Effect of Surface Freezing of a Cationic Surfactant and n-Alkane Mixed Adsorbed Film on Counterion Distribution and Surface Dilational Viscoelasticity Studied by Total Reflection XAFS and Surface Quasi-Elastic Light Scattering.

When liquid alkane droplets are placed on a surfactant solution surface having a proper surface density, alkane molecules penetrated into the surfactant-adsorbed film to form a mixed monolayer. Such a mixed monolayer undergoes a thermal phase transition from two-dimensional liquid to solid monolayers upon cooling when surfactant tail and alkane have similar chain lengths. We applied the total-reflection XAFS spectroscopy and surface quasi-elastic light scattering to the mixed adsorbed film of cetyltrimethylammonium bromide and hexadecane to elucidate the impact on the surface phase transition on the counterion distribution of the mixed monolayer. The EXAFS analysis verified that a higher percentage of counter Br- ions were localized in the Stern layer than in the diffuse double layer in the surface solid film compared to the surface liquid film, which resulted in a reduction in the surface elasticity measured by the SQELS. The finding that the surface phase transition accompanies the change in the counterion distribution will be important to consider the future applications of the colloidal systems, in which the coexistence of a surfactant and alkane molecules is essential, such as foams and emulsions.

Decreased thymic output predicts progression of chronic kidney disease.

BACKGROUND: Chronic kidney disease (CKD) is age-related disease, and decreased renal function is associated with the premature aging of T cells and increased incidence of other age-related diseases. However, the relationship between T cell senescence and CKD progression remains unclear. Here, we investigated the relationship between T cell senescence, as indicated by decreased thymic output and increased proportion of highly differentiated CD28- T cells, and CKD progression. RESULTS: A total of 175 patients with non-dialysis-dependent CKD were enrolled in this study. Thymic output was assessed based on the CD45RA+CD31+CD4+ cell (recent thymic emigrant [RTE]) counts (RTEs) (/mm3) and the proportion of RTE among CD4+ T cells (RTE%). Highly differentiated T cells were assessed based on the proportion of CD28- cells among CD4+ T cells (CD28-/CD4+) and CD28- cells among CD8+ T cells (CD28-/CD8+). The primary outcome was estimated glomerular filtration rate (eGFR) decline of ≥40% or initiation of renal replacement therapy. The association between T cell senescence and renal outcomes was examined using Cox proportional hazards models and restricted cubic splines. The median age was 73 years, 33% were women, and the median eGFR was 26 mL/min/1.73 m2. The median RTEs, RTE%, CD28-/CD4+, and CD28-/CD8+ were 97.5/mm3, 16.2, 5.3, and 49.7%, respectively. After a median follow-up of 1.78 years, renal outcomes were observed in 71 patients. After adjusting for age, sex, eGFR, proteinuria, diabetes, and cytomegalovirus seropositivity, decreased RTEs, which corresponded to decreased thymic output, significantly and monotonically increased the risk of poor renal outcome (p = 0.04), and decreased RTE% and increased highly differentiated CD28-/CD4+ T cells also tended to monotonically increase the risk (p = 0.074 and p = 0.056, respectively), but not CD28-/CD8+ T cells. CONCLUSIONS: Decreased thymic output in CKD patients, as well as increased highly differentiated CD4+ T cells, predicted renal outcomes. Thus, the identification of patients prone to CKD progression using T cell senescence, particularly decreased RTE as a biomarker, may help to prevent progression to end-stage kidney disease.

A multicenter, phase II study of full-dose THP-COP therapy for elderly patients with newly diagnosed, advanced-stage, aggressive non-Hodgkin lymphoma.

The cyclophosphamide, doxorubicin, vincristine, and prednisolone (CHOP) regimen, containing doxorubicin (DXR), which is a key drug for aggressive non-Hodgkin lymphoma (NHL), is a standard chemotherapeutic regimen; however, its administration in elderly patients is often intolerable. Pirarubicin (tetrahydropyranyl adriamycin [THP]) is an anthracycline developed in Japan. We have conducted a phase II trial of a full-dose THP-COP (modified CHOP regimen with DXR replaced by THP) regimen for elderly patients with newly diagnosed, advanced-stage, aggressive NHL. Patients aged 70-79 years old with previously untreated NHL according to the Working Formulation (D through H and J), disease stage I with a bulky mass or stage II-IV, and performance status of 0-1 were eligible. The THP-COP regimen, which consisted of 750-mg/m2 cyclophosphamide, 50-mg/m2 THP, 1.4-mg/m2 vincristine (capped at 2.0 mg) on day 1, and 100-mg prednisolone daily on days 1 to 5, was delivered every 3 weeks for 6 cycles. The primary endpoint was complete response (CR) rate. Twenty-nine patients were enrolled in the study. The CR rate was 65.5% (95% confidence interval, 45.7-82.1%). The 3-year failure-free and overall survival rates were 54.1% and 53.9%, respectively. The most frequent observed grade 3 or 4 toxicity was neutropenia, which occurred in 80% of the patients. Grade 3 cardiac dysfunction was observed in one patient. The full-dose THP-COP regimen exhibited similar efficacy and safety, and a tendency for less cardiac toxicity, when compared with the standard CHOP regimen in elderly Japanese patients with newly diagnosed, advanced-stage, aggressive NHL.

Gitelman syndrome with a novel frameshift variant in SLC12A3 gene accompanied by chronic kidney disease and type 2 diabetes mellitus.

Gitelman syndrome is an autosomal recessive genetic disease caused by pathogenic variants in SLC12A3 resulting in the loss of function of the Na-Cl co-transporter (NCC) in the distal tubules. Hypokalemia and diuretic effects can cause secondary type 2 diabetes and renal function decline. Here, we present the case of a 49-year-old male patient with chronic persistent treatment-resistant hypokalemia for the past 13 years who had been receiving treatment for type 2 diabetes mellitus for 6 years. He was referred to our department due to the presence of urinary protein, impaired renal function, high renin activity, and hyperaldosteronism. Laboratory test results showed hypokalemia, hypomagnesemia, hypocalciuria, and metabolic alkalosis. Using next-generation and Sanger sequencing, we identified a novel stop-gain variant (NM_000339.3:c.137del [p.His47fs]) and a missense variant (NM_000339.3:c.2927C > T [p.Ser976Phe]) in the SLC12A3 gene. This novel pathogenic variant was located at the intracellular N-terminus of the NCC. Based on these findings, the patient was diagnosed with Gitelman syndrome. The use of next-generation sequencing facilitated the exclusion of diseases with similar clinical symptoms.

Listening to Sutra-Chanting Reduces Bereavement Stress in Japan.

Buddhist Chaplains chanting sutras after the Great East Japan Earthquake in 2011 often encountered survivors who felt that hearing sutra chanting itself ameliorated their bereavement grief. This research is the first experimental examination of the effects of sutra chanting on listeners' bereavement stress. Prior research demonstrates that sudden pet loss causes bereavement stress in students and that physiological stress can be noninvasively measured by salivary alpha-amylase. We asked Japanese college students to raise pet goldfish until they developed an attachment to them, then confiscated the fish, and told the students that they had to be killed. To compare the bereavement stress of groups listening and not listening to sutra chanting, we used psychological and salivary analyses. Perceived Stress Scale (PSS), Multidimensional Empathy Scale (MES), and State half of the State Trait Anxiety Inventory (STAI) psychological scales showed no statistically significant differences between sutra and control groups, but salivary analyses indicated measurable stress reduction in the sutra-listening group only. This pilot study tentatively confirmed the hypothesis that listening to Buddhist sutra chanting reduces Japanese bereavement stress. Further research is needed both to verify these stress-reduction effects and to determine whether such effects are primarily musical or cultural/spiritual.

Condensed Film Formation and Molecular Packing in Cationic Surfactant-Cholesterol and Zwitterionic Surfactant-Cholesterol Systems at the Hexane/Water Interface.

A condensed film formation of surfactants with a charged head group at the oil/water interface was achieved by mixing surfactants of different geometric shapes to control molecular packing at the interface. The adsorbed films of mixed tetradecyltrimethylammonium bromide (C14TAB)-cholesterol (Chol) and tetradecylphosphocholine (C14PC)-Chol systems at the hexane/water interface were examined by interfacial tension and X-ray reflectivity measurements. The interfacial tension versus Chol concentration curves have break points because of the expanded-condensed phase transition of the adsorbed film. A two dimensional (2D) phase diagram, phase diagram of adsorption, indicated that 1:1 mixing in the condensed film is energetically favorable because of stronger mutual interaction between different molecules than between the same ones. The electron density profile normal to the interface manifested that the packing of C14TAB (or C14PC) and Chol molecules is like a 2D solid in the condensed state. As C14TAB and C14PC molecules take a corn shape with a large head group (critical packing parameter: CPP ≈ 1/3) and Chol takes an inverted corn shape with a bulky sterol ring (CPP > 1), the mixing of corn shape and inverted corn shape molecules produces well-ordered packing to promote solid-like molecular packing at the interface by energy gain because of vdW interaction between hydrophobic chains in addition to attractive ion-dipole interaction between head groups. Furthermore, the heterogeneous feature in the adsorbed film of the C14TAB-Chol system is explained by an interplay between contact energy and dipole interaction, which contribute to line tension at the domain boundary.

Safety and efficacy of anti-programmed cell death-1 monoclonal antibodies before and after allogeneic hematopoietic cell transplantation for relapsed or refractory Hodgkin lymphoma: a multicenter retrospective study.

We conducted a multicenter study on anti-programmed cell death-1 monoclonal antibodies (anti-PD-1 mAbs) before/after allogeneic hematopoietic cell transplantation (allo-HCT) for Hodgkin lymphoma. Anti-PD-1 mAbs were administered to 25 patients before allo-HCT and to 20 after allo-HCT. In pre-allo-HCT setting, the median interval from the last administration to allo-HCT was 59 days. After allo-HCT, 12 patients developed non-infectious febrile syndrome requiring high-dose corticosteroid. The cumulative incidences of grade II-IV acute graft-versus-host disease (aGvHD) were 47.1%. Eight patients who had GvHD prophylaxis with post-transplant cyclophosphamide (PTCy) had less frequent aGvHD (grade II-IV, 14.6% versus 58.8%; P = 0.086). The 1 year overall survival (OS), relapse/progression, and non-relapse mortality rates were 81.3%, 27.9%, and 8.4%. In post-allo-HCT setting, the median interval from allo-HCT to the first administration was 589 days. The overall and complete response rates were 75% and 40%. At 100 days after anti-PD-1 therapy, the cumulative incidences of grade II-IV aGvHD, moderate-to-severe chronic GvHD, and grade 3-4 immune-related toxicity were 15.0%, 30.0%, and 30.0%. While the 1 year relapse/progression rate was 47.4%, the 1 year OS probability was 89.7%. In conclusion, immune-related complications were frequent despite modifications of GvHD prophylaxis or anti-PD-1 mAb dosing. In anti-PD-1-mAb-pretreated patients, PTCy-based GvHD prophylaxis may be effective.

The 31-nucleotide Y4-RNA fragment in plasma is a potential novel biomarker.

The 31- and 32-nt 5'-fragment of Y4-RNA (Y4RNAfr) exists abundantly in human peripheral blood plasma. Although physiological roles of the plasma Y4RNAfr are not well established, its potential utility as a diagnostic/prognostic marker for acute coronary syndrome was suggested. In this paper, to establish a normal range of the Y4RNAfr level in plasma, we measured plasma Y4RNAfr levels of 40 healthy persons using the method we have developed, and compared them with other blood test data. From the obtained data, we tentatively regarded <0.1 fmol/ng as normal for the Y4RNAfr level in peripheral blood plasma. And the white blood cell count (WBC) and the C-reactive protein (CRP) level showed moderate positive correlations with the Y4RNAfr level, suggesting that Y4RNAfr could be a potential novel inflammatory marker. We also measured the Y4RNAfr level in peripheral blood plasma from four multiple myeloma patients. The plasma Y4RNAfr level was abnormal in all four myeloma patients, and the levels for two patients were far beyond the normal level. The WBC for each patient was normal and the CRP levels for two patients were normal. These observations together suggest that a high level of Y4RNAfr in peripheral blood plasma and a normal WBC could be indicative of multiple myeloma.

Solid Film Formation at the Tetradecane/Aqueous Hexadecyltrimethylammonium Bromide Solution Interface Studied by Interfacial Tensiometry and X-ray Reflectometry.

The effect of oil on condensed film formation in the adsorbed film of hexadecyltrimethylammonium bromide (C16TAB) at the tetradecane (C14)/water (W) interface was examined by interfacial tension and X-ray reflectivity measurements. The interfacial tension vs temperature curves have break point due to the expanded?condensed phase transition of the adsorbed film. The partial molar entropy of C16TAB at the interface changes discontinuously, whereas the interfacial density changes almost continuously at the phase transition point. The electron density profile normal to the interface manifested that the condensed film is regarded as a two-dimensional (2D) solid rotator phase in which C16TAB and C14 molecules are densely packed with perpendicular orientation. Combining the interfacial tension and X-ray reflectivity data, the mixing ratio of C16TAB to C14 in the solid film was determined to be 2:3 and thus the film is enriched in oil molecules than surfactant ones. Furthermore, the partial molar entropy change of C14 associated with solid film formation was found to be largely negative and very close to that of surface freezing of liquid alkane, manifesting that C14 molecules are well ordered to form a 2D solid film by mixing with C16TAB molecules at the interface. The solid film formation of the present system is driven by effective vdW interactions between adsorbed C16TAB and intercalated C14 molecules. The morphology of the condensed domain observed during phase transition suggested that the contact energy is more predominant than the dipole repulsion at the domain boundary, which promotes coalescence of small domains into large ones during phase transition.

Parathyroid hormone and premature thymus ageing in patients with chronic kidney disease.

Premature immune ageing, including thymic atrophy, is observed in patients with chronic kidney disease (CKD). Parathyroid hormone (PTH) and fibroblast growth factor 23 (FGF23), which are mineral and bone disorder (MBD)-related factors, affect immune cells and possibly cause thymic atrophy. We examined the cross-sectional association between thymic atrophy, evaluated as the number of CD3+CD4+CD45RA+CD31+ cells [recent thymic emigrants (RTE)/µL], and MBD-related factors [(serum PTH, FGF23, and alkaline phosphatase (ALP) level] in 125 patients with non-dialysis dependent CKD. Median estimated glomerular filtration rate (eGFR) was 17 mL/min/1.73 m2. Older age (r = -0.46), male sex (r = -0.34), lower eGFR (r = 0.27), lower serum-corrected calcium (r = 0.27), higher PTH (r = -0.36), and higher ALP level (r = -0.20) were identified as determinants of lower number of RTE. In contrast, serum concentrations of FGF23 and phosphorus were not correlated with RTE. Multivariate non-linear regression analysis indicated a negative association between serum PTH and log-transformed RTE (P = 0.030, P for non-linearity = 0.124). However, the serum levels of FGF23 and ALP were not associated with RTE. In patients with CKD, serum PTH concentrations were related to thymic atrophy which contributes to immune abnormality.

Assessment of CPM reliability: quantification of the within-subject reliability of 10 different protocols.

Background and aims Conditioned Pain Modulation (CPM) is a well-established phenomenon and several protocols have shown acceptable between-subject reliability [based on intraclass correlation coefficient (ICC) values] in pain-free controls. Recently, it was recommended that future CPM test-retest reliability studies should explicitly report CPM reliability based on CPM responders and non-responders (within-subject reliability) based on measurement error of the test stimulus. Identification of reliable CPM paradigms based on responders and non-responders may be a step towards using CPM as a mechanistic marker in diagnosis and individualized pain management regimes. The primary aim of this paper is to investigate the frequency of CPM responders/non-responders, and to quantify the agreements in the classification of responders/non-responders between 2 different days for 10 different CPM protocols. Methods Data from a previous study investigating reliability of CPM protocols in healthy subjects was used. In 26 healthy men, the test-stimuli used on both days were: Pain thresholds to electrical stimulation, heat stimulation, manual algometry, and computer-controlled cuff algometry as well as pain tolerance to cuff algometry. Two different conditioning stimuli (CS; cold water immersion and a computer-controlled tourniquet) were used in a randomized and counterbalanced order in both sessions. CPM responders were defined as a larger increase in the test stimulus response during the CS than the standard error of measurement (SEM) for the test-stimuli between repeated baseline tests without CS. Results Frequency of responders and non-responders showed large variations across protocols. Across the studied CPM protocols, a large proportion (from 11.5 to 73.1%) of subjects was classified as CPM non-responders when the test stimuli standard error of measurements (SEM) was considered as classifier. The combination of manual pressure algometry and cold water immersion induced a CPM effect in most participants on both days (n=16). However, agreement in the classification of CPM responders versus non-responders between days was only significant when assessed with computer-controlled pressure pain threshold as test-stimulus and tourniquet cuff as CS (κ=0.36 [95% CI, 0.04-0.68], p=0.037). Conclusions and implications Agreements in classification of CPM responders/non-responders using SEM as classifier between days were generally poor suggesting considerable intra-individual variation in CPM. The most reliable paradigm was computer-controlled pressure pain threshold as test-stimulus and tourniquet cuff as conditioning stimulus. However while this CPM protocol had the greatest degree of agreement of classification of CPM responders and non-responders across days, this protocol also failed to induce a CPM response in more than half of the sample. In contrast, the commonly used combination of manual pressure algometry and cold water immersion induced a CPM effect in most participants however it was inconsistent in doing so. Further exploration of the two paradigms and classification of responders and non-responders in a larger heterogeneous sample also including women would further inform the clinical usefulness of these CPM protocols. Future research in this area may be an important step towards using CPM as a mechanistic marker in diagnosis and in developing individualized pain management regimes.

Bortezomib maintenance therapy in transplant-ineligible myeloma patients who plateaued after bortezomib-based induction therapy: a multicenter phase II clinical trial.

In the era of novel therapeutic agents for multiple myeloma (MM), both the significance of achieving the plateau phase and the efficacy of subsequent maintenance therapy remain unclear. In the present study, we evaluated the efficacy and safety of bortezomib maintenance therapy (biweekly for 1 year) in transplant-ineligible MM patients who plateaued after bortezomib-based induction therapy. Of 36 evaluable patients, the overall response rate during induction therapy was 61%, with a stringent complete response in 6%, a complete response in 6%, a very good partial response in 17%, and a partial response in 33%. Twenty patients achieved the plateau phase and subsequently received bortezomib maintenance therapy. Median progression-free survival from the induction and maintenance therapies was 13.8 months (95% confidence interval, 11.4-23.7 months) and 10.7 months (95% confidence interval, 3.7-10.7 months), respectively. During maintenance therapy, there were no cases with grade ≥ 2 peripheral neuropathy, nor was there any improvement in the quality of the response. In conclusion, although maintenance therapy with biweekly bortezomib for up to 1 year was feasible, plateau-oriented bortezomib induction therapy followed by bortezomib maintenance therapy was not adequate in newly diagnosed transplant-ineligible MM patients.

Effect of Hydrophobic Chain Structure on Phase Transition and Domain Formation of Hybrid Alcohol Films Adsorbed at the Hexane/Water Interface.

The phase transition and domain formation of the adsorbed film of two kinds of hybrid alcohols (CF3(CF2)m-1(CH2)nOH, FmHnOH), 2-perfluorooctylethanol (F8H2OH) and 2-perfluorohexylhexanol (F6H6OH), as a mixture at the hexane/water interface was investigated by interfacial tensiometry and X-ray reflection. The interfacial tension γ versus total molality m curve of pure F8H2OH has a break point at high concentration, and thus, the mean area per molecule A changes discontinuously at high interfacial pressure π, corresponding to the phase transition between expanded and condensed films. The Fresnel divided reflectivity R/RF versus Qz plots in the expanded state was well-fitted by the domain model for incoherent interference to determine the interfacial coverage, which is the fraction of the interface covered by the condensed phase. This indicates that the expanded film is heterogeneous and consists of a condensed F8H2OH domain, the size of which is larger than the X-ray coherence length (∼5 µm). In the mixed system, the discontinuous change in A at the phase transition point becomes small with increasing the bulk composition of F6H6OH X2 in the mixture, and eventually the A value changes continuously; i.e, the phase transition becomes obscure in X2 ≥ 0.6. This behavior was linked to an increase in interfacial coverage with X2. Furthermore, the R/RF versus Qz plot was fitted by the domain model for coherent interference, suggesting that the size of the domain is smaller than 5 µm. These results are probably due to the reduction of domain line tension by preferential adsorption of F6H6OH at the F8H2OH domain boundary.

Computational analysis of water residence on ceramide and sphingomyelin bilayer membranes.

Many physical chemical properties of lipid membranes, for example, the thickness, phase state, order parameter, and fluidity, can be understood straightforwardly. Water residence on a membrane is, however, an exception. To tackle this problem, we have performed molecular dynamics simulations of the distribution of water normal to the surface of several lipid membranes and from this deduced the associated water residence time. Our analysis of the results clearly indicates that lipid membranes have hydration shells on their surface, just as a solute in an aqueous solution does, and that the water residence time can be estimated from the potential for the mean force field derived from the distribution function of the water. We have done this atomic-scale analysis for ceramide bilayers and contrasted the calculation results with those for sphingomyelin bilayers, revealing that sphingomyelin bilayers can retain water molecules longer than ceramide bilayers and that the total number of water molecules retained on the membrane surface of sphingomyelin is larger than that for ceramide. In addition, we find that not only polar atoms of lipid molecules, such as oxygen, but also non-polar atoms, such as carbon, influence the motion of water on the membranes.

Surface-induced phase separation of a sphingomyelin/cholesterol/ganglioside GM1-planar bilayer on mica surfaces and microdomain molecular conformation that accelerates Abeta oligomerization.

Ganglioside GM1 mediates the amyloid beta (Abeta) aggregation that is the hallmark of Alzheimer's disease (AD). To investigate how ganglioside-containing lipid bilayers interact with Abeta, we examined the interaction between Abeta40 and supported planar lipid bilayers (SPBs) on mica and SiO(2) substrates by using atomic force microscopy, fluorescence microscopy, and molecular dynamics computer simulations. These SPBs contained several compositions of sphingomyelin, cholesterol, and GM1 and were treated at physiological salt concentrations. Surprisingly high-speed Abeta aggregation of fibril formations occurred at all GM1 concentrations examined on the mica surface, but on the SiO(2) surface, only globular agglomerates formed and they formed slowly. At a GM1 concentration of 20mol%, unique triangular regions formed on the mica surface and the rapidly formed Abeta aggregations were observed only outside these regions. We have found that some unique surface-induced phase separations are induced by the GM1 clustering effects and the strong interactions between the GM1 head group and the water layer adsorbed in the ditrigonal cavities on the mica surface. The speed of Abeta40 aggregation and the shape of the agglomerates depend on the molecular conformation of GM1, which varies depending on the substrate materials. We identified the conformation that significantly accelerates Abeta40 aggregation, and we think that the detailed knowledge about the GM1 molecular conformation obtained in this work will be useful to those investigating Abeta-GM1 interactions.

[Prophylaxis with acyclovir for herpes zoster infection during bortezomib-dexamethasone combination therapy].

A novel molecular targeting drug, a proteasome inhibitor, bortezomib (Bor), has been reported to be highly effective for relapsed/refractory, as well as for newly diagnosed multiple myeloma, but is also associated with a high frequency of herpes zoster (HZ) infection (13%). We conducted a retrospective survey on HZ infection (profile) after Bor therapy in our hospital. Six of 30 patients developed HZ infection during bortezomib-dexamethasone treatment (BD therapy). Age, performance status, and stem cell transplantation were not related risk factors for HZ infection. HZ developed when acyclovir (ACV) was not administrated to all six cases. Continuous administration of ACV decreased the incidence of HZ infection. Based on these results, we started an anti- HZ prophylaxis program using ACV for all patients receiving BD therapy. Further study is warranted to establish the optimal dose and duration of ACV for appropriate prophylaxis of HZ infection.

Monitoring of hormonal drug effect in a single breast cancer cell using an estrogen responsive GFP reporter vector delivered by a nanoneedle.

In this study, we have evaluated a sensor system for a hormonal drug effect in a single cell level using a novel low invasive single cell DNA delivery technology using a nanoneedle. An estrogen responsive GFP reporter vector (pEREGFP9) was constructed and its estrogenic response activity was confirmed in breast cancer cells (MCF-7) using lipofection as the means of transferring the vector to the cells. The pEREGFP9 vector was delivered to a single MCF-7 using a nanoneedle and the effect of ICI 182,780, which is an antagonist of estrogen, was observed using the GFP expression level. By ICI 182,780 treatment, the fluorescence intensity of the GFP was decreased by 30-50% within 24h. This technology is the very first trial of single cell diagnosis and we are looking forward to applying it to precious single cell diagnosis in medical fields.

Feasibility and efficacy of high-dose melphalan, cyclophosphamide, etoposide, and dexamethasone (LEED) chemotherapy with or without rituximab followed by autologous stem cell transplantation for aggressive and relapsed non-Hodgkin's lymphoma.

To investigate the feasibility and efficacy of high-dose chemotherapy (HDCT) followed by autologous stem cell transplantation (ASCT) for patients with newly diagnosed aggressive and relapsed non-Hodgkin's lymphoma (NHL), we administered LEED, a drug-only HDCT regimen consisting of melphalan, cyclophosphamide, etoposide, and dexamethasone, followed by ASCT in this single-institution trial. Furthermore, rituximab was added to the LEED regimen (R-LEED) for patients with CD20+ NHL. Twenty-six patients in the LEED group and 24 patients in the R-LEED group were enrolled and assessed for this study. All patients achieved complete engraftment after ASCT. As for the nonhematologic toxicities, infection toxicities of grades 3 and 4 were observed in 9 patients (34.6%) of the LEED group and 12 patients (50%) of the R-LEED group. Four patients (15.4%) in the LEED group and 5 (20.8%) in the R-LEED group developed grade 3 toxicity in the elevation of aspartate aminotransferase/alanine aminotransferase levels. Other grade 4 toxicities were rare in both groups. With a median follow-up time from the date of ASCT of 30 months in the LEED group and 18 months in the R-LEED group, the overall survival rates were 66.5% and 78.2%, respectively. These results suggested that LEED, as well as R-LEED, was a safe and feasible high-dose regimen for aggressive and relapsed NHL.

A tandemly repeated thyroglobulin core promoter has potential to enhance efficacy for tissue-specific gene therapy for thyroid carcinomas.

Recombinant adenoviruses, carrying herpes simplex virus thymidine kinase (HSVtk) genes, were developed to evaluate the possibility of tissue-specific gene therapy for thyroid carcinomas. The HSVtk gene was driven by a minimal thyroglobulin (TG) promoter (AdTGtk) and a tandemly repeated minimal TG promoter (Ad2 x TGtk) to obtain thyroid-specific cell killing ability. The transduction of HSVtk genes by infection with Ad2 x TGtk followed by ganciclovir (GCV) treatment showed more powerful cytotoxicity for TG-producing FRTL5 cells, a rat normal thyroid cell line, and FTC-133 cells, a human follicular thyroid carcinoma cell line, than when infected with AdTGtk in vitro. The cell killing ability of Ad2 x TGtk was 10- to 30-fold higher than that of AdTGtk and similar to that of AdCMVtk, which carries HSVtk under the control of CMV promoter. Whereas after treatment with adenovirus/GCV to non-TG-producing cell lines (undifferentiated thyroid carcinoma cell lines and carcinoma cell lines from other tissues), Ad2 x TGtk and AdTGtk needed more than 100-fold concentrated GCV to reach IC(50) compared to AdCMVtk. We confirmed the enhanced efficacy of Ad2 x TGtk for tissue-specific cytotoxicity in vivo. After adenovirus/GCV treatment for FTC-133 tumor-bearing nude mice, Ad2 x TGtk enhanced tumor growth inhibition and survival rates compared to AdTGtk. Tumor growth inhibition and survival rates by Ad2 x TGtk were similar to that by AdCMVtk. Moreover, any toxic effect for rat normal tissues was not revealed after intravenous injections with Ad2 x TGtk and intraperitoneal administrations with GCV in vivo, whereas severe liver damages were observed after treatment with AdCMVtk/GCV. These data indicate a beneficial effect of Ad2 x TGtk for tissue-specific gene therapy for TG-producing thyroid carcinomas without toxicity for normal tissues.