The effect of inbreeding constraints and offspring distribution on time to the most recent common ancestor.

The expected time to the most recent common ancestor (MRCA) of two alleles in a diploid individual is 4N+2 under random mating with a Poisson progeny distribution, but 8N-2 under maximum avoidance of inbreeding, which entails two progeny per mating pair. (N is the number of mating pairs, hence 2N is the number of individuals, hence 4N is the number of alleles.) The interrelationship of inbreeding constraints and offspring distribution is investigated by varying the level of sib mating: prohibiting sib mating increases the time to MRCA by four generations and decreases the variance of the offspring distribution by 2/N. With two progeny per mating pair, the expected time to the MRCA is 8N-2 under both random mating and sib mating prohibited, as well as under maximum avoidance of inbreeding, but this result does not hold for all mating structures with two progeny per mating pair.

Intranasal administration of glial-derived neurotrophic factor (GDNF) rapidly and significantly increases whole-brain GDNF level in rats.

Previous studies have shown that glial cell line-derived neurotrophic factor (GDNF) exerts significant neuroprotective effects on substantia nigra (SN) neurons in the rat 6-hydroxydopamine (6-OHDA) model of Parkinson's disease (PD). In this study we used enzyme-linked immunosorbent assay (ELISA) to determine GDNF brain levels and distribution to target regions (i.e. striatum and SN) following intranasal administration of GDNF at different time points after administration. Brain levels increased significantly within 1h following a single 50-µg dose of GDNF in a liposomal formulation, returning to baseline by 24h. In a second study, different doses of GDNF (10-150 µg) in phosphate-buffered saline (PBS) were studied at the 1-h time point. Dose-dependent increases in brain GDNF levels were observed with apparent saturation of uptake at doses above 100 µg. Liposomes delivered 10-fold more GDNF to brain than PBS despite yielding similar neuroprotective efficacy in the 6-OHDA model, suggesting incomplete release of GDNF from liposomes in tissue. In a third study, autoradiography was performed on brain sections taken 1h after intranasal (125)I-labeled GDNF. Radioactivity was detected throughout the brain along the rostral-to-caudal axis, indicating that nasally administered GDNF can reach target areas. Collectively, these results demonstrate that intranasal administration of GDNF in liposomes or PBS achieves significant increases in GDNF in target brain areas, supporting use of intranasal administration as a non-invasive means of delivering GDNF to the brain to protect dopamine neurons and arrest disease progression in PD.

Neurotrophic and neuroprotective efficacy of intranasal GDNF in a rat model of Parkinson's disease.

Glial cell line-derived neurotrophic factor (GDNF) exerts neurotrophic and neuroprotective effects on substantia nigra (SN) dopamine neurons and has great therapeutic potential for Parkinson's disease (PD). Hindering this potential is the fact that GDNF cannot cross the blood-brain barrier. The aim of this study was to assess the effects of GDNF administered by the intranasal route in normal rats, and in the unilateral 6-hydroxydopamine (6-OHDA) model of PD. In the first study, rats received single intranasal doses of 50-µg GDNF in phosphate-buffered saline (PBS) or cationic liposomes, but no 6-OHDA. In the second study, rats were nasally administered 10, 50 or 150 µg of GDNF in PBS or cationic liposomes 1h before injection of 6-OHDA. All groups were sacrificed 3-4 weeks later. Both intranasal GDNF treatments induced a neurotrophic effect in the SN insofar as the number of tyrosine hydroxylase (TH)-positive neurons was significantly higher than in controls given intranasal PBS liposomes. Dopamine cell counts were also higher in the intact SN of 6-OHDA-lesioned rats compared to controls given PBS liposomes. Most importantly, intranasal GDNF provided significant neuroprotective efficacy indicated by greater TH immunostaining density in the lesioned versus intact SN of rats given single 50-µg doses of GDNF in PBS, or 150-µg doses of liposomal GDNF, compared to lesioned rats given PBS liposomes. Three 50-µg doses given at daily intervals (1 day before, 1h before, and 1 day after 6-OHDA) provided even greater protection than single 150-µg doses. Multiple doses at short intervals may therefore provide greater neuroprotection than single bolus doses. These results demonstrate both a neurotrophic effect of intranasal GDNF in the intact SN as well as neuroprotective efficacy in the unilateral 6-OHDA model, supporting pursuit of this approach as a potential treatment for PD.

Mucin impedes cytotoxic effect of 5-FU against growth of human pancreatic cancer cells: overcoming cellular barriers for therapeutic gain.

Mucins are high molecular weight glycoproteins expressed on the apical surface of normal epithelial cells. In cancer disease mucins are overexpressed on the entire cellular surface. Overexpression of MUC1 mucin in pancreatic tumours has been correlated with poor patient survival. Current chemotherapeutic approaches such as 5-fluorouracil (5-FU) has produced limited clinical success. In this study we investigated the role of mucin in cytotoxic drug treatment to determine whether the extracellular domain of mucin impedes cytotoxic drug action of 5-FU. Human pancreatic cancer cells revealed high and relatively moderate MUC1 levels for Capan-1 and HPAF-II, respectively, compared to MUC1 negative control (U-87 MG glioblastoma) that showed relatively non-specific anti-MUC1 uptake. Benzyl-alpha-GalNAc (O-glycosylation inhibitor) was used to reduce mucin on cell surfaces, and neuraminidase was used to hydrolyse sialic acid at the distal end of carbohydrate chains. Benzyl-alpha-GalNAc had no effect on cell morphology or proliferation at the concentrations employed. The inhibition of O-glycosylation resulted in significant 5-FU antiproliferative activity against Capan-1 and HPAF-II, but not against U-87 MG. However, the exposure of cells to neuraminidase failed to improve the cytotoxic action of 5-FU. Our experimental findings suggest that the overexpression of mucin produced by human pancreatic tumours might limit the effectiveness of chemotherapy.

Coalescent size versus coalescent time with strong selection.

This work studies the coalescent (ancestral pedigree, genealogy) of the entire population. The coalescent structure (topology) is robust, but selection changes the rate of coalescence (the time between branching events). The change in the rate of coalescence is not uniform, rather the reduction in the time between branching events is greatest when the coalescent is small (immediately after the common ancestor is the only member of the coalescent) with little change when the coalescent is large (immediately preceding when that common ancestor becomes fixed and the size of the coalescent is N). This provides that the reduction in the coalescent time due to selection is much greater than the reduction in the cumulative size of the coalescent (total number of ancestors of the present population after and including the most recent common ancestor) due to selection. If Ns>>1, the coalescent and fixation times are approximately equal to [Formula: see text] , which is much less than the value N which would result from neutral drift (N rather than the canonical haploid neutral fixation time 2N is the appropriate comparison for the model considered here), in particular, it is 70% less for Ns=10 and 95% less for Ns=100. However, for those values of Ns, and N ranging between 10(3) and 10(6), the reduction in the cumulative size of the coalescent of the entire population compared to the neutral case ranges from 17% to 65% (depending on the values of N and s). The coalescent time for two individuals for Ns of 10 and 100 is reduced by approximately 70% and 94%, respectively, compared with the neutral case. Because heterozygosity is proportional to the coalescent time for two individuals and the number of segregating alleles is proportional to the cumulative size of the coalescent, selection reduces heterozygosity more than it reduces the number of segregating alleles.

Simulation of heating-compressed fast-ignition cores by petawatt laser-generated electrons.

We report on unique particle-in-cell simulations to understand the relativistic electron beam thermalization and subsequent heating of highly compressed plasmas. The simulations yield heated core parameters in good agreement with the GEKKO-PW experimental measurements, given reasonable assumptions of laser-to-electron coupling efficiency and the distribution function of laser-produced electrons. The classical range of the hot electrons exceeds the mass density-core diameter product rhoL by a factor of several. Anomalous stopping appears to be present and is created by the growth and saturation of an electromagnetic filamentation mode that generates a strong back-EMF impeding hot electrons on the injection side of the density maxima.

A logistic branching process for population genetics.

A logistic (regulated population size) branching process population genetic model is presented. It is a modification of both the Wright-Fisher and (unconstrained) branching process models, and shares several properties including the coalescent time and shape, and structure of the coalescent process with those models. An important feature of the model is that population size fluctuation and regulation are intrinsic to the model rather than externally imposed. A consequence of this model is that the fluctuation in population size enhances the prospects for fixation of a beneficial mutation with constant relative viability, which is contrary to a result for the Wright-Fisher model with fluctuating population size. Explanation of this result follows from distinguishing between expected and realized viabilities, in addition to the contrast between absolute and relative viabilities.

Influence of cationic lipids on the stability and membrane properties of paclitaxel-containing liposomes.

Paclitaxel (taxol) is a poorly soluble anticancer agent that is in widespread clinical use. Liposomes provide a less toxic vehicle for solubilizing the drug and increasing the therapeutic index of paclitaxel in model tumor systems. The role of liposome membrane composition in the stability of paclitaxel-containing formulations is understood partially for neutral and anionic liposomes, but poorly for other compositions. We investigated the effect of dialkyl cationic lipids on the stability and physical properties of paclitaxel-containing liposomes, using circular dichroism (CD), fluorescence spectroscopy, and differential interference contrast microscopy (DIC). DOTAP (1,2-dioleoyl-3-trimethylammonium propane), a cationic lipid used frequently for gene delivery, was combined at various ratios with dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), or distearoylphosphatidylcholine (DSPC). In the absence of DOTAP, the stability of liposomes containing > or =3 mol% paclitaxel was observed to follow the following rank order: DPPC >DSPC > DMPC. Increasing concentrations of DOTAP increased the physical stability of all compositions, and maximal stabilization was achieved at 30-50 mol% DOTAP, depending on the paclitaxel concentration and the acyl chain length of the phosphatidylcholine. The relationship between stability and mole fraction of DOTAP was complex for some compositions. DOTAP exerted a major fluidizing effect on DMPC, DPPC, and DSPC membranes, and the addition of paclitaxel at 3-8 mol% did not increase fluidity further. Studies of membrane phase domain behavior using the probe Laurdan (6-dodecanoyl-2-dimethylaminonaphthalene) indicated that both paclitaxel and DOTAP were miscible with the phosphatidylcholine phase. The physical events leading to destabilization of formulations are hypothesized to arise from concentration-dependent paclitaxel self-association rather than immiscibility of the membrane lipids. Given the increased incorporation and stability of paclitaxel in DOTAP-containing membranes and the potential for enhanced interaction with cells, cationic liposomes may provide a therapeutic advantage over previously described liposome formulations.

John Graunt, John Arbuthnott, and the human sex ratio.

John Graunt was the first person to compile data that showed an excess of male births over female births. He also noticed spatial and temporal variation in the sex ratio, but the variation in his data is not significant. John Arbuthnott was the first person to demonstrate that the excess of male births is statistically significant. He erroneously concluded that there is less variation in the sex ratio than would occur by chance, and asserted without a basis that the sex ratio would be uniform over all time and space.

In vivo measurement of gene expression, angiogenesis and physiological function in tumors using multiphoton laser scanning microscopy.

Intravital microscopy coupled with chronic animal window models has provided stunning insight into tumor pathophysiology, including gene expression, angiogenesis, cell adhesion and migration, vascular, interstitial and lymphatic transport, metabolic microenvironment and drug delivery. However, the findings to date have been limited to the tumor surface (< 150 microm). Here, we show that the multiphoton laser-scanning microscope can provide high three-dimensional resolution of gene expression and function in deeper regions of tumors. These insights could be critical to the development of novel therapeutics that target not only the tumor surface, but also internal regions.

Phospholipid-cationic lipid interactions: influences on membrane and vesicle properties.

Liposomes composed of synthetic dialkyl cationic lipids and zwitterionic phospholipids such as dioleoylphosphatidylethanolamine have been studied extensively as vehicles for gene delivery, but the broader potentials of these cationic liposomes for drug delivery have not. An understanding of phospholipid-cationic lipid interactions is essential for rational development of this potential. We evaluated the effect of the cationic lipid DOTAP (N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium) on liposome physical properties such as size and membrane domain structure. DSC (differential scanning calorimetry) showed progressive decrease and broadening of the phase transition temperature of dipalmitoylphosphatidylcholine (DPPC) with increasing fraction of DOTAP, in the range of 0.4-20 mol%. Laurdan (6-dodecanolyldimethylamino-naphthalene), a fluorescent probe of membrane domain structure, showed that DOTAP and DPPC remained miscible at all ratios tested. DOTAP reduced the size of spontaneously-forming PC-containing liposomes, regardless of the acyl chain length and degree of saturation. The anionic lipid DOPG (dioleoylphosphatidylglycerol) had similar effects on DPPC membrane fluidity and size. However, DOTAP/DOPC (50/50) vesicles were taken up avidly by OVCAR-3 human ovarian tumor cells, in contrast to DOPG/DOPC (50/50) liposomes. Overall, DOTAP exerts potent effects on bilayer physical properties, and may provide advantages for drug delivery.

Role of tumor-host interactions in interstitial diffusion of macromolecules: cranial vs. subcutaneous tumors.

The large size of many novel therapeutics impairs their transport through the tumor extracellular matrix and thus limits their therapeutic effectiveness. We propose that extracellular matrix composition, structure, and distribution determine the transport properties in tumors. Furthermore, because the characteristics of the extracellular matrix largely depend on the tumor-host interactions, we postulate that diffusion of macromolecules will vary with tumor type as well as anatomical location. Diffusion coefficients of macromolecules and liposomes in tumors growing in cranial windows (CWs) and dorsal chambers (DCs) were measured by fluorescence recovery after photobleaching. For the same tumor types, diffusion of large molecules was significantly faster in CW than in DC tumors. The greater diffusional hindrance in DC tumors was correlated with higher levels of collagen type I and its organization into fibrils. For molecules with diameters comparable to the interfibrillar space the diffusion was 5- to 10-fold slower in DC than in CW tumors. The slower diffusion in DC tumors was associated with a higher density of host stromal cells that synthesize and organize collagen type I. Our results point to the necessity of developing site-specific drug carriers to improve the delivery of molecular medicine to solid tumors.

The coalescent time in the presence of background fertility selection.

Selection ultimately entails differential reproductive success over several generations. This can be measured as a correlation of the number of progeny an individual has with the number of progeny its parent had. This correlation could have a genetic or a cultural basis. The effect of such a correlation is to multiply the single generation sampling variance (Vdeltap) in the diffusion approximation for fixation time by (1-b)+bx(1+r)/(1-r), where bxrn is the correlation between the number of progeny of an individual and its ancestor n generations ago (e.g., b is the heritability and br is the resultant parent-offspring progeny number correlation if the progeny number is genetically determined). This results in a reduction of the fixation or coalescent time by division by this factor. Sex differences in this correlation have been observed, and this provides an explanation for the difference of coalescent times of y-chromosomes and mitochondria.

Exercise without dietary restriction as a means to long-term fat loss in the obese cardiac patient.

BACKGROUND: To examine the effects of a 12-month daily walking program without dietary restriction on the metabolic rate, body composition and blood lipid profile of overweight and moderately obese patients following myocardial infarction. DESIGN: longitudinal training (preliminary study). SETTING: out-patient cardiac rehabilitation program. PARTICIPANTS: twelve consecutive volunteers (8M, 4F) with a body mass index of 25-40 kg/m2. Relative to average cardiac patients, the men but not the women were significantly heavier (100.8 vs 77.4 kg [M], 70.7 vs 74.2 kg [F]) and fatter (hydrostatic estimate of body fat 34.0% vs 23.1% [M]; 38.3% vs 36.3% [F]) than the general cardiac patient. MEASURES: body mass, hydrostatic weighing, triglycerides, total, HDL- and LDL-cholesterol, resting and peak oxygen intake, one week food intake diaries. RESULTS: Daily walking increased progressively from 20 min to 43 min over 3 months, and was then held constant for 9 months. Peak aerobic power increased 24%, from 19.9 to 24.6 ml/[kg.min] (p < 0.001). Resting oxygen intake rose from 3.1 to 3.4 ml/[kg.min], (p < 0.05). Energy intake increased from 6.10 to 6.57 MJ/day, but body mass decreased by an average of 4.5 kg (p < 0.05, 4.1 kg [M], 5.1 kg [F]), and body fat content diminished from 35.4 to 33.2% (p < 0.02, 1.8% [M], 3.2% [F]), with no change in lean body mass (57.7 vs 57.8 kg). Triglycerides diminished from 2.63 to 2.28 mmol/L (p < 0.005). Total and LDL-cholesterol also tended to change favorably (from 6.15 to 5.80 and 4.44 to 3.80 mmol/L respectively, but HDL-cholesterol was unchanged). CONCLUSIONS: A daily walking program without dietary restriction induces a favorable change in body composition and lipid profile in moderately obese cardiac patients. An exercise-induced increase of resting metabolism apparently makes an important contribution to this outcome.

Influence of steroid injection on ligament healing in the rat.

The effect of a single local injection of long acting corticosteroid on the healing of acute rat medial collateral ligament injuries was studied. The medial collateral ligaments of 81 adult female rats were exposed surgically. In 32 rats, the ligament was transected sharply, the overlying muscle was closed, and a human equivalent dose of dexamethasone was injected under the muscle layer, bathing the injured ligament. The identical operation with no corticosteroid injection was done in 32 additional rats: in the remaining 17 animals, the incision was closed without ligament transection or injection. The rats were divided into 3 groups of 25. Each group consisted of 10 rats that were injected, 10 that were not injected, and 5 that underwent sham operations. One group was euthanized 6 days after surgery, 1 group after 10 days, and 1 group after 20 days. Histologic evaluation and biomechanical testing were performed for each subgroup. A cellular pathologist examined a smaller group of 6 rats (2 from each group) for histologic changes 40 days after surgery. No histologic differences were noted between the injected and noninjected ligaments 6, 10, or 20 days after injury. At 40 days, the injected specimens showed a slightly more mature crimp pattern than the noninjected specimens. Mechanical testing demonstrated no significant difference in ultimate load or ultimate stress between the injected and noninjected groups. There were no detrimental effects of a single dose administration of dexamethasone on the histologic appearance or biomechanical strength of healing rat medial collateral ligaments.

The effect of mating structure and progeny distribution on heterozygosity versus the number of alleles as measures of variation.

Heterozygosity and the number of alleles are both measures of the genetic variation of a population. They are qualitatively similar if the distribution of the number of progeny is Poisson, but not necessarily for other distributions. In particular, selfing decreases heterozygosity and also decreases the number of alleles when the progeny distribution is Poisson, but decreases heterozygosity and increases the number of alleles when there are two progeny per individual. This is because heterozygosity is closely related to the breeding structure (inbreeding) of the population, whereas the number of alleles is more influenced by the variation of the sampling process. Branching processes are employed to model the dynamics of mutant alleles, with drift and subsequent mutation contributing to extinction. In populations of four individuals, double first cousin mating has greater heterozygosity, but fewer alleles, than half-sib mating.

Genetic interactions and dosage effects of Polycomb group genes of Drosophila.

The Polycomb (Pc) group of genes are required for maintenance of cell determination in Drosophila melanogaster. At least 11 Pc group genes have been described and there may be up to 40; all are required for normal regulation of homeotic genes, but as a group, their phenotypes are rather diverse. It has been suggested that the products of Pc group genes might be members of a heteromeric complex that acts to regulate the chromatin structure of target loci. We examined the phenotypes of adult flies heterozygous for every pairwise combination of Pc group genes in an attempt to subdivide the Pc group functionally. The results support the idea that Additional sex combs (Asx), Pc, Polycomblike (Pcl), Posterior sex combs (Psc), Sex combs on midleg (Scm), and Sex combs extra (Sce) have similar functions in some imaginal tissues. We show genetic interactions among extra sex combs (esc) and Asx, Enhancer of Pc, Pcl, Enhancer of zeste E(z), and super sex combs and reassess the idea that most Pc group genes function independently of esc. Most duplications of Pc group genes neither exhibit anterior transformations nor suppress the extra sex comb phenotype of Pc group mutations, suggesting that not all Pc group genes behave as predicted by the mass-action model. Surprisingly, duplications of E(z) enhance homeotic phenotypes of esc mutants. Flies with increasing doses of esc+ exhibit anterior transformations, but these are not enhanced by mutations in trithorax group genes. The results are discussed with respect to current models of Pc group function.

Interactions of polyhomeotic with Polycomb group genes of Drosophila melanogaster.

The Polycomb (Pc) group genes of Drosophila are negative regulators of homeotic genes, but individual loci have pleiotropic phenotypes. It has been suggested that Pc group genes might form a regulatory hierarchy, or might be members of a multimeric complex that obeys the law of mass action. Recently, it was shown that polyhomeotic (ph) immunoprecipitates in a multimeric complex that includes Pc. Here, we show that duplications of ph suppress homeotic transformations of Pc and Pcl, supporting a mass-action model for Pc group function. We crossed ph alleles to all members of the Polycomb group, and to E(Pc) and Su(z)2 to look for synergistic effects. We observed extragenic noncomplementation between ph503 and Pc, Psc1 and Su(z)2(1) in females, and between ph409 and Sce1, ScmD1 and E(z)1 mutations in males, suggesting that these gene products might interact directly with ph. Males hemizygous for a temperature-sensitive allele, ph2, are lethal when heterozygous with mutants in Asx, Pc, Pcl, Psc, Sce and Scm, and with E(Pc) and Su(z)2. Mutations in trithorax group genes were not able to suppress the lethality of ph2/Y; Psc1/+ males. ph2 was not lethal with extra sex combs, E(z), super sex combs (sxc) or l(4)102EFc heterozygotes, but did cause earlier lethality in embryos homozygous for E(z), sxc and l(4)102EFc. However, ph503 did not enhance homeotic phenotypes of esc heterozygotes derived from homozygous esc- mothers. We examined the embryonic phenotypes of ph2 embryos that were lethal when heterozygous or homozygous for other mutations. Based on this phenotypic analysis, we suggest that ph may perform different functions in conjunction with differing subsets of Pc group genes.

Antagonism of estrogenic effects on feeding behavior by central implants of anisomycin.

The following experiment determined whether the estrogenic suppression of food intake is dependent upon changes in protein synthesis within neurons of the paraventricular nucleus of the hypothalamus (PVN). Ovariectomized rats were treated centrally with anisomycin-filled or empty (control) cannulae in the PVN. Females were injected with either 2.0 micrograms of estradiol benzoate (EB) or the oil vehicle and the inner cannulae were removed 2 h later. EB injections significantly lowered food and water intake in the central control group but not in animals given PVN implants of anisomycin. Body weight gain decreased for all females. EB induced comparable levels of female sexual behavior in both groups, demonstrating that anisomycin implants did not affect the ability of estradiol to stimulate lordosis. These findings indicate that the effects of estradiol on food intake require the activation of protein synthesis in estrogen-sensitive PVN neurons.

Half-sib mating structures.

All ways in which all matings in a population can be between half-sibs under a generalization of regular systems of inbreeding are characterized for both finite and infinite populations. A model of random half-sib mating is developed and analyzed, and the asymptotic configuration of populations subject to it is described. The classical model of half-sib mating which ensues from the standard definition of regular systems of inbreeding is only one of many ways a population can propagate by half-sib mating, and a wide range of genetic identity is possible dependent on which half-sib mating structure governs a population.