A clinally varying promoter polymorphism associated with adaptive variation in wing size in Drosophila.

Body size often shows adaptive clines in many ectotherms across altitude and latitude, but little is known about the genetic basis of these adaptive clines. Here we identify a polymorphism in the Dca (Drosophila cold acclimation) gene in Drosophila melanogaster that influences wing size, affects wing:thorax allometry and also controls a substantial proportion of the clinal wing-size variation. A polymorphism in the promoter region of Dca had two common alleles showing strong reciprocal clinal variation in frequency with latitude along the east coast of Australia. The Dca-237 allele increased towards the tropics where wing size is smaller. A within-population association study highlighted that an increase in the frequency of this allele decreased wing size but did not influence thorax size. A manipulated increase in the level of expression of Dca achieved through UAS-GAL4 was associated with a decrease in wing size but had no effect on thorax size. This was consistent with higher Dca expression levels in family lines with higher frequency of the Dca-237 allele. Genetic variation in the promoter region of the Dca gene appears to influence adaptive size variation in the eastern Australian cline of Drosophila melanogaster and accounts for more than 10% of the genetic variation in size within and between populations.

Latitudinal and cold-tolerance variation associate with DNA repeat-number variation in the hsr-omega RNA gene of Drosophila melanogaster.

An 8-bp deletion in the hsr-omega heat-stress gene of Drosophila melanogaster has previously been associated with latitude, and with heat tolerance that decreases with latitude. Here we report a second polymorphic site, at the 3'-end of hsr-omega, at which multiple alleles segregate in natural populations for copy number of a approximately 280 bp tandem repeat. On each of 3 consecutive years (2000, 2001 and 2002) among populations sampled along the Australian eastern coast, repeat number was negatively associated with latitude. Neither altitudinal association was detected in 2002 when five high-altitude sites were included, nor was a robust association detected with local temperature or rainfall measures. Although in a large number of family lines, derived from a population located centrally in the latitudinal transect, no association between hsr-omega repeat number and heat tolerance occurred, a negative association of repeat number with cold tolerance was detected. As cold tolerance also exhibits latitudinal clines we examined a set of cold-tolerant populations derived by selection and found both reduced repeat number and low constitutive levels of the omega-n repeat-bearing transcript. In a sample from the central population, linkage disequilibrium was measured between repeat number and linked markers that also cline latitudinally. However, such disequilibrium could not account for the cline in repeat number or tolerance associations. Finally, during adult recovery from cold exposure a large increase occurred in tissue levels of the omega-c transcript. Together these data suggest that a latitudinal cline in hsr-omega repeat number influences cold-tolerance variation in this species.

Ribosomal protein S14 is encoded by a pair of highly conserved, adjacent genes on the X chromosome of Drosophila melanogaster.

We describe a Drosophila DNA clone of tandemly duplicated genes encoding an amino acid sequence nearly identical to human ribosomal protein S14 and yeast rp59. Despite their remarkably similar exons, the locations and sizes of introns differ radically among the Drosophila, human, and yeast (Saccharomyces cerevisiae) ribosomal protein genes. Transcripts of both Drosophila RPS14 genes were detected in embryonic and adult tissues and are the same length as mammalian S14 message. Drosophila RPS14 was mapped to region 7C5-9 on the X chromosome. This interval also encodes a previously characterized Minute locus, M(1)7C.

PARP co-activates B-MYB through enhanced phosphorylation at cyclin/cdk2 sites.

PARP is a multifunctional protein that can affect genome stability, transcription control, telomere length and cell death. Recently we have reported that PARP binds to and enhances B-MYB transactivating potential. B-MYB is a potentially oncogenic transcription factor involved in mammalian cell proliferation, survival and differentiation. B-MYB gene expression is growth regulated and B-MYB protein is phosphorylated during S phase by cyclin A or E/cdk2 kinase, resulting in augmented transactivating potential. Here we show that PARP induces phosphorylation of B-MYB protein at cdk2 phosphorylation sites, since a B-MYB protein with mutated cdk2 phosphorylation sites is refractory to PARP-induced phosphorylation and co-activation in mammalian cells. We propose that PARP functions as a B-MYB co-factor by promoting cyclin/cdk2-dependent B-MYB phosphorylation. These results highlight a novel role for PARP as a factor that integrates cyclin-dependent kinases signaling with gene transcription.

Analysis of the Cut Locus of DROSOPHILA MELANOGASTER.

Mutants of the cut (ct) locus can be divided into two classes: viable and lethal. Most of the viable alleles are characterized by varying degrees of scalloping and notching of the wings. One mutant, kinked femur, exhibits kinking of the femurs and failure of wing expansion, but no other changes in wing structure. In heterozygous combination with the other viable alleles, it exhibits complete complementation, but it fails to complement with lethal ct alleles with respect to its viable phenotype. Similarly, all of the other viable ct alleles express a mutant wing phenotype when heterozygous with lethal ct alleles.-Mapping experiments indicate that the lethal alleles, which comprise the majority of all ct mutations recovered, are confined to a small region at the right end of the locus. That this restriction is real and not an artifact imposed by the limited number of lethal mutations mapped in the locus is supported by an examination of the mutant ct(JC20), a presumptive deficiency for the left-most third of the locus. Despite its behavior as a deletion, ct(JC20) is viable, though mutant, in combination with the lethal alleles. The restriction of the noncomplementary lethals to a small part of the locus, distinct from the other ct mutants, suggests a polarity that may define a segment that functions only in cis within the complex.-Based on the comparison of the data with the prediction of several models, we suggest that the left portion of the locus, which contains the viable alleles, defines a regulatory region controlling the expression of the locus, while the segment encoding a polypeptide product is at the right end and only it is capable of mutating to a lethal state.

Evidence for a Sex-Linked Haplo-Inviable Locus in the Cut-Singed Region of DROSOPHILA MELANOGASTER.

Many cytologically normal and rearranged cut mutants have been reported, but no known deficiency involves both ct and its close neighbor, singed. This fact prompted an investigation of the mutational response of the ct-sn interval. Approximately 24,000 F(1) female progeny of 7-day-old males given 2000 or 3000r X-ray exposures were examined for the presence of newly induced mutations at the Notch, carmine, ct, and sn loci. One sn, 2 cm-ct, 31 N, and 33 ct mutants were found, indicating that the frequency of recovery of ct mutants is much greater than that of either cm or sn, as high even as N. Among the F(1) female progeny were two deficiency mutants that expressed both cm and ct (separated by 21 bands), but none expressed both ct and sn (separated by only 14 bands). Of the 18 cytologically analyzed ct mutants, two proved to be deficiencies; neither extended farther to the right than 7C1. No reported ct deficiency extends with certainty farther to the right than 7C4. This fact, together with the scarcity of sn deficiencies, suggests the presence of a haplo-insufficient locus between ct and sn that prevents recovery of ct-sn deficiencies. The analysis of the deficiency component of Tp sn(S93), a short transposition which moves most of the ct-sn interval from 7BC to 8D, proved the existence, just to the left of sn, of a haplo-inviable locus that prevents the development of females heterozygous for its deficiency.-A marked similarity between mutants at the N and ct loci was noted.

Pilocarpine tablets for the treatment of dry mouth and dry eye symptoms in patients with Sjögren syndrome: a randomized, placebo-controlled, fixed-dose, multicenter trial. P92-01 Study Group.

BACKGROUND: Patients with Sjögren syndrome (SS) experience slowly progressive infiltration of lacrimal and salivary glands by mononuclear cells. This leads to diminished secretions, with resultant symptoms of xerostomia and xerophthalmia. Although pilocarpine hydrochloride tablets are currently indicated for the treatment of radiation-induced xerostomia, their effects on dry mouth or dry eyes in patients with SS are unclear. OBJECTIVE: To assess the safety and efficacy of pilocarpine (Salagen) tablets as symptomatic treatment for dry mouth and dry eyes caused by SS in a multicenter, doubleblind, placebo-controlled trial. METHODS: After providing written informed consent, 373 patients with primary or secondary SS and clinically significant dry mouth and dry eyes were randomized to receive 2.5-mg pilocarpine, 5-mg pilocarpine, or placebo tablets 4 times daily for 12 weeks. Symptoms were assessed by questionnaires with visual analog scales or categorical checkboxes. Whole-mouth salivary flow rates were measured. RESULTS: A significantly greater proportion of patients in the 5-mg pilocarpine group showed improvement compared with the placebo group (P< or =.01) in global assessments of dry mouth, dry eyes, and other symptoms of dryness (P< or =.05). Salivary flow was significantly increased 2- to 3-fold (P<.001) after administration of the first dose and was maintained throughout the 12-week study. The most common adverse effect was sweating, and no serious drug-related adverse experiences were reported. CONCLUSION: Administration of 5-mg pilocarpine tablets 4 times daily (20 mg/d) was well tolerated and produced significant improvement in symptoms of dry mouth and dry eyes and other xeroses in patients with SS.

An improved radiolabeling technique of ivalon and its use for dynamic monitoring of complications during therapeutic transcatheter embolization.

Transcatheter embolization by Ivalon particles for treatment of arteriovenous malformations has been an accepted therapeutic technique for many years. We describe a new and efficient radiolabeling technique of Ivalon particles using [99mTc]sulfur colloid. Continuous and dynamic monitoring of injected radiolabeled Ivalon particles is made possible by viewing the persistence scope of a portable gamma camera whose head is positioned over the patient undergoing therapeutic embolization. Therefore, if inadvertent pulmonary embolism or reflux migration of radiolabeled Ivalon particles has occurred, the angiographer is immediately aware of this potentially serious or fatal complication and can take corrective action. We describe two patients, each with an arteriovenous malformation, who had therapeutic embolization with radiolabeled Ivalon particles, one resulting in reflux migration and the other resulting in inadvertent pulmonary embolism.

Photon contribution to tumor dose from considerations of 131I radiolabeled antibody uptake in liver, spleen, and whole body.

The contribution of penetrating photon energy to tumor dose is usually ignored because of the difficulty in calculating absorbed fractions and because it is frequently assumed to represent a small proportion of the total energy. The MABDOSE software--written explicitly to simulate photon transport for the calculation of penetrating radiation absorbed fractions--was used to simulate the 131I photon spectrum originating from the liver, spleen, and whole body source organs. Specific absorbed fractions were calculated for tumors of radius 1.0, 1.5, and 2.0 cm placed near the liver and spleen in the Reference Man geometry. Cumulated activities were estimated using values reported from the literature. Dosimetry estimates from the combined cumulated activity and specific absorbed fractions indicate that neglecting the photon contribution underestimates the tumor dose by 10%-25%.

Pharmacokinetic modeling.

For radiation dosimetry calculations of radiolabeled monoclonal antibodies, (MAB), pharmacokinetics are critical. Specifically, pharmacokinetic modeling is a useful component of estimation of cumulated activity in various source organs in the body. It is thus important to formulate general methods of pharmacokinetic modeling and of pharmacokinetic data reduction, leading to cumulated activities. In this paper different types of models are characterized as "empirical," "analytical," and "compartmental" pharmacokinetic models. There remains a pressing need for quantitative studies in man for a proper understanding of the pharmacokinetics of MAb. Pharmacokinetic modeling of radiolabeled MAb in vivo has relied on relatively limited studies in man and complementary detailed measurements in animals. In either case, any model chosen for analysis of such data is inevitably based on measurements of limited accuracy and precision as well as assumptions regarding human physiology. Very few macroscopic compartmental pharmacokinetic models for MAb, have been tested over a range of conditions to determine their predictive ability. Extracorporeal immunoadsorption represents one approach for drastically altering the biokinetics of antibody distribution, and may serve to validate a given pharmacokinetic model. In addition to macroscopic modeling, the microscopic evaluation of the time-dependent distribution of radiolabeled MAb in tissues is of utmost importance for a proper understanding of the kinetics and radiobiologic effect. Many tumors do not exhibit homogeneous uptake. A mathematical understanding of that distribution is thus essential for accurate tumor dosimetry estimates. This review summarizes methodologies for pharmacokinetic modeling, critically reviews specific pharmacokinetic models and demonstrates the capability of modeling for predictive calculations of altered pharmacokinetics, emphasizing its use in dosimetric calculations.

MABDOSE. I: Characterization of a general purpose dose estimation code.

The MABDOSE software represents a general tool to perform internal radiation dosimetry. It uses a three-dimensional lattice in which to conduct radiation transport, scoring energy deposition in discrete voxels. The dosimetry system currently relies on the same algorithm used by the MIRD committee for photon transport, and assumes local deposition of particulate energy. A mathematical description of the dosimetry system's implementation is given. Additionally, a characterization of the system with respect to memory requirements, data libraries used, and simulation timing benchmarks, is presented. Validation of the software is presented in a companion manuscript.

MABDOSE. II: Validation of a general purpose dose estimation code.

UNLABELLED: The MABDOSE software represents a general tool to assess internal radiation dose. A suite of tests are described that validate the dosimetry system's implementation. METHODS: The validation suite is divided among tests that verify target digitalization, tumor digitalization and organ replacement, cumulated activity calculation, random number generation, radiation transport, and dose calculation. RESULTS: A comparison between Reference Man organ volumes and MABDOSE organ volumes at (5 mm)3 resolution demonstrates volume correspondence within 10% save for ten organs having dimensions smaller than the target lattice resolution. An accounting of normal organ volume replaced by an arbitrary tumor volume indicates mass is conserved. A comparison between cumulated activities generated by MABDOSE and solutions obtained analytically demonstrates exact correspondence for curve-fitting algorithms. For mathematical modeling algorithms, cumulated activity solutions converge to their correct values provided sufficient data of high precision are input, accompanied by reasonable initial estimates of rate constants. A comparison of MABDOSE results with the MIRD 3 report demonstrates good agreement (<8% difference) in absorbed fractions for spheres at energies from 20 keV to 2.75 MeV. A comparison of MABDOSE results with the Cristy-Eckerman report demonstrates marginal agreement (specific absorbed fractions within a factor of 2 for all Reference Man organs) at simulation energies of 20, 50, and 100 keV. Lack of exact correspondence is attributed to volume digitalization errors, and to differences in cross-section libraries, interpolation schemes between cross-section data points, and random number generators. Finally, the doses reported by MABDOSE correspond to the correct algebraic combination of paired cumulated activities and "S" values. CONCLUSIONS: The MABDOSE program has been validated as a general purpose computation tool for use in internal radionuclide dosimetry.

Dosimetry of solid tumors.

Dosimetry data arising from a decade of radioimmunotherapy are summarized along with techniques utilized to arrive at the reported dose estimates. Generality of the MIRD methodology allows it to serve as a vehicle for the calculation of solid tumor dosimetry although several limitations exist. Nonstandard geometries of solid tumors will ultimately necessitate determination of absorbed fractions for the individual tumors. Several approaches currently under investigation are described. For reasons of practicality, solid tumor dosimetry estimates continue to use the assumption of homogeneous activity distribution in a source organ, accounting for either all radiation or only nonpenetrating radiation. As computation tools become available for incorporating inhomogeneous cellular level data, the currently used "average dose" as an index of tumor sterilization will likely be replaced with a statistical distribution based on the number of viable cells in the tumor volume. Estimates of a tumor control dose would be based upon a linear extension of dose coupled with a threshold dose for cell sterilization.

Abdominal organ segmentation using texture transforms and a Hopfield neural network.

Abdominal organ segmentation is highly desirable but difficult, due to large differences between patients and to overlapping grey-scale values of the various tissue types. The first step in automating this process is to cluster together the pixels within each organ or tissue type. We propose to form images based on second-order statistical texture transforms (Haralick transforms) of a CT or MRI scan. The original scan plus the suite of texture transforms are then input into a Hopfield neural network (HNN). The network is constructed to solve an optimization problem, where the best solution is the minima of a Lyapunov energy function. On a sample abdominal CT scan, this process successfully clustered 79-100% of the pixels of seven abdominal organs. It is envisioned that this is the first step to automate segmentation. Active contouring (e.g., SNAKE's) or a back-propagation neural network can then be used to assign names to the clusters and fill in the incorrectly clustered pixels.

A new method for the correction of gamma camera nonuniformity due to spatial distortion.

A methodology for correcting scintillation camera nonuniformity resulting from spatial linearity distortion is described. The method simultaneously corrects for both variations in count density and distortions in spatial linearity without altering the intrinsic sensitivity or resolution of the imaging system. This approach to linearity correction requires only a single flood image from which the spatial shift vectors are derived. The algorithm has been implemented on a PC-based, EISA bus microcomputer, and reduces the measurement of integral uniformity for 201Tl from 25% to 5%. Combined with regional correction for photopeak pulse-heights shifts (i.e. energy correction), the algorithm represents a novel technique for implementing uniformity correction completely within software.

Late life schizophrenia and its treatment: pharmacologic issues in older schizophrenic patients.

Schizophrenia in late life includes both chronic early-onset schizophrenia and late-onset schizophrenia. Treatment with neuroleptics in relatively low doses is often useful in controlling psychotic symptoms in these patients. The value of neuroleptics is limited, however, by the risks of side effects such as tardive dyskinesia.

MABDOS: a generalized program for internal radionuclide dosimetry.

Dosimetry calculations of monoclonal antibodies (MABs) are made difficult because the focus of radioactivity is targeted for a nonstandard volume in a nonstandard geometry, precluding straightforward application of the MIRD formalism. MABDOS, a program for performing internal radionuclide dosimetry, addresses this shortcoming. It accounts for the perturbations introduced by the inclusion of tumor(s) as additional source/target of radiation. Biodistribution data are conformed to a compartmental model, allowing the forward simulation of compartmental activity. Integration of the resultant time-activity curves yields cumulative activities for each compartment. Allowing compartments to represent physical organ spaces, cumulative activities are loaded into a column vector representing radionuclide source organs. A matrix multiplication between column vector and the appropriate S table is carried out. S factors for tumors as source and target organ are calculated by treating the tumor as a spherical perturbation to Standard Man geometry, and carrying out a Monte Carlo simulation on-the-fly. The software presents an integrated modeling/dosimetry environment and offers a viable methodology for performing prospective treatment planning.

On the possibility of 'real-time' Monte Carlo calculations for the estimation of absorbed dose in radioimmunotherapy.

Dosimetry calculations of monoclonal antibodies (MABs) are made difficult because the focus of radioactivity is targeted for a nonstandard volume in a nonstandard geometry, precluding straightforward application of the MIRD formalism. The MABDOS software addresses this shortcoming by interactive placement of a spherical perturbation into the Standard Man geometry for each tumor focus. S tables are calculated by a Monte Carlo simulation of photon transport for each organ system (including tumor) that localizes activity. Performance benchmarks are reported that measure the time required to simulate 60,000 photons for each penetrating radiation in the spectrum of 99mTc and 131I using the kidney as source organ. Results indicate that calculation times are probably prohibitive on current microcomputer platforms. Mini and supercomputers offer a realistic platform for MABDOS patient dosimetry estimates.

Ambulatory surgery: next-generation strategies for physicians and hospitals.

Physicians' interest in investing and practicing in independent ambulatory surgery centers (ASCs) has grown. In the face of physician involvement in ASC development, healthcare organizations must contend with possible loss of surgical volume and revenues as well as decreased physician support and loyalty to the organization. Healthcare organizations can encourage physicians to remain in the organization by addressing physicians' concerns about the financial prospects and efficiency of independent and hospital-based ambulatory surgical arenas.