogenes reverse these processes are not fully understood, although a general requirement for oncoproteins such as human papillomavirus E6 and E7 has suggested that the p53 and Rb pathways are targeted. Expression of the catalytic component of telomerase, hTERT, alone significantly extends the lifespan of human fibroblasts. Here we show that telomerase activity is not sufficient for immortalization of human keratinocyte or mammary epithelial cells: we find that neither addition of hTERT nor induction of telomerase activity by E6, both of which are active in maintaining telomere length, results in immortalization. Inactivation of the Rb/p16 pathway by E7 or downregulation of p16 expression, in combination with telomerase activity, however, is able to immortalize epithelial cells efficiently. Elimination of p53 and of the DNA-damage-induced G1 checkpoint is not necessary for immortalization, neither is elimination of p19ARF.

Alendronate/interleukin-1beta cotreatment increases interleukin-6 in bone and UMR-106 cells: dose dependence and relationship to the antiresorptive effect of alendronate.

Aminobisphosphonates inhibit bone resorption but have been shown to elicit acute-phase-like elevations in interleukin-6 (IL-6) in bone in vitro. The current studies were carried out to determine the relationship between the antiresorptive effects of the aminobisphosphonate alendronate and its effects on IL-6. Resorption was elicited in cultured 19-day fetal rat limb bones by 72 h treatment with interleukin-1beta (IL-1beta). Bone mass was quantitated at the end of the culture period to assess resorption. IL-6 was determined by bioassay (7TD1 cell proliferation). IL-1beta (18 and 180 pM) stimulated bone resorption and increased IL-6. Alendronate (70 microM) inhibited the IL-1beta-stimulated resorption. Alendronate alone did not affect IL-6 production by the bones. The IL-6 production from bones stimulated with 18 pM IL-1beta was not significantly affected by alendronate, but the IL-6 production from bones stimulated with 180 pM IL-1beta plus alendronate (21 and 70 microM) was higher than with IL-1beta alone. Indomethacin (1 mM) inhibited the IL-6 increase elicited by 180 pM IL-1beta and the enhanced IL-6 production elicited by cotreatment with IL-1beta and alendronate. Since bone cultures contain multiple cell types, further experiments were carried out to determine whether alendronate could increase IL-1beta-stimulated IL-6 production in an osteoblast cell line, UMR-106. Alendronate alone did not affect IL-6 in UMR-106 cells. Alendronate (70 microM) in combination with IL-1beta (180, 1.8, or 8 nM), or 7 microM alendronate, in combination with 8 nM IL-1beta, significantly increased IL-6 in 48 h cell cultures. The results from the bone organ cultures show that alendronate can enhance IL-6 production elicited by higher concentrations of the cytokine IL-1beta in bone, but that this effect on IL-6 does not prevent the inhibitory actions of alendronate on bone resorption. The results with the UMR106 cells indicate that one cellular site at which this enhancement of IL-6 production can occur is the osteoblast.

Inactivation of p16 in human mammary epithelial cells by CpG island methylation.

Proliferation of human mammary epithelial cells (HMEC) is limited to a few passages in culture due to an arrest in G1 termed selection or mortality stage 0, M0. A small number of cells spontaneously escape M0, continue to proliferate in culture, and then enter a second mortality stage, M1, at which they senesce. Evidence that M0 involves the Rb pathway comes from the observation that expression of human papillomavirus type 16 E7 alleviates the M0 proliferation block, and we further show that the Rb-binding region of E7 is required to allow cells to bypass M0. In contrast, E6 does not prevent HMEC from entering M0 but, rather, is involved in M1 bypass. Here we show that inactivation of the D-type cyclin-dependent kinase inhibitor p16INK4A is associated with escape from the M0 proliferation block. Early-passage HMEC express readily detectable amounts of p16 protein, whereas normal or E6-expressing HMEC that escaped M0 expressed markedly reduced amounts of p16 mRNA and protein. This initial reduction of p16 expression was associated with limited methylation of the p16 promoter region CpG island. At later passages, a further reduction in p16 expression occurred, accompanied by increased CpG island methylation. In contrast, reduction of p16 expression did not occur in E7-expressing HMEC that bypassed M0, due to inactivation of Rb. These observations in the E6-expressing HMEC correlate well with the finding that CpG island methylation is a mechanism of p16 inactivation in the development of human tumors, including breast cancer.

Human papillomavirus type 16 E7 alleviates a proliferation block in early passage human mammary epithelial cells.

Human mammary epithelial cells (HMEC) isolated from reduction mammoplasty tissue are proliferative for several passages but then enter a period termed ¿selection' or M0 during which the majority of the cells become larger, flattened, and less proliferative. Early passage HMEC (prior to M0) were transduced with human papillomavirus type-16 E6, E7, or E6/E7 by using recombinant retroviral vectors. E7 alone or E6/E7, but not E6 alone, alleviated the M0 proliferation block, suggesting a possible role for Rb or Rb-related proteins, but not p53, in M0. In addition, cells in M0 did not have increased levels of p53 or p21 proteins, further indicating that induction of these proteins is not required for the M0 proliferation block. Early passage cells contained Rb which was primarily hyperphosphorylated while cells in M0 contained Rb protein which was predominantly underphosphorylated. Cells in M0 accumulated in G1 or B0 and expressed reduced levels of cyclin A and CDK2 proteins.