'Odds Are: They Win': a disruptive messaging innovation for challenging harmful products and practices of the gambling industry.

OBJECTIVE: This paper presents an evidence informed rationale for focussing on harmful gambling products and industry practices in public health messaging through the example of a recent innovation called 'Odds Are: They Win'. METHODS: 'Odds Are: They Win' was initially developed through coproduction involving public health professionals and people with lived experience of gambling harms and implemented across a city-region area. A review of relevant evidence was undertaken, upon which the research team reflected to draw out the implications of 'Odds Are: They Win' for gambling harms messaging. RESULTS: Evidence is mounting that safer gambling campaigns framed in terms of individual responsibility are ineffective and can generate stigma. 'Odds Are: They Win' presents an alternative focus that is not anti-gambling but raises awareness of industry manipulation of the situational and structural context of gambling. This is in-keeping with historical lessons from the stop smoking field and emerging research in critical health literacy. The latter highlights the potential of education on the social and commercial determinants of health to stimulate behaviour change and collective action. CONCLUSION: 'Odds Are: They Win' is a potentially disruptive innovation for the gambling harms field. Research is required to robustly evaluate this intervention across diverse criteria, target audiences, and delivery settings.

Gene expression in a starch synthase IIa mutant of barley: changes in the level of gene transcription and grain composition.

The barley shrunken grain mutant M292 has a novel high-amylose starch phenotype caused by a mutation in the starch synthase IIa gene (SsIIa) located at the starch excess-6 (sex6) locus on chromosome 7H of barley. The loss of SSIIa enzyme activity leads to a decrease in amylopectin synthesis to less than 20% of the levels found in wild-type grains. Detailed composition analysis indicates that the contents of protein, non-starch polysaccharides, lipid, sucrose and hexoses, and fructo-oligosaccharides are increased in mature M292 grain compared to wild type. Using a microarray analysis, we characterize the differences between the transcription profiles of wild-type and mutant barley endosperms at mid-grain fill. The expression changes include genes involved in carbon storage, stress-related genes, and a number of transcripts with unassigned function. The changes in gene expression are discussed in terms of the altered grain composition of the mutant seed.

The effect of osmotic stress on the metaphase II spindle of human oocytes, and the relevance to cryopreservation.

BACKGROUND: Knowing osmotic tolerance limits is important in the design of optimal cryopreservation procedures for cells. METHODS: Mature human oocytes were exposed to anisosmotic sucrose solutions at concentrations of 35, 75, 150, 600, 1200, or 2400 (+/-5) milliosmolal (mOsm) at 37 degrees C. A control treatment at 290 mOsm was also utilized. Oocytes were randomly allocated to each experimental treatment. After the treatment, the oocytes were cultured for 1 h, then fixed in cold methanol. Immunocytochemical staining and fluorescence microscopy were used to assess the morphology of the metaphase II (MII) spindle. Logistic regression was used to determine if media osmolality had a significant effect on spindle structure. RESULTS: Osmolality was a significant predictor of spindle morphology. Hyposmotic effects at 35, 75, and 150 mOsm resulted in 100, 67, and 56% of oocytes having abnormal spindles, respectively. Hyperosmotic effects at 600, 1200, and 2400 mOsm resulted in 44, 44, and 100% of the spindles with abnormal structure, respectively. CONCLUSIONS: Anisosmotic conditions lead to disruption of the MII spindle in human oocytes. Applying this fundamental knowledge to human oocyte cryopreservation should result in increased numbers of cells maintaining viability.

The doctor on the patient's turf: assessing patient satisfaction with physician home visit programs.

We evaluated a physician home visit program (n = 23 patients) focusing on program implementation and quality. Quality was measured by evaluating patient satisfaction with services using a patient satisfaction scale and interviews with patients, caregivers, and providers. Scale results showed patients expressed the highest satisfaction with access to routine care and physician consideration. Patients expressed less satisfaction with access to emergency care and continuity of care. Physician communication and integration with home- and community-based service providers were other areas of concern. Recommendations include enhancing physician communication skills in the home, providing care for urgent medical conditions, improving chart documentation, and incorporating community-based chronic care experts into the program.

Difficulty or dependency? Effects of measurement scales on disability prevalence among older Americans.

This study examines how the prevalence of disability among subgroups of older persons is influenced by the activity of daily living (ADL) scale selected. Using the 1993 Survey of Asset and Income Dynamics of the Oldest Old, we construct four disability scales, ranging from difficulty performing an ADL to receiving help with an ADL, or dependency. In all instances, the difficulty scales produce substantially higher estimates of disability than the dependency scales. Larger percentages of women, Blacks, and less educated persons are classified disabled under all scales compared with men, Whites, and more educated persons. Specific subgroups are more sensitive to the scale used. The policy implications of using ADL scales to establish program eligibility are discussed.

Resource effects on access to long-term care for frail older people.

Access to long-term care depends primarily on personal resources, including family members and income, and on external resources, including Medicaid and Medicare. This study investigates how resources affect frail older individuals' access to long-term care, with a focus on Black and White widows. Data from the 1989 National Long-Term Care Survey is used, in conjunction with state-level Medicaid and Medicare reimbursement rates for nursing home and home health care, to estimate the likelihood of five types of care arrangements. Results show that children are a primary resource for unmarried individuals in maintaining access to informal care. Income effects are nonlinear in relation to nursing home care: increasing incomes below the mean income are associated with decreasing probabilities of nursing home care, while increasing incomes above the mean are associated with increasing probabilities of nursing home care. Income and Medicaid effects are interrelated, with nonlinearities associated with income having the potential to adversely affect some older persons' ability to access nursing home care.

Care arrangement decisions for frail older women: quantitative and qualitative perspectives.

This study uses both quantitative and qualitative methods to investigate how a range of care arrangement decisions for frail older unmarried women are made. Quantitative data from the 1989 National Long-Term Care Survey provides information concerning factors that predict the probability of five categories of care arrangements, including self only care, nursing home care, informal only care, formal only care, and a mix of formal and informal care. Qualitative interview data provides information on what occurs during care arrangement decision-making processes. Results are combined to explain the choice of care arrangements for a small sample of chronically disabled older unmarried women. Results show that need factors, such as age and disability, are strong predictors of the need for assistance. Family members played a central role in determining care arrangements and often helped an older woman to avoid an unwanted care arrangement. The use of a broad measure of impairment resulted in high levels of disability for the sample participants. Both disability status and care arrangements were transitory in nature.

Enhancing inter-network cooperation among organizations providing mental health services to older persons.

The authors examine how organizations in several networks voluntarily cooperate to provide mental health services to older persons and recommend how organizational coordination and the delivery system can be improved. Information from mail surveys and in-depth interviews was analyzed. Little evidence of formal coordination across networks was found. Established organizations and agencies providing a larger number of services were more integrated. Providers recommended that organizational coordination be increased. Providers suggested that a more centralized delivery system and proactive education for clients and providers would be effective strategies to enhance service delivery.

Chemical mechanism of DNA cleavage by the homing endonuclease I-PpoI.

Homing endonucleases are distinguished by their ability to catalyze the cleavage of double-stranded DNA with extremely high specificity. I-PpoI endonuclease, a homing endonuclease from the slime mold Physarum polycephalum, is a small enzyme (2 x 20 kDa) of known three-dimensional structure that catalyzes the cleavage of a long target DNA sequence (15 base pairs). Here, a detailed chemical mechanism for catalysis of DNA cleavage by I-PpoI endonuclease is proposed and tested by creating six variants in which active-site residues are replaced with alanine. The side chains of three residues (Arg61, His98, and Asn119) are found to be important for efficient catalysis of DNA cleavage. This finding is consistent with the proposed mechanism in which His98 abstracts a proton from an attacking water molecule bound by an adjacent phosphoryl oxygen, Arg61 and Asn119 stabilize the pentavalent transition state, and Asn119 also binds to the essential divalent metal cation (e.g., Mg(2+) ion), which interacts with the 3'-oxygen leaving group. Because Mg(2+) is required for cleavage of a substrate with a good leaving group (p-nitrophenolate), Mg(2+) likely stabilizes the pentavalent transition state. The pH-dependence of k(cat) for catalysis by I-PpoI reveals a macroscopic pK(a) of 8.4 for titratable groups that modulate product release. I-PpoI appears to be unique among known restriction endonucleases and homing endonucleases in its use of a histidine residue to activate the attacking water molecule for in-line displacement of the 3'-leaving group.

Molecular characterisation and expression of a wound-inducible cDNA encoding a novel cinnamyl-alcohol dehydrogenase enzyme in lucerne (Medicago sativa L.)

A lucerne (alfalfa, Medicago sativa) stem cDNA library was screened with a cinnamyl-alcohol dehydrogenase (CAD) cDNA probe from tobacco (Nicotiana tabacum cv. Samsun). Two distinctly different cDNA clones (54% identical) were isolated and identified as putative CAD-encoding cDNAs by comparison of their nucleotide sequences with those of CAD-encoding DNA sequences from other plant species. One of the cDNAs, MsaCad2, was found to be 99.4% identical at the nucleotide level to the previously isolated lucerne cad cDNA which encodes a CAD isoform involved in lignin biosynthesis. The other cDNA, MsaCad1, has not been reported previously in lucerne, and encodes a protein related to the ELI3 class of elicitor-inducible defence-related plant proteins. The MsaCad1- and MsaCad2-encoded proteins were expressed in Escherichia coli and CAD1 was shown to be active with a range of cinnamyl, benzyl and aliphatic aldehyde substrates, while CAD2 was specific for the cinnamyl aldehydes only. Each of the respective genes is present as one or two copies. The MsaCad1 gene is expressed most actively in stem and floral tissue, whereas MsaCad2 is most actively expressed in stem, hypocotyl and root tissue. In stem tissue, expression of both genes occurs predominantly in internodes 4 and 5 (from the apex). MsaCad2, in contrast to MsaCad1, is not significantly expressed in the top three internodes of the stem. Both MsaCad1 and MsaCad2 are wound-inducible, and the wound-responsiveness of each gene is modulated by salicylic acid.

Molecular cloning of two different cDNAs for maize acetyl CoA carboxylase.

Acetyl CoA carboxylase (EC 6.4.1.2) in plants is a chloroplast-localized, biotin-containing enzyme that catalyses the carboxylation of acetyl CoA to malonyl CoA, the first committed step of the fatty acid biosynthesis pathway. Acetyl CoA carboxylase is the target site for the monocotyledon-specific aryloxyphenoxypropionate and cyclohexanedione groups of herbicides. We have purified a herbicide-sensitive acetyl CoA carboxylase from maize leaves to homogeneity (specific activity 7 mumol min-1 mg-1), separating it during the purification from a minor herbicide-resistant acetyl CoA carboxylase. The purified enzyme is a dimer of 230 kDa subunits. Antibodies raised to the purified acetyl CoA carboxylase detected three cross-reacting clones in a maize leaf cDNA expression library, each having an insert of 4-4.5 kb. Restriction analysis and sequencing showed that the cDNAs were derived from two different transcripts. Comparison of the deduced amino acid sequences with those of chicken and yeast acetyl CoA carboxylases confirmed that both types encoded acetyl CoA carboxylase, corresponding to the C-terminal half of the enzyme. The overall identity of the maize and chicken sequences was 37% (58% similarity) but for some shorter regions was much higher. Analysis of six other acetyl CoA carboxylase clones recovered from the maize cDNA library showed four belonged to one type and two to the other. The nucleotide sequence similarity between the two types of cDNA was approximately 95% in the coding region but considerably less in the 3'-untranslated region. Northern blot analysis of maize RNA showed a single band of 8.2-8.5 kb for acetyl CoA carboxylase mRNA. Southern blot hybridisations indicated that there are probably no more than two genes in maize for acetyl CoA carboxylase. The possible significance of two different cDNAs for acetyl CoA carboxylase is discussed.

Attenuation of nursing-related ovarian suppression and high fertility in well-nourished, intensively breast-feeding Amele women of lowland Papua New Guinea.

Intense, sustained nursing lengthens inter-birth intervals and is causally linked with low natural fertility. However, in traditional settings, the effects of such nursing on fertility are difficult to disentangle from those of nutrition. Results from a prospective, direct observational study of reproductive function in well-nourished Amele women who nurse intensively and persistently but who also have high fertility are here presented. Endocrine measures show that ovarian activity resumes by median 11.0 months postpartum. Median duration of postpartum amenorrhoea is 11.3 months, time to next conception is 19.0 months, and the inter-birth interval is 28.0 months. Average life time fertility is 6.8. High fertility in Amele women is due both to refractoriness of reproductive function to suckling stimuli, and to maintenance of equivalent age-specific fertility rates across the reproductive life span.

PIP: Evidence is presented, from a study of 52 breast-feeding women with an infant under 5 years of age from 12 Amele villages in lowland Papua New Guinea, that women with good nutritional status experienced less variation in ovarian suppression than reported elsewhere among less well-nourished populations. Direct observations of nursing behavior were recorded during 660 hours. The average number of observations per child, which were conducted at different developmental stages, was 1.9. 1549 nursing events were recorded. 38 women had serum samples drawn to determine prolactin levels. Saliva samples were collected from 51 women for measurement of gonadal steroids. Women reported menstrual patterns in the preceding month. Measures were also taken of weight, skinfolds of the triceps and subscapular area, and mid-upper-arm circumference. Birth interval, completed fertility, and age at introduction of solid foods and denial of breast were obtained from ongoing demographic surveillance on nutrition in 1982-84 and 1982-83. Laboratory methods were described for serum prolactin and progesterone analysis. The results showed skewed results for the hormonal and some nursing variables, which were transformed by logarithmic functions. All Amele mothers practice indulgent demand feeding schedules. Supplementary foods are introduced at the median age of 7.4 methods. Early supplementation is liquids or semiliquids, followed by mashed starchy staples, and finally a standard diet. Cessation of breast feeding occurs at a subsequent pregnancy of the advanced age of the child (5 years). The median age of cessation was 36.3 months. The breast feeding pattern during the first 18 months was 1.5-3 times per hour for 1-3 minutes. Infant age was not linearly related to nursing frequency declines or feeding intervals. Nursing frequency was unrelated to morbidity. Prolactin declined strongly when bout frequency and interval were stable. Multiple regression findings were that duration of postpartum amenorrhea was explained by mother's parity and age, feeding interval, and time postpartum. Time of supplementary feeding was unrelated to duration of amenorrhea, nursing frequency, or feeding duration. 16% of interval variance was explained by length of postpartum amenorrhea, which was correlated with length of subsequent birth interval. 45% of the variance in prolactin was explained by triceps skinfolds and bout length.

Failure to isolate human T cell lymphotropic virus type I and to detect variant-specific genomic sequences by polymerase chain reaction in Melanesians with indeterminate western immunoblot.

The controversy over the endemicity of human T cell lymphotropic virus type I (HTLV-I) in Melanesia has been settled recently by the isolation of genetically distinct, highly divergent sequence variants of HTLV-I from unrelated inhabitants of Papua New Guinea and the Solomon Islands. Still at issue, however, is the significance of the high frequency of indeterminate HTLV-I Western blots (defined as reactivity to only gag-encoded proteins) among Melanesians. To investigate whether this indeterminate seroreactivity reflects specific reactivity to the Melanesian HTLV-I variants, 27 seroindeterminate Melanesians from Papua New Guinea and the Solomon Islands were studied for evidence of HTLV-I infection. Although antibodies against Melanesian variant-specific env gene products and variant-specific env gene sequences were detected by Western blot analysis and polymerase chain reaction, respectively, in all 11 HTLV-I Western blot-positive Melanesians, none of the 27 seroindeterminate Melanesians had such variant-specific antibodies or HTLV-I proviral sequences. In addition, attempts to isolate HTLV-I from seroindeterminate individuals were unsuccessful. These data indicate that HTLV-I infection is not the cause of the indeterminate Western blot reactivity seen in Melanesia.

Highly divergent molecular variants of human T-lymphotropic virus type I from isolated populations in Papua New Guinea and the Solomon Islands.

To determine the molecular genetic relationship between Melanesian strains of human T-lymphotropic virus type I (HTLV-I) and cosmopolitan prototype HTLV-I, we amplified by PCR, then cloned, and sequenced a 522-base-pair region of the HTLV-I env gene in DNA extracted from uncultured (fresh) and cultured peripheral blood mononuclear cells obtained from six seropositive Melanesian Papua New Guineans and Solomon Islanders, including a Solomon Islander with HTLV-I myeloneuropathy. Unlike isolates of HTLV-I from Japan, the West Indies, the Americas, and Africa, which share greater than or equal to 97% sequence homology, the Melanesian strains of HTLV-I were only 91.8%-92.5% identical with a prototype Japanese HTLV-IATK-1. The nucleotide sequence of proviral DNA from the Solomon Islander with HTLV-I myeloneuropathy also diverged markedly from that of HTLV-I isolated from Japanese patients with HTLV-I-associated myelopathy and from Jamaican patients with tropical spastic paraparesis, suggesting that these variant viruses are capable of causing disease. The HTLV-I variants from Papua New Guineans, in turn, differed by nearly 4% from the Melanesian variants from Solomon Islanders, indicating the existence of another HTLV-I quasi-species. By contrast, HTLV-I strains from two residents of Bellona Island, a Polynesian Outlier within the Solomon Islands, were closely related to cosmopolitan prototype HTLV-I (greater than or equal to 97% sequence identity), suggesting recent introduction, possibly during this century. These findings are consistent with a proto-Melanesian HTLV-I strain of archaic presence, which evolved independently of contemporary cosmopolitan strains, and pose new questions about the origin and global dissemination of HTLV-I.

Characterization of a variant of human T-lymphotropic virus type I isolated from a healthy member of a remote, recently contacted group in Papua New Guinea.

We report the characterization of a variant of human T-lymphotropic virus type I (HTLV-I) isolated from an interleukin 2-dependent, CD8+ T-cell line derived from peripheral blood mononuclear cells of a healthy member of a remote, recently contacted hunter-horticulturalist group (Hagahai) in Madang province of Papua New Guinea. Antigenic characterization of this variant, designated PNG-1, by immunofluorescence, indicated no expression of gag-encoded proteins p19 and p24 (even after incubation with 5-bromo-2'-deoxyuridine), using monoclonal and polyclonal antibodies against HTLV-I gag gene products. Virus-specific proteins of 15, 19, 46, 53, and 61/68 kDa were demonstrated by Western blot analysis, using sera from patients with serologically and/or virologically confirmed HTLV-I myeloneuropathy, sera from HTLV-I-infected rabbits, and antibodies prepared against the C terminus of the major envelope glycoprotein gp46. Restriction endonuclease maps of PNG-1 proviral DNA differed from that of a prototype strain of HTLV-I (MT-2), but, as verified by polymerase chain reaction, PNG-1 was definitely HTLV-I, not HTLV-II. Nucleotide sequencing and further molecular genetic studies of this variant may provide insights into the origin and evolution of HTLV-I.