Exploring the energetics of histone H1.1 and H1.4 duplex DNA interactions.

H1.1 and H1.4 bind tightly to both short DNA oligomers and to CT-DNA (Ka≈1×10(7)). Binding is accompanied by an unfavorable enthalpy change (∆H≈+22 kcal/mol) and a favorable entropy change (-T∆S≈-30 kcal/mol). The Tm for the H1.4/CT-DNA complex is increased by 9 °C over the Tm for the free DNA. H1.4 titrations of the DNA oligomers yield stoichiometries (H1/DNA) of 0.64, 0.96, 1.29, and 2.04 for 24, 36, 48, and 72-bp DNA oligomers. The stoichiometries are consistent with a binding site size of 37±1 bp. CT-DNA titration data are consistent with binding site sizes of 32 bp for H1.1 and 36 bp for H1.4. The heat capacity changes, ΔCp, for formation of the H1.1 and H1.4/CT-DNA complexes are -160 cal mol(-1) K(-1) and -192 cal mol(-1)K(-1) respectively. The large negative ΔCp values indicate the loss of water from the protein DNA interface in the complex.

Calorimetric studies of the interactions of linker histone H1(0) and its carboxyl (H1(0)-C) and globular (H1(0)-G) domains with calf-thymus DNA.

Histone H1 is a chromatin protein found in most eukaryotes. ITC and CD have been used to study the binding of H1(0) and its C-terminal, H1(0)-C, and globular, H1(0)-G, domains to a highly polymerized DNA. ITC results indicate that H1(0) and H1(0)-C bind tightly to DNA (Ka≈1×10(7)), with an unfavorable ΔH (ΔH≈+22kcal/mol) and a favorable ΔS (-TΔS≈-30kcal/mol). Binding H1(0)-G to DNA at 25°C is calorimetrically silent. A multiple independent site model fits the ITC data, with the anomaly in the data near saturation attributed to rearrangement of bound H1, maximizing the number of binding sites. CD experiments indicate that H1(0)/DNA and H1(0)-C/DNA complexes form with little change in protein structure but with some DNA restructuring. Salt dependent ITC experiments indicate that the electrostatic contribution to binding H1(0) or H1(0)-C is small ranging from 6% to 17% of the total ΔG.

Computed tomography of humans and bowed stringed instruments. Some interesting similarities.

We have used computed tomography to evaluate bowed stringed instruments and have noted interesting analogies with CT scans of humans. In humans, CT commonly detects a broad range of normal anatomic variations. Similarly, CT of violins and cellos demonstrates a wide range of normal structural variations. CT, often used to detect infections in humans, also detects defects from wood infestations. In humans, the unrelenting effect of gravity causes deformity of the demineralized spine. In old stringed instruments, plastic deformity of wood results from unrelenting string pressure. Trauma causes bone fractures in humans and wood fractures in bowed stringed instruments, and repairing fractures in both humans and stringed instruments requires various splitting devices. In summary, CT provides the physician and the luthier with a unique, noninvasive tool that can characterize the broad range of normal structures, pathological conditions, and repair.

Phase I and pharmacokinetic analysis of high-dose tamoxifen and chemotherapy in normal and tumor-bearing dogs.

PURPOSE: To determine whether tamoxifen plasma concentrations capable of blocking P-glycoprotein (Pgp) in vitro can be safely achieved in dogs and whether doxorubicin pharmacokinetic alterations occur when tamoxifen is coadministered. METHODS: Tamoxifen dose escalation studies were conducted in 7 normal dogs and in 19 tumor-bearing dogs receiving full-dose chemotherapy. Plasma tamoxifen and serum doxorubicin disposition were analyzed for putative drug interactions. RESULTS: Steady-state plasma concentrations of tamoxifen and N-desmethyl tamoxifen (NDMT) were 5-10 microM following oral tamoxifen administration at 600 mg/m2 every 12 h for 7 days to normal and tumor-bearing dogs. Mild-moderate gastrointestinal toxicity (diarrhea, anorexia) and reversible neurotoxicity were observed in dogs receiving chemotherapy plus high-dose tamoxifen. Myelosuppression was not affected by combined treatment in tumor-bearing dogs. High-dose tamoxifen decreased the clearance and volume of distribution of full-dose doxorubicin. CONCLUSIONS: Concentrations of tamoxifen/ NDMT sufficient to inhibit Pgp may be achieved in dogs receiving full-dose chemotherapy with a moderate but acceptable increase in gastrointestinal toxicity. Tamoxifen affects doxorubicin metabolism in dogs at high doses resulting in increased serum exposure. Pharmacologic manipulation of Pgp expression or function in normal and tumor tissue in dogs may facilitate investigation of novel anticancer treatment strategies in humans.

Use of CT in detection of internal damage and repair and determination of authenticity in high-quality bowed stringed instruments.

Computed tomography (CT) was used to evaluate 17 high-quality violins and cellos crafted between 1633 and 1872 by master craftsmen such as Guarneri, Amati, and Stradivari. Multiple high-resolution CT scans were obtained in each instrument, and additional scans were obtained when defects or repair was detected. Varying degrees of internal damage (eg, wormholes, air gaps, plastic deformities of wood) or repair (eg, glue lines, filler material, wooden cleats and patches) not seen at visual inspection were detected in all 17 instruments. In addition, CT allowed noninvasive identification of the internal wood grain pattern unique to each instrument, thereby facilitating verification of authenticity to help protect against loss, theft, or forgery. The information provided by CT analysis of valuable bowed stringed instruments may prove useful to prospective buyers or to insurance companies that specialize in insuring such instruments against accidental loss or damage.

CT analysis of bowed stringed instruments.

PURPOSE: To determine the utility of computed tomography (CT) for the noninvasive evaluation of bowed stringed instruments. MATERIALS AND METHODS: Thirty-seven instruments that ranged in quality from student instruments to exquisite Stradivarius violins were analyzed with CT. Accuracy of thickness measurements was determined from 24 measurements of cross-sectional pieces sawed from a student violin. Accuracy of density measurements was determined from 328 CT attenuation measurements of 16 woods used in stringed instruments. RESULTS: Substantial differences of normal structure were noted between the masterpieces crafted in Cremona, Italy, and factory-produced student instruments. Unexpected defects were detected in nine of 14 instruments older than 100 years and ranged from a few wormholes (eight instruments) to many wormholes and extensive repair (one violin). CT thickness and attenuation measurements correlated well to the line of identity with actual measurements (P < .0001). Two cellos and a viola have been constructed from CT-derived information. The viola was awarded a gold medal at a recent international competition. CONCLUSION: CT provides the modern luthier and acoustic scientist with a unique tool for characterization of normal structure, defects, and repair and for accurate measurements of wood thickness and density. CT-derived information aids in the replication of original masterpieces. CT evaluation may have an important role in the valuation, insurance, and identification of valuable stringed instruments.