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Pengetahuan dan intensi yang baik diperlukan untuk mengatasi dampak buruk dari gempa bumi. Peningkatan pengetahuan dapat dilakukan dengan memberikan edukasi kesiapsiagaan bencana dengan metode yang digunakan yaitu ceramah, tanya jawab dan diskusi bersama, serta pemberian buku saku mengenai kesiapsiagaan bencana gempa bumi. Pelaksanaan edukasi kesiapsiagaan bencana gempa bumi diberikan kepada mahasiswa Program Studi Administrasi Rumah Sakit dan Pendidikan Matematika Universitas Alma Ata Yogyakarta, yang berjumlah 45 peserta. Sebelum dan sesudah penyampaian materi edukasi kesiapsiagaan bencana diukur pengetahuan peserta melalui pengisian pretest dan posttest. Dari hasil pretest dan posttest menunjukkan bahwa masih adanya peserta yang masuk kategori lemah terhadap kesiapsiagaan dalam mengantisipasi bencana gempa bumi. Hal tersebut, dipengaruhi oleh pengetahuan dan intensi peserta yang masih kurang. Berdasarkan hasil, diharapkan dapat disusun program edukasi berkala tentang kesiapsiagaan bencana gempa bumi agar seluruh mahasiswa dapat berpartisipasi.

The original online version of this article was revised because a number of authors had the wrong affiliation number next to their names. © 2021, Springer Nature B.V.

Background & aims: Genome-wide association studies in primary biliary cholangitis (PBC) have failed to find X chromosome (chrX) variants associated with the disease. Here, we specifically explore the chrX contribution to PBC, a sexually dimorphic complex autoimmune disease. Methods: We performed a chrX-wide association study, including genotype data from 5 genome-wide association studies (from Italy, United Kingdom, Canada, China, and Japan; 5244 case patients and 11,875 control individuals). Results: Single-marker association analyses found approximately 100 loci displaying P < 5 × 10-4, with the most significant being a signal within the OTUD5 gene (rs3027490; P = 4.80 × 10-6; odds ratio [OR], 1.39; 95% confidence interval [CI], 1.028-1.88; Japanese cohort). Although the transethnic meta-analysis evidenced only a suggestive signal (rs2239452, mapping within the PIM2 gene; OR, 1.17; 95% CI, 1.09-1.26; P = 9.93 × 10-8), the population-specific meta-analysis showed a genome-wide significant locus in East Asian individuals pointing to the same region (rs7059064, mapping within the GRIPAP1 gene; P = 6.2 × 10-9; OR, 1.33; 95% CI, 1.21-1.46). Indeed, rs7059064 tags a unique linkage disequilibrium block including 7 genes: TIMM17B, PQBP1, PIM2, SLC35A2, OTUD5, KCND1, and GRIPAP1, as well as a superenhancer (GH0XJ048933 within OTUD5) targeting all these genes. GH0XJ048933 is also predicted to target FOXP3, the main T-regulatory cell lineage specification factor. Consistently, OTUD5 and FOXP3 RNA levels were up-regulated in PBC case patients (1.75- and 1.64-fold, respectively). Conclusions: This work represents the first comprehensive study, to our knowledge, of the chrX contribution to the genetics of an autoimmune liver disease and shows a novel PBC-related genome-wide significant locus.

Two approaches are available for the analysis of the size and size distribution of the domains in a crystalline powder: pattern decomposition (or fitting) and pattern modelling. In the first case, the information is obtained from some fitting parameter and no direct connection exists between the model and the data; in the second case, a microstructure model is employed to simulate the peak profile and the model parameters are directly refined against the experimental data. The most common methods for the analysis are discussed and the formulae for the simulation of the size-broadened profile are provided.

This article introduces a new procedure to model two cases related to the simulation of the movements of the Tower of Pisa: installation of micropiles and soil extraction. Standard FEM requires geometry definition in the initial configuration. In this case the initial configuration is much different from the final one. m-PISE procedure overcomes this limit. Previous models based these two simulations on a back-analysis procedure. In m-PISE the volume changes are the same as the extracted or injected in-situ, providing a forecasting of the movements. The results obtained through this new procedure show very good agreement with the measured data.

Rosacea is a common and chronic disorder, characterized by flushing and persistent erythema in the central facial area.1,2 The disease onset is typically between the ages of 20 and 50 years, and women are more often affected than men.3,4 Rosacea has considerable psychosocial impact and causes embarrassment, anxiety and low self-esteem.5,6 Erythema is the primary feature of rosacea and presents ubiquitously among patients. Other cutaneous signs such as telangiectasia, papules, pustules and oedema may also present.7,8 Although several medications are approved for the treatment of inflammatory lesions of rosacea, there is currently no approved medication directly targeting erythema of rosacea, making it a key unmet medical need.4 In the absence of effective treatment, patients are usually advised to avoid environmental and lifestyle triggers that can exacerbate erythema.9–11 While the exact cause of erythema of rosacea is not known, it is hypothesized that erythema results from dysregulation in the cutaneous vasomotor responses, which leads to abnormal dilation of facial blood vessels upon various stimuli.12–14 Therefore, agents with vasoconstrictive activity may have a symptomatic effect on erythema. Transcriptomic studies suggest the involvement of adrenergic receptors in the neurovascular regulation pathway.15 Brimonidine tartrate (BT) is a highly selective α2-adrenergic receptor agonist, with potent vasoconstrictive activity.16 It is currently approved for the treatment of open-angle glaucoma, with well-documented efficacy and safety.17,18 BT applied topically to the face is hypothesized to reduce erythema of rosacea. In the present two Phase II studies, we aimed to determine the optimal dose regimen of BT in the treatment of moderate to severe erythema of rosacea, and to evaluate the efficacy and safety of the treatment using two specifically developed novel scales for erythema.

Smectite clays suspended in a platinum deposition solution are here shown to increase the specific surface area of electrodeposited Pt on gold and porous carbon paper supports, and to enhance the specific activity of Pt towards methanol oxidation under steady-state conditions. Organomodified natural bentonite shows a more pronounced effect with respect to natural montmorillonite and synthetic laponite. The effect of clay, as evidenced by microscopy and X-ray diffraction, is to decrease the average size of the Pt crystallites and simultaneously the degree of their mutual coalescence. This effect is associated with suppression of Pt secondary nucleation observed in deposition current transients, and the electrocatalytic activity is suggested to be affected by the intergrain boundaries.

Particles of Fe2Al5 (60–211 μm diameter) were leached at 10, 30 and 50 °C in 3 M NaOH containing varying amounts of sodium chromate in the range 0.005–0.05 M. The kinetics of leaching was fitted to solid-state kinetic models. The model best fitting the data was the Avrami–Erofeev model −ln(1 − α)1/1.5 = kt. Apparent activation energies were in the range 60–63 kJ mol−1. The BET surface areas of the promoted materials were high, with values in the range 57–91 m2 g−1. Catalysts were tested for the water–gas-shift (WGS) reaction at 350 °C and it was found that the chromium content of the catalysts had a significant effect on the WGS activity, which went through a maximum for a catalyst containing around 2 wt% chromium.

Mineralogical and mechanical characterization of Venice lagoon soils is required for design and construction of movable floodgates that aim to safeguard the city of Venice against recurrent floods. An instrumented circular test embankment was constructed in the lagoon area, enabling accurate measurement of relevant ground displacements. In situ stress-strain-time measurements were carried out in order to investigate the viscous behaviour of Venice lagoon soils during and after embankment loading. Site monitoring was kept up also during embankment removal so that information on soil behaviour in unloading is available, too. This paper illustrates key results from embankment monitoring and also focuses on modelling of creep behaviour. A recently developed anisotropic constitutive model was calibrated for Venice lagoon soils and is used in back analysis of the embankment construction and removal process. The constitutive parameters of the model were calibrated from in situ and laboratory tests.

Bendamustine has demonstrated substantial efficacy in the treatment of hematologic malignancies and continues to distinguish itself from other alkylating agents with regard to its activity in tumor cells. The mechanistic and clinical differences associated with bendamustine may be directly related to its unique structural features. Although the precise mechanisms of action are still poorly understood, bendamustine is associated with extensive and durable DNA damage. The increased potency of bendamustine may be due to secondary mechanisms such as inhibition of mitotic checkpoints, inefficient DNA repair, and initiation of p53-dependent DNA-damage stress response, all of which lead to mitotic catastrophe and apoptosis. It has also been hypothesized that the presence of a benzimidazole ring in addition to the nitrogen mustard group may influence the way bendamustine interacts with DNA and/or confer antimetabolite properties. Further elucidation of the mechanisms of action for bendamustine and the signaling pathways involved in the response to bendamustine-induced DNA damage is essential to maximize its therapeutic potential, identify biomarkers for response, and understand the potential for synergy with other agents involved in DNA damage and inhibition of DNA repair. This review will discuss the current understanding and hypotheses regarding bendamustine mechanisms of action and suggest future investigations that would shed light on the many unanswered questions.

OBJECTIVE • To study the pharmacokinetic and toxicity profile of intravesically administered TMX-101, with its active ingredient R-837, a synthetic Toll-like receptor (TLR)-7 agonist, in a pig model. MATERIALS AND METHODS • TLR-7 expression was determined by immunohistochemistry in human and pig bladder tissue. • Four groups of six pigs received a 1-h intravesical instillation with R-837 of different formulations. • Pharmacokinetic analysis was performed on plasma. Toxicity evaluation included monitoring the well-being of the animals, peripheral blood cell counts, and interleukin-6 and creatinine measurements. Urine was collected for R-837 measurement and dipstick analysis. • In total, three pigs per group were sacrificed 24 h post-treatment, and the remaining animals were sacrificed after 1 week. Histopathological examination of the bladder wall was performed. RESULTS • TLR-7 was homogeneously expressed in human and pig urothelium. • R-837 and vehicle were well tolerated without deterioration in animal well-being. • Systemic R-837 absorption was low. • Mean maximum plasma concentration of R-837 differed depending on the formulation. Post-treatment, plasma levels were negligible at 24 h. • Histopathological examination of the bladders did not show significant abnormalities, apart from the intended inflammatory reaction in the R-837 treated groups. CONCLUSION • Intravesically administered R-837 in pigs, which showed a similar TLR-7 distribution in bladder tissue as humans, is well tolerated and causes no bladder wall toxicity, and formulations with poloxamer and hydroxypropyl-β-cyclodextrin showed less systemic absorption.

Caustic leaching of fine particles of Fe2Al5 alloy to produce skeletal Fe catalysts was studied using a 24 factorial experimental design, in which alloy particle size, aqueous NaOH concentration, temperature and stirrer speed were varied. Analysis of the results from the design showed that the BET surface area of the skeletal iron increased with decreases in temperature, caustic concentration and particle size according to SBET = 222.7 − 0.461 · T − 2.35 · cNaOH − 0.0245 · D p. An Avrami–Erofeev model −ln(1 − α) = kt with an activation energy of 55 ± 5 kJ mol−1 and a shrinking core model for volume contraction 1 − (1 − α)1/3 = kt and an activation energy of 56 ± 5 kJ mol−1 provided the best fit to the kinetic data for leaching.

Objective: To study the immune response caused by the intravesical administration of the immunomodulator R-837 in various formulations and to estimate its therapeutic potential for bladder cancer. Methods: Female C57BL/6 mice were intravesically treated with different formulations of R-837, a Toll-like receptor 7 agonist used for treating genital warts and skin malignancy. The tested formulation mixtures contained different ratios of lactic acid, a thermosensitive poloxamer polymer (Lutrol F127) and 2-(hydroxypropyl)-β-cyclodextrin (HPβCD). Induction of tumor necrosis factor α (TNFα) and keratinocyte-derived chemokine (KC) was analyzed by Luminex microbeads assay. The therapeutic potential of intravesical administration of R-837 was assessed in an orthotopic, syngeneic mouse model of bladder cancer using MB49 cells. Results: Intravesical administration of R-837 in lactic acid alone induced systemic and bladder TNFα and KC in a dose-dependent manner. Formulations including poloxamer decreased systemic absorption of R-837 and significantly reduced systemic and local induction of KC. Addition of HPβCD in the poloxamer formulation particularly reversed levels of systemic and local levels of TNFα and KC. Histological examination showed that poloxamer-HPβCD formulation allowed infiltration of mononuclear cells into urothelium and lamina propria. In studies using orthotopic mouse bladder cancer, the tumor loads in R-837-treated mice were significantly lower than those in vehicle-treated or non-treated mice. Conclusion: The optimized poloxamer-HPβCD formulation of R-837 shows therapeutic potential for bladder cancer while avoiding adverse side-effects.

The higher stiffness of Pb-free SAC solders makes Pb-free assemblies more sensitive to drop impact. In order to be able to optimize the drop test performance, it is necessary to have better insight into the crack propagation in the Pb-free solder joints. This study combines crack-front mapping using the dye and pry method and electrical FE simulation to establish a relation between DC electrical resistance and cracked area, and hence monitor the initiation and propagation of cracks in individual solder joints as the PCB assemblies are subjected to JEDEC type mechanical shock and high speed cyclic bending. The carrier in the study is a ball grid array (BGA), a critical component family for drop impact. Combinations of solder alloys and pad finishes, SnPb on organic solderable preservative (OSP), SAC305 on electroless nickel/immersion gold (ENIG), SAC101 on OSP, and SAC101(d) on ENIG are studied regarding the failure mode and crack propagation. This paper demonstrates that, for the large majority of Pb-free solder joints, there is a negligible initiation period; cracks can start forming at the first PCB bending cycle. The presence of large cracks, especially at both sides can increase the compliance of the joint and slow down crack growth. Even if large cracks are present, the resistance increase is less than 1 m? per interconnect, which is far from the 100 ? that is often taken as a failure criterion. Brittle joints as found with SAC305 on ENIG have erratic propagation rates while ductile joints are much more predictable. Therefore, the way to optimize the drop test performance of a Pb-free BGA assembly is to prolong the crack propagation within the ductile bulk solder material.

The crystal structure and the fault structure of the intermetallic Nb2Co7 phase were determined using DIFFaX+ refinements on the basis of synchrotron powder-diffraction data. The crystal structure was confirmed to be of the monoclinic Zr2Ni7 type. The layer faulting can be understood as partial occurrence of structural features of the Yb2Ag7-type structure, which contains the same “triple layers” as the Zr2Ni7-type structure does, but stacked in a different fashion. Introduction The binary Nb-Co system [1] contains, among other phases, the intermetallic Nb2Co7 compound with the recently identified crystal structure described in ref. [2]. For an intermetallic, Nb2Co7 exhibits an unusually high plastic deformability [3, 4]. Consequently, the question arises, whether there is a relation between the actual crystal structure and the high deformability. The X-ray powder-diffraction patterns used of Nb2Co7 for structural studies exhibit strong line broadening, which can be ascribed to faulting with respect to the layered structure of Nb2Co7. Such defects are expected to play a role in the deformability of this phase, but the traditional Rietveld approach is not able to handle fully the diffraction phenomena associated with faulting. Therefore, DIFFaX+ refinements are provided here, which depart from the layered nature of the material and from previous qualitative considerations concerning the character of the faulting [2]. The results provide useful inside into the defect structure of the material. Crystal structure and faulting model Structure/microstructure analyses were conducted on a Nb2Co7 powder sample containing tiny amounts of the cubic C15-NbCo2 Laves phase (equilibrium phase with Nb2Co7 in a twophases region at the final annealing temperature of 900°C [1]). An X-ray powder-diffraction pattern recorded on the powder diffractometer installed at the ID31 beamline of the ESRF synchrotron (Grenoble, F) using a wavelength of 0.400094 A was available for the specimen. 424 European Powder Diffraction Conference, EPDIC 11 The average (defect free) crystal structure of Nb2Co7 was previously identified [2] to be similar to monoclinic Zr2Ni7 [5, 6], figure 1a. Within the crystal structure one can identify triple-layers parallel to (001) with a complex internal atomic structure: an inner Kagometype layer (KA) of “Co3” sandwiched by two outer “close-packed” layers (CL) “NbCo2” (with Nb-Co ordering), leading to a layer composition of Nb2Co7 corresponding also to the overall phase composition. The triple layers (CL-KA-CL) by themselves ideally exhibit an hexagonal layer symmetry, but in view of the final crystal structure, a centred rectangular unit cell with basis vectors a and b should be considered, identical to the lattice-basis vectors of the monoclinic crystal structure (figure 1b). (a) a tri pl e la ye r

Dislocations are known for their peculiar effects on the diffraction profiles. Each given dislocation type, produces specific anisotropic line broadening basically determined by the so-called contrast factor (CF). Owing to the lack of a suitable mathematical frame to deal with dislocations in crystal of any symmetry, CFs have been so far determined for high symmetry materials. In this work we present the main points of a general approach for determining CF, as well as a specific application to scheelite CaWO4.

Drop impact reliability has become an important criterion when assessing the reliability of portable electronics. Finite element software such as ABAQUS is used to simulate solder joints during drop-impact tests and determine the drop-impact reliability of the product. This modelling requires the rate-dependent constitutive properties of the solder in the range of strain rates between 0.1 s− 1 to 300 s− 1. Unfortunately, the constitutive properties of most solder materials in this range of strain rates have not been characterized. This paper presents the mechanical properties of four solder alloys — 63Sn37Pb, Sn1.0Ag0.1, Sn3.5Ag, and Sn3.0Ag0.5Cu at strain rates between 0.005 s− 1 and 300 s− 1. The lead-free solder alloys tested exhibited greater strain rate sensitivity compared to 63Sn37Pb. In addition, both the lead-free solders with high Ag content (≥ 3%) exhibited much higher flow stresses compared to 63Sn37Pb. The higher flow stress could explain the lower reliability of these joints in drop tests as a result of shifting of the failure site from the ductile solder to the brittle intermetallic compound [Wong EH, Rajoo R, Mai Y-W, Seah SKW, Tsai KT, Yap LM. Proc. 55th Elec. Comp. & Tech. Conf. 2005;1202–9]. Based on the data obtained, Sn1.0Ag0.1 seems to be the most suitable lead-free alloy to replace eutectic tin-lead in products where drop-impact performance is required.

The stress–strain properties of eutectic Sn-Pb and lead-free solders at strain rates between 0.1 s−1 and 300 s−1 are required to support finite-element modeling of the solder joints during board-level mechanical shock and product-level drop-impact testing. However, there is very limited data in this range because this is beyond the limit of conventional mechanical testing and below the limit of the split Hopkinson pressure bar test method. In this paper, a specialized drop-weight test was developed and, together with a conventional mechanical tester, the true stress–strain properties of four solder alloys (63Sn-37Pb, Sn-1.0Ag-0.1Cu, Sn-3.5Ag, and Sn-3.0Ag-0.5Cu) were generated for strain rates in the range from 0.005 s−1 to 300 s−1. The sensitivity of the solders was found to be independent of strain level but to increase with increased strain rate. The Sn-3.5Ag and the Sn-3.0Ag-0.5Cu solders exhibited not only higher flow stress at relatively low strain rate but, compared to Sn-37Pb, both also exhibited higher rate sensitivity that contributes to the weakness of these two lead-free solder joints when subjected to drop impact loading.

Purpose: Bendamustine has shown clinical activity in patients with disease refractory to conventional alkylator chemotherapy. The purpose of this study was to characterize the mechanisms of action of bendamustine and to compare it with structurally related compounds.Experimental Design: Bendamustine was profiled in the National Cancer Institute in vitro antitumor screen. Microarray-based gene expression profiling, real-time PCR, immunoblot, cell cycle, and functional DNA damage repair analyses were used to characterize response to bendamustine and compare it with chlorambucil and phosphoramide mustard.Results: Bendamustine displays a distinct pattern of activity unrelated to other DNA-alkylating agents. Its mechanisms of action include activation of DNA-damage stress response and apoptosis, inhibition of mitotic checkpoints, and induction of mitotic catastrophe. In addition, unlike other alkylators, bendamustine activates a base excision DNA repair pathway rather than an alkyltransferase DNA repair mechanism.Conclusion: These results suggest that bendamustine possesses mechanistic features that differentiate it from other alkylating agents and may contribute to its distinct clinical efficacy profile.

SDX-308 and SDX-309 are potent indole-pyran analogues of SDX-101 (R-etodolac) which has anti-tumour activity unrelated to cyclooxygenase-2 inhibition. Their cytotoxic activity was further studied herein using a well-characterized human tumour cell-line panel containing ten cell lines, as well as in 58 primary tumour cell samples from a variety of diagnoses. The indole-pyran analogues of SDX-101 were in general considerably more active in both cancer cell lines and primary tumour samples. Low cross-reactivity with standard agents was observed, indicating a unique mechanism of action. No apparent influence on efficacy was observed via classical mechanisms of multidrug-resistance. SDX-101 and SDX-309 showed higher relative activity in haematological compared to solid tumour samples, while SDX-308 had pronounced solid-tumour activity. High SDX-308 cytotoxic efficacy was observed in non-small cell lung cancer, renal cancer and ovarian cancer samples, and also in chronic lymphocytic leukaemia. In conclusion, the indole-pyran analogues showed a favourable pharmacological profile and represent a potentially important new class of drugs for cancer treatment.

Deep Mixing Method (DMM) for the Rio Puerto Nuevo Project in San Juan was chosen by the Corps of Engineers to improve the soils adjacent to a 92" forced sewer main relocation and to improve the soils for the base of a new drainage channel. The soils consisted of sandy fill, soft clays, organics and stiff, plastic clays. Production comprised over 3,000 DMM elements, installed to a maximum depth of 57 feet deep. Each element consisted of two 5.5' diameter overlapping columns installed simultaneously. The layout of the elements was such to meet the 100% Area Replacement Ratio required by the design. Wet grab sampling and UCS testing of the retrieved specimens were designed as the main QC/QA method. Upon completion of production, approximately 3,000 specimens were cast. The contract also required continuous coring of at least 12 randomly selected DMM elements and the testing of 8 cored specimens per bore for strength assessment. This paper will illustrate means and methods used in field to collect the wet grab and the cored samples, focusing on the significant differences between the two UCS data sets.

We conducted a prospective randomized clinical trial to assess the mobilizing efficacy of filgrastim, lenograstim and molgramostim following a disease-specific chemotherapy regimen. Mobilization consisted of high-dose cyclophosphamide in 45 cases (44%), and cisplatin/ifosfamide/etoposide or vinblastine in 22 (21%), followed by randomization to either filgrastim or lenograstim or molgramostim at 5 microg/kg/day. One hundred and three patients were randomized, and 82 (79%) performed apheresis. Forty-four (43%) patients were chemonaive, whereas 59 (57%) were pretreated. A median number of one apheresis per patient (range, 1-3) was performed. The median number of CD34+ cells obtained after mobilization was 8.4 x 10(6)/kg in the filgrastim arm versus 5.8 x 10(6)/kg in the lenograstim arm versus 4.0 x 10(6)/kg in the molgramostim arm (P=0.1). A statistically significant difference was observed for the median number of days of growth factor administration in favor of lenograstim (12 days) versus filgrastim (13 days) and molgramostim (14 days) (P

In this study we report that R-etodolac (SDX-101), at clinically relevant concentrations, induces potent cytotoxicity in drug-sensitive multiple myeloma (MM) cell lines, as well as in dexamethasone (MM.1R)-, doxorubicin (Dox40/RPMI8226)-, and bortezomib (DHL4)-resistant cell lines. Immunoblot analysis demonstrates that R-etodolac induces apoptosis characterized by caspase-8, -9, and -3 and PARP (poly-ADP [adenosine diphosphate]-ribose polymerase) cleavage and down-regulation of cyclin D1 expression. Subcytotoxic doses of R-etodolac up-regulate myeloid cell leukemia-1 proapoptotic variant (Mcl-1S), while enhancing dexamethasone (Dex)-induced caspase activation and apoptosis. The combination of R-etodolac with Dex results in a highly synergistic cytotoxic effect. R-etodolac also induces apoptosis against primary cells isolated from patients with MM refractory to chemotherapy. Although interleukin 6 (IL-6) and insulin-like growth factor-1 (IGF-1) abrogate Dex-induced MM cell cytotoxicity, neither IL-6 nor IGF-1 protects against R-etodolac-induced cytotoxicity in MM cells. R-etodolac also inhibits viability of MM cells adherent to bone marrow stromal cells (BMSCs), thereby overcoming a mechanism of drug resistance commonly observed with other conventional chemotherapeutic agents. Our data, therefore, indicate that R-etodolac circumvents drug resistance in MM cells at clinically relevant concentrations, targets Mcl-1, and can be synergistically combined with Dex. (Blood. 2005;106:706-712)

The p16INK4A/CDKN2A (p16) gene on chromosome 9p21 is inactivated in >90% of invasive pancreatic cancers. In 40% of pancreatic cancers the p16 gene is inactivated by homozygous deletion, in 40% by an intragenic mutation coupled with loss of the second allele, and in 10-15% by hypermethylation of the p16 gene promoter. Immunohistochemical labeling for the p16 gene product parallels gene status, but does not provide information of the mechanism of p16 gene inactivation. The methylthioadenosine phosphorylase gene (MTAP) gene also resides on chromosome 9p21, approximately 100 kb telomeric to the p16 gene. The MTAP gene is frequently contained within p16 homozygous deletions, producing concordant loss of both p16 and MTAP gene expression. Concordant loss of both p16 and MTAP protein expression can therefore be used as a surrogate marker for p16 homozygous deletion. Here we immunolabeled a series of pancreatic intraepithelial neoplasia (PanIN) lesions of various histologic grades for the p16 and MTAP gene products using a high-throughput PanIN tissue microarray (TMA) format. We demonstrate concordant loss of p16 and MTAP protein expression in 6/73 (8%) PanINs, including five high-grade lesions and one low-grade lesion. Immunolabeling for both p16 and MTAP protein expression provides a tool to evaluate tissues with intact morphology for p16 gene homozygous deletions. The concordant loss of expression of both genes in PanIN lesions demonstrates that homozygous deletions of the p16 tumor suppressor gene can occur in noninvasive precursor lesions.

Background Insulin resistance, a frequent finding in hypertensive patients, leads to accelerated cardiovascular damage. It has been suggested that a crosstalk between angiotensin II and insulin signaling pathways may provoke insulin resistance, and may contribute to the development of cardiovascular damage. To identify a common pathophysiologic pathway between metabolic disorders and cardiovascular remodeling, we investigated the effect of angiotensin II and insulin on extracellular signal regulated kinases 1 and 2 (ERK1/2), isoforms of mitogen-activated protein kinases (MAPK) involved in cellular proliferation and extracellular matrix deposition. Methods Skin fibroblasts from normotensive subjects, insulin sensitive hypertensive subjects, and insulin resistant hypertensive subjects were cultured and used after four passages. The ERK1/2 expression and phosphorylation were measured by Western blot using specific antibodies, respectively anti-ERK1/2 and anti-pERK1/2. Expression of AT 1 receptor for angiotensin II was determined by reverse transcriptase–polymerase chain reaction in real time. Results The ERK1/2 were similarly expressed in skin fibroblasts from all groups; ERK1/2 phosporylation evoked by angiotensin II was significantly higher in fibroblasts from hypertensive patients in comparison to normotensive subjects, but the increase was observed only in insulin resistant hypertensive subjects. The effect of insulin on ERK1/2 phosphorylation was not significantly different in the three groups. Treatment with the combination of insulin and angiotensin II increased ERK1/2 phosphorylation to a greater extent in comparison to the single agonists in normotensive subjects and in insulin sensitive but not in insulin resistant hypertensive subjects. Conclusions Angiotensin II stimulated ERK1/2 activation is increased in insulin resistant hypertensive subjects, and it may play a role in the pathogenesis of insulin resistance and accelerated cardiovascular damage.

R-etodolac, a nonsteroidal anti-inflammatory drug, inhibits the progression of CWRSA6 androgen-independent and LuCaP-35 androgen-dependent prostate cancer xenograft growth through downregulation of cyclin D1 expression via the PPARgamma pathway. PPARgamma protein degradation, observed post-R-etodolac treatment, resulted from phospho-MAP kinase (p44/42) induction by R-etodolac negatively regulating PPARgamma function. Negative regulation of PPARgamma was overcome by a combination regimen of R-etodolac with the HER-kinase axis inhibitor, rhuMab 2C4, which demonstrated an additive antitumor effect. We further show that the inhibition of HER-kinase activity by rhuMab 2C4 is sufficient to inhibit PPARgamma protein degradation. This study introduces a novel concept of an in vivo crosstalk between the HER-kinase axis and PPARgamma pathways, ultimately leading to negative regulation of PPARgamma activity and tumor growth inhibition.

B cell chronic lymphocytic leukemia (CLL) is characterized by an accumulation of mature, functionally incompetent B cells. Wnts are a large family of secreted glycoproteins involved in cell proliferation, differentiation, and oncogenesis. The classical Wnt signaling cascade inhibits the activity of the enzyme glycogen synthase kinase-3beta, augmenting beta-catenin translocation to the nucleus, and the transcription of target genes. Little is known about the potential roles of Wnt signaling in CLL. In this study, we quantified the gene expression profiles of the Wnt family, and their cognate frizzled (Fzd) receptors in primary CLL cells, and determined the role of Wnt signaling in promoting CLL cell survival. Wnt3, Wnt5b, Wnt6, Wnt10a, Wnt14, and Wnt16, as well as the Wnt receptor Fzd3, were highly expressed in CLL, compared with normal B cells. Three lines of evidence suggested that the Wnt signaling pathway was active in CLL. First, the Wnt/beta-catenin-regulated transcription factor lymphoid-enhancing factor-1, and its downstream target cyclin D1, were overexpressed in CLL. Second, a pharmacological inhibitor of glycogen synthase kinase-3 beta, SB-216763, activated beta-catenin-mediated transcription, and enhanced the survival of CLL lymphocytes. Third, Wnt/beta-catenin signaling was diminished by an analog of a nonsteroidal antiinflammatory drug (R-etodolac), at concentrations that increased apoptosis of CLL cells. Taken together, these results indicate that Wnt signaling genes are overexpressed and are active in CLL. Uncontrolled Wnt signaling may contribute to the defect in apoptosis that characterizes this malignancy.

Molecular targeting therapies for hematological malignant diseases such as monoclonal antibodies and small molecules have been reviewed. Imatinib mesylate (STI571) targets the tyrosine kinase activity of the bcr-abl fusion protein in CML, and was superior to IFN-α plus low-dose cytarabine in newly diagnosed chronic-phase CML in a phase III randomized study. Imatinib induced apoptosis in bcr-abl-positive cells in vitro, and activates several signaling pathways such as PI3K/Akt, STAT5 and Ras/MAPK. Combination therapies with imatinib and new strategies for downregulation of intracellular Bcr-Abl protein levels have also been investigated from the phenomenon of resistance to imatinib. Anti-CD20 (rituximab) became the first monoclonal antibody approved for the treatment of a relapsed/refractory follicular/low-grade NHL and promising results were obtained from a phase III randomized study. Although antibody-dependent cell-mediated cytotoxicity and complement-mediated cytotoxicity are likely to be the major effectors of B-cell depletion in vivo, direct cytotoxicity by CD20 monoclonal antibody on B-cell lines in vitro has been reported. Anti-CD33 (Mylotarg) and FLT3 inhibitors for AML have also been used in clinical trials and signaling pathways induced by these agents are under intensive investigation. Arsenic trioxide, like all-trans-retinoic acid (ATRA), downregulates promyelocytic leukemia protein/retinoic acid receptor-α (PML/RARα) fusion protein and induced apoptosis in APL cells, and promising results were obtained from ATRA-resistant APL patients. Finally we show our promising in vitro and in vivo data of R-etodolac (a non-steroidal anti-inflammatory drug lacking cyclooxygenase inhibitor activity) against chronic lymphocytic leukemia (CLL) cells.