Computation of maximal local (un)stable manifold patches by the parameterization method

In this work we develop some automatic procedures for computing high order polynomial expansions of local (un)stable manifolds for equilibria of differential equations. Our method incorporates validated truncation error bounds, and maximizes the size of the image of the polynomial approximation relative to some specified constraints. More precisely we use that the manifold computations depend heavily on the scalings of the eigenvectors: indeed we study the precise effects of these scalings on the estimates which determine the validated error bounds. This relationship between the eigenvector scalings and the error estimates plays a central role in our automatic procedures. In order to illustrate the utility of these methods we present several applications, including visualization of invariant manifolds in the Lorenz and FitzHugh-Nagumo systems and an automatic continuation scheme for (un)stable manifolds in a suspension bridge problem. In the present work we treat explicitly the case where the eigenvalues satisfy a certain non-resonance condition.Comment: Revised version, typos corrected, references adde

The Relationship of Visceral Adipose Tissue with Markers of Energy Homeostasis Following Weight-Loss

Excess levels of adipose tissue, in particular visceral adipose tissue (VAT), is closely associated with the metabolic syndrome and dysregulation of energy homeostasis. It is hypothesized that leptin resistance results in overconsumption of calories and reduced satiety. Recently, brain derived neurotrophic factor (BDNF), beyond functioning in learning and memory, is shown to play a role in energy homeostasis via its positive satiety effects on the hypothalamus. However, it remains to be elucidated how changes in visceral adipose tissue are associated with changes in circulating leptin and BDNF after weight-loss. PURPOSE: To identify changes in adiposity and circulating leptin and BDNF following a 3-month weight-loss program. METHODS: Sixty-five obese (mean±SEM; age=47.9±1.1 years; BMI=34.5±0.8 kg/m2;), men and women completed a 3-month weight-loss program that consisted of a reduced energy intake of 1200-1500 kcals/day using a high-volume low-calorie diet combined with a progressive walking program to target 300 min/wk. Fasted (12 hr) blood samples were collected at baseline and post-weight-loss (3 months) and assayed for concentrations of glucose, insulin, BDNF, and leptin. Using DXA, total VAT and subcutaneous (SubQ) adipose tissue mass were measured at baseline and post-weight-loss (3 months). To identify significant changes over time, ANOVA with repeated measures was performed with significance set at p\u3c0.05. RESULTS: Following the 3-month weight-loss program, both BMI and HOMA-IR were significantly reduced 9.3% and 49%, respectively. The reduction in BMI and HOMA-IR were matched by a significant reduction in both VAT (-658 g; -33%, p\u3c0.001) and SubQ (-367 g; -17%, p\u3c0.001). Interestingly, leptin was reduced and BDNF was increased by 43% (p\u3c0.001) and 42% (p=0.011), respectively. Linear regression revealed that changes in VAT were associated with changes in leptin (b=0.298, p=0.026), but not with BDNF (b=0.027, p=0.896). CONCLUSION: This study shows that the reduction in VAT, by caloric restriction and physical activity, was associated with the reduction in circulating leptin concentrations, but not with changes in BDNF. Changes in leptin and BDNF may be in part responsible for the normalization of the energy homeostasis observed after weight-loss; however, changes in BDNF may be independent of VAT

The Effect of Gender on Circulating Adipokines during Weight Loss and Weight Maintenance

During obesity, the altered release of adipokines, leptin and adiponectin, have been strongly associated with development of the metabolic syndrome. Treatment with weight loss has been shown to increase adiponectin, in particular high molecular weight (HMW) adiponectin, and reduce circulating leptin levels indicating an increased leptin sensitizing effect. Interestingly, a gender dichotomy has been identified with women generally possessing higher plasma concentrations of both adipokines. Weight loss effects have been well established; however, it remains to be determined how the gender differences in adiponectin and leptin will affect these adipokines during prolonged weight maintenance. PURPOSE: To identify gender differences in adiponectin and leptin concentrations following a 6-month weight loss and weight maintenance program. METHODS: Sixty-five obese (mean±SEM; age=47.9±1.1 years; BMI=34.3±0.7 kg/m2;) adults (M=20, F=45) completed a 3-month weight loss program that consisted of a reduced energy intake of 1200-1500 kcals/day using a high-volume low-calorie diet combined with a progressive walking program to target 300 min/wk. During the 3-month weight maintenance program, participants consumed sufficient calories to maintain weight loss with continued walking to target 300 min/wk. Fasted (12 hr) blood samples were collected at baseline, post- weight loss (3 months), and weight maintenance (6 months) and assayed for glucose, insulin, total and HMW adiponectin, and leptin. To identify significant changes over time and between gender, a repeated measures (time x gender) ANOVA was performed with significance set at P\u3c0.05. Results: At baseline, no significant difference in BMI or HOMA-IR were observed between genders. Following 3 months of weight loss, BMI was significantly reduced 9.9% and 8.5% in men and women, respectively, and BMI remained unchanged through the weight maintenance program. Interestingly, only men demonstrated a significant reduction in HOMA-IR following weight loss. Following weight maintenance in women, HOMA-IR increased slightly such that it was not significantly different than the baseline or weight loss time points. At baseline, women had significantly higher circulating levels of total and HMW adiponectin, and leptin. No significant changes in total or HMW adiponectin were observed over time for either gender. Following weight-loss, leptin concentrations were reduced 49.6% and 39.2% in men and women, respectively. Interestingly, only women demonstrated a transient reduction in leptin through the weight maintenance program. Conclusions: At baseline, we identified the presence of a clear gender dichotomy for total and HMW adiponectin, and leptin concentrations. Despite these significant differences in circulating adipokines at baseline, both men and women responded similarly to a 6-month weight loss and weight maintenance program

The molecular basis of color vision in colorful fish: Four Long Wave-Sensitive (LWS) opsins in guppies (Poecilia reticulata) are defined by amino acid substitutions at key functional sites

<p>Abstract</p> <p>Background</p> <p>Comparisons of functionally important changes at the molecular level in model systems have identified key adaptations driving isolation and speciation. In cichlids, for example, long wavelength-sensitive (LWS) opsins appear to play a role in mate choice and male color variation within and among species. To test the hypothesis that the evolution of elaborate coloration in male guppies (<it>Poecilia reticulata</it>) is also associated with opsin gene diversity, we sequenced long wavelength-sensitive (LWS) opsin genes in six species of the family Poeciliidae.</p> <p>Results</p> <p>Sequences of four LWS opsin genes were amplified from the guppy genome and from mRNA isolated from adult guppy eyes. Variation in expression was quantified using qPCR. Three of the four genes encode opsins predicted to be most sensitive to different wavelengths of light because they vary at key amino acid positions. This family of LWS opsin genes was produced by a diversity of duplication events. One, an intronless gene, was produced prior to the divergence of families Fundulidae and Poeciliidae. Between-gene PCR and DNA sequencing show that two of the guppy LWS opsins are linked in an inverted orientation. This inverted tandem duplication event occurred near the base of the poeciliid tree in the common ancestor of <it>Poecilia </it>and <it>Xiphophorus</it>. The fourth sequence has been uncovered only in the genus <it>Poecilia</it>. In the guppies surveyed here, this sequence is a hybrid, with the 5' end most similar to one of the tandem duplicates and the 3' end identical to the other.</p> <p>Conclusion</p> <p>Enhanced wavelength discrimination, a possible consequence of opsin gene duplication and divergence, might have been an evolutionary prerequisite for color-based sexual selection and have led to the extraordinary coloration now observed in male guppies and in many other poeciliids.</p

Stromal and therapy-induced macrophage proliferation promotes PDAC progression and susceptibility to innate immunotherapy

Tumor-associated macrophages (TAMs) are abundant in pancreatic ductal adenocarcinomas (PDACs). While TAMs are known to proliferate in cancer tissues, the impact of this on macrophage phenotype and disease progression is poorly understood. We showed that in PDAC, proliferation of TAMs could be driven by colony stimulating factor-1 (CSF1) produced by cancer-associated fibroblasts. CSF1 induced high levels of p21 in macrophages, which regulated both TAM proliferation and phenotype. TAMs in human and mouse PDACs with high levels of p21 had more inflammatory and immunosuppressive phenotypes. p21 expression in TAMs was induced by both stromal interaction and/or chemotherapy treatment. Finally, by modeling p21 expression levels in TAMs, we found that p21-driven macrophage immunosuppression in vivo drove tumor progression. Serendipitously, the same p21-driven pathways that drive tumor progression also drove response to CD40 agonist. These data suggest that stromal or therapy-induced regulation of cell cycle machinery can regulate both macrophage-mediated immune suppression and susceptibility to innate immunotherapy

Involvement of Complexin 2 in Docking, Locking and Unlocking of Different SNARE Complexes during Sperm Capacitation and Induced Acrosomal Exocytosis

Acrosomal exocytosis (AE) is an intracellular multipoint fusion reaction of the sperm plasma membrane (PM) with the outer acrosomal membrane (OAM). This unique exocytotic event enables the penetration of the sperm through the zona pellucida of the oocyte. We previously observed a stable docking of OAM to the PM brought about by the formation of the trans-SNARE complex (syntaxin 1B, SNAP 23 and VAMP 3). By using electron microscopy, immunochemistry and immunofluorescence techniques in combination with functional studies and proteomic approaches, we here demonstrate that calcium ionophore-induced AE results in the formation of unilamellar hybrid membrane vesicles containing a mixture of components originating from the two fused membranes. These mixed vesicles (MV) do not contain the earlier reported trimeric SNARE complex but instead possess a novel trimeric SNARE complex that contained syntaxin 3, SNAP 23 and VAMP 2, with an additional SNARE interacting protein, complexin 2. Our data indicate that the earlier reported raft and capacitation-dependent docking phenomenon between the PM and OAM allows a specific rearrangement of molecules between the two docked membranes and is involved in (1) recruiting SNAREs and complexin 2 in the newly formed lipid-ordered microdomains, (2) the assembly of a fusion-driving SNARE complex which executes Ca2+-dependent AE, (3) the disassembly of the earlier reported docking SNARE complex, (4) the recruitment of secondary zona binding proteins at the zona interacting sperm surface. The possibility to study separate and dynamic interactions between SNARE proteins, complexin and Ca2+ which are all involved in AE make sperm an ideal model for studying exocytosis

Different Pattern of Immunoglobulin Gene Usage by HIV-1 Compared to Non-HIV-1 Antibodies Derived from the Same Infected Subject

A biased usage of immunoglobulin (Ig) genes is observed in human anti-HIV-1 monoclonal antibodies (mAbs) resulting probably from compensation to reduced usage of the VH3 family genes, while the other alternative suggests that this bias usage is due to antigen requirements. If the antigen structure is responsible for the preferential usage of particular Ig genes, it may have certain implications for HIV vaccine development by the targeting of particular Ig gene-encoded B cell receptors to induce neutralizing anti-HIV-1 antibodies. To address this issue, we have produced HIV-1 specific and non-HIV-1 mAbs from an infected individual and analyzed the Ig gene usage. Green-fluorescence labeled virus-like particles (VLP) expressing HIV-1 envelope (Env) proteins of JRFL and BaL and control VLPs (without Env) were used to select single B cells for the production of 68 recombinant mAbs. Ten of these mAbs were HIV-1 Env specific with neutralizing activity against V3 and the CD4 binding site, as well as non-neutralizing mAbs to gp41. The remaining 58 mAbs were non-HIV-1 Env mAbs with undefined specificities. Analysis revealed that biased usage of Ig genes was restricted only to anti-HIV-1 but not to non-HIV-1 mAbs. The VH1 family genes were dominantly used, followed by VH3, VH4, and VH5 among anti-HIV-1 mAbs, while non-HIV-1 specific mAbs preferentially used VH3 family genes, followed by VH4, VH1 and VH5 families in a pattern identical to Abs derived from healthy individuals. This observation suggests that the biased usage of Ig genes by anti-HIV-1 mAbs is driven by structural requirements of the virus antigens rather than by compensation to any depletion of VH3 B cells due to autoreactive mechanisms, according to the gp120 superantigen hypothesis

Heritable Differences in Schooling Behavior among Threespine Stickleback Populations Revealed by a Novel Assay

Identifying the proximate and ultimate mechanisms of social behavior remains a major goal of behavioral biology. In particular, the complex social interactions mediating schooling behavior have long fascinated biologists, leading to theoretical and empirical investigations that have focused on schooling as a group-level phenomenon. However, methods to examine the behavior of individual fish within a school are needed in order to investigate the mechanisms that underlie both the performance and the evolution of schooling behavior. We have developed a technique to quantify the schooling behavior of an individual in standardized but easily manipulated social circumstances. Using our model school assay, we show that threespine sticklebacks (Gasterosteus aculeatus) from alternative habitats differ in behavior when tested in identical social circumstances. Not only do marine sticklebacks show increased association with the model school relative to freshwater benthic sticklebacks, they also display a greater degree of parallel swimming with the models. Taken together, these data indicate that marine sticklebacks exhibit a stronger tendency to school than benthic sticklebacks. We demonstrate that these population-level differences in schooling tendency are heritable and are shared by individuals within a population even when they have experienced mixed-population housing conditions. Finally, we begin to explore the stimuli that elicit schooling behavior in these populations. Our data suggest that the difference in schooling tendency between marine and benthic sticklebacks is accompanied by differential preferences for social vs. non-social and moving vs. stationary shelter options. Our study thus provides novel insights into the evolution of schooling behavior, as well as a new experimental approach to investigate the genetic and neural mechanisms that underlie this complex social behavior

Comparison of Antibody Repertoires Produced by HIV-1 Infection, Other Chronic and Acute Infections, and Systemic Autoimmune Disease

Background Antibodies (Abs) produced during HIV-1 infection rarely neutralize a broad range of viral isolates; only eight broadly-neutralizing (bNt) monoclonal (M)Abs have been isolated. Yet, to be effective, an HIV-1 vaccine may have to elicit the essential features of these MAbs. The V genes of all of these bNt MAbs are highly somatically mutated, and the VH genes of five of them encode a long (≥20 aa) third complementarity-determining region (CDR-H3). This led us to question whether long CDR-H3s and high levels of somatic mutation (SM) are a preferred feature of anti-HIV bNt MAbs, or if other adaptive immune responses elicit them in general. Methodology and Principal Findings We assembled a VH-gene sequence database from over 700 human MAbs of known antigen specificity isolated from chronic (viral) infections (ChI), acute (bacterial and viral) infections (AcI), and systemic autoimmune diseases (SAD), and compared their CDR-H3 length, number of SMs and germline VH-gene usage. We found that anti-HIV Abs, regardless of their neutralization breadth, tended to have long CDR-H3s and high numbers of SMs. However, these features were also common among Abs associated with other chronic viral infections. In contrast, Abs from acute viral infections (but not bacterial infections) tended to have relatively short CDR-H3s and a low number of SMs, whereas SAD Abs were generally intermediate in CDR-H3 length and number of SMs. Analysis of VH gene usage showed that ChI Abs also tended to favor distal germline VH-genes (particularly VH1-69), especially in Abs bearing long CDR-H3s. Conclusions and Significance The striking difference between the Abs produced during chronic vs. acute viral infection suggests that Abs bearing long CDR-H3s, high levels of SM and VH1-69 gene usage may be preferentially selected during persistent infection

In vivo pharmacological evaluations of novel olanzapine analogues in rats: a potential new avenue for the treatment of schizophrenia

Olanzapine (Olz) is one of the most effective antipsychotic drugs commonly used for treating schizophrenia. Unfortunately, Olz administration is associated with severe weight gain and metabolic disturbances. Both patients and clinicians are highly interested in the development of new antipsychotics which are as effective as atypical antipsychotics but which have a lower propensity to induce metabolic side effects. In the present study, we examined two new derivatives of Olz; OlzEt (2-ethyl-4-(4′-methylpiperazin-1′-yl)-10Hbenzo[b]thieno[2,3-e][1,4]diazepine), and OlzHomo (2-ethyl-4-(4′-methyl-1′,4′-diazepan-1′-yl)-10H-benzo[b]thieno[2,3-e] [1,4]diazepine), for their tendency to induce weight gain in rats. Weight gain and metabolic changes were measured in female Sprague Dawley rats. Animals were treated orally with Olz, OlzEt, OlzHomo (3 or 6 mg/kg/day), or vehicle (n = 8), three times daily at eight-hour intervals for 5 weeks. Furthermore, a phencyclidine (PCP)-treated rat model was used to examine the prevention of PCP-induced hyperlocomotor activity relevant for schizophrenia therapy. Male Sprague Dawley rats were pre-treated with a single dose (3 mg/kg/day) of Olz, OlzEt, OlzHomo, or vehicle (n = 12), for 2 weeks. Locomotor activity was recorded following a subcutaneous injection with either saline or PCP (10 mg/kg). Olz was found to induce weight gain, hyperphagia, visceral fat accumulation, and metabolic changes associated with reduced histamatergic H1 receptor density in the hypothalamus of treated rats. In contrast, OlzEt and OlzHomo presented promising antipsychotic effects, which did not induce weight gain or fat deposition in the treated animals. Behavioural analysis showed OlzEt to attenuate PCP-induced hyperactivity to a level similar to that of Olz; however, OlzHomo showed a lower propensity to inhibit these stereotyped behaviours. Our data suggest that the therapeutic effectiveness of OlzHomo may be delivered at a higher dose than that of Olz and OlzEt. Overall, OlzEt and OlzHomo may offer a better pharmacological profile than Olz for treating patients with schizophrenia. Clinical trials are needed to test this hypothesis