A Nuclear and Cytoplasmic Characterization of Bovine Oocytes Reveals That Cysteamine Partially Rescues the Embryo Development in a Model of Low Ovarian Reserve.

Decreased oocyte quality is a major determinant of age-associated fertility decline. Similarly, individuals affected by early ovarian aging carry low-quality oocytes. Using an established bovine model of early ovarian aging, we investigated key features of 'quality' oocyte maturation, associated with the onset of egg aneuploidy and reproductive aging, such as histone modifications, mitochondria distribution and activity, reduced glutathione (GSH) content, and gap junction functionality. Bovine ovaries were classified according to the antral follicle count (AFC), and the retrieved oocytes were processed immediately or matured in vitro. We observed alterations in several cellular processes, suggesting a multifactorial etiology of the reduced oocyte quality. Furthermore, we performed a rescue experiment for one of the parameters considered. By adding cysteamine to the maturation medium, we experimentally increased the free radical scavenger ability of the 'low competence' oocytes and obtained a higher embryo development. Our findings show that adopting culture conditions that counteract the free radicals has a positive impact on the quality of 'compromised' oocytes. Specifically, cysteamine treatment seems to be a promising option for treating aging-related deficiencies in embryo development.

Rationale and pre-clinical evidences for the use of autologous cartilage micrografts in cartilage repair.

BACKGROUND: The management of cartilage lesions is an open issue in clinical practice, and regenerative medicine represents a promising approach, including the use of autologous micrografts whose efficacy was already tested in different clinical settings. The aim of this study was to characterize in vitro the effect of autologous cartilage micrografts on chondrocyte viability and differentiation and perform an evaluation of their application in racehorses affected by joint diseases. MATERIALS AND METHODS: Matched human chondrocytes and micrografts were obtained from articular cartilage using Rigenera® procedure. Chondrocytes were cultured in the presence or absence of micrografts and chondrogenic medium to assess cell viability and cell differentiation. For the pre-clinical evaluation, three racehorses affected by joint diseases were treated with a suspension of autologous micrografts and PRP in arthroscopy interventions. Clinical and radiographic follow-ups were performed up to 4 months after the procedure. RESULTS: Autologous micrografts support the formation of chondrogenic micromasses thanks to their content of matrix and growth factors, such as transforming growth factor ß (TGFß) and insulin-like growth factor 1 (IGF-1). On the other hand, no significant differences were observed on the gene expression of type II collagen, aggrecan, and SOX9. Preliminary data in the treatment of racehorses are suggestive of a potential in vivo use of micrografts to treat cartilage lesions. CONCLUSION: The results reported in this study showed the role of articular micrografts in the promoting chondrocyte differentiation suggesting their potential use in the clinical practice to treat articular lesions.

Ultra-low Doses of Follicle Stimulating Hormone and Progesterone Attenuate the Severity of Polycystic Ovary Syndrome Features in a Hyperandrogenized Mouse Model.

BACKGROUND: Polycystic-ovary syndrome (PCOS) is a reproductive illness characterized by hyperandrogenism and anovulation. Using hyperandrogenized mice, it was demonstrated that the oral administration of incremental dose of follicle stimulating hormone (FSH) attenuated some of PCOS characteristics. This work aimed to study the effect of ultra-low doses of combined FSH and progesterone orally administered on PCOS murine model. Moreover, the effect of sequential kinetic activation of administered hormones was tested. METHODS: Thirty-two female mice were used as animal model (four groups of eight animals each). Mice were hyperandrogenized by injection of dehyidroepiandrosterone diluted in sesame oil. Control group received only oil. Simultaneously, each animal daily received per os an activated or a not-activated combination of FSH (0.44 pg) plus progesterone (0.44 pg) or saline solution as control. Serum testosterone, estradiol, progesterone and luteinizing hormone were analyzed as endocrine markers and a morphological study of antral follicle was conducted. Data were analyzed by one-way ANOVA, followed by multiple comparison test. The p<0.05 was considered significant. RESULTS: Dehyidroepiandrosterone treatment increased both estradiol and progesterone serum levels, besides testosterone, while reduced luteinizing hormone (p<0.05); histological examination revealed an increase of cystic follicles (p<0.05). Irrespective of activation, the combined FSH and progesterone treatments restored estradiol level (p>0.05 vs. control group) and reduced cystic signs in the follicles (p<0.05 vs. dehyidroepiandrosterone treatment). CONCLUSION: This study indicate that ultra-low doses of FSH and progesterone orally administrated can reduce the sternness of PCOS in the mouse model and open a route for the study of innovative approaches for PCOS treatment.

Age-related modulation of angiogenesis-regulating factors in the swine meniscus.

An in-depth knowledge of the native meniscus morphology and biomechanics in its different areas is essential to develop an engineered tissue. Meniscus is characterized by a great regional variation in extracellular matrix components and in vascularization. Then, the aim of this work was to characterize the expression of factors involved in angiogenesis in different areas during meniscus maturation in pigs. The menisci were removed from the knee joints of neonatal, young and adult pigs, and they were divided into the inner, intermediate and outer areas. Vascular characterization and meniscal maturation were evaluated by immunohistochemistry and Western blot analysis. In particular, expression of the angiogenic factor Vascular Endothelial Growth Factor (VEGF) and the anti-angiogenic marker Endostatin (ENDO) was analysed, as well as the vascular endothelial cadherin (Ve-CAD). In addition, expression of Collagen II (COLL II) and SOX9 was examined, as markers of the fibro-cartilaginous differentiation. Expression of VEGF and Ve-CAD had a similar pattern in all animals, with a significant increase from the inner to the outer part of the meniscus. Pooling the zones, expression of both proteins was significantly higher in the neonatal meniscus than in young and adult menisci. Conversely, the young meniscus revealed a significantly higher expression of ENDO compared to the neonatal and adult ones. Analysis of tissue maturation markers showed an increase in COLL II and a decrease in SOX9 expression with age. These preliminary data highlight some of the changes that occur in the swine meniscus during growth, in particular the ensemble of regulatory factors involved in angiogenesis.

Effect of oral administration of low-dose follicle stimulating hormone on hyperandrogenized mice as a model of polycystic ovary syndrome.

BACKGROUND: Polycystic Ovary Syndrome (PCOS) is a widespread reproductive disorder characterized by a disruption of follicular growth and anovulatory infertility. In women with PCOS, follicular growth and ovulation can be induced by subcutaneous injections of low doses of follicle stimulating hormone (FSH). The aim of this study was to determine the effect of oral administration of recombinant human FSH (rhFSH) on follicle development in a PCOS murine model. Moreover, since it is unlikely that intact rhFSH is present into the circulation after oral administration, the biological activity of a peptide fragment, derived from the predicted enzymatic cleavage sites with the FSH molecule, was investigated in vitro on cumulus-enclosed oocytes (COCs). METHODS: Female peripubertal mice were injected with dehydroepiandrosterone (DHEA) diluted in sesame oil for 20 consecutive days and orally treated with a saline solution of rhFSH. A control group received only sesame oil and saline solution. At the end of treatments, blood was analyzed for hormone concentrations and ovaries were processed for morphological analysis. The presumptive bioactive peptide was added during in vitro maturation of bovine COCs and the effects on cumulus expansion and on maturation rate were evaluated. RESULTS: DHEA treatment increased serum levels of testosterone, estradiol and progesterone as well as the percentage of cystic follicles. Orally administered rhFSH restored estradiol level and reduced the percentage of cystic follicles. Despite these results indicating a reduction of the severity of PCOS in the mouse model, the presumptive bioactive peptide did not mimic the effect of rhFSH and failed to induce bovine cumulus expansion and oocyte maturation in vitro. CONCLUSIONS: Although further studies are needed, the present data supports the concept that orally administrated FSH could attenuate some of the characteristic of PCOS in the mouse model.

The adenosine salvage pathway as an alternative to mitochondrial production of ATP in maturing mammalian oocytes.

Although the oocyte is the largest cell in the body and an unavoidable phase in life, its physiology is still poorly understood, and other cell types provide little insight into its unique nature. Even basic cellular functions in the oocyte such as energy metabolism are not yet fully understood. It is known that the mitochondria of the female gamete exhibit an immature form characterized by limited energy production from glucose and oxidative phosphorylation. We show that the bovine oocyte uses alternative means to maintain ATP production during maturation, namely, the adenosine salvage pathway. Meiosis resumption is triggered by destruction of cyclic AMP by phosphodiesterases producing adenosine monophosphate that is converted into ATP by adenylate kinases and creatine kinases. Inhibition of these enzymes decreased ATP production, and addition of their substrates restored ATP production in denuded oocytes. Addition of phosphocreatine to the oocyte maturation medium influenced the phenotype of the resulting blastocysts. We propose a model in which adenylate kinases and creatine kinases act as drivers of ATP production from added AMP during oocyte maturation.

Reductions in the number of mid-sized antral follicles are associated with markers of premature ovarian senescence in dairy cows.

High-producing dairy cows are subfertile; however, the mechanisms responsible for the decreased fertility are unknown. The aim of the present study was to test the hypothesis that culled dairy cows (4-8 years old) characterised by 'Lo' ovaries (i.e. those with <10 mid-antral follicles) are affected by premature ovarian senescence. Cows in which both ovaries were 'Lo' ovaries represented 5% of the total population analysed, and exhibited reduced ovarian size (P<0.001) and increased perifollicular stroma (P<0.05) compared with age-matched controls (i.e. cows in which both ovaries had >10 mid-antral follicles; 'Hi' ovaries). The total number of follicles, including healthy and atretic primordial, primary, secondary and small antral follicles, was lower in Lo ovaries (P<0.01). Interestingly, the primordial follicle population in Lo ovaries was lower (P<0.05) than in the control. Finally, the follicular fluid of mid-antral follicles from Lo ovaries had reduced oestradiol and anti-Müllerian hormone levels (P<0.05), but increased progesterone concentrations (P<0.05). Together, these data account for the reduced fertility of cows with Lo ovaries and are in agreement with previous observations that oocytes isolated from Lo ovaries have reduced embryonic developmental competence. Cows with a specific Lo ovary condition may represent a suitable model to address the causes of low fertility in high-yielding dairy cows, as well as the condition of premature ovarian aging in single-ovulating species.

Process of diffusing cancer survivorship care into oncology practice.

The LIVESTRONG Centers of Excellence were funded to increase the effectiveness of survivorship care in oncology practice. This study describes the ongoing process of adopting and implementing survivorship care using the framework of the diffusion of innovation theory of change. Primary data collection included telephone interviews with 39 members from the eight centers and site visits. Organizational characteristics, overall progress, and challenges for implementation were collected from proposals and annual reports. Creating an awareness of cancer survivorship care was a major accomplishment (relative advantage). Adoption depended on the fit within the cancer center (compatibility), and changed over time based on trial and error (trialability). Implementing survivorship care within the existing culture of oncology and breaking down resistance to change was a lengthy process (complexity). Survivorship care became sustainable as it became reimbursed, and more new patients were seen (observability). Innovators and early adopters were crucial to success. Diffusion of innovation theory can provide a strategy to evaluate adoption and implementation of cancer survivorship programs into clinical practice.

The effect of cilostamide on gap junction communication dynamics, chromatin remodeling, and competence acquisition in pig oocytes following parthenogenetic activation and nuclear transfer.

In the pig, the efficiency of in vitro embryo production and somatic cell nuclear transfer (SCNT) procedures remains limited. It has been suggested that prematuration treatments (pre-IVM) based on the prolongation of a patent, bidirectional crosstalk between the oocyte and the cumulus cells through gap junction mediate communication (GJC), with the maintenance of a proper level of cAMP, could improve the developmental capability of oocytes. The aim of this study was to assess: 1) dose-dependent effects of cilostamide on nuclear maturation kinetics, 2) the relationship between treatments on GJC functionality and large-scale chromatin configuration changes, and 3) the impact of treatments on developmental competence acquisition after parthenogenetic activation (PA) and SCNT. Accordingly, cumulus-oocyte complexes were collected from 3- to 6-mm antral follicles and cultured for 24 h in defined culture medium with or without 1 µM cilostamide. GJC functionality was assessed by Lucifer yellow microinjection, while chromatin configuration was evaluated by fluorescence microscopy after nuclear staining. Cilostamide administration sustained functional coupling for up to 24 h of culture and delayed meiotic resumption, as only 25.6% of cilostamide-treated oocytes reached the pro-metaphase I stage compared to the control (69.7%; P < 0.05). Moreover, progressive chromatin condensation was delayed before meiotic resumption based upon G2/M biomarker phosphoprotein epitope acquisition using immunolocalization. Importantly, cilostamide treatment under these conditions improved oocyte developmental competence, as reflected in higher blastocyst quality after both parthenogenetic activation and SCNT.

Adult cancer survivorship care: experiences from the LIVESTRONG centers of excellence network.

BACKGROUND: The objectives of this study were to characterize survivorship models of care across eight LIVESTRONG Survivorship Center of Excellence (COE) Network sites and to identify barriers and facilitators influencing survivorship care. METHODS: Using the framework of the Chronic Care Model (CCM), quantitative and qualitative methods of inquiry were conducted with the COEs. Methods included document reviews, key informant telephone interviews with 39 participants, online Assessment of Chronic Illness Care (ACIC) surveys with 40 participants, and three site visits. RESULTS: Several overarching themes emerged in qualitative interviews and were substantiated by quantitative methods. Health system factors supporting survivorship care include organization and leadership commitment and program champions at various levels of the health care team. System barriers include reimbursement issues, lack of space, and the need for leadership commitment to support changes in clinical practices as well as having program "champions" among clinical staff. Multiple models of care include separate survivorship clinics and integrated models as well as consultative models. COEs' scores on the ACIC survey showed overall "reasonable support" for survivorship care; however, the clinical information system domain was least developed. Although the ACIC findings indicated "reasonable support" for self-management, the qualitative analysis revealed that self-management support was largely limited to health promotion provided in clinic-based education and counseling sessions, with few COEs providing patients with self-management tools and interventions. CONCLUSIONS: The CCM framework captured experiences and challenges of these COEs and provided insight into the current state of survivorship care in the context of National Cancer Institute-designated comprehensive cancer centers. Findings showed that cancer patients and providers could benefit from clinical information systems that would better identify candidates for survivorship care and provide timely information. In addition, a crucial area for development is self-management support outside of clinical care. IMPLICATIONS FOR CANCER SURVIVORS: Cancer survivors may benefit from learning about the experience and challenges faced by the eight LIVESTRONG Centers of Excellence in developing programs and models for cancer survivorship care, and these findings may inform patient and caregiver efforts to seek, evaluate, and advocate for quality survivorship programs designed to meet their needs.

Self-reported and clinical measurement of three chronic disease risks among low-income women in West Virginia.

BACKGROUND: This study assessed the validity of several self-reported cardiovascular risk factors among low-income women aged 40-64 years in West Virginia. METHODS: A cross-sectional survey was conducted of 733 women participating in the Well Integrated Screening and Evaluation for Women Across the Nation (WISEWOMAN) project in West Virginia to examine agreement between self-report and clinical screenings in the prevalence of risk factors related to coronary heart disease (CHD). Women participating in the study were interviewed face-to-face before administration of clinical screenings that assessed height, weight, Quetelet's index, high blood pressure (systolic > or =140 mm Hg or diastolic > or =90 mm Hg), and elevated total cholesterol concentrations (> or =200 mg/dL and > or =240 mg/dL). RESULTS: The overall results showed high sensitivity and specificity for each of the risk factors examined; for overweight/obesity, the sensitivity was 96% and specificity was 93%; for cholesterol > or =240 mg/dL, sensitivity was 85% and specificity was 67%; for hypertension, sensitivity was 77% and specificity was 86%. Using a threshold value of > or =240 mg/dL for hypercholesterolemia led to higher sensitivity but a lower specificity than for a value of > or =200 mg/dL. CONCLUSIONS: This study found that among low-income women at higher risk for cardiovascular disease (CVD), self-reported values for high body mass index (BMI), hypercholesterolemia, and hypertension were well correlated with clinical measures, as indicated by high sensitivity values. Thus, self-reported values can be used for surveillance, targeted screenings, and health promotion activities, including lifestyle changes.

Isolation, growth and differentiation of equine mesenchymal stem cells: effect of donor, source, amount of tissue and supplementation with basic fibroblast growth factor.

Mesenchymal stem cells (MSC) are increasingly used as therapeutical aid for the orthopaedic injuries in the horse. MSC populate different tissues but the most commonly used for clinical purposes are isolated from bone marrow or adipose tissue. The first objective of this study was to investigate if the donor animal, the tissue of origin and the technique of isolation could influence the number of MSC available for transplantation after a short-term expansion. The second aim was to devise a culture system capable of increasing MSC lifespan and we tested the effect of basic fibroblast growth factor (bFGF). Results indicate that MSC can be efficiently isolated from both sources and supplementation of bFGF enhances proliferation rate maintaining differentiation potential. In addition, this study shows that collection, expansion and storage of frozen MSC can be performed for later therapeutic use.

Perceived barriers to physical activity according to stage of change and body mass index in the west virginia wisewoman population.

PURPOSE: Low-income Appalachian women aged 40-64 years are an understudied population relative to physical activity barriers. To inform a cardiovascular risk factor intervention targeting this population through the West Virginia WISEWOMAN program, we determined at baseline participants' perceptions about the presence of 6 barriers to physical activity and respective differences according to stage of readiness for change (SOC). METHODS: Low-income women aged 40-64 who had participated in a state cancer screening program were recruited through 13 clinics. Participants were interviewed at clinics using a health risk/behavioral survey that included questions about barriers to and SOC for physical activity. Weight, height, and waist circumference were measured. PRINCIPAL FINDINGS: Baseline screening was completed by 733 women; 84% were overweight (body mass index [BMI] 25-29.9 [24.1%]) or obese (BMI > or = 30 [59.8%]). The prevalence of barriers was greatest for lack of support (52.4%) and lack of willpower (50.8%); lack of time was reported by only 30.1%. On chi(2) test, significant differences were revealed across SOC for time (p = .026), don't need more physical activity (p = .003), and willpower (p < 001); and for time (p = .006) and don't need more (p < .000) across BMI categories. Time increased from precontemplation (21.1%) to maintenance (35%). Obese participants reported time less frequently (25.7%) than participants of normal weight (40.2%). CONCLUSIONS: Lack of willpower and support-not time-seem to be the dominant physical activity barriers in the West Virginia WISEWOMAN population. Interventions targeting willpower in early SOC and in obese participants may be promising toward increasing physical activity.

Knowledge, barriers, and predictors of colorectal cancer screening in an Appalachian church population.

INTRODUCTION: This study examined knowledge about and barriers to colorectal cancer screening and predictors of screening adherence among members from 16 Appalachian churches as part of a larger study on the prevention and early detection of colorectal cancer. METHODS: Baseline data were collected on 839 respondents aged 50 years and older through a self-administered survey, and 23 focus groups were conducted with 205 church members RESULTS: Survey results showed that older age, male sex, being current for other cancer screening, being physically active, having perceived support from others for screening, better provider communication, knowledge about screening guidelines, greater perceived susceptibility to colorectal cancer, and a family history of the disease were predictors of screening adherence. Major barriers to screening in both surveys and focus groups were failure of providers to recommend screening, lack of knowledge about the need for screening, and the belief that screening was not necessary without symptoms. Fear of cancer, lack of knowledge about screening methods other than colonoscopy, reliance on physicians for screening information, and the need for people to feel at risk for screening to occur were other findings from the focus groups. Focus groups supported survey findings and provided further insights. CONCLUSION: Several factors predictive of colorectal cancer screening in this study can be modified through educational interventions. Recognizing and changing risk factors for colorectal cancer, raising awareness of screening guidelines, and encouraging adults aged 50 years and older to discuss screening with their health care provider could increase colorectal cancer screening.

CDC-funded intervention research aimed at promoting colorectal cancer screening in communities.

BACKGROUND: Although strong scientific evidence has shown that screening for colorectal cancer saves lives, most U.S. adults who are at the recommended age are not being screened. Prior studies suggest that barriers to routine screening vary by race, ethnicity, socioeconomic status, urban/rural residence, health insurance status, and factors related to health care providers and the health care environment. Relatively few studies, however, have identified and tested intervention approaches to promote routine colorectal cancer screening among diverse populations. METHODS: The Division of Cancer Prevention and Control at CDC has funded ongoing projects to develop and test interventions to promote routine colorectal cancer screening among medically underserved populations in Appalachia, the Lower Rio Grande Valley in Texas, the High Plains region of Colorado, and other U.S. communities. RESULTS: This article provides an overview of colorectal cancer screening intervention studies currently funded by CDC that focus on a wide range of populations, including medically underserved persons who live in predominately rural areas, Hispanic and non-Hispanic persons, urban African Americans, persons with low health literacy, and persons enrolled in managed care organizations. CONCLUSIONS: These CDC-funded intervention research projects are likely to contribute importantly to evidence about what works to promote colorectal cancer screening in diverse U.S. communities. .

Correlates of physical activity in a community sample of older adults in Appalachia.

Although much has been learned about the global determinants of physical activity in adults, there has been a lack of specific focus on gender, age, and urban/rural differences. In this church-based community sample of Appalachian adults (N = 1,239), the primary correlates of physical activity included age, gender, obesity, and self-efficacy. Overall, 42% of all participants and 31% of adults age 65 years or older met recommended guidelines for physical activity, which suggests that most participants do not engage in adequate levels of physical activity. Of participants who met physical activity guidelines, the most common modes of moderate and vigorous activity were walking briskly or uphill, heavy housework or gardening, light strength training, and biking. These particular activities that focus on building self-efficacy might be viable targets for intervention among older adults in rural communities.

Improving multiple behaviors for colorectal cancer prevention among african american church members.

The WATCH (Wellness for African Americans Through Churches) Project was a randomized trial comparing the effectiveness of 2 strategies to promote colorectal cancer preventive behaviors among 587 African American members of 12 rural North Carolina churches. Using a 2 X 2 factorial research design, the authors compared a tailored print and video (TPV) intervention, consisting of 4 individually tailored newsletters and targeted videotapes, with a lay health advisor (LHA) intervention. Results showed that the TPV intervention significantly improved (p <.05) fruit and vegetable consumption (0.6 servings) and recreational physical activity (2.5 metabolic task equivalents per hour) and, among those 50 and older (n = 287), achieved a 15% increase in fecal occult blood testing screening (p =.08). The LHA intervention did not prove effective, possibly because of suboptimal reach and diffusion.

Effects of a tailored health promotion program for female blue-collar workers: health works for women.

BACKGROUND: This study assessed the effects of the Health Works for Women (HWW) intervention on improving multiple behaviors including nutrition and physical activity among rural female blue-collar employees in North Carolina. METHODS: Nine small to mid-size workplaces were randomly assigned to either intervention or delayed intervention conditions. After a baseline survey, an intervention consisting of two computer-tailored magazines and a natural helpers program was conducted over 18 months. Delayed worksites received one tailored magazine. Approximately 77 and 76% of baseline respondents completed follow-up surveys at 6 and 18 months, respectively, and 538 women (63%) completed all three surveys. RESULTS: At the 18-month follow-up, the intervention group had increased fruit and vegetable consumption by 0.7 daily servings compared to no change in the delayed group (P < 0.05). Significant differences in fat intake were observed at 6 months (P < 0.05) but not at 18 months. The intervention group also demonstrated improvements in strengthening and flexibility exercise compared to the delayed group. The rates of smoking cessation and cancer screening did not differ between study groups. CONCLUSIONS: The HWW project was a successful model for achieving certain health behavior changes among blue-collar women.