The WWOX gene in brain development and pathology.

IMPACT STATEMENT: WW domain-containing oxidoreductase encoded by the WWOX gene is a transcription regulator and a key player in a number of cellular and biological processes such as tumor suppression, cell proliferation, apoptosis induction, steroid metabolism, and central nervous system development. This review provides a comprehensive summary of currently known roles and discusses the importance of WWOX gene for CNS development and functioning.

Boundary spanning at the science-policy interface: the practitioners' perspectives.

Cultivating a more dynamic relationship between science and policy is essential for responding to complex social challenges such as sustainability. One approach to doing so is to "span the boundaries" between science and decision making and create a more comprehensive and inclusive knowledge exchange process. The exact definition and role of boundary spanning, however, can be nebulous. Indeed, boundary spanning often gets conflated and confused with other approaches to connecting science and policy, such as science communication, applied science, and advocacy, which can hinder progress in the field of boundary spanning. To help overcome this, in this perspective, we present the outcomes from a recent workshop of boundary-spanning practitioners gathered to (1) articulate a definition of what it means to work at this interface ("boundary spanning") and the types of activities it encompasses; (2) present a value proposition of these efforts to build better relationships between science and policy; and (3) identify opportunities to more effectively mainstream boundary-spanning activities. Drawing on our collective experiences, we suggest that boundary spanning has the potential to increase the efficiency by which useful research is produced, foster the capacity to absorb new evidence and perspectives into sustainability decision-making, enhance research relevance for societal challenges, and open new policy windows. We provide examples from our work that illustrate this potential. By offering these propositions for the value of boundary spanning, we hope to encourage a more robust discussion of how to achieve evidence-informed decision-making for sustainability.

WWOX--the FRA16D cancer gene: expression correlation with breast cancer progression and prognosis.

AIMS: WWOX is a tumour suppressor gene involved in various tumours including breast cancer. High chromosomal abnormalities in a genomic region spanned by WWOX are associated with the fact that this gene covers approximately 1 million base pairs of the second most affected among common chromosomal fragile sites FRA16D. We evaluated WWOX expression levels in breast cancer samples in association with diagnostics-prognostics markers. METHODS: We performed quantitative real-time RT-PCR to analyse levels of expression of WWOX in 132 cases of breast cancer. We evaluated the relationship between WWOX mRNA levels, clinico-pathological factors, expression of aberrant WWOXDelta6-8 mRNA and other cancer related genes. RESULTS: Expression of WWOX was higher in patients younger than 50 years old, in ER and PR positive tumours vs negative for those receptors and tumours without lymph node metastasis vs LN+. WWOX mRNA levels were also higher in tumours with higher apoptotic index (Bcl2/Bax ratio). Negative associations were found between WWOX expression and cytokeratins 5/6 and 17 (P<0.05). High level expression of WWOX was also associated with better disease free survival. Presence of WWOXDelta6-8 transcripts were accompanied with lower WWOX wild type mRNA level. CONCLUSIONS: Reduced WWOX expression commonly observed in various neoplasias in cases of breast cancer is associated with markers of bad prognosis. Our findings reveal additional evidence that WWOX may be involved in steroid (estrogens) metabolism and signaling pathways. WWOX can be considered as a new target for gene therapy development due to the association of high WWOX expression with improved disease free survival.

Cyclin E expression in breast cancer correlates with negative steroid receptor status, HER2 expression, tumor grade and proliferation.

The main objective of this retrospective study was to investigate relations between cyclin E and pathoclinical factors in patients with operable breast cancer. Expression of cyclin E was analyzed by immunohistochemistry in specimens of invasive ductal breast cancer tissue obtained from 189 women during radical mastectomy. Overall, 110 tumor samples were regarded to be cyclin E positive. Cyclin E expression was more often seen in tumors with: negative steroid receptor status (p<0.0001), higher proliferative index (p=0.0014), higher tumor grade (p=0.0017), and presence of HER2 (p=0.0171). With a median follow-up of 58 months, expression of cyclin E together with negative steroid receptor status determined poor prognosis with a 5-year cancer-specific survival rate of 58%. It differed significantly from a survival curve of cyclin E negative and steroid receptor positive patients (87%, p=0.0005). No significant difference was observed in comparison with survival of cyclin E positive and steroid receptor positive patients (68%, p=0.221). We demonstrated that cyclin E expression in breast cancer cells was associated with negative steroid receptor status, HER2 presence, higher tumor grade and higher proliferation index. Expression of cyclin E together with lack of steroid receptors determined poor prognosis.

Increased telomerase activity in mouse skin premalignant progression.

It has been postulated that the expression of the ribonucleoprotein telomerase is necessary to overcome cellular senescence and that malignant tumors must express telomerase to maintain their immortality. In most human adult tissues, telomerase activity is not detected. In contrast, several murine tissues express various levels of telomerase. Mouse skin however, does not show telomerase activity. Using the mouse skin chemical carcinogenesis system, a well-characterized model for studying premalignant and malignant progression, we assayed telomerase activity at various stages of premalignant papilloma progression by means of the recently developed telomeric repeat amplification protocol. We observed that at 10 weeks of promotion, only one mouse skin papilloma of 11 analyzed showed high levels of telomerase activity. The number of papillomas showing higher levels of telomerase activity increased at 20 weeks, and at 30 weeks of promotion, 100% of papillomas expressed significantly higher levels of telomerase. We learned from previous studies that early papillomas are diploid, well-differentiated lesions, whereas late papillomas are aneuploid and very dysplastic. It appears that the progressive increase in telomerase activity is associated with the increased level of genomic instability and the phenotypic progression of these premalignant tumors. It is also possible, however, that the increase in telomerase activity could be in part a consequence of an increase in the proportion of proliferating cells. Nevertheless, the mouse skin system may be a very useful in vivo model for the study and development of anti-telomerase therapeutic strategies.

WWOX, a novel WW domain-containing protein mapping to human chromosome 16q23.3-24.1, a region frequently affected in breast cancer.

Studies were conducted with the final goal of identifying genes of interest mapping to the chromosome region 16q23.3-24.1, an area commonly affected by allelic losses in breast cancer. To this end we generated a detailed physical map of the genomic region spanning between sequence-tagged site markers D16S518 and D16S516. To identify candidate genes, we used shotgun genomic sequencing as well as isolation and analysis of transcripts mapping to the area of interest. We identified and cloned a novel gene, the genomic structure of which spans the whole region of interest. We named this gene WWOX because it contains two WW domains coupled to a region with high homology to the short-chain dehydrogenase/reductase family of enzymes. The ORF of WWOX is 1245 bp long, encoding a 414-amino acid protein. This gene is composed of nine exons. We performed a mutation screening of WWOX exons in a panel of breast cancer lines, most of which are hemizygous for the 16q genomic region indicated. We found no evidence of mutations, thus indicating that WWOX is probably not a tumor suppressor gene. However, we observed that one case of homozygous deletion as well as two previously described translocation breakpoints map to intronic regions of this gene. We speculate that WWOX may span the yet uncharacterized common fragile site FRA16D region. In expression studies we found overexpression of WWOX in breast cancer cell lines when compared with normal breast cells and tissues. The highest normal expression of WWOX was observed in hormonally regulated tissues such as testis, ovary, and prostate. This expression pattern and the presence of a short-chain dehydrogenase/reductase domain and specific amino acid features suggest a role for WWOX in steroid metabolism. Interestingly, the presence of WW domains in the structure of WWOX indicate the likelihood that this protein physically interacts with other proteins. The unique features of WWOX and its possible association with cancer processes make it an interesting target for further investigation.

Effects of estrogen on global gene expression: identification of novel targets of estrogen action.

The important role played by the sex hormone estrogen in disease and physiological processes has been well documented. However, the mechanisms by which this hormone elicits many of its normal as well as pathological effects are unclear. To identify both known and unknown genes that are regulated by or associated with estrogen action, we performed serial analysis of gene expression on estrogen-responsive breast cancer cells after exposure to this hormone. We examined approximately 190,000 mRNA transcripts and monitored the expression behavior of 12,550 genes. Expression levels for the vast majority of those transcripts were observed to remain constant upon 17beta estradiol (E2) treatment. Only approximately 0.4% of the genes showed an increase in expression of > or =3-fold by 3 h post-E2 treatment. We cloned five novel genes (E2IG1-5), which were observed up-regulated by the hormonal treatment. Of these the most highly induced transcript, E2IG1, appears to be a novel member of the family of small heat shock proteins. The E2IG4 gene is a new member of the large family of leucine-rich repeat-containing proteins. On the basis of architectural and domain homology, this gene appears to be a good candidate for secretion in the extracellular environment and, therefore, may play a role in breast tissue remodeling and/or epithelium-stroma interactions. Several interesting genes with a potential role in the regulation of cell cycle progression were also identified to increase in expression, including Pescadillo and chaperonin CCT2. Two putative paracrine/autocrine factors of potential importance in the regulation of the growth of breast cancer cells were identified to be highly up-regulated by E2: stanniocalcin 2, a calcium/phosphate homeostatic hormone; and inhibin-beta B, a TGF-beta-like factor. Interestingly, we also determined that E2IG1 and stanniocalcin 2 were exclusively overexpressed in estrogen-receptor-positive breast cancer lines, and thus they have the potential to serve as breast cancer biomarkers. This data provides a comprehensive view of the changes induced by E2 on the transcriptional program of human E2-responsive cells, and it also identifies novel and previously unsuspected gene targets whose expression is affected by this hormone.

WWOX, the FRA16D gene, behaves as a suppressor of tumor growth.

We recently reported the cloning of WWOX, a gene that maps to the common fragile site FRA16D region in chromosome 16q23.3-24.1. It was observed that the genomic area spanned by WWOX is affected by chromosomal translocations and homozygous deletions. Furthermore, the high incidence of allelic loss in breast, ovarian, prostate, and other cancers affecting this region suggests that WWOX is a candidate tumor suppressor gene. Expression of WWOX is highly variable in breast cancer cell lines, with some cases showing low or undetectable levels of expression. In this report, we demonstrate that ectopic WWOX expression strongly inhibits anchorage-independent growth in soft agar of breast cancer cell lines MDA-MB-435 and T47D. Additionally, we observed that WWOX induces a dramatic inhibition of tumorigenicity of MDA-MB-435 breast cancer cells when tested in vivo. We also detected the common occurrence of aberrant WWOX transcripts with deletions of exons 5-8 or 6-8 in various carcinoma cell lines, multiple myeloma cell lines, and primary breast tumors. These aberrant mRNA forms were not detected in normal tissues. Interestingly, we further observed that proteins encoded by such aberrant transcripts display an abnormal nuclear localization in contrast to the wild-type WWOX protein that localizes to the Golgi system. Our data indicate that WWOX behaves as a potent suppressor of tumor growth and suggest that abnormalities affecting this gene at the genomic and transcriptional level may be of relevance in carcinogenesis.

The influence of the WWOX gene on the regulation of biological processes during endometrial carcinogenesis.

The purpose of the present study was to investigate the role of WW domain containing oxidoreductase (WWOX) downregulation in biological cancer-related processes in normal (non-malignant) and cancer endometrial cell lines. We created an in vitro model using the normal endometrial cell line, THESC, and 2 endometrial cancer cell lines with varying degrees of differentiation, the Ishikawa (well-differentiated) and the MFE296 (moderately differentiated) cells, in which the WWOX tumor suppressor gene was silenced using Gipz lentiviral shRNA. In this model, we examined the changes in invasiveness via biological assays, such as zymography, migration through a basement membrane, the adhesion of cells to extracellular matrix proteins, anchorage-independent growth and colony formation assay. We also evaluated the correlation between the mRNA expression of the WWOX gene and genes involved in the processes of carcinogenesis, namely catenin beta-1 (CTNNB1) and zinc finger E-box binding homeobox 1 (ZEB1) (gene transcription), cadherin 1 (CDH1) and ezrin (EZR) (cell adhesion), vimentin (VIM) (structural proteins), as well as phosphatase and tensin homolog (PTEN) (tumor suppression) and secreted protein, acidic, cysteine-rich (osteonectin) (SPARC) (SPARC) (cell growth regulation) by RT-qPCR. Downregulation of the WWOX gene in the moderately differentiated MFE296 cell line caused decreased migratory capacity, and a reduction of matrix metalloproteinase-2 (MMP-2) activity. However, these cells grew in semisolid medium and exhibited higher expression of CDH1 and EZR (cell adhesion) and secreted protein, acidic, cysteine-rich (osteonectin) (SPARC) (cell growth regulation). Moreover, in the well-differentiated endometrial cancer (Ishikawa) cell line, WWOX gene silencing resulted in an increased ability of the cells to proliferate indefinitely. Additionally, WWOX regulated changes in adhesion potential in both the normal and cancer cell lines. Our results suggest that the WWOX tumor suppressor gene modulated the processes of cell motility, cell adhesion, gene expression and remodeling in endometrial cell lines.

Correlation between depression and burden observed in informal caregivers of people suffering from dementia with time spent on caregiving and dementia severity.

OBJECTIVE: The aim of the study is to compare data on the examined population of informal caregivers of people suffering from dementia with previous studies, as well as to assess the correlation between (i) depression determined on the basis of the Center for Epidemiologic Studies Depression Scale and (ii) caregiver burden measured by means of the Zarit Caregiver Burden Scale and some chosen parameters, such as total time devoted to caregiving, time of caregiving in hours per week and level of dementia severity measured by Global Deterioration Scale. PATIENTS AND METHODS: 41 informal caregivers of people suffering from dementia from different backgrounds were evaluated using the Zarit Caregiver Burden Scale and the Center for Epidemiologic Studies Depression Scale. Demographic data about the time devoted to caregiving and the number of hours spend on caregiving weekly were gathered. The type of dementia and its stage were registered using the Global Deterioration Scale (GDS). With the aid of the Statistica StatSoft program, mutual correlations between the parameters were measured. The study was conducted within the framework of AAL UnderstAID--a platform that supports and helps to understand and assist caregivers in the care of a relative with dementia. The international project is co-founded by the Joint Programme Ambient Assisted Living (Grant code: ESR-aal 2012 5 107). RESULTS: No significant correlations between the level of depression severity evaluated in caregivers and the total time of taking care of a demented person or time of caregiving in hours per week were observed. Similarly, no significant correlation between depression severity level and dementia severity level measured on the GDS scale were noted. There was also no significant correlation between Zarit Caregiver Burden Scale scores and the above-mentioned parameters. CONCLUSIONS: The level of depression among caregivers do not depend on socio-demographic factors.

Constitutive telomerase activity in cells with tissue-renewing potential from estrogen-regulated rat tissues.

To investigate the role of telomerase in estrogen-regulated rodent tissues, we assayed the activity levels of this enzyme and measured cell proliferation and indicators of cellularity in vagina, mammary gland, and uterus from virgin, pregnant, ovariectomized, and ovariectomized estradiol-treated rats. No association was observed between telomerase activity and increased cell proliferation. Telomerase activity was significantly higher (P=0.003) in vagina obtained from ovariectomized rats (very low proliferation) than in vagina from ovariectomized and estradiol-treated rats (high proliferation, high differentiation). The high telomerase levels observed in vagina from ovariectomized rats indicates that the same epithelial compartment (i.e., basal layer) that has the potential to reconstitute the epithelium also contains the cells that express telomerase. The lower telomerase activity in the keratinized (differentiated) vagina was probably due to dilution of the number of telomerase-producing cells by the terminally differentiated non-telomerase-producing cells. Similar results were observed in uterus from ovariectomized versus ovariectomized and estradiol-treated rats. Telomerase activity was highest in uterus from pregnant rats. Telomerase levels in samples from total mammary gland fat pads varied considerably between groups and appeared to be representative of the amount of epithelium present in the sample. Interestingly, when mammary gland samples from the same animals were obtained from pure epithelial organoid preparations, no differences in telomerase activity could be distinguished between animals or groups. Overall these data suggest that telomerase activity, particularly in rat vagina and uterus, appears to be associated with a cell subpopulation showing proliferative and tissue reconstitution potential and not directly associated with proliferation status per se.

Bovine 1.709 satellite. Recombination hotspots and dispersed repeated sequences.

The nucleotide sequence of 3800 base-pair repeated unit of bovine 1.709 satellite was determined. The 3800 base-pair unit is not internally repeated and contains members of at least three different families of elements that are dispersed in the bovine genome. Two of three elements are associated with extensive length polymorphism within the satellite repeat unit. One of these comprises the 3' end of the bovine Alu-like sequences; the second is composed of C-A dinucleotides.

GADD45A and EPB41 as tumor suppressor genes in meningioma pathogenesis.

Deletions of 1p occur in approximately 30% of meningiomas. Based on loss of heterozygosity (LOH) analysis, two regions on 1p have been suspected to be carriers of tumor suppressor genes. We chose the GADD45A and EPB41 genes as tumor suppressor candidates based on their function and chromosomal localization. We analyzed 19 cases of meningioma with LOH of 1p by means of sequencing of the GADD45A gene and Western blotting of the GADD45a protein. Twenty cases of meningioma without 1p LOH were also analyzed by Western blotting to find out if changes of the GADD45a protein expression occurred. Nineteen samples with 1p LOH (12 grade I; 7 grade II, WHO classification) and 20 samples without 1p LOH (18 grade I; 2 grade II) were also analyzed by means of real-time polymerase chain reaction to find abnormalities in EPB41 mRNA levels in meningioma. LOH analysis was performed using seven microsatellite markers: D1S508 (1p36.2), D1S199 (1p36.1) D1S2734 (1p36.1), D1S2720 (1p34), D1S197 (1p32), D1S162 (1p32), D1S429 (1p11). LOH analysis confirmed previously described localization of putative tumor suppressor genes on 1p and involvement in meningioma pathogenesis (1p36 and 1p32). The open reading frame of GADD45A and intron splicing sites showed neither mutations nor polymorphisms. GADD45a protein molecular weight and expression level were unaltered in meningiomas with and without 1p LOH. We conclude that the GADD45A gene is not involved in meningioma tumorigenesis. EPB41 gene expression was unchanged in all analyzed meningiomas. This suggests that involvement of the EPB41 gene (4.1R protein) in meningioma pathogenesis should be reconsidered.

Analysis of telomerase activity levels in breast cancer: positive detection at the in situ breast carcinoma stage.

Telomerase activity has been implicated to be associated with most human malignant tumors, including breast cancer. To evaluate possible associations with well-known prognostic factors in breast cancer, we performed a semi-quantitative analysis of telomerase activity levels using the very sensitive PCR-mediated telomeric repeat amplification protocol. Telomerase activity was detected in 99 of 104 breast cancer samples analyzed (95.2%), whereas no activity was detected in 10 of 10 adjacent nonmalignant breast tissues. Analysis of five breast fibroadenoma samples revealed telomerase activity in one (20%). In contrast to previous observations, we observed that 100% of stage I breast tumors were positive for telomerase activity. More interestingly, we detected telomerase activity in six of six ductal carcinoma in situ samples (i.e., stage 0). In our semiquantitative analysis of levels of enzymatic activity, we found no statistically significant correlation at the P < h 0.05 level between telomerase levels and lymph node metastasis status, estrogen and progesterone receptor status, tumor size, S-phase fraction, and ploidy. The only statistically significant correlation was found with patient age (rho = -0.3; P = 0.03). We observed no statistically significant difference in the telomerase activity levels of early tumors (stages 0 and 1) versus more advanced lesions (stages II to IV). Nevertheless, stage IV tumors displayed a tendency for higher telomerase activity levels. In summary, no clear association was observed between telomerase levels and known breast cancer prognostic indicators. However, telomerase detection by the telomeric repeat amplification protocol method, due to its high sensitivity, may be of value in early breast cancer diagnosis and detection, because our data indicate that telomerase reactivation appears to constitute a relatively early event in breast carcinogenesis.

WWOX, the common chromosomal fragile site, FRA16D, cancer gene.

Gross chromosomal rearrangements and aneuploidy are among the most common somatic genomic abnormalities that occur during cancer initiation and progression, in particular in human solid tumor carcinogenesis. The loss of large chromosomal regions as consequence of gross rearrangements (e.g. deletions, monosomies, unbalanced translocations and mitotic recombination) have been traditionally associated with the existence of tumor suppressor genes within the areas affected by the loss of genetic material. The long arm of chromosome 16 was identified as being frequently associated with structural abnormalities in multiple neoplasias, that led us to focus attention on the detailed genetic dissection of this region resulting in the cloning of the putative tumor suppressor gene, WWOX (WW domain containing Oxidoreductase). Interestingly, the WWOX gene resides in the very same region as that of the common chromosomal fragile site 16D (FRA16D). The WWOX gene encodes a protein that contains two WW domains, involved in protein-protein interactions, and a short chain dehydrogenase (SDR) domain, possibly involved in sex-steroid metabolism. We have identified the WWOX WW domain ligand as the PPXY motif confirming the biochemical activity of this domain. WWOX normally resides in the Golgi and we will demonstrate that Golgi localization requires an intact SDR. Inactivation of the WWOX gene during tumorigenesis can occur by homozygous deletions and possibly mutation, however, aberrantly spliced forms of WWOX mRNA have been observed even when one allele is still intact. The aberrantly spliced mRNAs have deletions of the exons that encode the SDR and these WWOX protein isoforms display abnormal intracellular localization to the nucleus possibly functioning as dominant negative inhibitors of full length WWOX. Thus, generation of aberrant transcripts of WWOX may represent a novel mechanism to functionally inactivate WWOX without genomic alteration of the remaining allele. In this article we will review the cloning and identification of WWOX as the target of FRA16D. In addition, we will discuss the possible biochemical functions of WWOX and present evidence that ectopic WWOX expression inhibits tumor growth.

Film badges for personal dosimetry of roentgen radiation.

The paper describes the calibration procedure for and data obtained from routine X-ray dosimetry service in Poland. In 1998 the number of readouts amounted to 152,000. The dosimetry concerned 31,281 workers in 3,113 enterprises. The annual collective dose was assessed to be 16.7 man.Sv. The analysis of the calibration quality is discussed as well. Only 2.9% of all points lay outside the quality curve while the number of the acceptable outliers was 5%.

Compatibility of soil amendments with entomopathogenic nematodes.

The impact of inorganic and organic fertilizers on the infectivity, reproduction, and population dynamics of entomopathogenic nematodes was investigated. Prolonged (10- to 20-day) laboratory exposure to high inorganic fertilizer concentrations inhibited nematode infectivity and reproduction, whereas short (1-day) exposures increased infectivity. Heterorhabditis bacteriophora was more sensitive to adverse effects than were two species of Steinernema. In field studies, organic manure resulted in increased densities of a native population of Steinernema feltiae, whereas NPK fertilizer suppressed nematode densities regardless of manure applications. Inorganic fertilizers are likely to be compatible with nematodes in tank mixes and should not reduce the effectiveness of nematodes used for short-term control as biological insecticides, but may interfere with attempts to use nematodes as inoculative agents for long-term control. Organic manure used as fertilizer may encourage nematode establishment and recycling.

[Sources of x-rays in school]. / Zródla promieniowania rentgenowskiego w szkole.

Exposure of teachers of physics to x-radiation emitted by instruments which are used for demonstrating rarefied gas discharges during physics lessons at secondary schools is discussed. The measurements performed provide an explicit evidence that an effective, anual exposure dose under the most unfavourable conditions does not exceed admissible levels according to Polish regulations pertaining to persons non-occupationally exposed to ionizing radiation.

[The incidence of virus respiratory infections in small children in the season 1994-95]. / Wystepowanie wirusowych zakazen dróg oddechowych u malych dzieci w sezonie 1994-1995.

As previously, by antibody fluorescent technique the etiology of viral respiratory infections was determinated in small children hospitalized because of respiratory disease in the epidemic season 1994-95. Data obtained were compared with the results from the preceding season. Among a total of 371 patients, virus infection was detected in 170 (45,8%). Data for the whole season did not distinctly differ from those obtained previously; however, some differences were found in the occurrence of the individual virus infections in consecutive months of the epidemic season. The second (spring) wave of RS virus infections in this season was lower and more extended in time than in the previous seasons. Highest incidence of adenoviruses was observed in July 1995. The incidence of parainfluenza type 4 virus was higher in the disease of small children. In the youngest (below 1 month) age group percent of virus infections was higher, as compared with data from the preceding season.

[Teaching schizophrenic patients self-care skills]. / Uczenie pacjentów chorych na schizofrenie umiejetnosci samoobslugi.

The level of knowledge and skills on self-care abilities has been examined among the group of 30 long-term patients suffering from schizophrenia. The research has been made four times during the period of 18 months after the training session had been completed. The final results show that three months duration of the training session exerts influence on the progress in learning and change of the view point. However, little change is observed as regards the daily functioning of patients.