Active by nature: exploring cancer survivors' exercise barriers, facilitators, preferences, and psychosocial benefits of engaging in outdoor physical activity.

INTRODUCTION: Large numbers of cancer survivors struggle with mental health after cancer diagnosis. Cancer survivors are encouraged to engage in physical activity in order to improve physical and mental health. Additional benefits to physical activity engagement in natural environments have been reported but this has not been explored in cancer survivors. METHODS: Study participants had to be over the age of 19, a Canadian resident, and have had a cancer diagnosis. Recruitment to complete an online survey occurred through social media and snowball sampling. The data collected included physical activity participation, preferences and location, barriers and facilitators of engagement in outdoor physical activity, nature-related questions, and measures of psychosocial health. The sample was split by the number of outdoor physical activity minutes (> 150 min per week). Correlations were computed to examine the role of outdoor physical activity minutes on measured psychosocial health outcomes. RESULTS: One hundred and fourteen (N = 114) cancer survivors completed the online questionnaire. More than half of the respondents indicated that an outdoor environment was central to their physical activity of choice with walking identified as the most common outdoor physical activity. Group support was the main expected facilitator of success in an outdoor walking program. Outdoor active participants were significantly more motivated and confident to be physically active and reported significantly more benefit and enjoyment in being physically active than outdoor inactive participants. Minutes of outdoor physical activity was significantly correlated with subjective happiness, nature relatedness, and higher quality of life. No significant correlations were found between minutes of outdoor physical activity and generalized anxiety. CONCLUSION: While future research is needed to further explore the role of nature in cancer survivor psychosocial health, we believe that our data suggests preference and benefit for outdoor physical activity in cancer survivors.

The patient advisor, an organizational resource as a lever for an enhanced oncology patient experience (PAROLE-onco): a longitudinal multiple case study protocol.

BACKGROUND: Quebec is one of the Canadian provinces with the highest rates of cancer incidence and prevalence. A study by the Rossy Cancer Network (RCN) of McGill university assessed six aspects of the patient experience among cancer patients and found that emotional support is the aspect most lacking. To improve this support, trained patient advisors (PAs) can be included as full-fledged members of the healthcare team, given that PA can rely on their knowledge with experiencing the disease and from using health and social care services to accompany cancer patients, they could help to round out the health and social care services offer in oncology. However, the feasibility of integrating PAs in clinical oncology teams has not been studied. In this multisite study, we will explore how to integrate PAs in clinical oncology teams and, under what conditions this can be successfully done. We aim to better understand effects of this PA intervention on patients, on the PAs themselves, the health and social care team, the administrators, and on the organization of services and to identify associated ethical and legal issues. METHODS/DESIGN: We will conduct six mixed methods longitudinal case studies. Qualitative data will be used to study the integration of the PAs into clinical oncology teams and to identify the factors that are facilitators and inhibitors of the process, the associated ethical and legal issues, and the challenges that the PAs experience. Quantitative data will be used to assess effects on patients, PAs and team members, if any, of the PA intervention. The results will be used to support oncology programs in the integration of PAs into their healthcare teams and to design a future randomized pragmatic trial to evaluate the impact of PAs as full-fledged members of clinical oncology teams on cancer patients' experience of emotional support throughout their care trajectory. DISCUSSION: This study will be the first to integrate PAs as full-fledged members of the clinical oncology team and to assess possible clinical and organizational level effects. Given the unique role of PAs, this study will complement the body of research on peer support and patient navigation. An additional innovative aspect of this study will be consideration of the ethical and legal issues at stake and how to address them in the health care organizations.

The differential effects of norepinephrine and dopamine on cerebrospinal fluid pressure and spinal cord perfusion pressure after acute human spinal cord injury.

STUDY DESIGN: Prospective vasopressor cross-over interventional studyObjectives:To examine how two vasopressors used in acute traumatic spinal cord injury (SCI) affect intrathecal cerebrospinal fluid pressure and the corresponding spinal cord perfusion pressure (SCPP). SETTING: Vancouver, British Columbia, Canada. METHODS: Acute SCI patients over the age of 17 with cervical or thoracic ASIA Impairment Scale (AIS). A, B or C injuries were enrolled in this study. Two vasopressors, norepinephrine and dopamine, were evaluated in a 'crossover procedure' to directly compare their effect on the intrathecal pressure (ITP). The vasopressor cross-over procedures were performed in the intensive care unit where ITP, mean arterial pressure (MAP) and heart rate were being continuously measured. The SCPP was calculated as the difference between MAP and ITP. RESULTS: A total of 11 patients were enrolled and included in our analysis. There were 6 patients with AIS A, 3 with AIS B and 2 with AIS C injuries at baseline. We performed 24 cross-over interventions in these 11 patients. There was no difference in MAP with the use of norepinephrine versus dopamine (84±1 mm Hg for both; P=0.33). Conversely, ITP was significantly lower with the use of norepinephrine than with dopamine (17±1 mm Hg vs 20±1 mm Hg, respectively, P<0.001). This decrease in ITP with norepinephrine resulted in an increased SCPP during the norepinephrine infusion when compared with dopamine (67±1 mm Hg vs 65±1 mm Hg respectively, P=0.0049). CONCLUSION: Norepinephrine was able to maintain MAP with a lower ITP and a correspondingly higher SCPP as compared with dopamine in this study. These results suggest that norepinephrine may be preferable to dopamine if vasopressor support is required post SCI to maintain elevated MAPs in accordance with published guidelines.

Quality and accuracy of publicly accessible cancer-related physical activity information on the Internet: a cross-sectional assessment.

In this study, we assessed the quality of publicly available cancer-related physical activity (PA) information appearing on reputable sites from Canada and other English-speaking countries. A cross-sectional Internet search was conducted on select countries (Canada, USA, Australia, New Zealand, UK) using Google to generate top 50 results per country for the keywords "'physical activity' AND 'cancer'". Top results were assessed for quality of PA information based on a coding frame. Additional searches were performed for Canadian-based sites to produce an exhaustive list. Results found that many sites offered cancer-related PA information (94.5%), but rarely defined PA (25.2%). Top 50 results from each country did not differ on any indicator examined. The exhaustive list of Canadian sites found that many sites gave information about PA for survivorship (78.3%) and prevention (70.0%), but rarely defined (6.7%) or referenced PA guidelines (28.3%). Cancer-related PA information is plentiful on the Internet but the quality needs improvement. Sites should do more than mention PA; they should provide definitions, examples and guidelines. With improvements, these websites would enable healthcare providers to effectively educate their patients about PA, and serve as a valuable resource to the general public who may be seeking cancer-related PA information.

Development and validation of a computerized algorithm for International Standards for Neurological Classification of Spinal Cord Injury (ISNCSCI).

STUDY DESIGN: Validation study. OBJECTIVES: To describe the development and validation of a computerized application of the international standards for neurological classification of spinal cord injury (ISNCSCI). SETTING: Data from acute and rehabilitation care. METHODS: The Rick Hansen Institute-ISNCSCI Algorithm (RHI-ISNCSCI Algorithm) was developed based on the 2011 version of the ISNCSCI and the 2013 version of the worksheet. International experts developed the design and logic with a focus on usability and features to standardize the correct classification of challenging cases. A five-phased process was used to develop and validate the algorithm. Discrepancies between the clinician-derived and algorithm-calculated results were reconciled. RESULTS: Phase one of the validation used 48 cases to develop the logic. Phase three used these and 15 additional cases for further logic development to classify cases with 'Not testable' values. For logic testing in phases two and four, 351 and 1998 cases from the Rick Hansen SCI Registry (RHSCIR), respectively, were used. Of 23 and 286 discrepant cases identified in phases two and four, 2 and 6 cases resulted in changes to the algorithm. Cross-validation of the algorithm in phase five using 108 new RHSCIR cases did not identify the need for any further changes, as all discrepancies were due to clinician errors. The web-based application and the algorithm code are freely available at www.isncscialgorithm.com. CONCLUSION: The RHI-ISNCSCI Algorithm provides a standardized method to accurately derive the level and severity of SCI from the raw data of the ISNCSCI examination. The web interface assists in maximizing usability while minimizing the impact of human error in classifying SCI. SPONSORSHIP: This study is sponsored by the Rick Hansen Institute and supported by funding from Health Canada and Western Economic Diversification Canada.

Evaluation of a pilot Pressure Ulcer Prevention Initiative (PUPI) for patients with traumatic spinal cord injury.

OBJECTIVE: The purpose of this study was to determine whether implementation of a Pressure Ulcer Prevention Initiative (PUPI) changed the assessment and treatment of patients with a traumatic spinal cord injury (SCI) in an acute care setting, and improved patient outcomes. METHOD: The success of implementation was evaluated by examining the percentage of patients with completed occupational therapist (OT) skin care assessments and prescriptions for therapeutic support surfaces (TSS; i.e., mattresses) before implementation (historical, cohort 1) and after implementation (experimental, cohort 2). Patient outcomes were evaluated by examining changes in PU incidence, severity, timing and recurrence, as well as PU prevalence and satisfaction with life in the community. RESULTS: Final analysis included 70 patients in cohort 1 and 73 in cohort 2. OT skin care assessment documentation (31% to 60%; p<0.001) and TSS prescriptions (31% to 60%; p=0.02) significantly increased following the implementation. The PU incidence based on patient charts (both nursing and OT assessments) did not increase significantly (26% to 36%; p=0.2). However, documented PU incidence according to OT assessments showed a substantial increase (14% to 33%; p=0.002). No effect of the PUPI was seen on immediate or long-term patient outcomes during the study period. CONCLUSION: PUPI was successful in changing clinical practice in PU prevention but no statistically significant improvements in PU-related patient outcomes were demonstrated. Results from this study identified facilitators and barriers to implementation and highlighted the complexity and difficulty of instituting effective preventative or therapeutic interventions for this population in an acute care setting. This information will assist with refinements of the PUPI and inform similar future initiatives.

A prospective evaluation of hemodynamic management in acute spinal cord injury patients.

STUDY DESIGN: Prospective observational study of acute spinal cord-injured (SCI) patients. OBJECTIVES: To determine how effectively mean arterial blood pressure (MAP) and spinal cord perfusion pressure (SCPP) are maintained at target levels in acute SCI patients. SETTING: Single-institution study at a Canadian level-one trauma center. METHODS: Twenty-one individuals with cervical or thoracic SCI were enrolled within 48 h of injury. A lumbar intrathecal drain was inserted for monitoring intrathecal cerebrospinal fluid pressure (ITP). The MAP was monitored concurrently with ITP, and the SCPP was calculated. Data was recorded hourly from the time of first assessment until at least the end of the 5th day post injury. RESULTS: All subjects had at least one recorded episode with a MAP below 80 mm Hg, and 81% had at least one episode with a MAP below 70 mm Hg. On average, subjects with cervical injuries had 18.4% of their pressure recordings below 80 mm Hg. Subjects with thoracic cord injuries had on average 35.9% of their MAP recordings <80 mm Hg. CONCLUSION: It is common practice to establish MAP targets for optimizing cord perfusion in acute SCI. This study suggests that even in an acute SCI referral center, when prospectively scrutinized, the actual MAP may frequently fall below the intended targets. Such results raise awareness of the vigilance that must be kept in the hemodynamic management of these patients, and the potential discrepancy between routinely setting target MAP according to 'practice guidelines' and actually achieving them.

Medical, demographic and social cognitive correlates of physical activity in a population-based sample of colorectal cancer survivors.

Physical activity (PA) improves quality of life in colorectal cancer survivors (CRC) and may reduce the risk of disease recurrence and early death. Few studies, however, have examined the correlates of PA in CRC survivors. Using the Alberta Cancer Registry, 2000 randomly selected CRC survivors were mailed a self-reported questionnaire assessing medical, demographic, behavioural and social cognitive variables from the theory of planned behaviour (TPB). Of the 600 survivors who responded, 33% were meeting public health PA guidelines and almost half were completely sedentary. Higher PA was reported by survivors who were younger, unmarried, better educated, wealthier, employed, non-smokers, social drinkers, not treated with radiation therapy, disease-free, in better health and less comorbidity. In multivariate path analysis, these variables were not directly associated with PA after controlling for the TPB variables. The TPB explained 34% (P < 0.001) of the variance in PA behaviour with direct associations for intention (ß= 0.22; P= 0.015) and planning (ß= 0.18; P= 0.001). Intention, in turn, had 62% (P < 0.001) of its variance explained by perceived behavioural control (ß= 0.43; P < 0.001), affective attitude (ß= 0.25; P < 0.001) and instrumental attitude (ß= 0.15; P < 0.001). The TPB may be a useful framework for developing population-based interventions to increase PA in CRC survivors.

[Sensitivity and specificity of the worry and anxiety questionnaire (WAQ) in a sample of health-care users in the province of Quebec]. / Sensibilité et spécificité du questionnaire sur les inquiétudes et l'anxiété (QIA) dans un échantillon d'utilisateurs de soins de santé au Québec.

INTRODUCTION: Generalized anxiety disorder (GAD) is a prevalent anxiety disorder characterized by persistent, excessive worrying. Even if GAD's ill consequences on health and quality of life are well documented, this disorder is still difficult to identify in primary care. The worry and anxiety questionnaire (WAQ) is a questionnaire assessing specific GAD symptoms, as defined by the Diagnostic and Statistical Manual of Mental Disorders-4th Edition (DSM-IV). OBJECTIVES: This study aimed at assessing the capacity of the French version of the WAQ's to identify individuals with GAD in a sample of individuals reporting a certain level of anxiety. A second objective was to identify which of its items better distinguish individuals with GAD from those without. According to these results, different scoring algorithms have been developed and their effect on the WAQ's sensitivity and specificity indicators has been explored. DESIGN OF THE STUDY: The sample was drawn from a mother study in which 1110 health-care users completed questionnaires while waiting for a medical consultation with a family physician. Of those, a subsample of 219 individuals reported anxiety symptoms typical of GAD, as assessed by the WAQ. Among those who agreed to participate in the study's second phase (n=176), 100 were randomly selected and invited within one to three months to a clinical interview assessing their anxiety symptoms more thoroughly. Thirty-three individuals accepted and thus formed the present sample. The clinical interview was the anxiety disorders interview schedule (ADIS). The ADIS is a semi-structured diagnostic interview following the DSM-IV criteria. It assesses all anxiety disorders and includes screening questions on mood, substance use and psychotic disorders. Participants also completed the WAQ for a second time at the time of the interview. RESULTS: Nineteen individuals received a diagnosis of GAD after completing the ADIS while 13 did not. Sixteen of the 19 individuals with GAD were correctly identified with the WAQ, compared to eight out of 13 for individuals without GAD. Sensitivity of the WAQ's actual scoring algorithm is thus of 84.2% and its specificity of 61.5%. The number of false negatives produced by the WAQ in this sample (3/19, 15.8%) was lower than the number of false positives (5/13, 38.5%). Positive and negative predictive power is thus of 76.2% and 72.7%, respectively. Receiver-operating characteristic (ROC) curves analyses indicated that the most useful items to identify individuals with GAD were those assessing the presence of excessive worrying, the number of days disturbed by worries, the degree with which worries interfere with daily functioning and the degree of control over worries. Knowing an individual's outcome on the WAQ increases the probability of correctly identifying an individual with or without GAD by 8.5 times compared to mere chance. A new scoring algorithm, where the cut-off score on the excessive worrying item was increased by one unit, considerably improves the WAQ's specificity (84.6%), without altering its sensitivity by much (78.9%). This new scoring algorithm thus increases the probability of correctly identifying individuals with and without GAD to 20.6 times (again compared to chance). CONCLUSION: The WAQ is thus a useful instrument in screening GAD, even in a sample of anxious individuals. Its original scoring algorithm shows excellent sensitivity, a valued quality in an instrument used for screening. On the other hand, it is possible to increase the specificity of the WAQ by raising the cut-off point on the excessive worrying item, making the instrument useful as a diagnostic aid or as a screening questionnaire for GAD, in particular among a sample displaying anxiety. This questionnaire is thus an easy-to-complete and adaptable instrument that can be used by family physicians to help them identifying individuals with GAD.

Repression of the alpha-fetoprotein gene promoter by progesterone and chimeric receptors in the presence of hormones and antihormones.

Using transient transfection assays, we showed that repression of the alpha-fetoprotein promoter by intact and deletion mutants of the progesterone receptor and by chimeric progesterone/glucocorticoid-estrogen receptors in the presence of their cognate hormones was closely correlated with their ability to bind to a progesterone/glucocorticoid-responsive element. This negative regulation was also observed in the presence of antihormones, providing evidence that receptor-antihormone complexes can bind to their responsive elements in vivo.

Enhancer and promoter elements directing activation and glucocorticoid repression of the alpha 1-fetoprotein gene in hepatocytes.

Mutations were introduced in 7 kilobases of 5'-flanking rat alpha 1-fetoprotein (AFP) genomic DNA, linked to the chloramphenicol acetyltransferase gene. AFP promoter activity and its repression by a glucocorticoid hormone were assessed by stable and transient expression assays. Stable transfection assays were more sensitive and accurate than transient expression assays in a Morris 7777 rat hepatoma recipient (Hepa7.6), selected for its strong AFP repression by dexamethasone. The segment of DNA encompassing a hepatocyte-constitutive chromatin DNase I-hypersensitive site at -3.7 kilobases and a liver developmental stage-specific site at -2.5 kilobases contains interacting enhancer elements sufficient for high AFP promoter activity in Hepa7.6 or HepG2 cells. Deletions and point mutations define an upstream promoter domain of AFP gene activation, operating with at least three distinct promoter-activating elements, PEI at -65 base pairs, PEII at -120 base pairs, and DE at -160 base pairs. PEI and PEII share homologies with albumin promoter sequences, PEII is a near-consensus nuclear factor I recognition sequence, and DE overlaps a glucocorticoid receptor recognition sequence. An element conferring glucocorticoid repression of AFP gene activity is located in the upstream AFP promoter domain. Receptor-binding assays indicate that this element is the glucocorticoid receptor recognition sequence which overlaps with promoter-activating element DE.

The alpha1-fetoprotein locus is activated by a nuclear receptor of the Drosophila FTZ-F1 family.

The alpha1-fetoprotein (AFP) gene is located between the albumin and alpha-albumin genes and is activated by transcription factor FTF (fetoprotein transcription factor), presumed to transduce early developmental signals to the albumin gene cluster. We have identified FTF as an orphan nuclear receptor of the Drosophila FTZ-F1 family. FTF recognizes the DNA sequence 5'-TCAAGGTCA-3', the canonical recognition motif for FTZ-F1 receptors. cDNA sequence homologies indicate that rat FTF is the ortholog of mouse LRH-1 and Xenopus xFF1rA. Rodent FTF is encoded by a single-copy gene, related to the gene encoding steroidogenic factor 1 (SF-1). The 5.2-kb FTF transcript is translated from several in-frame initiator codons into FTF isoforms (54 to 64 kDa) which appear to bind DNA as monomers, with no need for a specific ligand, similar KdS (approximately equal 3 x 10(-10) M), and similar transcriptional effects. FTF activates the AFP promoter without the use of an amino-terminal activation domain; carboxy-terminus-truncated FTF exerts strong dominant negative effects. In the AFP promoter, FTF recruits an accessory trans-activator which imparts glucocorticoid reactivity upon the AFP gene. FTF binding sites are found in the promoters of other liver-expressed genes, some encoding liver transcription factors; FTF, liver alpha1-antitrypsin promoter factor LFB2, and HNF-3beta promoter factor UF2-H3beta are probably the same factor. FTF is also abundantly expressed in the pancreas and may exert differentiation functions in endodermal sublineages, similar to SF-1 in steroidogenic tissues. HepG2 hepatoma cells seem to express a mutated form of FTF.

Functional analysis of developmentally regulated chromatin-hypersensitive domains carrying the alpha 1-fetoprotein gene promoter and the albumin/alpha 1-fetoprotein intergenic enhancer.

During liver development, the tandem alpha 1-fetoprotein (AFP)/albumin locus is triggered at the AFP end and then asymmetrically enhanced; this is followed by autonomous repression of the AFP-encoding gene. To understand this regulation better, we characterized the two early developmental stage-specific DNase I-hypersensitive (DH) sites so far identified in rat liver AFP/albumin chromatin: an intergenic DH-enhancer site and the AFP DH-promoter site. Mutation-transfection analyses circumscribed the DH-enhancer domain to a 200-bp DNA segment stringently conserved among species. Targeted mutations, DNA-protein-binding assays, and coexpression experiments pinpointed C/EBP as the major activatory component of the intergenic enhancer. Structure-function relationships at the AFP DH-promoter site defined a discrete glucocorticoid-regulated domain activated cooperatively by HNF1 and a highly specific AFP transcription factor, FTF, which binds to a steroid receptor recognition motif. The HNF1/FTF/DNA complex is deactivated by glucocorticoid receptors or by the ubiquitous factor NF1, which eliminates HNF1 by competition at an overlapping, high-affinity binding site. We propose that the HNF1-NF1 site might serve as a developmental switch to direct autonomous AFP gene repression in late liver development. We also conclude that the intergenic enhancer is driven by C/EBP alpha primarily to fulfill albumin gene activation functions at early developmental stages. Factor FTF seems to be the key regulator of AFP gene-specific functions in carcinoembryonic states.

Efficacy of a single oral dose of a live bivalent E. coli vaccine against post-weaning diarrhea due to F4 and F18-positive enterotoxigenic E. coli.

F4- and F18-positive enterotoxigenic E. coli strains (F4-ETEC and F18-ETEC) are important causes of post-weaning diarrhea (PWD) in pigs. F4 (antigenic variant ac) and F18 (ab and ac) fimbriae are major antigens that play an important role in the early stages of infection. Herein, the efficacy of a live oral vaccine consisting of two non-pathogenic E. coli strains, one F4ac- and one F18ac-positive, was evaluated using F4ac-ETEC and F18ab-ETEC challenge models. A randomized, masked, placebo-controlled, block design, parallel-group confirmatory study with two different vaccination-challenge intervals (7 and 21 days) was conducted for each challenge model. The vaccine was administered in one dose, to ≥18-day-old piglets via drinking water. Efficacy was assessed by evaluating diarrhea, clinical observations, weight gain and fecal shedding of F4-ETEC or F18-ETEC. Anti-F4 and anti-F18 immunoglobulins in blood were measured. The vaccination resulted in significant reductions in clinical PWD and fecal shedding of F4-ETEC and F18-ETEC after the 7- and 21-day-post-vaccination heterologous challenges, except for after the 21-day-post-vaccination F4-ETEC challenge, when the clinical PWD was too mild to demonstrate efficacy. A significant reduction of mortality and weight loss by vaccination were observed following the F18-ETEC challenge. The 7-day protection was associated with induction of anti-F4 and anti-F18 IgM, whereas the 21-day protection was mainly associated with anti-F4 and anti-F18 IgA. The 7-day onset and 21-day duration of protection induced by this vaccine administered once in drinking water to pigs of at least 18days of age were confirmed by protection against F4-ETEC and F18-ETEC, and induction of F4 and F18-specific immunity. Cross protection of the vaccine against F18ab-E. coli was demonstrated for both the 7- and 21-day F18-ETEC challenges.

Dietary fat in relation to prognostic indicators in breast cancer.

The relation of diet, especially fat intake, to recognized prognostic indicators for breast cancer was investigated in 666 women with a newly diagnosed infiltrating breast carcinoma. Diet during the year preceding diagnosis was assessed by interview using a food frequency questionnaire covering the intake of 114 food items. Prognostic indicators included axillary node involvement at diagnosis, estrogen receptor status, and selected histologic features of the primary tumor such as nuclear grade, histologic grade, tubule formation, mitotic activity, and nuclear size of tumor cells. After adjustment for total energy intake, age, body weight, and tumor size at diagnosis, an increase in saturated fat intake was related to an increased frequency of node involvement at diagnosis among postmenopausal patients. In contrast, an elevation in polyunsaturated fat intake was related to a reduction of the percentage of patients with positive nodes at diagnosis. This relation was observed among both premenopausal and postmenopausal patients. Dietary fat was not related to the estrogen receptor status of tumors. No association was found between dietary habits and histologic features of the primary tumor. These data suggest that dietary fat may have an effect on the growth or spread of breast cancer during the preclinical phase of the disease and that this effect may vary according to the type of fat considered.

Isolation of rat alpha1-fetoprotein messenger RNA from Morris hepatoma 7777.

A double-antibody procedure has been developed for the isolation of alpha1-fetoprotein (AFP)-synthesizing polysomes from Morris hepatoma 7777. The polyadenylic acid-containing RNA, subsequently purified by differential sedimentation on sucrose gradient and oligodeoxythymidylic acid-cellulose chromatography, migrates as a single 21S component in polyacrylamide gel electrophoresis; in a cell-free translation system, it yields a peptide product immunoprecipitable by anti-rat AFP antiserum, but not by anti-rat albumin, and which migrates slightly faster than serum AFP on sodium dodecyl sulfate-urea-polyacrylamide gels. This messenger RNA fraction was used for the synthesis of a radioactive complementary DNA. In hybridization assays, the complementary DNA reassociated with its purified template at a Cr0t1/2 [product of RNA concentration (mol of nucleotides per liter) X half-time (sec)] of 1.5 X 10(-2). By constitute 3,2, and less than 0.01% of total polyadenylic acid-containing polysomal RNA of Morris hepatoma 7777, 10-day-old-rat liver, and adult rat liver, respectively. The high specificity of the polysome immunoprecipitation system, the electrophoretic homogeneity of the isolated messenger RNA fraction, its selective translation into AFP, and the specificity of the hybridization probe indicate that the procedure described yields a highly purified rat AJP messenger RNA.

Differential regulation of normal and tumor alpha 1-fetoprotein genes in fetal hepatocyte x hepatoma hybrids.

Fetal rat hepatocytes and mouse hepatoma cells actively expressing alpha 1-fetoprotein (AFP) and albumin genes were fused with the use of Sendai virus, and the expression of normal (rat) and tumor (mouse) AFP and albumin genes was analyzed in hybrid clones. The tumor AFP gene and both albumin genes were active in 103 hybrids. Expression of the normal fetal rat AFP gene, however, was maintained in only 3 hybrids, and it was frequently lost or decreased selectively upon subcloning. Furthermore, the normal AFP gene, when expressed, was more reactive than the tumor AFP gene to repression by a glucocorticosteroid hormone. These results suggest constitutive differences in the manner an oncofetal gene is activated and regulated in normal and neoplastic states. AFP gene expression in normal hepatocytes appears to be subordinated to a differentiation program degenerated and bypassed in hepatoma cells.

Tandem arrangement of the human serum albumin multigene family in the sub-centromeric region of 4q: evolution and chromosomal direction of transcription.

The albumin gene family is comprised of four genes encoding: serum albumin (ALB), alpha-fetoprotein (AFP), alpha-albumin (ALF), and vitamin D-binding protein (DBP; also known as GC). The genes are regulated developmentally, expressed in the liver, and the proteins are secreted into the bloodstream. The GC gene, and the tandemly linked ALB and AFP genes, have been previously localized to human chromosome 4q11-13. Using techniques of fluorescence in situ hybridization to chromatin fibres, chromosome walking and DNA sequencing of genomic clones, we now report on the chromosomal location of the ALF gene and the organization of the entire gene family. The four genes are tandemly linked in the 4q sub-centromeric region: 5'ALB-5'AFP-5'ALF-5'GC3'-centromere, and hence are transcribed in the same, centromere-bound, direction. The linear arrangement of the four genes along the chromosome is not correlated with their temporal expression in the human ontogeny. It appears that GC is very close (and may be the gene proximal) to the centromere. The linear chromosomal arrangement of the four genes and the structural differences between them are congruent with the following evolutionary divergence of the gene family. Starting with the first duplication of an ancestral progenitor gene, a single evolutionary line led to the contemporary GC, leaving ALB/AFP/ALF on the other line of descent. The second duplication occurred in this ALB lineage, giving rise to ALB and the AFP/ALF progenitor, and the third, most recent one, gave rise to the AFP-ALF pair.

The nuclear receptor fetoprotein transcription factor is coexpressed with its target gene HNF-3beta in the developing murine liver, intestine and pancreas.

During organogenesis, the winged helix hepatocyte nuclear factor 3beta (HNF-3beta) protein participates in regulating gene transcription in the developing esophagus, trachea, liver, lung, pancreas, and intestine. Hepatoma cell transfection studies identified a critical HNF-3beta promoter factor, named UF2-H3beta, and here, we demonstrate that UF2-H3beta is identical to the fetoprotein transcription factor (FTF). In situ hybridization studies of mouse embryos demonstrate that FTF expression initiates in the foregut endoderm during liver and pancreatic morphogenesis (day 9) and that earlier expression of FTF is observed in the yolk sac endoderm, branchial arch and neural crest cells (day 8). Abundant FTF hybridization signals are observed throughout morphogenesis of the liver, pancreas, and intestine and its expression continues in the epithelial cells of these adult organs. In day 17 mouse embryos and adult pancreas, however, expression of FTF becomes restricted to the exocrine acinar and ductal epithelial cells.