Dysregulated STAT1-SOCS1 control of JAK2 promotes mammary luminal progenitor cell survival and drives ERα(+) tumorigenesis.

We previously reported that STAT1 expression is frequently abrogated in human estrogen receptor-α-positive (ERα(+)) breast cancers and mice lacking STAT1 spontaneously develop ERα(+) mammary tumors. However, the precise mechanism by which STAT1 suppresses mammary gland tumorigenesis has not been fully elucidated. Here we show that STAT1-deficient mammary epithelial cells (MECs) display persistent prolactin receptor (PrlR) signaling, resulting in activation of JAK2, STAT3 and STAT5A/5B, expansion of CD61(+) luminal progenitor cells and development of ERα(+) mammary tumors. A failure to upregulate SOCS1, a STAT1-induced inhibitor of JAK2, leads to unopposed oncogenic PrlR signaling in STAT1(-/-) MECs. Prophylactic use of a pharmacological JAK2 inhibitor restrains the proportion of luminal progenitors and prevents disease induction. Systemic inhibition of activated JAK2 induces tumor cell death and produces therapeutic regression of pre-existing endocrine-sensitive and refractory mammary tumors. Thus, STAT1 suppresses tumor formation in mammary glands by preventing the natural developmental function of a growth factor signaling pathway from becoming pro-oncogenic. In addition, targeted inhibition of JAK2 may have significant therapeutic potential in controlling ERα(+) breast cancer in humans.

Characterization of BMS-911543, a functionally selective small-molecule inhibitor of JAK2.

We report the characterization of BMS-911543, a potent and selective small-molecule inhibitor of the Janus kinase (JAK) family member, JAK2. Functionally, BMS-911543 displayed potent anti-proliferative and pharmacodynamic (PD) effects in cell lines dependent upon JAK2 signaling, and had little activity in cell types dependent upon other pathways, such as JAK1 and JAK3. BMS-911543 also displayed anti-proliferative responses in colony growth assays using primary progenitor cells isolated from patients with JAK2(V617F)-positive myeloproliferative neoplasms (MPNs). Similar to these in vitro observations, BMS-911543 was also highly active in in vivo models of JAK2 signaling, with sustained pathway suppression being observed after a single oral dose. At low dose levels active in JAK2-dependent PD models, no effects were observed in an in vivo model of immunosuppression monitoring antigen-induced IgG and IgM production. Expression profiling of JAK2(V617F)-expressing cells treated with diverse JAK2 inhibitors revealed a shared set of transcriptional changes underlying pharmacological effects of JAK2 inhibition, including many STAT1-regulated genes and STAT1 itself. Collectively, our results highlight BMS-911543 as a functionally selective JAK2 inhibitor and support the therapeutic rationale for its further characterization in patients with MPN or in other disorders characterized by constitutively active JAK2 signaling.

Purification and characterization of a tumor lipid-mobilizing factor.

Cancer patients with weight loss showed urinary excretion of a lipid-mobilizing factor (LMF), determined by the ability to stimulate lipolysis in isolated murine epididymal adipocytes. Such bioactivity was not detectable in the urine of cancer patients without weight loss or in normal subjects. The LMF was purified using a combination of ion exchange, exclusion, and hydrophobic interaction chromatographies to give a single component of apparent Mr 43,000, which showed homology in amino acid sequence with human plasma Zn-alpha2-glycoprotein. Both substances showed the same mobility on denaturing and nondenaturing gels and the same chymotrypsin digestion pattern, both stained heavily for carbohydrate, and they showed similar immunoreactivity. Polyclonal antisera to human plasma Zn-alpha2-glycoprotein was also capable of neutralization of the bioactivity of human LMF in vitro. Using competitive PCR to quantify expression of Zn-alpha2-glycoprotein, we found that only those tumors that were capable of producing a decrease in carcass lipid expressed mRNA for Zn-alpha2-glycoprotein. These results provide strong evidence to suggest that tumor production of Zn-alpha2-glycoprotein is responsible for the lipid catabolism seen in cancer patients.

Catabolic factors in cancer cachexia.

A lipid mobilizing factor has been purified from a cachexia-inducing mouse colon adenocarcinoma (MAC16) using a combination of ion exchange (Mono Q), exclusion (Superose) and reverse phase hydrophobic chromatography. The purification process led to a 3,500-fold increase in the specific activity. Serum from mice bearing the MAC16 tumour contained antibodies reactive with fractions containing lipid mobilizing activity and detectable as a 24 kDa immunoreactive band on Western blotting. Serum from mice transplanted with a related tumour, MAC13, not producing cachexia, did not contain antibodies. A similar immunoreactive band was detectable in the urine of patients with cancer cachexia, but was absent from the urine of normal subjects. A monoclonal antibody produced by fusion of splenocytes from mice bearing the MAC16 tumour with mouse Balb/c myeloma cells attenuated the development of cachexia in mice transplanted with the MAC16 tumour and inhibited tumour growth. These results suggest that the M(r) 24 kDa antigen may be important in tumour growth and cachexia.

Induction of muscle protein degradation and weight loss by a tumor product.

Splenocytes from mice bearing a cachexia-inducing tumor (MAC16) have been fused with mouse myeloma cells to produce hybridomas, which have been cloned to produce antibody reactive to a material which copurified with a lipid-mobilizing factor isolated from the same tumor. The monoclonal antibody has been used to investigate factors potentially involved in the development of cachexia. The major protein detectable by immunoprecipitation of a partially purified lipid-mobilizing factor was M(r) 69,000, whereas Western blotting showed two bands of M(r) 69,000 and M(r) 24,000. Although the monoclonal antibody did not neutralize lipid-mobilizing activity in an in vitro assay, it did neutralize a serum factor capable of protein degradation in isolated gastrocnemius muscle. Affinity purification of MAC16 tumor homogenates using the monoclonal antibody yielded two immunoreactive bands of M(r) 69,000 and M(r) 24,000, which were further fractionated on a hydrophobic column (C8). This material was capable of inducing tyrosine release from isolated gastrocnemius muscle, and the effect could be blocked with the monoclonal antibody. The two immunoreactive bands from the hydrophobic column were capable of inducing weight loss in mice, whereas nonimmunoreactive fractions had no effect on body weight. The M(r) 24,000 species had a unique amino acid sequence, whereas the M(r) 69,000 species gave the same sequence as the M(r) 24,000 material, together with that for albumin. The M(r) 24,000 species contained carbohydrate, and lectin blotting showed a strong reaction with wheat germ and Erythrina crystagalli agglutinins. This suggests that the material is a glycoprotein or proteoglycan that shows strong binding affinity for albumin, possibly through the carbohydrate residues.

Purification and characterization of a lipid-mobilizing factor associated with cachexia-inducing tumors in mice and humans.

A scheme is described for the purification of a lipid-mobilizing factor from a cachexia-inducing murine tumor (MAC16) using a combination of ion exchange (Mono Q), exclusion (Superose), and hydrophobic (C8) chromatography. This process yields an active material with an apparent molecular weight of 24,000 with an overall purification of 3,500 from the tumor homogenate and representing 0.005% of the total protein present. The material tends to aggregate to high molecular mass, is acidic (pI < 4), and displays heterogeneity of charge as evidenced by a broad elution profile on ion exchange and exclusion chromatography and multiple peaks on hydrophobic columns. The purified material was heat and alkali (pH 10.4) labile and activity could be completely inhibited by sulfatase, suggesting that the negative charge could arise from sulfate residues. There was no evidence that the material possessed triglyceride lipase activity. Animals transplanted with the MAC16 tumor and with a delayed weight loss contained in their serum antibodies that recognized a M(r) 24,000 band on Western blots. This material copurified with the lipid-mobilizing factor. Such antibodies were not present in the serum of mice transplanted with the MAC13 tumor, which does not induce cachexia, suggesting that the antibodies were directed to the induction of cachexia rather than the tumor itself. Urine from patients with cancer cachexia also contained a lipid-mobilizing factor which adhered to DEAE-cellulose and gave an apparent M(r) of 24,000 by exclusion chromatography. Western blotting using serum from MAC16 tumor-bearing animals showed the presence of a band of M(r) 24,000 in such fractions, which was not detected using serum from mice bearing the MAC13 tumor. This band was not present in Western blots of urine from normal subjects. The fact that serum from mice bearing the MAC16 tumor can detect the human lipid-mobilizing activity suggests a high degree of structural similarity between the two and raises the possibility that cachexia in humans may be caused by the same species as in the mouse.

Tumour-associated hypoglycaemia in a murine cachexia model.

Animals bearing a cachexia-inducing tumour, the MAC16 adenocarcinoma, showed a progressive decrease in blood glucose levels with increasing weight loss, while animals bearing a histologically similar tumour, the MAC13 adenocarcinoma, showed no change in either body weight or blood glucose levels with growth of the tumour. The effect of the MAC16 tumour on blood glucose levels appeared to be unrelated to food intake, glucose consumption by the tumour, or to the production of increased levels of IGF-I and IGF-II mRNA by the tumour cells. The relationship between the induction of cachexia and alteration in blood glucose levels remains unknown.

An examination of change in reports of AIDS-related knowledge and attitudes in 1986 and 1988.

This study employed a cross-sectional design to examine change in AIDS-related attitudes, knowledge, and behaviors among students at a western university between 1986 and 1988. Analysis indicates that knowledge of how AIDS is transmitted improved. There was high association between attitudes to AIDS patients and attitudes towards homosexuals. Although attitudes towards individuals with AIDS did not change, attitudes towards homosexuals became more negative. Men reported more negative attitudes towards AIDS patients and towards homosexuals than did women. Most importantly, more students in 1988 reported changing their behavior than in 1986. These results are congruent with those from other investigations and suggest that AIDS-education campaigns and media attention to AIDS are working to change students' high-risk behaviors.

Children's beliefs about listening: is it enough to be still and quiet?

This study examined children's conception of listening and their performance as listeners, 7-, 9-, and 11-year-old children were interviewed for their conceptions of good listening, their beliefs about appropriate actions for confused listeners to take, their attributions of responsibility for a listener's confusion, their reports of how speakers and listeners feel during communication breakdown, and their ability to detect inconsistencies during a comprehension-monitoring task. Results indicated that older children relied less on behavioral orientation and more on attempts to comprehend and other criteria in their definitions of good listening. Children believed that appropriate listening responses depend on the situation, and there was a developmental increase in asking the speaker a question and listening more carefully. With age, children also tended to report more complex negative emotions for listeners and speakers experiencing a breakdown in understanding. Children recalled incongruent material more than congruent material on the comprehension-monitoring task.

The impact of origin community characteristics on rural-urban out-migration in a developing country.

It is widely believed that structural variables such as inequitable land distribution, lack of rural employment opportunities, and rural-urban wage and amenity gaps influence population movements in developing countries. Yet quantitative evidence is scant. In this paper a multilevel model is used to investigate the effects of individual-, household-, and areal-level factors on rural-urban out-migration in the Ecuadorian Sierra. Data from a detailed survey carried out in 1977-1978 and from government macro-areal statistics are used to investigate factors affecting the out-migration of youths aged 12-25. Preliminary conclusions are presented on the usefulness of multilevel models in studying migration and policy implications for Ecuador.

Migration plans of the rural populations of the Third World countries: a probit analysis of micro-level data from Asia, Africa, and Latin America.

PIP: This study 1) examines the extent to which a given set of microlevel factors has predictive value in different socioeconomic settings and 2) demonstrates the utility of a probit estimation technique in examining plans of rural populations to migrate. Data were collected in 1977-1979 in Thailand, Egypt, and Colombia, 3 countries which differ in culture, extent of urbanization, and proportion of labor force engaged in nonextractive industries. The researchers used identical questionnaires and obtained interviews in 4 rural villages with the "migration shed" of each country's capital city. There were 1088 rural-resident men and women interviewed in Thailand, 1088 in Colombia, and 1376 in Egypt. The researchers gathered information about year-to-year changes in residence, marital status, fertility, housing, employment status, occupation, and industry. While in all 3 countries return moves are relatively frequent, especially among males, the proportions of migrants who have moved 3 or more times do not rise above 10%. The model used portrays the formation of migration intentions of the individual as the outcome of a decision process involving the subjective weighing of perceived differentials in well-being associated with current residence and 1 or more potential destinations, taking into account the direct relocation costs and ability to finance a move. The researchers used dichotomous probit and ordinal probit techniques and 4 variations on the dependant variable to generate some of the results. The only expectancy variable significant in all countries is age. Education is also positively and significantly associated with intentions to move for both sexes in Colombia and Egypt. Marital status is a deterrent to migration plans for males in Colombia and both sexes in Egypt. Previous migration experience fails to show any significant relationship to propensity to move. Conclusions drawn from the data include: 1) the effects of age and economic status appear to increase, both in strength and significance, for males in countries as the likelihood of a move increases; and 2) the effect of the kin and friend contract variable in Colombia appears to be related to its usefulness in explaining th initial consideration of a move rather than the plans that carry a probability or certainty of implementation. The careful measurement of strength of migration intentions and the application of ordinal probit estimation methods to the analysis of prospective migration may contribute to the refinement of our understanding of the process of migration decision making across a range of geographical, cultural, and developmental contexts.^ieng

Infant mortality estimates for Papua New Guinea.

PIP: When a country's census data are either incomplete or subject to common error patterns, serious bias can be introduced by using direct methods of estimating infant mortality rates (IMRs). Indirect census based methods have been developed to provide crude, but reasonable estimates of IMRs in these situations. Indirectly estimated IMRs were derived from child survivorship data from the 1966 and 1971 Papua New Guinea census. IMR trends were estimated as well for both urban and rural sectors of the New Guinea region, the Papua region, and the nation as a whole. An extension of the Brass method as described by Feeny in 1976 was used for the estimations. Combined urban and rural sectors of Papua New Guinea showed a 3.98 rate of decline of IMR from 1966 to 1971 (in infant deaths per thousand live births per year). IMRs were extrapolated from a 1960.58 mean IMR of 144.68 (infant deaths per thousand live births) to 101.2 in 1971.5. An IMR estimate based only on the 1971 census was 116.4. 2 of Feeny's 5 implicit assumptions were violated in the application of this indirect estimation method to Papua New Guinea data, i.e. that proportions of deceased children must be accurately reflected in reported numbers born and deceased, and that reporting of women's ages must be reasonably accurate. The author discusses the probable error introduced into the trend estimates because of this. Comparisons between the indirectly estimated IMR series and both previously published IMR series and information on the introduction of health services in the country was made in an attempt to corroborate the indirect estimates.^ieng

Further evidence on family change in Papua New Guinea.

PIP: This paper attempts to elaborate on previous papers on family change in postwar urban Papua New Guinea and to point out the implications of the emerging patterns as they relate to demands likely to be made on existing government programs in future years. There has been rapid growth of an educated, urban resident population, with the use of wages and cash much more prevalent than before. There is some attenuation of family ties with more emphasis on the nuclear family, which is likely to be smaller than the traditional, large family. In rural areas people prefer large families. There continues to be use of contraception and family planning, an extension of traditional means of controlling reproductive behavior. The impact of social implications of weakened family ties on government programs is marginal. It is clear, however, that there is likely to be substantial growth in the urban population (3.8%/year based on 1971 mortality levels) because of rapid declines in death rates. This will make it more difficult for government to provide health care, education and employment for the population, especially young people. Tables give statistics on 1) family type by education for male heads of households, 2) preference of male respondents for a large or small family, 3) desired number of children by education, male and female respondents, 4) attitudes toward family limitation by education, male respondents, 5) use of family planning by education, female respondents, and 6) population characteristics implied by 1971 levels of fertility and mortality.^ieng