Nonadherent Population Research Articles

215 - Adherence to a Telehealth Heart Failure Program was Associated Fewer Hospitalizations

Adherence of patients plays an important role in the ability of patients to manage their heart failure (HF). As the utilization of telehealth increases, the effect of the technology and remote care on management of disease remains unknown. It is anticipated that those patients who are more adherent with the telehealth program will exhibit greater management of their HF. This study assessed the impact of a care management program which utilized telehealth technology (Medtronic Care Management Services) to offer improved heart failure care remotely. Methods: A prospective, single-arm, quality initiative program evaluating the expansion of a telehomecare network into rural areas was performed. Following discharge for a HF admission, high-risk patients with HF were enrolled into a telehealth heart failure monitoring program which consisted of daily weight and symptom questions transmitted via telehealth technology for evaluation by a site HF nurse. Follow-up visits were performed at 1 week and 30, 60 and 90 days post-discharge. Adherence was defined as the number of transmissions per week with adherent patients having 5 or more transmissions per week for at least 10 out of 14 weeks and nonadherent patients having less than 5 transmissions per week for 10 out of 14 weeks. The number of hospitalizations, both unscheduled all-cause and all-cause, was evaluated to determine overall disease management. Results: 844 patients (459 males, average age 70.2 ± 13.6 years) were followed for 9 months. Of those patients, at 31 days, 478 were found to be adherent and 238 nonadherent, whereas at 90 days, 477 were adherent and 234 were non-adherent. The unscheduled all-cause hospitalization rate for the adherent population was 7.7% and 12.6% and for the non-adherent population 12.6% and 36.3% at 31 (P = .05) and 90 (P = < 0.001) days, respectively. The all-cause rehospitalization rate was also lower in the adherent group at 31 (adherent = 9.0%, non-adherent 14.3%; P = .04) and 90 days (adherent = 21.6%, nonadherent = 44.0%; P < .001). Conclusion: Patients with the highest frequency of transmissions had fewer hospitalizations. Patients more adherent to transmissions were associated with better HF outcomes.

High Rate of Treatment Completion in Program Settings With 12-Dose Weekly Isoniazid and Rifapentine for Latent Mycobacterium tuberculosis Infection.

Randomized controlled trials have demonstrated that the newest latent tuberculosis (LTBI) regimen, 12 weekly doses of directly observed isoniazid and rifapentine (3HP), is as efficacious as 9 months of isoniazid, with a greater completion rate (82% vs 69%); however, 3HP has not been assessed in routine healthcare settings. Observational cohort of LTBI patients receiving 3HP through 16 US programs was used to assess treatment completion, adverse drug reactions, and factors associated with treatment discontinuation. Of 3288 patients eligible to complete 3HP, 2867 (87.2%) completed treatment. Children aged 2-17 years had the highest completion rate (94.5% [155/164]). Patients reporting homelessness had a completion rate of 81.2% (147/181). In univariable analyses, discontinuation was lowest among children (relative risk [RR], 0.44 [95% confidence interval {CI}, .23-.85]; P = .014), and highest in persons aged ≥65 years (RR, 1.72 [95% CI, 1.25-2.35]; P < .001). In multivariable analyses, discontinuation was lowest among contacts of patients with tuberculosis (TB) disease (adjusted RR [ARR], 0.68 [95% CI, .52-.89]; P = .005) and students (ARR, 0.45 [95% CI, .21-.98]; P = .044), and highest with incarceration (ARR, 1.43 [95% CI, 1.08-1.89]; P = .013) and homelessness (ARR, 1.72 [95% CI, 1.25-2.39]; P = .001). Adverse drug reactions were reported by 1174 (35.7%) patients, of whom 891 (76.0%) completed treatment. Completion of 3HP in routine healthcare settings was greater overall than rates reported from clinical trials, and greater than historically observed using other regimens among reportedly nonadherent populations. Widespread use of 3HP for LTBI treatment could accelerate elimination of TB disease in the United States.

Abstract 13150: The Economic Implications of Non-adherence to Aspirin in Patients With Prior Primary Cardiovascular Events

Introduction: Adherence to medications for secondary prevention of cardiovascular events has been shown to be poor in both general and high risk populations. Non-adherent patients are at higher risk of experiencing another CV event, leading to adverse clinical events and high costs. Few prior studies have explored the economic and consequences of non-adherence to aspirin in relation to cardiovascular health. Hypothesis: Improved adherence to aspirin will lead to lower healthcare costs and improved long-term survival rates. Methods: We developed a generalizable Markov model to estimate the cost-effectiveness of aspirin adherence. The rates of aspirin adherence, events and costs were gathered from published literature. We applied the model separately to a general (low-risk) population with no comorbidities and a high-risk population with type II diabetes. Outcomes of interest included fatal and nonfatal events, including secondary myocardial infarction (MI), stroke, other clinical CV events including PAD-related amputation as well as aspirin-related side effects (gastrointestinal bleeding). All costs were inflated to 2016 U.S. dollars to reflect the current cost of treatment. Results: The aspirin adherence rates for the low-risk and high-risk model were 85.8% and 76.5%, respectively. Both models demonstrated neutral average 5-year cost per member when comparing adherent to non-adherent population. The average 5-year survival was 0.26 years or 3.1 months longer amongst the adherent population in the low-risk model and 0.37 years or 4.4 months longer in the high-risk model. As a result, the ICER is $23 for low risk population while adherent strategy is dominant (lower cost, higher effectiveness) for high risk population. Conclusions: Adherence to aspirin therapy after experiencing a primary CV event is critical for patients’ well-being and can facilitate better health outcomes. This study demonstrates that aspirin adherence is essentially cost neutral and extends survival in low and high-risk patient populations, with a potentially cost saving outcome for patients with comorbid conditions such as diabetes.

Non Compliance Pattern of Anti Hypertensive Treatment

A cross-sectional type of observational study was conducted on hypertensive patients attending at medicine department (OPD and admitted) of Z. H. Sikder Women’s Medical College Hospital, Dhaka, from January to June 2014, to see the patient compliance of anti-hypertensive drugs. The sample size was 185. Patients with evidence of hypertension as per Seventh Joint National Committee Criteria’ (defined as systolic blood pressure &gt;140 mm oh Hg or diastolic blood pressure &gt;90 mm of Hg or use any anti-hypertensive medication, regardless of blood pressure. Time from initiation of antihypertensive treatment was at least 3 months and patients between 18-70 years were included in this study. A pre-tested questionnaire was used to gather sociodemographical and relevant epidemiological data. Standard blood pressure measurement was recorded as the average of 2 measurements to detect anti-hypertension. Patients were asked for their reasons of noncompliance, knowledge regarding importance of compliance, side effects of drugs and family support. Mean age of the patients was 47.40(±5.09) years. 75.9% of the non adherent population is in the 41-50 years age group. 85% percent of the patients were found non-adherent to treatment. Out of 158 non adherent population, 54.43% were male and 45.57% were female. 38.6% of the non-adherent people have no education or a primary education, only 7.6% were graduate and above. 42.41% of the non-adherent people were poor and earning &lt; 5000 taka/month. Majority duration of disease ranged from 1 to 15 years which was 50%, with an average of 3.99(±3.47) years. The most commonly prescribed antihypertensive medicine was beta blockers. 49.4% of the non-adherent population was advised to take relatively costly antihypertensive drug (Calcium channel blocker, ACEI, ARB). The main reason for not taking the medication was forgetfulness (48.1%) and the economic reason for non-compliance was only 7.59%. About 63% patients felt hypertension is not a curable disease, 42% 0f the study subjects were not sure about the signs and symptoms of HTN and another 30.6% said that HTN had no definite sign symptom.77.4% of the respondent said that preservation of previous prescription is important to understand HTN status. The overall Knowledge on complication of uncontrolled hypertension of the adherent people was relatively higher than that of nonadherent people. 96.4% of the adherent people knew that uncontrolled hypertension can cause brain stroke, whereas only 81.65% of the non-adherent people knew that. Most of the respondent of the both adherent and non-adherent group informed that hypotension, severe weakness, anorexia, nausea and impotency were the important side effect of antihypertensive drugs. Most of the (87.5%) participant went for follow up when they felt hypertensive, 35% change medicine for expenses and 27% stop taking drug when BP is under control. 78% did not know whether only regular anti-hypertensive can prevent complication or not, 62% of the respondent did not think that to buy drug is wastage of money. One-third of the respondent said that they were never asked to take medicines or visit physician by their family members. And half of the respondents had to depend on monetary support of the family member for treatment. 99.36% of non-adherent missed their follow up.J Dhaka Medical College, Vol. 24, No.2, October, 2015, Page 108-116

Tailored message interventions versus typical messages for increasing participation in colorectal cancer screening among a non-adherent population: A randomized controlled trial.

BackgroundThe purpose of this study was to examine the effectiveness and cost-efficiency of a tailored message intervention compared with a non-tailored message intervention for increasing colorectal cancer (CRC) screening rates among a non-adherent population, in a community-based client reminder program.MethodsAfter a baseline survey for psychological segmentation, 2140 eligible individuals were randomly assigned either to a group with a tailored matched-message condition (N = 356), a group with a non-tailored unmatched-message condition (N = 355), or to two control groups, one using a typical message with a professional design (N = 717) and one without a professional design (N = 712). The main outcome measure was attendance rates in a community-organized CRC screening program within five months of receiving a print reminder.ResultsThere was a significant difference in fecal occult blood test (FOBT) attendance rates at follow-up assessments between the tailored matched-message condition (14.0 %) and the control (9.9 %; OR = 1.48, p = 0.026), while there was no significant difference between the unmatched-message condition (11.0 %) and the control (OR = 1.12, p = 0.558), and between the matched-message condition and the unmatched-message condition (OR = 1.32, p = 0.219). The cost of a one-person increase in FOBT screening was 3,740 JPY for the tailored matched-message condition, while it was 2,747 JPY for the control.ConclusionsA tailored-message intervention for segmented individuals designed to increase CRC screening rates in a community-based client reminder program was significantly effective compared to a usual reminder, but not more effective than an unmatched message in a randomized controlled trial, and was not sufficiently effective to highlight its value from a cost perspective. Therefore, the tailored intervention including target segmentation needs to be improved for future implementation in a CRC screening program for a non-adherent population.Trial registrationUMIN Clinical Trials Registry UMIN000004384. Date of Registration: March 2011.Electronic supplementary materialThe online version of this article (doi:10.1186/s12889-016-3069-y) contains supplementary material, which is available to authorized users.

The Information and Motivation and Behavioral Skills Model of ART Adherence among HIV-Positive Adults in Mexico.

Middle-income countries are in need of research that uses theoretical-based models to assess factors that predict adherence to antiretroviral therapy (ART) and help in the design, implementation, and evaluation of interventions for nonadherent populations. In Mexico, the Information and Motivation and Behavioral Skills (IMB) Model of ART Adherence constructs is useful in describing and predicting adherence behaviors in various samples but has not been articulated to people living with HIV (PLWH) on ART. The aim of this was to characterize the IMB core constructs and identify correlates of ART adherence in an HIV-positive clinic sample in Mexico. A convenience sample of 109 HIV-positive patients attending their monthly visits at a local public hospital were interviewed with the Spanish version of the LifeWindows IMB ART Adherence Questionnaire (LW-IMB-AAQ) as well as a sociodemographic questionnaire. All participants were recruited from a hospital-based outpatient clinical care site. Partial confirmation of the relationships proposed by the IMB Model of ART Adherence was found. As predicted by the model, only behavioral skills had direct association with all measures of self-reported adherence, and motivation was associated with behavioral skills. Information did not demonstrate significant relations to either motivation or behavioral skills, nor did it directly associate with adherence. Self-reported adherence did not associate with CD4 counts, nor did any of the IMB model core constructs. Applicability of the IMB Model of ART Adherence in this setting is discussed. The IMB Model of ART Adherence offered promise in this population and could help tailor population-specific interventions to promote high rates of ART adherence.

Adherence to highly active antiretroviral therapy in Hyderabad, India: barriers, facilitators and identification of target groups.

We assessed the barriers and facilitators to highly active antiretroviral therapy adherence and determined their prevalence among HIV/AIDS patients in Hyderabad, India. We conducted a cross-sectional study among HIV-infected adults prescribed highly active antiretroviral therapy and receiving care from nine clinics. Depression was screened using Patient Health Questionnaire 9 and facilitators of HIV medication adherence were assessed using an 11-item scale which yielded a total positive attitude to disease score. Prevalence ratios of non-adherence between different categories of potential risk factors were calculated. We compared mean 'facilitators to adherence' scores between the adherent and non-adherent population. Multivariable Poisson regression with robust variance was used to identify independent risk factors. Among the 211 respondents, nearly 20% were non-adherent, approximately 8% had either moderately severe or severe depression and mean score for combined facilitators to medication adherence was 33.35 (±7.88) out of a possible 44 points. Factors significantly associated with non-adherence included older age, female sex worker, moderate-to-severe depression and the combined facilitators to medication adherence score. These data from a broad range of clinical settings in Hyderabad reveal that key groups to focus on for adherence intervention are female sex workers, older persons and those with depression.

Effect of providing risk information on undergoing cervical cancer screening: a randomized controlled trial.

BackgroundIn Japan, the cervical cancer screening rate is extremely low. Towards improving the cervical cancer screening rate, encouraging eligible people to make an informed choice, which is a decision-making process that relies on beliefs informed by adequate information about the possible benefits and risks of screening, has attracted increased attention in the public health domain. However, there is concern that providing information on possible risks of screening might prevent deter from participating.MethodsIn total, 1,912 women aged 20–39 years who had not participated in screening in the fiscal year were selected from a Japanese urban community setting. Participants were randomly divided into 3 groups. Group A received a printed reminder with information about the possible benefits of screening, group B received a printed reminder with information about possible benefits and risks, and group C received a printed reminder with simple information only (control group).ResultsOut of 1,912 participants, 169 (8.8%) participated in cervical cancer screening. In the intervention groups, 137 (10.9%) participated in cervical cancer screening, compared to only 32 (4.9%) of the control group (p < 0.001). In addition, logistic regression analysis revealed that there was no significant difference in screening rate between group A and group B (p = 0.372).ConclusionsProviding information on the possible risks of screening may not prevent people from taking part in cervical cancer screening among a Japanese non-adherent population.

Defining the role of medication adherence in poor glycemic control among a general adult population with diabetes.

AimsThis study assesses the attributable impact of adherence to oral glucose medications as a risk factor for poor glycemic control in population subgroups of a large general population, using an objective medication adherence measure.MethodsUsing electronic health records data, adherence to diabetes medications over a two-year period was calculated by prescription-based Medication Possession Ratios for adults with diabetes diagnosed before January 1, 2010. Glycemic control was determined by the HbA1c test closest to the last drug prescription during 2010–2012. Poor control was defined as HbA1c>75 mmol/mol (9.0%). Medication adherence was categorized as “good” (>80%), “moderate” (50–80%), or “poor” (<50%). Logistic regression models assessed the role medication adherence plays in the association between disease duration, age, and poor glycemic control. We calculated the change in the attributable fraction of glucose control if the non-adherent diabetic medication population would become adherent by age-groups.ResultsAmong 228,846 diabetes patients treated by oral antiglycemic medication, 46.4% had good, 28.8% had moderate, and 24.8% had poor adherence. Good adherence rates increased with increasing disease duration, while glycemic control became worse. There was a strong inverse association between adherence level and poor control (OR = 2.50; CI = 2.43–2.58), and adherence was a significant mediator between age and poor control.ConclusionsA large portion of the diabetes population is reported to have poor adherence to oral diabetes medications, which is strongly associated with poor glycemic control in all disease durations. While poor adherence does not mediate the poorer glycemic control seen in patients with longer-standing disease, it is a significant mediator of poor glycemic control among younger diabetes patients. A greater fraction of poorly controlled younger patients, compared to older patients, could be prevented if at least 80% adherence to their medications was achieved. Therefore, our results suggest that interventions to improve adherence should focus on this younger sub-group.

Effect of Niclosamide on Basal-like Breast Cancers

Basal-like breast cancers (BLBC) are poorly differentiated and display aggressive clinical behavior. These tumors become resistant to cytotoxic agents, and tumor relapse has been attributed to the presence of cancer stem cells (CSC). One of the pathways involved in CSC regulation is the Wnt/β-catenin signaling pathway. LRP6, a Wnt ligand receptor, is one of the critical elements of this pathway and could potentially be an excellent therapeutic target. Niclosamide has been shown to inhibit the Wnt/β-catenin signaling pathway by causing degradation of LRP6. TRA-8, a monoclonal antibody specific to TRAIL death receptor 5, is cytotoxic to BLBC cell lines and their CSC-enriched populations. The goal of this study was to examine whether niclosamide is cytotoxic to BLBCs, specifically the CSC population, and if in combination with TRA-8 could produce increased cytotoxicity. Aldehyde dehydrogenase (ALDH) is a known marker of CSCs. By testing BLBC cells for ALDH expression by flow cytometry, we were able to isolate a nonadherent population of cells that have high ALDH expression. Niclosamide showed cytotoxicity against these nonadherent ALDH-expressing cells in addition to adherent cells from four BLBC cell lines: 2LMP, SUM159, HCC1187, and HCC1143. Niclosamide treatment produced reduced levels of LRP6 and β-catenin, which is a downstream Wnt/β-catenin signaling protein. The combination of TRA-8 and niclosamide produced additive cytotoxicity and a reduction in Wnt/β-catenin activity. Niclosamide in combination with TRA-8 suppressed growth of 2LMP orthotopic tumor xenografts. These results suggest that niclosamide or congeners of this agent may be useful for the treatment of BLBC.

Purified Dendritic Cell-Tumor Fusion Hybrids Supplemented with Non-Adherent Dendritic Cells Fraction Are Superior Activators of Antitumor Immunity

BackgroundStrong evidence supports the DC-tumor fusion hybrid vaccination strategy, but the best fusion product components to use remains controversial. Fusion products contain DC-tumor fusion hybrids, unfused DCs and unfused tumor cells. Various fractions have been used in previous studies, including purified hybrids, the adherent cell fraction or the whole fusion mixture. The extent to which the hybrids themselves or other components are responsible for antitumor immunity or which components should be used to maximize the antitumor immunity remains unknown.MethodsPatient-derived breast tumor cells and DCs were electro-fused and purified. The antitumor immune responses induced by the purified hybrids and the other components were compared.ResultsExcept for DC-tumor hybrids, the non-adherent cell fraction containing mainly unfused DCs also contributed a lot in antitumor immunity. Purified hybrids supplemented with the non-adherent cell population elicited the most powerful antitumor immune response. After irradiation and electro-fusion, tumor cells underwent necrosis, and the unfused DCs phagocytosed the necrotic tumor cells or tumor debris, which resulted in significant DC maturation. This may be the immunogenicity mechanism of the non-adherent unfused DCs fraction.ConclusionsThe non-adherent cell fraction (containing mainly unfused DCs) from total DC/tumor fusion products had enhanced immunogenicity that resulted from apoptotic/necrotic tumor cell phagocytosis and increased DC maturation. Purified fusion hybrids supplemented with the non-adherent cell population enhanced the antitumor immune responses, avoiding unnecessary use of the tumor cell fraction, which has many drawbacks. Purified hybrids supplemented with the non-adherent cell fraction may represent a better approach to the DC-tumor fusion hybrid vaccination strategy.

Microfluidic device capable of medium recirculation for non-adherent cell culture.

We present a microfluidic device designed for maintenance and culture of non-adherent mammalian cells, which enables both recirculation and refreshing of medium, as well as easy harvesting of cells from the device. We demonstrate fabrication of a novel microfluidic device utilizing Braille perfusion for peristaltic fluid flow to enable switching between recirculation and refresh flow modes. Utilizing fluid flow simulations and the human promyelocytic leukemia cell line, HL-60, non-adherent cells, we demonstrate the utility of this RECIR-REFRESH device. With computer simulations, we profiled fluid flow and concentration gradients of autocrine factors and found that the geometry of the cell culture well plays a key role in cell entrapping and retaining autocrine and soluble factors. We subjected HL-60 cells, in the device, to a treatment regimen of 1.25% dimethylsulfoxide, every other day, to provoke differentiation and measured subsequent expression of CD11b on day 2 and day 4 and tumor necrosis factor-alpha (TNF-α) on day 4. Our findings display perfusion sensitive CD11b expression, but not TNF-α build-up, by day 4 of culture, with a 1:1 ratio of recirculation to refresh flow yielding the greatest increase in CD11b levels. RECIR-REFRESH facilitates programmable levels of cell differentiation in a HL-60 non-adherent cell population and can be expanded to other types of non-adherent cells such as hematopoietic stem cells.

Characteristics and response of mouse bone marrow derived novel low adherent mesenchymal stem cells acquired by quantification of extracellular matrix.

PURPOSEThe aim of present study was to identify characteristic and response of mouse bone marrow (BM) derived low-adherent bone marrow mesenchymal stem cells (BMMSCs) obtained by quantification of extracellular matrix (ECM).MATERIALS AND METHODSNon-adherent cells acquired by ECM coated dishes were termed low-adherent BMMSCs and these cells were analyzed by in vitro and in vivo methods, including colony forming unit fibroblast (CFU-f), bromodeoxyuridine (BrdU), multi-potential differentiation, flow cytometry and transplantation into nude mouse to measure the bone formation ability of these low-adherent BMMSCs. Titanium (Ti) discs with machined and anodized surfaces were prepared. Adherent and low-adherent BMMSCs were cultured on the Ti discs for testing their proliferation.RESULTSThe amount of CFU-f cells was significantly higher when non-adherent cells were cultured on ECM coated dishes, which was made by 7 days culturing of adherent BMMSCs. Low-adherent BMMSCs had proliferation and differentiation potential as adherent BMMSCs in vitro. The mean amount bone formation of adherent and low-adherent BMMSCs was also investigated in vivo. There was higher cell proliferation appearance in adherent and low-adherent BMMSCs seeded on anodized Ti discs than machined Ti discs by time.CONCLUSIONLow-adherent BMMSCs acquired by ECM from non-adherent cell populations maintained potential characteristic similar to those of the adherent BMMSCs and therefore could be used effectively as adherent BMMSCs in clinic.

Single-cell analysis of K562 cells: An imatinib-resistant subpopulation is adherent and has upregulated expression of BCR-ABL mRNA and protein

In chronic myeloid leukemia (CML) cells from different stages of maturation may have differential expression of BCR-ABL at both messenger RNA (mRNA) and protein level. However, the significance of such differential expression to clinical disease behavior is unknown. Using the CML-derived, BCR-ABL expressing cell line, K562, distinct plastic-adherent (K562/Adh) and nonadherent (K562/NonAdh) subpopulations were established and then analyzed both as single cells and as bulk cell populations. BCR-ABL mRNA was upregulated in K562/Adh compared with K562/NonAdh cells in both single cell and bulk population analyses (p < 0.0001). Similarly, phosphorylation of BCR protein was upregulated in K562/Adh, compared with K562/NonAdh cells (63.42% vs. 23.1%; p = 0.007), and these two K562 subpopulations were found to express significantly different microRNA species. Furthermore, treatment with the BCR-ABL tyrosine kinase inhibitor, imatinib, reduced cell viability more rapidly in K562/NonAdh compared with K562/Adh cells (p < 0.005) both at single and bulk cell levels. This discovery of an adherent subpopulation of K562 cells with increased BCR-ABL mRNA, increased phosphorylated BCR protein expression, differential microRNA expression, and increased imatinib resistance suggests that a similar subpopulation of cells can also mediate clinical resistance to imatinib during treatment of patients with CML.

Factors Predicting Intentional Non-Adherence In Chronic Myeloid Leukemia: A Multivariate Analysis On 2546 Patients By The CML Advocates Network

Factors Predicting Intentional Non-Adherence In Chronic Myeloid Leukemia: A Multivariate Analysis On 2546 Patients By The CML Advocates Network

NCAM polysialylation during adherence transitions: Live cell monitoring using an antibody‐mimetic EGFP‐endosialidase and the viability dye DRAQ7

Polysialylation of neural cell adhesion molecule (NCAM) in small-cell lung cancer (SCLC) is thought to regulate NCAM-mediated cell-surface interactions, imparting antiadhesive properties to cells. However, SCLC cells in culture demonstrate anchorage-independent growth and spontaneously generate adherent forms. Here, the ability of polySia-NCAM to influence cell proliferation and adherence is unclear. We analyzed live SCLC cell polySia-NCAM expression by flow cytometry, using the novel combination of a polySia antibody-mimetic eGFP-tagged endosialidase and the viability dye DRAQ7. Enrichment for adherence (<30 population doublings) in SCLC cell lines resolved populations with increased (SHP-77 and COR-L279) or negligible (NCI-H69) polysialylation compared with nonadherent parent populations. Adherent forms retained NCAM expression as confirmed by immunofluorescence and immunoblotting. Initial transition to adherence and loss of polysialylation in NCI-H69 was linked to a reduced proliferation rate with no increase in cell death. This reduced proliferation rate was reiterated in vivo as determined by the growth of noninvasive subcutaneous xenografts in mice. Continued selection for enhanced substrate adherence in NCI-H69 (>150 population doublings) resolved cells with stable re-expression of polySia and increased growth rates both in vitro and in vivo. Endoneuraminidase removal of polySia from re-expressing cells showed that rapid adherence to extracellular matrix components was functionally independent of polySia. PolySia expression was not altered when isolated adherent forms underwent enforced cell-cell contact in three-dimensional culture. Coculture of polySia expression variants modulated overall polySia expression profiles indicating an influence of SCLC microcommunity composition independent of substrate adherence potential. We conclude that an obligatory linkage between substrate adherence potential and polySia expression is rejected for SCLC cells. We suggest that a degree of homeostasis operates to regulate polysialylation within heterogeneous cell populations. The findings suggest a new model for SCLC progression while the application of live cell profiling of polysialylation could be used to assess polySia-NCAM-targeted therapies.

A negative selection methodology using a microfluidic platform for the isolation and enumeration of circulating tumor cells

Circulating tumor cells (CTCs) exist in the peripheral blood stream of metastatic cancer patients at rates of approximately 1 CTC per billion background cells. In order to capture and analyze this rare cell population, various techniques exist that range from antibody-based surface marker positive selection to methods that use physical properties of CTCs to negatively exclude background cells from a CTC population. However, methods to capture cells for functional downstream analyses are limited due to inaccessibility of the captured sample or labeling techniques that may be prohibitive to cell function. Here, we present a negative selection method that leverages a Microfluidic Cell Concentrator (MCC) to allow collection and analysis of this rare cell population without needing cell adhesion or other labeling techniques to keep the cells within the chamber. Because the MCC is designed to allow collection and analysis of non-adherent cell populations, multiple staining steps can be applied in parallel to a given CTC population without losing any of the population. The ability of the MCC for patient sample processing of CTCs for enumeration was demonstrated with five patient samples, revealing an average of 0.31CTCs/mL. The technique was compared to a previously published method – the ELISPOT – that showed similar CTC levels among the five patient samples tested. Because the MCC method does not use positive selection, the method can be applied across a variety of tumor types with no changes to the process.

CXCL12/CXCR4 signaling in the osteoblast regulates the mesenchymal stem cell and osteoclast lineage populations

The chemokine CXCL12 and its receptor CXCR4 play a key role in regulation of hematopoietic stem cells and cell migratory function during morphogenesis. Osteoblasts express both the ligand and the receptor, but little is known about the role of CXCL12-CXCR4 signaling in maintaining skeletal homeostasis. Using Cre-Lox technology to delete CXCR4 in mature osteoblasts in mice, we show here a significant decrease in bone mass and alterations in cancellous bone structure. CXCR4 gene ablation increased the number of colony-forming units (CFU), CFU-positive for alkaline phosphatase (CFU-AP(+)), and mineralizing nodules in bone marrow stromal cell (BMSC) cultures. The adipocyte precursor population decreased in BMSCs harvested from the KO animals. The nonadherent population of BMSCs harvested from the long bone diaphysis of KO animals formed more osteoclasts, a finding that was associated with increased circulatory levels of pyridinoline, a marker of bone resorption. Our data show that osteoblast-specific CXCR4 deletion has profound effects on the mesenchymal stem cell pool and allocation to the osteoblastic and adipocytic cell lineages. They also show that CXCL12/CXCR4 signaling in the mature osteoblast can feedback to regulate the osteoclast precursor pool size and play a multifunctional role in regulating bone formation and resorption.

CPC-029 Characterization of Patients Non-Adherent to Highly Active Antiretroviral Therapy (HAART) Between 2010–2011

BackgroundThanks to the introduction of Highly Active Antiretroviral Therapy (HAART), a decrease has been observed in the number of hospital admissions, the incidence of opportunistic infections and mortality associated with...

Human dental pulp stem cells cultured in serum-free supplemented medium

Growing evidence show that human dental pulp stem cells (DPSCs) could provide a source of adult stem cells for the treatment of neurodegenerative pathologies. In this study, DPSCs were expanded and cultured with a protocol generally used for the culture of neural stem/progenitor cells.Methodology: DPSC cultures were established from third molars. The pulp tissue was enzymatically digested and cultured in serum-supplemented basal medium for 12 h. Adherent (ADH) and non-adherent (non-ADH) cell populations were separated according to their differential adhesion to plastic and then cultured in serum-free defined N2 medium with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). Both ADH and non-ADH populations were analyzed by FACS and/or PCR.Results: FACS analysis of ADH-DPSCs revealed the expression of the mesenchymal cell marker CD90, the neuronal marker CD56, the transferrin receptor CD71, and the chemokine receptor CXCR3, whereas hematopoietic stem cells markers CD45, CD133, and CD34 were not expressed. ADH-DPSCs expressed transcripts coding for the Nestin gene, whereas expression levels of genes coding for the neuronal markers β-III tubulin and NF-M, and the oligodendrocyte marker PLP-1 were donor dependent. ADH-DPSCs did not express the transcripts for GFAP, an astrocyte marker. Cells of the non-ADH population that grew as spheroids expressed Nestin, β-III tubulin, NF-M and PLP-1 transcripts. DPSCs that migrated out of the spheroids exhibited an odontoblast-like morphology and expressed a higher level of DSPP and osteocalcin transcripts than ADH-DPSCs.Conclusion: Collectively, these data indicate that human DPSCs can be expanded and cultured in serum-free supplemented medium with EGF and bFGF. ADH-DPSCs and non-ADH populations contained neuronal and/or oligodendrocyte progenitors at different stages of commitment and, interestingly, cells from spheroid structures seem to be more engaged into the odontoblastic lineage than the ADH-DPSCs.