Multiple myeloma cells expressing low levels of CD138 have an immature phenotype and reduced sensitivity to lenalidomide

CD138 expression is a hallmark of plasma cells and multiple myeloma cells. However, decreased expression of CD138 is frequently observed in plasma cells of myeloma patients, although the clinical significance of this is unclear. To evaluate the significance of low expression of CD138 in MM, we examined the phenotypes of MM cells expressing low levels of CD138. Flow cytometric analysis of primary MM cells revealed a significant decrease in CD138 expression in patients with relapsed/progressive disease compared with untreated MM patients. Patients with low levels of CD138 had a worse overall survival compared with patients with high levels of CD138, in newly diagnosed patients and patients receiving high-dose chemotherapy followed by autologous stem-cell transplantation. Two MM cell lines, KYMM-1 (CD138- low) and KYMM-2 (CD138- high), were established from a single MM patient with decreased CD138 expression. High expression of BCL6 and PAX5, and downregulation of IRF4, PRDM1 and XBP1 was observed in KYMM-1 compared with KYMM-2 cells, indicative of the immature phenotype of KYMM-1. KYMM-1 was less sensitive to lenalidomide than KYMM-2, while no difference in sensitivity to bortezomib was observed. KYMM-2 cells were further divided in CD138+ and CD138- fractions using anti-CD138-coated magnetic beads. CD138- cells sorted from the KYMM-2 cell line also showed high BCL6, low IRF4 expression and decreased sensitivity to lenalidomide compared with CD138+ cells. Our observations suggest that low CD138 expression relates to i) poor prognosis, ii) immature phenotype and iii) low sensitivity to lenalidomide. The observed distinct characteristics of CD138 low MM cells, suggest this should be recognized as a new clinical entity. Establishment of a treatment strategy for MM cells expressing low levels of CD138 is needed to improve their poor outcome.     

Cancer specific promoter CpG Islands hypermethylation of homeodomain only protein X gene and its potential tumor suppressive role in pancreatic carcinogenesis

ABSTRACT: BACKGROUND: We have recently identified homeodomain only protein X (HOPX) as a tumor suppressor gene candidate, characterized by tumor-specific promoter DNA hypermethylation in human cancers, and it can remarkably inhibit tumors' aggressive phenotypes. In this current study, we for the first time examined methylation level of HOPX and tested the functional relevance in pancreatic cancer (PC). METHODS: Clinical features of HOPX promoter hypermethylation was investigated in 89 PC tissues, and immunohistochemistry was added. We also examined its functional relevance in phenotype assays such as soft agar, proliferation, invasion, and cell cycle analysis. RESULTS: PC tissues had HOPX gene hypermethylation as compared to the corresponding normal pancreas tissues, and its uniqueness was robust to discriminate tumor from normal tissues (AUC = 0.85, P < 0.0001). Unexpectedly, HOPX was increased in expression in tumor tissues, and immunohistochemistry revealed its predominant expression in the Langerhans islet cells, where HOPX is reduced in expression for PC cells with promoter hypermethylation. HOPX transfectants exhibited G1 arrest with subG1 accumulation, and inhibit tumor forming and invasive ability. CONCLUSION: Defective expression of HOPX which is consistent with promoter DNA hypermethylation may explain aggressive phenotype of pancreatic cancer, and intense expression of HOPX in the Langerhans cells may in turn uniquely contribute to pancreatic carcinogenesis.     

Epigenetic Silencing of HOPX Promotes Cancer Progression in Colorectal Cancer

Homeodomain-only protein X (HOPX)-β promoter methylation was recently shown to be frequent in human cancers and was suggested as tumor suppressor gene in esophageal and gastric cancer. The aim of this study was to investigate the mechanistic roles of HOPX-β promoter methylation and its clinical relevance in colorectal cancer (CRC). HOPX-β promoter methylation was assessed in human CRC cell lines and 294 CRC tissues. HOPX mRNA and protein levels were measured in relation to HOPX-β promoter methylation. The effects of forced HOPX expression on tumorigenesis were studied using in vitro and in vivo assays. The association between HOPX-β promoter methylation and clinical relevance of CRC patients was determined. HOPX-β promoter methylation is cancer-specific and frequently found in CRC cell lines and tissues, resulting in the down-regulation of HOPX mRNA and protein levels. In CRC cell lines, forced expression of HOPX suppressed proliferation, invasion, and anchorage-independent growth. DNA microarray analyses suggested critical downstream genes that are associated with cancer cell proliferation, invasion or angiogenesis. In a mouse xenograft model, HOPX inhibited tumorigenesis and angiogenesis. Finally, HOPX-β promoter methylation was associated with worse prognosis of stage III CRC patients (hazard ratio= 1.40, P = .035) and also with poor differentiation (P = .014). In conclusion, HOPX-β promoter methylation is a frequent and cancer-specific event in CRC progression. This epigenetic alteration may have clinical ramifications in the diagnosis and treatment of CRC patients.     

Antimicrobial peptides in human skin disease

The skin continuously encounters microbial pathogens. To defend against this, cells of the epidermis and dermis have evolved several innate strategies to prevent infection. Antimicrobial peptides are one of the primary mechanisms used by the skin in the early stages of immune defense. In general, antimicrobial peptides have broad antibacterial activity against gram-positive and negative bacteria and also show antifungal and antiviral activity. The antimicrobial activity of most peptides occurs as a result of unique structural characteristics that enable them to disrupt the microbial membrane while leaving human cell membranes intact. However, antimicrobial peptides also act on host cells to stimulate cytokine production, cell migration, proliferation, maturation, and extracellular matrix synthesis. The production by human skin of antimicrobial peptides such as defensins and cathelicidins occurs constitutively but also greatly increases after infection, inflammation or injury. Some skin diseases show altered expression of antimicrobial peptides, partially explaining the pathophysiology of these diseases. Thus, current research suggests that understanding how antimicrobial peptides modify susceptibility to microbes, influence skin inflammation, and modify wound healing, provides greater insight into the pathophysiology of skin disorders and offers new therapeutic opportunities.     

Effect of prior acid etching on bonding durability of single-step adhesives

This study investigated the effect of prior phosphoric acid etching on the enamel bond strength of five single-step self-etch adhesive systems: Absolute, Clearfil tri-S Bond, Fluoro Bond Shake One, G-Bond and One-Up Bond F Plus. Bovine mandibular incisors were mounted in self-curing resin, and the facial surfaces were wet ground with #600 silicon carbide paper. Adhesives were applied to the enamel surfaces with or without prior phosphoric-acid etching and light irradiated. The resin composites were condensed into a mold and light irradiated. In total, 40 specimens were tested per adhesive system with and without prior acid etching and were further divided into two groups: those stored in water at 37 degrees C for 24 hours without cycling and those stored in water at 37 degrees C for 24 hours followed by thermal cycling between 5 degrees C and 55 degrees C with 10,000 repeats. After storage under each set of conditions, the specimens were tested in shear mode at a crosshead speed of 1.0 mm/minute. Two-way analysis of variance, the Student's t-test and the Tukey HSD test were used to analyze the data at a significance level of 0.05. For the specimens without prior acid etching, the mean bond strengths to enamel ranged from 11.0 to 14.6 MPa after 24-hour storage in water, while the corresponding values for specimens with prior acid etching ranged from 15.2 to 19.3 MPa. When these specimens were subjected to thermal cycling, the mean bond strengths ranged from 11.3 to 17.0 MPa without prior acid etching and from 12.3 to 23.2 MPa with prior acid etching. The changes in enamel bond strengths differed among the adhesive systems tested. After 24-hour storage in water, the most common failure modes were adhesive failure and mixed failure for specimens with and without prior acid etching, respectively. Thus, through a careful choice of adhesive system, prior acid etching can increase the bond strengths of single-step self-etch adhesive systems.     

Ultrasonic monitoring of the setting of glass-ionomer luting cements

This study used ultrasonic measurements to monitor the setting behaviour, and changes in the elastic modulus, of glass–ionomer cements. The ultrasonic equipment comprised a pulser–receiver, transducers, and an oscilloscope. The two-way transit time through the mixing cement disk was divided by two, in order to account for the down-and-back travel path, and then multiplied by the sonic velocity within the material. The sonic velocities of the longitudinal and shear waves were used to determine the elastic modulus. In the earliest stages of the setting process, most of the ultrasound energy was absorbed by the cements and the second echoes were relatively weak. As the cements hardened, the sound velocities increased until they reached a plateau. The changes in sound velocities differed among the glass–ionomer cements tested. The mean elastic moduli of the specimens ranged from 2.6 to 6.2 GPa after 15 min, from 13.4 to 20.4 GPa after 24 h, and from 11.4 to 22.4 GPa after 1 month. The ultrasonic method used in this study has considerable potential for determining the setting processes of luting cements.     

Setting behaviour of luting cements monitored by an ultrasonic method

The purpose of this study was to monitor the setting behaviour and elastic modulus of luting cements using an ultrasonic device. The ultrasonic equipment comprised a pulser-receiver, transducers and an oscilloscope. The transit time through the cement disk was multiplied by the thickness of the specimen, and the sonic velocity within the material was then calculated. The sonic velocities of the longitudinal and shear waves were used to determine the elastic modulus. Analysis of variance and the Tukey HSD test were used to compare the elastic moduli of the set cements. In the earliest stages of the setting process, most of the ultrasound energy was absorbed by the cements and the sound waves were relatively weak. As the cements hardened, the sound velocities increased and this tendency differed among the luting cements used. The mean elastic moduli of the specimens ranged from 2.9 to 9.9 GPa after 15 min, from 14.4 to 20.3 GPa after 24 h and from 12.1 to 15.9 GPa after 1 month. The setting processes of the luting cements were thus clearly defined by using the present ultrasonic method.     

Glioblastoma treated with postoperative radio-chemotherapy: Prognostic value of apparent diffusion coefficient at MR imaging

Purpose

To retrospectively evaluate whether the mean, minimum, and maximum apparent diffusion coefficient (ADC) of glioblastomas obtained from pretreatment MR images is of prognostic value in patients with glioblastoma.

Materials and methods

The institutional review board approved our study and waived the requirement for informed patient consent. Between February 1998 and January 2006, 33 patients (24 males, 9 females; age range 10-76 years) with supratentorial glioblastoma underwent pretreatment magnetic resonance (MR) imaging. The values of the mean, minimum, and maximum ADC (ADC_mean, ADC_MIN, and ADC_MAX, respectively) of each tumor were preoperatively determined from several regions of interest defined in the tumors. After surgical intervention, all patients underwent irradiation and chemotherapy performed according to our hospital protocol. The patient age, symptom duration, Karnofsky performance scale score, extent of surgery, and ADC were assessed using factor analysis of overall survival. Prognostic factors were evaluated using Kaplan-Meier survival curves, the log-rank test, and multiple regression analysis with the Cox proportional hazards model.

Results

Likelihood ratio tests confirmed that ADC_MIN was the strongest among the three prognostic factors. Total surgical removal was the most important predictive factor for overall survival (P < 0.01). ADC_MIN was also statistically correlated with overall survival (P < 0.05) and could be used to classify patients into different prognostic groups. Interestingly, ADC_MIN was also the strongest prognostic factor (P < 0.01) in the group of patients in whom total tumor removal was not possible.

Conclusion

The ADC_MIN value obtained from pretreatment MR images is a useful clinical prognostic biomarker in patients with glioblastoma.     

Radiosynthesis and evaluation of [11C]YM-202074 as a PET ligand for imaging the metabotropic glutamate receptor type 1

IntroductionDeveloping positron emission tomography (PET) ligands for imaging metabotropic glutamate receptor type 1 (mGluR1) is important for studying its role in the central nervous system. N-cyclohexyl-6-{[N-(2-methoxyethyl)-N-methylamino]methyl}-N-methylthiazolo[3,2-a]benzimidazole-2-carboxamide (YM-202074) exhibited high binding affinity for mGluR1 (K_i=4.8 nM), and selectivity over other mGluRs in vitro. The purpose of this study was to label YM-202074 with carbon-11 and to evaluate in vitro and in vivo characteristics of [¹¹C]YM-202074 as a PET ligand for mGluR1 in rodents.Methods[¹¹C]YM-202074 was synthesized by N-[¹¹C]methylation of its desmethyl precursor with [¹¹C]methyl iodide. The in vitro and in vivo brain regional distributions were determined in rats using autoradiography and PET, respectively.Results[¹¹C]YM-202074 (262–630 MBq, n=5) was obtained with radiochemical purity of >98% and specific activity of 27–52 GBq/μmol at the end of synthesis, starting from [¹¹C]CO₂ of 19.3–21.5 GBq. In vitro autoradiographic results showed that the high specific binding of [¹¹C]YM-202074 for mGluR1 was presented in the cerebellum, thalamus and hippocampus, which are known as mGluR1-rich regions. In ex vivo autoradiography and PET studies, the radioligand was specifically distributed in the cerebellum, although the uptake was low. Furthermore, the regional distribution was fairly uniform in the whole brain by pretreatment with JNJ16259685 (a mGluR1 antagonist). However, radiometabolite(s) was detected in the brain.ConclusionsFrom these results, especially considering the low brain uptake and the influx of radiometabolite(s) into brain, [¹¹C]YM-202074 may not be a useful PET ligand for in vivo imaging of mGluR1 in the brain.