A novel activating function of c-Src and Stat3 on HGF transcription in mammary carcinoma cells.

In the normal breast, hepatocyte growth factor (HGF) is primarily expressed by stromal cells, and stimulates in a paracrine manner epithelial cells expressing the HGF receptor (Met). In invasive human breast carcinomas, HGF and Met are frequently overexpressed, possibly establishing an autocrine HGF/Met loop that promotes tumour cell invasion. However, the mechanisms leading to autocrine HGF expression in carcinoma cells are not known. We previously demonstrated a cooperative effect between c-Src and Stat3 in the activation of HGF transcription in mammary carcinoma cells. The present report defines a novel Stat3 consensus site at nt -95 in the HGF promoter that is highly conserved in human and mouse, and is required for c-Src and Stat3 to activate HGF transcription in breast epithelial cells. DNA-protein binding studies demonstrated high affinity binding of a Stat3-containing complex to the nt -95 site. Endogenous Stat3 binding to this region of the HGF promoter in carcinoma cells expressing HGF was demonstrated using a chromatin immunoprecipitation assay. In addition, coexpression of Stat3 and activated c-Src caused increased expression of endogenous HGF mRNA and protein and marked cell scattering in breast epithelial cells. Our results delineate a novel c-Src/Stat3-dependent mechanism that regulates HGF promoter activity, and is linked to transformation of mammary epithelial cells.

Cross-hybridization of closely related genes on high-density macroarrays.

DNA macroarrays are used in many areas of molecular biology research for applications ranging from gene discovery to gene expression profiling. As an increasing number of specialized macroarrays containing genes related by function or pathway are becoming available, a question that needs to be addressed is the level of hybridization signal specificity between highly similar genes that can be achieved. We have examined the ability of our LifeGrid macroarrays to distinguish hybridization signals between closely related genes. We determined the level of cross-hybridization among genes ranging from 52% to 94% sequence identity. Fragments of genes fromfive protein families were arrayed onto nylonfilters. Thefilters were subsequently hybridized with a 33P-labeled probe prepared from a pool of synthetic mRNA transcripts containing a representative of each protein family. We found that fragments containing sequences with up to 94% sequence identity displayed relatively little cross-hybridization. We conclude that this macroarray system is very specific and that hybridization signals from closely related genes can be reliably measured.

Synthetic extracellular matrix supports healing of mucogingival surgery donor sites.

Palatal donor sites are commonly used in periodontal surgery. Their healing speed determines the patient's comfort. This may be enhanced by the use of collagen sponges, supplemented with various components of the extracellular matrix, which display chemotactic properties and enhance the proliferation and synthesis activity of fibroblasts. A collagen-based dressing supplemented with such extracellular matrix components, including chondroitin 4 sulfate, heparan sulfate, and fibronectin was used, in an attempt to facilitate the healing of donor sites in 10 periodontal patients. Immunohistological techniques were used on biopsy samples from the margin of these volunteer's healing donor sites to appreciate tissue reconstruction around the synthetic material. Our results indicate a fast epithelial growth, neovascularization, and spatial organization of the new matrix. Limited and topographically selective inflammatory reactions, characterized by polymorphonuclears and mononuclear cells seen near the wound's margin, could be responsible for the production of soluble factors supporting reconstruction.

Successful treatment of maternal septicemia due to Listeria monocytogenes at 26 weeks' gestation.

A case of successful therapy with ampicillin of maternal septicemia due to Listeria monocytogenes at 26 weeks' gestation with resultant delivery at term of a normal neonate is reported. The ability to positively influence the outcome of hematogenously acquired maternal-fetal infection is stressed.

Medicaid 2176 home and community-based care waivers: the first ten years.

Budget reconciliation legislation in 1981 created a waiver program whereby states could provide home and community-based care under Medicaid for certain populations. States spent nearly $1.7 billion on services delivered under these waivers, known as 2176 waivers, in 1991, compared with $3.8 million in 1982, when only six states were participating in the program. Although these programs have not been rigorously evaluated for effectiveness, they continue to be a popular approach to delivering care outside of institutions for various groups. Across all states, states spent 13.4 percent of their Medicaid long-term care dollars on care outside of institutions (which includes the 2176 waiver program).

Use of Medicaid 1915(c) home- and community-based care waivers to reconfigure state long-term care systems.

Since Congressional authorization in 1981, Medicaid 1915(c) home- and community-based care waivers have influenced states' efforts to transform their long-term care systems. In 1997, every state participated in the 1915(c) waiver program, while waiver expenditures, at $8.1 billion, represented 59.6 percent of all Medicaid community-based care expenditures. To explore state-level factors that appear related to these expenditures, the authors turn to a body of work on Medicaid resource allocation. They compare the influence of five factors--sociodemographic, supply, economic, programmatic, and political environment--on states' allocations to long-term care expenditures and 1915(c) waiver expenditures. The state economic environment was an important influence on total, as well as waiver expenditures. State regulation of long-term care supply demonstrated the most substantive relationship, increasing the share of dollars supporting 1915(c) waivers from 11.6 to 20.0 over the study period, all else equal.

Patient centered long-term care.

Drawing upon an individual's needs, values, and expectations to guide decisionmaking and care giving is integral to long-term care (LTC). Articles in this issue demonstrate that client values and preferences can be elicited and used to guide decisionmaking about LTC. Service delivery and payment features can be shaped to support the patient/consumer, as well as to support and strengthen her or his informal caregivers. Significant constraints to making LTC more client centered are also identified. Key issues relate to the availability of and methods to process information as well as pressures on provider staff that impede their ability to support clients and their families. More broadly, access to appropriate LTC services is being shaped by programmatic shifts and legal forces that may enhance or impede the ability to place patients/clients at the center of LTC.

A profile of home health users in 1992.

Recently, the use of home health services by Medicare beneficiaries has been growing. From 1987 to 1992, the percentage of all enrollees receiving home health rose from 4.8 to 7.2 percent, while the average number of visits among users increased from 23 to 54. This article uses the 1992 Medicare Current Beneficiary Survey (MCBS) to profile home health users. In addition to providing descriptive information about who uses Medicare home health, Tobit models are estimated to determine the factors that predict home health utilization and reimbursement. Various policy options for redesigning the home health benefit are also discussed.

The Medicare home health initiative.

This article describes the Medicare home health benefit and summarizes growth and change in the use of the benefit and in the industry providing home health care. The article also details the organization and goals of the Home Health Initiative, describes its four key components--quality assurance (QA), administration, policy, and research-and concludes with a discussion of the status of the Initiative.

Trends and issues in the Medicaid 1915(c) waiver program.

Over the past 15 years, Medicaid 1915(c) home and community-based waivers have made a substantial contribution to States' efforts to transform their long-term care (LTC) systems from largely institutional to community-based systems. By 1997, every State had implemented a waiver program for at least some subgroups of individuals with disabilities, and expenditures increased from $3.8 million in 1982 to more than $8.1 billion in 1997. Emerging, as well as long-standing, policy issues related to the waiver program include concerns with access, variation in availability by disability group, State decisions related to the provision of community-based LTC, and evidence on effectiveness.

The changing face of long-term care.

In this article, we present population estimates of individuals with disabilities and discuss the manner in which the composition of this population is changing. We then highlight aspects of service delivery systems that are evolving in response to the changing long-term care (LTC) population. Following a summary of financing issues, we discuss several cross-cutting issues related to the organization of service delivery, quality assurance (QA), and financing. Current and future Health Care Financing Administration (HCFA) research and demonstrations emerging from these issues are then described.

Qualitative analysis of the Program of All-inclusive Care for the Elderly (PACE).

The innovative model of capitated acute and chronic care for nursing-home-eligible elderly persons, which was developed at On Lok in San Francisco's Chinatown and stresses using community care in lieu of institutional care, has been replicated at eight sites around the country. The early experience in developing these sites tests the extent to which the principles of this approach, based on day health care, can be reproduced in a variety of other settings. Four of the eight sites have begun formal capitated care; enrollment has been less active than anticipated, apparently due to reluctance to change physicians and resistance to day care.

Analysis of the Community Periodontal Index of Treatment Needs--study on adults in France. IV. The significance of gingival recession.

A total of 1005 persons were examined using the CPITN criteria which were recorded for every tooth. All the teeth were also measured on both their buccal and lingual aspects to assess the amount of gingival recession. The combination of pocket depth and gingival recession was computed using a specially written program: 93.8 per cent of the teeth had 1 mm or less gingival recession; 82.5 per cent of the teeth with gingival recession did not present pockets; 26.6 per cent of all subjects had at least one tooth with gingival recession of 2 mm or more but only 9.9 per cent had at least one tooth with 2 mm or more gingival recession and a periodontal pocket.

An analysis of the community periodontal index of treatment needs. Studies on adults in France. V. Presentation of CPITN data in cross-tabulations.

A total of 1005 subjects were examined using the CPITN and DFT indices. CPITN data were modified in their presentation in order to be suitable for cross-tabulation. Two different methods were considered. One, described previously by Roland et al. (1984), classified subjects according to the combination of their highest CPITN score and the mean of the highest scores of every nonedentulous sextant. The other consisted of multiplying the above mentioned mean by the individual's highest CPITN score. A critical analysis of the methodology showed that the two number system developed by Roland et al. (1984) was impractical. The new method provides a linear array of values from 0 to 16. Very little overlapping of values was observed, which means that a relatively precise periodontal condition is characterised by each value. With this transformation the CPITN can be used as an index that quantifies periodontal conditions instead of treatment needs.

An analysis of the Community Periodontal Index of Treatment Needs. Studies on adults in France. III--Partial examinations versus full-mouth examinations.

1005 persons were examined using the Community Periodontal Index of Treatment Needs. Presence or absence of every clinical sign was registered for each tooth. A specially written program computed CPITN values corresponding to full mouth or partial examinations. Results compare the prevalence and treatment needs obtained through either full mouth or partial examinations. Cross tabulation analysis of the number of cases detected by full mouth and partial examinations shows differences in detection rates of 23.5 per cent for deep pockets, 17.6 per cent for moderate pockets, 13.4 per cent for calculus, 3.2 per cent for bleeding and 53.0 per cent for health. When determining treatment needs using the partial examination procedure approximations of the number of subjects needing hygiene education and scaling are acceptable but the evaluation of the number of individuals needing complex therapy is greatly underestimated. The CPITN is most accurate using full mouth examinations.

Antibody to Coxsackie B virus in diagnosing postviral fatigue syndrome.

OBJECTIVE: To study the association between coxsackie B virus infection and the postviral fatigue syndrome and to assess the immunological abnormalities associated with the syndrome. DESIGN: Case-control study of patients with the postviral fatigue syndrome referred by local general practitioners over one year. SETTING: General practitioner referrals in Dunbartonshire, Scotland. PATIENTS: 254 Patients referred with the postviral fatigue syndrome (exhaustion, myalgia, and other symptoms referable to postviral fatigue syndrome of fairly recent onset--that is, several months) and age and sex matched controls obtained from same general practitioner; 11 patients were rejected because of wrong diagnoses, resolution of symptoms, and refusal to participate, leaving 243 patients and matched controls. MAIN OUTCOME MEASURES: Detailed questionnaire (patients and controls) and clinical examination (patients) and blind analysis of blood sample at entry and after six months for determination of coxsackie B virus IgM and IgG antibodies and other variables (including lymphocyte protein synthesis, lymphocyte subsets, and immune complexes). RESULTS: Percentage positive rates for coxsackie B virus IgM at entry were 24.4% for patients and 22.6% for controls and for coxsackie B virus IgG 56.2% and 55.3% respectively; there were no significant differences between different categories of patients according to clinical likelihood of the syndrome nor any predictive value in a fourfold rise or fall in the coxsackie B virus IgG titre in patients between entry and review at six months. The rates of positive antibody test results in patients and controls showed a strong seasonal variation. Of the numerous immunological tests performed, only a few detected significant abnormalities; in particular the mean value for immune complex concentration was much higher in 35 patients and 35 controls compared with the normal range and mean value for total IgM was also raised in 227 patients and 35 controls compared with the normal range. CONCLUSIONS: Serological tests available for detecting coxsackie B virus antibodies do not help diagnose the postviral fatigue syndrome. Percentage positive rates of the antibodies in patients simply reflect the background in the population as probably do the raised concentrations of total IgM and immune complexes.

Joining forces to improve point-of-care testing.

Point-of-care (POC) testing takes a "laboratory" test directly to the patient's bedside. Although bedside testing is not new, the compliance standards are relatively new. Regulations ensure that all personnel are trained to properly perform the test. The central laboratory and nursing of this hospital merge the best skills of both units to create a successful POC testing program.

Exposure of prepubertal beef bulls to cycling females does not enhance sexual development.

The objective of this study was to determine whether continuous, long-term, fenceline exposure of prepubertal beef bulls to cycling beef females reduced age at puberty and influenced the percentage of bulls that passed an initial breeding soundness examination (BSE). Bulls (Angus, n = 37; Simmental, n = 22; Hereford, n = 10; Simmental × Angus, n = 8) at an average age of 202 ± 21.5 days were given either continuous fenceline and visual exposure to cycling females (exposed, n = 41) or no exposure (control, n = 36). Estrus was induced in cycling beef females so at least three females were in standing estrus each week during the 182 days of exposure to bulls. Scrotal circumference (SC), body weight, and blood samples were collected every 28 days. When bulls had SC of 26 cm or more, semen samples were obtained monthly via electroejaculation until puberty was achieved (≥50 × 10(6) sperm/mL with at least 10% progressive motility). Behavioral observations were conducted twice monthly: once when females were in estrus and once during diestrus. Homosexual mounting, flehmen responses, and number of times near penned females were recorded for each observation period. Breeding soundness examinations were conducted when the average age of bulls was 364 ± 21.5 days. Normal sperm morphology of at least 70% and sperm motility of at least 30% were required to pass the BSE. Age, body weight, and SC at puberty did not differ between exposed and control bulls (320 ± 28 and 311 ± 29 days; 466.2 ± 12.2 and 437.7 ± 13.5 kg; and 34.4 ± 2.5 and 34.9 ± 2.5 cm, respectively). Percentage of bulls passing their initial BSE did not differ between treatments (exposed, 87.8%; control, 75.0%). Treatment, month, and female estrous stage interacted (P = 0.05) to affect the number of mount attempts and flehmen responses. Exposed bulls entered the cow area more times (P < 0.001) during estrus than diestrus in Months 1, 2, and 3. We concluded that bulls given continuous, long-term, fenceline exposure to cycling beef females do not have enhanced sexual development.

Role of Nek2 on centrosome duplication and aneuploidy in breast cancer cells.

Breast cancer is the most common solid tumor and the second most common cause of death in women. Despite a large body of literature and progress in breast cancer research, many molecular aspects of this complex disease are still poorly understood, hindering the design of specific and effective therapeutic strategies. To identify the molecules important in breast cancer progression and metastasis, we tested the in vivo effects of inhibiting the functions of various kinases and genes involved in the regulation/modulation of the cytoskeleton by downregulating them in mouse PyMT mammary tumor cells and human breast cancer cell lines. These kinases and cytoskeletal regulators were selected based on their prognostic values for breast cancer patient survival. PyMT tumor cells, in which a selected gene was stably knocked down were injected into the tail veins of mice, and the formation of tumors in the lungs was monitored. One of the several genes found to be important for tumor growth in the lungs was NIMA-related kinases 2 (Nek2), a cell cycle-related protein kinase. Furthermore, Nek2 was also important for tumor growth in the mammary fat pad. In various human breast cancer cell lines, Nek2 knockdown induced aneuploidy and cell cycle arrest that led to cell death. Significantly, the breast cancer cell line most sensitive to Nek2 depletion was of the triple negative breast cancer subtype. Our data indicate that Nek2 has a pivotal role in breast cancer growth at primary and secondary sites, and thus may be an attractive and novel therapeutic target for this disease.