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Apparent molar volumes Vφof aqueous KCl, KOH, and NaOH and apparent molar heat capacities Cp, φof aqueous HCl, KCl, KOH, and NaOH have been determined at the pressure p=0.35 MPa, and at molalities 0.015 ⩽m/mol · kg−1⩽0.5. Densities were measured using a vibrating-tube densimeter (DMA 512, Anton Paar, Austria) at temperatures 278.15 ⩽T/K⩽368.15. These values were used to calculate the apparent molar volumes. A fixed-cell, differential-output, power-compensating, temperature-scanning calorimeter (NanoDSC model 6100, Calorimetry Sciences Corporation, Spanish Fork, UT, U.S.A.) was used to measure the heat capacities of the same solutions at temperatures 278.15 ⩽T/K⩽393.15. Results were fitted by using equations that describe the surfaces (m, T, Vφ) and (m, T, Cp, φ). Using these equations, we have calculated the surfaces (m, T, ΔrVm), (m, T, ΔrCp, m), (m, T, ΔrHm), (m,T, p Qa), and (m, T,ΔrSm) for the ionization of water in the presence of combinations of the above electrolytes. The last three surfaces were calculated by integration using our (m,T, ΔrCp, m) surface and literature values for the molality dependence of ΔrHmand pQaat T=298.15 K.

Apparent molar volumes Vφand apparent molar heat capacitiesCp, φwere determined for individual solutions of aqueousl-histidine, of aqueousl-histidine with equimolal HCl, and of aqueousl-histidine with equimolal NaOH at molalities m=(0.015 to 0.66 )mol · kg−1, at temperatures T=(278.15 to 393.15 ) K, and at the pressure p=0.35 MPa. Apparent molar volumes were generated from density measurements obtained with a vibrating-tube densimeter. Apparent molar heat capacities were generated from heat capacity measurements with a twin fixed-cell, power-compensation, differential-output, temperature-scanning calorimeter. These results were then fitted by regression to empirical equations to describe the (Vφ,m, T) and (Cp, φ, m, T) surfaces for each of the three systems. These regression equations were then used to calculate the changes in partial molar volumeΔrVmand partial molar heat capacityΔrCp, mas functions of mand Tfor both the first and second proton dissociation reactions for protonated aqueousl-histidine. The changes in enthalpy ΔrHmand entropyΔrSmand the acid dissociation molality quotientQawere then obtained as functions of mand Tfor each proton dissociation reaction by integration, using our (ΔrCp, m, m,T) results and literature values for ΔrHmandQa. Our results illustrate the unique thermodynamic properties of the cationic, neutral zwitterionic, and anionic forms ofl-histidine in aqueous solution.

A vibrating-tube densimeter (DMA 512P, Anton Paar, Austria) was used to investigate the densities and volumetric properties of aqueous potassium hydrogen phthalate (KHP) and potassium sodium phthalate (KNaP). Measurements were made at molalities mfrom (0.006 to0.66)mol · kg−1, at temperatures from 278.15 K to 368.15 K and at the pressure 0.35 MPa. The densimeter was calibrated through measurements on pure water and on 1.0 mol · kg−1NaCl(aq). We also used a twin fixed-cell, power-compensation, differential-output, temperature-scanning calorimeter (NanoDSC 6100, Calorimetry Sciences Corporation, Spanish Fork, UT, U.S.A.) to measure solution heat capacities. This was accomplished by scanning temperature and comparing the heat capacities of the unknown solutions to the heat capacity of water. Apparent molar volumes Vφand apparent molar heat capacities Cp, φof the solutions were calculated and fit by regression to equations that describe the surfaces (Vφ, T, m) and (Cp, φ, T, m). Standard state partial molar volumesV2oand heat capacities Cp,2owere estimated by extrapolation to the m=0 plane of the fitted surfaces. Previously determinedCp, φfor HCl(aq) and NaCl(aq) were used to obtain (ΔrCp, m, T, m) for the proton dissociation reaction of aqueous hydrogen phthalate. This (ΔrCp,m, T, m) surface was created by subtracting Cp,φfor KHP(aq) and for NaCl(aq) from the sum of Cp,φfor KNaP(aq) and for HCl(aq). Surfaces representing (ΔrHm, T, m) and (pQa, T, m), where pQadenotes the molality equilibrium quotient, were created by integration of our (ΔrCp,m, T, m) surface using values for (ΔrHm, m) and (pKa, m) at T=308.15 K from the literature as integration constants.

We have used a vibrating-tube densimeter to investigate the densities and volumetric properties of aqueous solutions, and a fixed-cell scanning calorimeter to measure aqueous solution heat capacities. Solutions investigated include sugars, neutral weak acids and bases, neutralized acids and bases, neutral and protonated amino acids, surfactants, and both strong and ion-paired electrolytes. We have made measurements at molalities m from 0.005 mol·kg−1 to 0.5 mol·kg−1, at temperatures T from 278.15 K to 393.15 K, and at the pressure p = 0.35 MPa. We have calculated the apparent molar volumes Vφ and apparent molar heat capacities Cp,φ of the solutions and fit them by regression to equations that describe the surfaces (Vφ, T, m) and (Cp,φ, T, m). These results reflect changes and differences in both solute-solute and solute-solvent interactions. Standard state partial molar volumes V2° and heat capacities Cp,2° were estimated by extrapolation to the m = 0 mol·kg−1 plane of the fitted surfaces. We have calculated (Δr,Cp,m, T, m) surfaces for various proton dissociation reactions, and we have created surfaces representing (ΔrHm, T, m) and (pQa, T, m) by integration of our (ΔrCp,m, T, m) surfaces using values for (ΔrHm, m) and (pKa, m) at a reference T as integration constants. We have also created surfaces representing (ΔrSm, T, m) and (ΔrVm, T, m) for these dissociation reactions.

Apparent molar volumes Vφand apparent molar heat capacitiesCp, φwere determined for aqueous solutions of magnesium chloride (MgCl2) and cadmium chloride (CdCl2) at molalities m=(0.01 to 1.0)mol · kg−1, at T=278.15 K to T=393.15 K, and at the pressure 0.35 MPa. Apparent molar volumes were calculated from densities obtained using a vibrating-tube densimeter. Apparent molar heat capacities were obtained using a twin fixed-cell, power-compensation, differential-output, temperature-scanning calorimeter. These results were used to create the surfaces (Vφ, m, T) and (Cp, φ,m, T) which were fitted by regression. Differences in ion–ion interactions for MgCl2(aq) and CdCl2(aq) are observed by comparison of the regression surfaces.

Apparent molar heat capacities Cp, φand apparent molar volumesVφwere determined for aqueous solutions of d-glucose and d-galactose at temperatures from 278.15 K to 393.15 K and at the pressure 0.35 MPa. The molalities minvestigated ranged from 0.05 mol · kg−1to 0.5 mol · kg−1. A vibrating tube densimeter (DMA 512P, Anton PAAR, Austria) was used to investigate the densities and volumetric properties. Heat capacities were obtained using a twin fixed-cell, power-compensation, differential-output, temperature-scanning calorimeter (NanoDSC 6100, Calorimetry Sciences Corporation, Provo, UT, U.S.A.). The calculatedVφand Cp, φof the solutions were fit to equations that describe the surfaces (Vφ, T, m) and (Cp,φ, T, m). These surfaces showed no dependence of VφorCp, φon M. Infinite dilution partial molar volumesV2oand heat capacities Cp,2owere obtained over the range of temperature. The Cp,2oandV2ovalues were found to be large, positive, and to increase with increasing temperature.

We used a vibrating tube densimeter (DMA 512P, Anton Paar, Austria) to investigate the densities and volumetric properties of aqueous 2-amino-2-hydroxymethyl-propan-1,3-diol (Tris or THAM) and THAM plus equimolal HCl. We made measurements at molalities mfrom (0.005 to 0.5)mol · kg−1and at temperatures from 278.15 K to 393.15 K. We calibrated the densimeter through measurements on pure water and on 1.0 mol · kg−1NaCl at the pressure 0.35 MPa. We used a fixed-cell, power-compensation, differential-output, temperature-scanning calorimeter (NanoDSC 6100, Calorimetry Sciences Corporation, Provo, UT, U.S.A.) to measure solution heat capacities at molalities from (0.005 to 0.5)mol · kg−1and at temperatures from 278.15 K to 368.15 K. This was accomplished by scanning temperature and comparing the heat capacities of the unknown solutions to the heat capacity of water. We calculated the apparent molar volumes Vφand apparent molar heat capacitiesCp, φof the solutions and fitted them to equations that describe the surfaces (Vφagainst Tagainst m) and (Cp,φagainst Tagainst m). Standard state partial molar volumesV2oand heat capacities Cp,2owere estimated by extrapolation to the m=0 plane of the fitted surfaces. We used previously determinedCp, φfor HCl(aq) to obtainΔrCp, mfor the proton dissociation reaction of THAM ·H+(aq). The (ΔrCp,magainst Tagainst m) surface was created by subtracting Cp,φof THAM(aq) and HCl(aq) from the heat capacity of THAM ·HCl(aq). We created surfaces representing ΔrHmandpQaby integration of our ΔrCp,msurface over Twhile using values forΔrHmand pQaatT=298.15 Kfrom the literature as integration constants.

Apparent molar volumes Vφand heat capacities Cp, φof aqueous solutions of acetic acid (CH3CO2H)and sodium acetate (CH3CO2Na), and Cp, φfor hydrochloric acid (HCl) and sodium hydroxide (NaOH) were determined at temperatures from T=278.15 K to T=393.15 K at the pressure 0.35MPa. Apparent molar volumes were obtained from density measurements using an Anton Paar vibrating-tube densimeter (DMA 512, Anton Paar, Austria). Heat capacities were obtained using a fixed-cell, power-compensation, differential-output, temperature-scanning calorimeter (NanoDSC model 5100, Calorimetry Sciences Corporation, Provo, UT). Infinite dilution partial molar volumes Voand heat capacities Cpowere obtained over the range of temperatures by extrapolation to m=0 of the fitted surfaces (Yφ, T, m). The volume results were combined with values of Vofrom the literature for aqueous NaCl, HCl, and NaOH to obtain ΔrVmofor ionization of acetic acid. Measured values of Cp, φwere extrapolated to m=0 for HCl(aq) and NaOH(aq) and used to obtain values of ΔrCp, mofor ionization of CH3CO2H(aq). The values of ΔrVmoand ΔrCp, moallow the determination by integration of the equilibrium constant Kaand other thermodynamic properties for ionization of CH3CO2H(aq) from T=278.15 K to T=393.15 K at the pressure 0.35MPa.

The most common recognized cytogenetic change in B-CLL is a deletion involving band 13q14.3. Dewald found a heterozygous or homozygous 13q– in 92% of CLL patient samples using the FISH probe D13S319 that hybridizes to 13q14.3 (BJH 121:287, 2003). This contrasts with the usually normal conventional chromosome analysis (CCA) results partly because CLL cells divide infrequently but also because the 13q– is often visualized only by FISH (Stockero, Ca Genet Cytogenet 166:152, 2006). Mayr recently showed that CpG stimulation can reveal a chromosomally abnormal CLL clone in metaphase cells (Blood 107:742, 2006). The purpose of the present study was to compare the incidence of microdeletion and visible 13q deletion in CLL using interphase FISH and CCA after CpG culture on the same peripheral blood specimens.

BACKGROUND: Green tea has long been touted as a health promoting substance. The active chemical compounds in green tea are called polyphenols or catechins. Epigallocatechin gallate(EGCG) is the major catechin in green tea. We previously reported the in vitroability of EGCG to induce apoptotic cell death in chronic lymphocytic leukemia(CLL) B-cells in vitro (Blood 104:788). After publication of our findings, clinical activity in individuals using over the counter green tea extracts were reported (Leuk Res 30:707). Based on this information, we opened a phase I/II trial of green tea extracts for patients with asymptomatic, early stage CLL in fall of 2005.

BACKGROUND: Green tea has long been touted as a health promoting substance. The active chemical compounds in green tea are called polyphenols or catechins. Epigallocatechin gallate(EGCG) is the major catechin in green tea. We previously reported the in vitro ability of EGCG to induce apoptotic cell death in chronic lymphocytic leukemia(CLL) B-cells in vitro (Blood 104:788). After publication of our findings, clinical activity in individuals using over the counter green tea extracts were reported (Leuk Res 30:707). Based on this information, we opened a phase I/II trial of green tea extracts for patients with asymptomatic, early stage CLL in fall of 2005. METHODS: The purpose of the phase I portion of this trial was to determine the optimal dose of EGCG in the Polyphenon E preparation for chronic daily administration and define tolerability in CLL patients. Previously untreated patients with asymptomatic, Rai stage 0–II CLL not currently meeting National Cancer Institute(NCI) Working Group(WG) Criteria for treatment were eligible for participation. Polyphenon E with a standardized dose of EGCG was obtained from NCI. The phase I portion of the trial was designed with 8 dose levels(range 400–2000 mg orally BID) using the standard 3 patient per dose level design. Patients remained on study up to 6 months. Grade 2 adverse events attributed to study treatment that did not respond to supportive care were considered dose limiting toxicity. The trial was designed to administer Polyphenon E in the fasting state. After accrual to dose levels 1 and 2, the U.S. FDA mandated all U.S. trials of Polyphenon E administer drug in the fed state. Accordingly, drug was administered in the fed state for dose levels 3–8. Trough plasma EGCG levels were measured 1 month after initiation of therapy. Response was classified using the NCI WG Criteria. RESULTS: As of August 2007, 33 patients have been accrued to dose levels 1–8. The maximum tolerated dose(MTD) has not been reached. Side effects have generally been mild. The most common toxicities were nausea(grade 1: 42%; grade 2: 3%), elevation in SGOT (42%; all grade 1), and abdominal pain (36%; all grade 1). To date, no patient has had a sustained 50% reduction in both absolute lymphocyte count (ALC) and lymphadenopathy that would meet the NCI WG criteria for partial response. A majority of patients have had a reduction in ALC(Table). Among the 10 patients who had palpable adenopathy at study enrollment, 7 patients experienced at least a 50% reduction in the sum of the products of all nodal areas at some point during treatment. Trough plasma EGCG levels after 1 month of treatment ranged from 2.93974 ng/mL(median 40.5 ng/mL). Plasma levels did not clearly relate to the degree of reduction in ALC suggesting sensitivity to Polyphenon E may relate more to characteristics of the leukemic clone than plasma EGCG levels. CONCLUSION: Daily oral EGCG in the Polyphenon E preparation was well tolerated by CLL patients in this phase I trial. The MTD has not been reached. As classified by the NCI WG criteria, no partial or complete remissions have been observed to date, however declines in ALC and lymphadenopathy have been observed in the majority of patients. The phase II portion of this trial will open at Mayo Clinic Fall 2007. Best reduction in ALC n % of patients At least 10% decline 25 76% At least 20% decline 14 42% At least 30% decline 8 24% At least 40% decline 4 12% At least 50% decline 2 6%

The most common recognized cytogenetic change in B-CLL is a deletion involving band 13q14.3. Dewald found a heterozygous or homozygous 13q– in 92% of CLL patient samples using the FISH probe D13S319 that hybridizes to 13q14.3 (BJH 121:287, 2003). This contrasts with the usually normal conventional chromosome analysis (CCA) results partly because CLL cells divide infrequently but also because the 13q– is often visualized only by FISH (Stockero, Ca Genet Cytogenet 166:152, 2006). Mayr recently showed that CpG stimulation can reveal a chromosomally abnormal CLL clone in metaphase cells (Blood 107:742, 2006). The purpose of the present study was to compare the incidence of microdeletion and visible 13q deletion in CLL using interphase FISH and CCA after CpG culture on the same peripheral blood specimens. METHOD: We compared the chromosome 13 results by interphase FISH (fresh uncultured cells) and by a 20 metaphase CCA from 5–day CpG cultures. Our unselected CLL cohort (n=40) represented a typical and relatively high risk population: median age 61 (range 36–75), 48% CD38–positive, 62% ZAP–70 positive, and 48% IgVH unmutated. In addition 70% were previously treated for progressive disease, with all Rai stages represented. CpG RESULTS: By CCA of 40 patients, CpG stimulation revealed a clonal (multiple abnormal cells) or nonclonal (one abnormal cell) abnormal karyotype in 32 (80%) and a normal karyotype in 8 of the 40 patients (20%). Among the 32 abnormal cases, each chromosome 13 pair appeared normal in 17 and was abnormal in 15 (one was monosomy 13, 8 were 13q–, and 6 had a 13q translocation). CpG did not reveal a homozygous 13q abnormality in any patient. In total, CpG revealed a 13q abnormality in 15 of 40 (38%) patients (8 had multiple abnormal metaphases and 7 had only one abnormal metaphase). FISH RESULTS: By interphase FISH, 29 of 40 (73%) patients had a 13q–. The deletion was heterozygous in 18 patients, homozygous in 7, and mixed homo– and heterozygous in 4. Of the 18 with a heterozygous 13q loss by FISH, CpG revealed an abnormal 13 in only 8. Of the 11 patients with homozygous or mixed homo– and heterozygous 13q– by FISH, CpG revealed a heterozygous 13q abnormality in only 6. Of the 8 patients with a normal CpG karyotype, by FISH 5 had a 13q– in one or both 13s. Of the 9 patients with heterozygous 13q loss by CpG, the FISH result was 13q– in 4 and homozygous or mixed homo– and heterozygous 13q– in 5. Of the 6 with a chromosome 13 translocation by CpG, FISH revealed heterozygous 13q– in 4, mixed homo– and heterozygous 13q– in one, and in one case the 13q FISH result was within normal limits. In this latter case, CpG stimulation followed by metaphase FISH analysis confirmed that the apparently balanced translocation harbored a microdeletion 13q. Interphase FISH analysis confirmed the presence of a clonal deletion 13q in all 7 of the patients with a CpG metaphase result of a non-clonal (one metaphase cell) 13q abnormality. CONCLUSIONS: Even though CpG induces CLL metaphases in cell culture and reveals many chromosome abnormalities that cannot be identified by interphase FISH, cytogenetic evaluation of CLL by CpG alone significantly underestimates the incidence of 13q deletions. When a 20-cell analysis after CpG stimulation reveals a 13q abnromality in only one (apparently nonclonal) metaphase cell, concurrent interphase FISH analysis often confirms clonality for the 13q defect.

Background: Angiogenesis has been found to be an important regulator in the disease progression of both solid tumors and hematologic malignancies. In solid tumors, the balance of pro and anti-angiogenic factors appears to be a critical factor in tumorigenesis, with disturbance in favor of angiogenesis promoting tumor growth and metastasis. This observation led to the concept of an “angiogenic switch” where pro-angiogenic influences outweigh anti-angiogenic factors. Patients with CLL have detectable levels of plasma and cellular pro and anti-angiogenic cytokines as well as abnormal neovascularization in the marrow and lymph nodes. Limited studies of pro-angiogenic cytokines have suggested that inter-patient variation in serum/plasma and cellular levels of these markers may have prognostic implications. We evaluated pro- and anti-angiogenic cytokines in a large sample of patients with CLL to evaluate the presence of angiogenic switching and its implications for disease progression.