Hetero-epitaxial approach by using labile coordination sites to prepare catenated metal-organic frameworks with high surface areas.

A solid-state approach that takes advantage of the ordered 3D arrangement of active secondary building units allows the preparation of new interlocked MOFs that grow hetero-epitaxially on the crystal faces of a precursor phase that acts as a "topological blueprint". The synthetic strategy is exemplified by using rigid acetylene-based ligands to produce highly augmented Cu(II) acetate-based MOFs.

Stat1 activation attenuates IL-6 induced Stat3 activity but does not alter apoptosis sensitivity in multiple myeloma.

BACKGROUND: Multiple myeloma (MM) is at present an incurable malignancy, characterized by apoptosis-resistant tumor cells. Interferon (IFN) treatment sensitizes MM cells to Fas-induced apoptosis and is associated with an increased activation of Signal transducer and activator of transcription (Stat)1. The role of Stat1 in MM has not been elucidated, but Stat1 has in several studies been ascribed a pro-apoptotic role. Conversely, IL-6 induction of Stat3 is known to confer resistance to apoptosis in MM. METHODS: To delineate the role of Stat1 in IFN mediated sensitization to apoptosis, sub-lines of the U-266-1970 MM cell line with a stable expression of the active mutant Stat1C were utilized. The influence of Stat1C constitutive transcriptional activation on endogenous Stat3 expression and activation, and the expression of apoptosis-related genes were analyzed. To determine whether Stat1 alone would be an important determinant in sensitizing MM cells to apoptosis, the U-266-1970-Stat1C cell line and control cells were exposed to high throughput compound screening (HTS). RESULTS: To explore the role of Stat1 in IFN mediated apoptosis sensitization of MM, we established sublines of the MM cell line U-266-1970 constitutively expressing the active mutant Stat1C. We found that constitutive nuclear localization and transcriptional activity of Stat1 was associated with an attenuation of IL-6-induced Stat3 activation and up-regulation of mRNA for the pro-apoptotic Bcl-2 protein family genes Harakiri, the short form of Mcl-1 and Noxa. However, Stat1 activation alone was not sufficient to sensitize cells to Fas-induced apoptosis. In a screening of > 3000 compounds including bortezomib, dexamethasone, etoposide, suberoylanilide hydroxamic acid (SAHA), geldanamycin (17-AAG), doxorubicin and thalidomide, we found that the drug response and IC50 in cells constitutively expressing active Stat1 was mainly unaltered. CONCLUSION: We conclude that Stat1 alters IL-6 induced Stat3 activity and the expression of pro-apoptotic genes. However, this shift alone is not sufficient to alter apoptosis sensitivity in MM cells, suggesting that Stat1 independent pathways are operative in IFN mediated apoptosis sensitization.

In vitro activity of 20 agents in different prognostic subgroups of chronic lymphocytic leukemia--rolipram and prednisolone active in cells from patients with poor prognosis.

BACKGROUND: There is a need for development of new drugs for treatment of B-cell chronic lymphocytic leukemia (CLL), especially for poor-prognostic subgroups resistant to conventional therapy. OBJECTIVE: The in vitro antileukemic activity of 20 different anticancer agents was characterized in tumor cells from CLL, aiming at identifying agents active in poor-prognostic subgroups. DESIGN AND METHODS: In tumor cells from 40 CLL patients and in peripheral blood mononuclear cells (PBMC) from three healthy controls, the activity of 20 substances was assessed using a non-clonogenic assay. The CLL samples were characterized regarding genomic aberrations by interphase fluorescence in situ hybridization and immunoglobulin heavy-chain variable (IGHV) gene mutational status. RESULTS: In line with clinical experience, cells from patients with unfavourable genomic aberrations [del(11q)/del(17p)] showed lower drug sensitivity to fludarabine and chlorambucil than cells from patients with favourable cytogenetics [del(13q)/no aberration]. Most investigated drugs demonstrated similar activity in CLL cells from patients with unmutated and mutated IGHV genes as well as in CLL cells vs. PBMC. Interestingly, prednisolone and rolipram displayed high CLL specificity, high activity in CLL cells with unmutated IGHV genes and retained the effect in several cases with 11q/17p deletion. Further studies on prednisolone and rolipram revealed a synergy when these agents were combined in CLL cells, and suggested correlation between drug sensitivity and difference in downstream signaling. CONCLUSION: Prednisolone and rolipram are interesting for further studies in CLL with inferior prognosis. The study can also be considered a basis for future efforts to find drugs active in subsets of CLL patients that are resistant to conventional therapy.

The GNAS1 T393C polymorphism and lack of clinical prognostic value in chronic lymphocytic leukemia.

Chronic lymphocytic leukemia (CLL) is a clinically heterogeneous disease with no known single predisposing genetic factor shown in all cases. Recently, a single nucleotide polymorphism (SNP) T393C in the GNAS1 gene has been reported to have a clinical impact on CLL progression and overall survival. In order to further investigate the T393C SNP in CLL, we have genotyped 279 CLL cases and correlated the genotypes to clinical outcome and other known prognostic factors such as the immunoglobulin heavy chain variable (IGHV) gene mutation status and CD38 expression. In the present study, no difference in overall survival or time to treatment was observed in the CLL patients with the different genotypes in contrast to the previous report. Furthermore, no correlation was observed with the T393C genotypes and IGHV mutational status, Binet stage or CD38 in this cohort. In summary, our data does not support the use of the T393C GNAS SNP as a clinical prognostic factor in CLL.

Thioredoxin, produced by stromal cells retrieved from the lymph node microenvironment, rescues chronic lymphocytic leukemia cells from apoptosis in vitro.

BACKGROUND AND OBJECTIVES: The redox-regulatory protein thioredoxin has several functions including transcriptional regulation, and antioxidant, cytokine, and chemokine activities. We have previously shown that extracellular thioredoxin protects B-cell chronic lymphocytic leukemia (CLL) cells from apoptosis in vitro. In this study we were interested to determine whether thioredoxin is produced by cells surrounding the CLL cells in the in vivo microenvironment and whether this cell-derived thioredoxin has any leukemia growth-promoting effect in vitro. DESIGN AND METHODS: Lymph nodes from CLL patients (n=25) were analyzed for thioredoxin expression by immunohistology. Stromal cells purified from the lymph nodes were analyzed for thioredoxin secretion at the single cell level using an ELIspot assay. The survival effect of the stromal-derived thioredoxin was tested by co-culturing stromal- and CLL cells with and without Fab-fragments of an anti-thioredoxin antibody. RESULTS: The results indicated that the thioredoxin production correlated with the amount of proliferating cells and was mainly localized to the proliferation centers (pseudofollicles) in the CLL lymph nodes. The leukemia cells per se showed minimal thioredoxin levels; in contrast, stromal cells strongly expressed thioredoxin. Purified primary stromal cells, which secreted extracellular thioredoxin, significantly protected the CLL cells from undergoing apoptosis in 72 h co-cultures. Interestingly, this anti-apoptotic effect could be abrogated by addition of Fab-fragments of an anti- thioredoxin antibody. INTERPRETATION AND CONCLUSIONS: In conclusion, we have shown that stromal cells in the lymph node microenvironment produce thioredoxin and that the thioredoxin production is localized to the proliferation centers of the CLL lymph nodes. In addition, thioredoxin produced by purified stromal cells rescued CLL cells from apoptosis in vitro.

The immunoglobulin genes: structure and specificity in chronic lymphocytic leukemia.

The rearrangement of the immunoglobulin genes (IG) provides a large diversity of B-cell receptors conformations and allows the immune system to respond differently to foreign antigens. In chronic lymphocytic leukemia (CLL), there are a restricted number of stereotyped B-cell receptors rearranged by the tumor B-cells between CLL patients. These subsets with stereotyped receptors appear to have clinical implications, for example cases that rearrange the IGHV3-21 gene display poor clinical prognosis. The number of subsets with stereotyped receptors has been reported at a frequency of over 20% of CLL cases; however, the specificities of these receptors are still not clearly defined. Reactivity to epitopes from bacterial antigen, cytoskeleton components such as vimentin, and antigens on viable and apoptotic T-cell have been proposed. The role of antigen in CLL development is currently being more clearly defined with identification of stereotyped receptors, and their antigen specificity and the continued role antigen stimulation plays in CLL disease will be an important question in the future.

Patients with chronic lymphocytic leukemia with mutated VH genes presenting with Binet stage B or C form a subgroup with a poor outcome.

BACKGROUND AND OBJECTIVES: The immunoglobulin VH gene mutation status is a strong prognostic indicator in B-cell chronic lymphocytic leukemia (CLL), since unmutated VH genes are correlated with short survival. However, the traditional cut-off level dividing mutated and unmutated cases, i.e. more or less than 2% mutations, has been questioned and other cut-offs have been suggested. We investigated whether an alternative cut-off should be applied and the relation of mutational status to another prognostic marker, Binet staging. DESIGN AND METHODS: VH gene mutation status was assessed in 332 CLL cases by polymerase chain reaction amplification and nucleotide sequencing and was further correlated with overall survival using different VH mutation cut-offs (1-7%) and Binet stage. RESULTS: After testing different mutation borders, the 2% cut-off remained the best discriminative level for determining prognosis. Interestingly, prognostic stratification was improved by combining the information on VH gene mutation status with that of Binet stage: unmutated cases (all stages, n=151, mutated cases with stage A (n=77), and mutated cases with stage B or C (n=37) had a median survival of 82, 179 and 74 months, respectively. INTERPRETATION AND CONCLUSIONS: CLL cases displaying mutated VH genes with Binet stage B or C had a survival similar to that of unmutated cases and significantly shorter than that of mutated stage A CLL. Our result reveals clinical heterogeneity within the VH mutated CLL group by inclusion of Binet stage data, a finding which is of importance when considering surrogate marker(s) for VH mutation status.

Prognostic usage of V(H) gene mutation status and its surrogate markers and the role of antigen selection in chronic lymphocytic leukemia.

B-cell chronic lymphocytic leukemia (CLL) is a clinically heterogeneous disease with many patients surviving for decades with minimal or no treatment, whereas others succumb rapidly to their disease despite therapy. In recent years, new molecular prognostic factors have emerged in CLL that have significantly improved the subgrouping of the disease. One of the most important molecular predictors, the immunoglobulin V(H) gene mutation status, divides CLL into two prognostic groups, depending on the presence or absence of somatic hypermutation, where unmutated V(H) genes are associated with considerably worse prognosis than mutated V(H) genes. An exception to this appears to be CLL patients utilizing the V(H)3-21 gene as they have poor outcome irrespective of mutation status. Surrogate markers for the VH gene mutation status have been suggested, such as CD38 and ZAP-70 expression. However, the CD38 level was later shown to display poor correlation to the mutation status, although it may still serve as an independent prognostic factor. More promising is the expression levels of ZAP-70, which appears to be both a strong surrogate marker for V(H) gene mutation status, although discrepancies have been reported, as well as an independent prognostic marker. Immunoglobulin gene analysis has also indicated the possibility of antigen selection in CLL considering the significant bias in V(H) gene usage. Intriguingly, the V(H)3-21+ group and several other CLL subsets using certain V(H) genes was recently reported to display strikingly restricted immunoglobulin gene features, in both their heavy and light chain gene rearrangements, thus further high-lighting the possible role of antigen involvement in CLL development.

The immunoglobulin genes and chronic lymphocytic leukemia (CLL).

The somatic hypermutation (SMH) status of the immunoglobulin (Ig) V(H) genes can divide chronic lymphocytic leukemia (CLL) into two prognostic subsets, with mutated V(H) genes display superior survival compared to unmutated cases. Biased V(H) gene usage has also been reported in CLL which may reflect antigen selection. In a V(H) gene analysis of 265 CLL cases we confirmed the prognostic impact of the V(H) mutation status and found preferential V(H) gene usage in both the mutated and unmutated subset. Interestingly, CLL cases rearranging one particular V(H) gene, V(H)3-21, displayed poor outcome despite that two-thirds showed mutated V(H) genes. Many of the V(H)3-21 utilizing cases expressed lambda light chains, rearranged a Vlambda2-14 gene, and had homologous complementarity determining region 3s (CDR3s), implying recognition of a common antigen epitope. We thus believe that the cases rearranging the V(H)3-21 gene comprises an additional CLL entity. We further analyzed the V(H) gene rearrangements and, specifically, the heavy chain CDR3 sequences in 346 CLL cases to investigate the role of antigens in CLL. We identified six new subgroups with similar HCDR3 features and restricted VL gene usage as in the V(H)3-21-using group. Our data indicate a limited number of antigen recognition sites in these subgroups and give further evidence for antigen selection in the development of CLL. Different mutational cutoffs have been used to distinguish mutated CLL in addition to the 2% cutoff. Using three levels of somatic mutations we divided 323 CLLs into subsets with divergent survival (<2%, 2-5% and >5% mutations). This division revealed a low-mutated subgroup (2-5%) with inferior outcome that would have been masked using the traditional 2% cutoff. A 1513A/C polymorphism in the P2X(7) receptor gene was reported to be more frequent in CLL, but no difference in genotype frequencies was revealed in our 170 CLL cases and 200 controls. However, CLL cases with the 1513AC genotype showed superior survival than 1513AA cases and this was in particular confined to CLL with mutated VH genes. In summary, we could define new prognostic subgroups in CLL using Ig gene rearrangement analysis. This also allowed us to gain insights in the biology and potential role of antigen involvement in the pathogenesis of CLL.

Towards multifunctional lanthanide-based metal-organic frameworks.

We report the synthesis, structure and physicochemical attributes of a new holmium(III)-based metal-organic framework whose 3D network structure gives rise to porosity; the reported structure-type can be varied using a range of different lanthanide ions to tune the photophysical properties and produce ligand-sensitised near-infrared (NIR) and visible light emitters.

Different gene expression in immunoglobulin-mutated and immunoglobulin-unmutated forms of chronic lymphocytic leukemia.

The mutation status of the immunoglobulin heavy chain variable regions (IgVH) has been found to be a good prognostic indicator for B-cell chronic lymphocytic leukemia (CLL) because unmutated VH genes are associated with rapid disease progression and shorter survival time. To study the differences in gene expression between the Ig-unmutated and Ig-mutated CLL subtypes, we performed gene expression profiling on 31 CLL cases and investigated the VH gene mutation status by sequencing. The array data showed that the greatest variances between the unmutated (20 cases) and the mutated (11 cases) group were in expressions of ZAP70, RAF1, PAX5, TCF1, CD44, SF1, S100A12, NUP214, DAF, GLVR1, MKK6, AF4, CX3CR1, NAFTC1, and HEX. ZAP70 was significantly more expressed in the Ig-unmutated CLL group, whereas the expression of all the other genes was higher in the Ig-mutated cases. These results corroborate a recent finding, according to which the expression of ZAP70 can predict the VH mutation status and suggest that RAF1, PAX5, and other differentially expressed genes may offer good markers for differentiating unmutated cases from mutated cases and thus serve as prognostic markers.

V(H)3-21 gene usage in chronic lymphocytic leukemia--characterization of a new subgroup with distinct molecular features and poor survival.

During recent years it has become evident that lymphoproliferative diseases of B-cell origin display preferential immunoglobulin (Ig) variable heavy chain (V(H)) gene usage. For instance, the V(H)1-69 and V(H)4-34 genes were early found to be overexpressed in B-cell chronic lymphocytic leukemia (CLL) and other B-cell lymphomas. The implications of biased V(H) gene usage have been speculated to be a result of stimulation of unknown antigens, which gives increased proliferation of B-cells with certain V(H) gene configuration and consequently higher probability to undergo transformation. Thus, V(H) gene usage may play a role in development of leukemias and lymphomas. Recently, we could confirm the over usage of the V(H)1-69 and V(H)4-34 genes in CLL, but a novel finding was that the V(H)3-21 gene was preferentially utilized in CLL patients with mutated V(H) genes. These V(H)3-21+ Ig rearrangements showed molecular peculiarities such as shorter lengths of the third complementarity determining region (CDR) and had similar amino acid composition of their CDR3s, implicating recognition of the same antigen in individual tumors. Most of the V(H)3-21+ patients also showed a predominance of lambda chain expression and biased usage of 1 specific V(lambda) gene, V2-14. Furthermore, overall survival appeared to correlate with V(H)3-21 usage and, regardless of V(H) gene mutation status, V(H)3-21+ patients had a poor outcome. All in all, it appears that V(H)3-21 gene usage define a new entity of CLL. The remaining question now to be clarified is if antigen(s) actually are involved in the pathogenesis of V(H)3-21+ CLL.

Similar humoral immunity parameters in chronic lymphocytic leukemia patients independent of VH gene mutation status.

Chronic lymphocytic leukemia (CLL) is a clonal B-cell disorder, which has recently been divided into 2 subtypes based on the somatic hypermutation status of the immunoglobulin heavy chain (IgVH) genes. In patients with unmutated tumor cells the survival time is approximately half of that in mutated cases, but the reason for this difference is poorly understood. Since infections are the major cause of mortality in CLL, we investigated the effect of the mutation status on host immunity and proneness to infections in patients with CLL. As expected, the disease progression seemed to be faster and the disease more advanced (Binet B and C) among unmutated patients than in the mutated ones. Surprisingly, no differences in humoral immunity [immunoglobulin G (IgG), IgM, IgA, IgG subclasses, anti-ABO blood group antibodies and mannan-binding lectin (MBL)] or immune responses (Haemophilus influenzae serotype b conjugate vaccination) were detected between these 2 patient groups. Furthermore, UM-patients were not more prone to infections compared to M-patients, and therapy had no impact on the incidence and pattern of infections in either of the patient groups. The current findings within this patient cohort reveal that the worse outcome in the unmutated subgroup is not caused by more severe defects in immunity and increased susceptibility to infections when compared with the hypermutated group. It is thus conceivable that active immunization procedures such as vaccination can successfully be applied on patients with unmutated IgVH gene and advanced disease stage.

Breaking bad news: a phenomenological exploration of Irish nurses' experiences of caring for patients when a cancer diagnosis is given in an acute care facility (part 1).

BACKGROUND: The focus of this article is on the meanings attributed by nurses who worked with patients receiving a cancer diagnosis within acute care settings in Ireland. OBJECTIVE: The aim of this article was to explore the nurses' perceptions of caring for patients who receive bad news in the form of a cancer diagnosis while in an acute care setting. METHODS: The article focuses on the perceptions of 20 nurses who formed the nurse participant group in a larger phenomenological study exploring giving and receiving a cancer diagnosis. Data were collected using unstructured in-depth interviews. Analysis was conducted using Koch's analytical framework. RESULTS: The nurses' narratives provided 2 emerging themes entitled, "connectedness: journeying as professional within the everyday world" and "connectedness: exclusion of professional within the everyday world." This article focuses on the first emerging theme and highlights the experiences of nurses as they reflect on their interactions with their patients before, during, and after the giving of a cancer diagnosis. CONCLUSIONS: This study highlights the importance of professional companionship. It provides insights into the nurse-patient challenges as a result of lack of information. IMPLICATIONS FOR PRACTICE: Lack of information and involvement affects the nurse's ability to be authentically present for the recipient and results in a fracture to the nurse-patient relationship. Understanding the experiences of nurses from acute care settings where the cancer diagnosis is often given will inform and enable the nurse working in oncology settings to engage patients in a more meaningful and focused way.

Extending the family of Zn-based MOFs: synthetic approaches to chiral framework structures and MOFs with large pores and channels.

Tri- and pentanuclear, kinetically stable SBUs were exploited for the preparation of the novel MOFs [Zn(3)(BTEB)(2)(DMF)(2)] and (Me(2)NH(2))[Zn(5)(BTEB)(3)(µ(3)-OH)(2)(DMF)(2)]. The applied synthetic approach results in topologies that are stabilised by tritopic benzene-trisethynylbenzoic acid (BTEB) linkers giving rise to chiral frameworks with large pores or channels.

Detection of explosive vapors with a charge transfer molecule: self-assembly assisted morphology tuning and enhancement in sensing efficiency.

Use of a fluorescent organic molecule consisting of binaphthyl functionalized with donor-acceptor substituted stilbenes for the detection of dinitrotoluene (DNT) and trinitrotoluene (TNT) vapors and enhancement in its sensing efficiency via self-assembly assisted morphology tuning are described.

Lipoprotein lipase is differentially expressed in prognostic subsets of chronic lymphocytic leukemia but displays invariably low catalytical activity.

Lipoprotein lipase (LPL) expression has been shown to correlate with IGHV mutational status and to predict outcome in chronic lymphocytic leukemia (CLL). We here investigated the prognostic impact of LPL expression in relation to other prognostic markers including IGHV3-21 usage in 140 CLL patients. Additionally, we studied the catalytic activity of LPL in CLL cells. A significant difference in LPL mRNA expression was detected in IGHV unmutated compared to mutated CLL patients (p<0.001). However, the poor-prognostic mutated/stereotyped IGHV3-21 patients did not differ from other mutated CLL cases. Clinical outcome was significantly different in CLL cases with high versus low LPL expression (p<0.001), and LPL expression exceeded mutation status/IGHV3-21 usage as an independent prognostic marker. Finally, LPL protein expression correlated significantly with mRNA expression and was higher in IGHV unmutated versus mutated CLL (p=0.018), although the majority of synthesized protein was catalytically inactive indicating a non-catalytical function in CLL.

Receiving bad news: a phenomenological exploration of the lived experience of receiving a cancer diagnosis.

This article explores the process of coming to a place of knowing one's diagnosis of cancer. The study was guided by the philosophy of hermeneutic phenomenology, with data collected via unstructured in-depth interviews. This article focuses on 10 people who received a cancer diagnosis (recipients). The analysis of the recipient narratives offered an interpretation of the phenomenon of receiving bad news as a process occurring over a period of time and not as a one-off event in time. The concept of bad news as a trajectory was clearly evident in the narratives and was represented through 3 themes: "disturbance of the everyday world," "surfacing within the lived world," and "embodiment within the lived world." The findings are consistent with the literature addressing diagnosis and end-of-life issues. Understanding the phenomenon of "knowing" is crucial in helping the healthcare professional recognize the changing information and psychosocial needs of the recipient as they experience the trajectory of bad news.

An evaluation of an attendance monitoring system for undergraduate nursing students.

Internationally the preparation and ongoing education of nurses continues to evolve in response the changing nature of both nursing and health care. The move into third level structures that has taken place in countries such as the UK and the Republic of Ireland, results in new challenges to the historical fabric of nurse education. One such challenge is monitoring of nursing students' attendance. Viewed by students as a patriarchal and draconian measure, the nursing profession historically value their ability to ensure the public and professional bodies that nursing students fully engage with educational programmes. University class sizes and the increased perception of student autonomy can negate against formalised monitoring systems. This paper reports on an evaluation of one such monitoring system. The findings revealed that attendance was recognised implicitly by nurse educators as an important learning activity within these programmes results and that current methods employed were less than reliable and so did little to appropriately control the phenomenon. Subsequent to the evaluation; a standardised approach to the measurement of absenteeism was employed. Deliberate short-term absence was a feature of this group. Reasons cited included travelling long distances, dissatisfaction with programme timetables and personal reasons. Preventative measures employed included improvement in student timetable delivery.