A global phylogeny of butterflies reveals their evolutionary history, ancestral hosts and biogeographic origins.

Butterflies are a diverse and charismatic insect group that are thought to have evolved with plants and dispersed throughout the world in response to key geological events. However, these hypotheses have not been extensively tested because a comprehensive phylogenetic framework and datasets for butterfly larval hosts and global distributions are lacking. We sequenced 391 genes from nearly 2,300 butterfly species, sampled from 90 countries and 28 specimen collections, to reconstruct a new phylogenomic tree of butterflies representing 92% of all genera. Our phylogeny has strong support for nearly all nodes and demonstrates that at least 36 butterfly tribes require reclassification. Divergence time analyses imply an origin ~100 million years ago for butterflies and indicate that all but one family were present before the K/Pg extinction event. We aggregated larval host datasets and global distribution records and found that butterflies are likely to have first fed on Fabaceae and originated in what is now the Americas. Soon after the Cretaceous Thermal Maximum, butterflies crossed Beringia and diversified in the Palaeotropics. Our results also reveal that most butterfly species are specialists that feed on only one larval host plant family. However, generalist butterflies that consume two or more plant families usually feed on closely related plants.

There is no evidence that Podoctidae carry eggs of their own species: Reply to Machado and Wolff (2017).

In our recent publication (Sharma et al., 2017), we tested the hypothesis that eggs attached to the legs of male Podoctidae (Opiliones, Laniatores) constituted a case of paternal care, using molecular sequence data in tandem with multiple sequence alignments to test the prediction that sequences of the eggs and the adults that carried them would indicate conspecific identity. We discovered that the sequences of the eggs belonged to spiders, and thus rejected the paternal care hypothesis for these species. Machado and Wolff (2017) recently critiqued our work, which they regarded as a non-critical interpretation and over-reliance on molecular sequence data, and defended the traditional argument that the eggs attached to podoctids are in fact harvestman eggs. Here we show that additional molecular sequence data also refute the identity of the eggs as conspecific harvestman eggs, using molecular cloning techniques to rule out contamination. We show that individual gene trees consistently and reliably place the egg and adult sequences in disparate parts of the tree topology. Phylogenetic methods consistently place all egg sequences within the order Araneae (spiders). We submit that evidence for the paternal care hypothesis based on behavioral, morphological, and natural history approaches is either absent or insufficient for concluding that the eggs of podoctids are conspecific.

A multilocus phylogeny of Podoctidae (Arachnida, Opiliones, Laniatores) and parametric shape analysis reveal the disutility of subfamilial nomenclature in armored harvestman systematics.

The taxonomy and systematics of the armored harvestmen (suborder Laniatores) are based on various sets of morphological characters pertaining to shape, armature, pedipalpal setation, and the number of articles of the walking leg tarsi. Few studies have tested the validity of these historical character systems in a comprehensive way, with reference to an independent data class, i.e., molecular sequence data. We examined as a test case the systematics of Podoctidae, a family distributed throughout the Indo-Pacific. We tested the validity of the three subfamilies of Podoctidae using a five-locus phylogeny, and examined the evolution of dorsal shape as a proxy for taxonomic utility, using parametric shape analysis. Here we show that two of the three subfamilies, Ibaloniinae and Podoctinae, are non-monophyletic, with the third subfamily, Erecananinae, recovered as non-monophyletic in a subset of analyses. Various genera were also recovered as non-monophyletic. As first steps toward revision of Podoctidae, the subfamilies Erecananinae Roewer, 1912 and Ibaloniinae Roewer, 1912 are synonymized with Podoctinae Roewer, 1912 new synonymies, thereby abolishing unsubstantiated subfamilial divisions within Podoctidae. We once again synonymize the genus Paralomanius Goodnight & Goodnight, 1948 with Lomanius Roewer, 1923 revalidated. We additionally show that eggs carried on the legs of male Podoctidae are not conspecific to the males, falsifying the hypothesis of paternal care in this group.

Aerobic training does not alter CRP in apparently healthy, untrained men.

AIM: Regular aerobic exercise may reduce cardiovascular disease (CVD) risk by lowering the concentration of inflammatory markers, such as C-reactive protein (CRP). While studies in diseased populations have shown significant decreases in CRP concentrations with regular aerobic training, little has been conclusively determined regarding the effects of aerobic training on CRP concentrations in apparently healthy, untrained populations. Aim of the study was to examine the effects of a 17-wk half marathon training program (TP) on CRP concentrations, aerobic fitness, and body composition in apparently healthy, untrained men. METHODS: Twenty men (29.3±1.0 y) enrolled as training subjects (TRN) in a 17-wk half marathon TP. An additional 22 men (27.8±1.4 y) served as controls (CON). Fasting blood samples were taken at four time points over the TP and were analyzed for CRP and interleukin-6 (IL-6) concentrations. Aerobic capacity (VO2max) and body fat percent (BF%) were measured before and after the TP. RESULTS: No significant post-training changes in CRP (P=0.70) or IL-6 concentrations (P=0.67) were seen in TRN as a result of the TP, despite significant improvements in VO2max (42.2±1.9 ml∙kg-1∙min⁻¹, P<0.0001) and significant reductions in resting heart rate (P=0.004), BF% (P=0.03), and body mass index (BMI, P=0.05). No significant changes in CRP, VO2max, BMI, or BF% were detected in CON over time. CONCLUSION: Regular aerobic training does not appear to affect CRP concentrations in apparently healthy, untrained men despite significant improvements in bodyweight, BF%, BMI, and VO2max.

Psychometric properties of a scale to measure menopause-related symptoms in two ethnicities.

OBJECTIVES: To evaluate the psychometric properties of a scale we constructed to measure menopause-related symptoms in Hispanic and White non-Hispanic women. METHODS: Items were generated from the literature and clinical experiences. Hispanic (n = 123, 51.34 +/- 5.14 years) and White (n = 210, 53.36 +/- 4.80 years) women completed the scale and other instruments used to validate respective constructs. A subgroup of 69 women completed our scale a second time. RESULTS: Following exploratory factor analysis using the entire sample, confirmatory factor analysis was conducted for Hispanic and White women, separately. The same four-factor model (vasomotor, psychological symptoms, relationship(s), and other symptoms) fit data for both ethnic groups. The coefficient alpha for internal consistency was 0.852 and 0.849 for the initial and second administration, respectively, and the test-retest reliability was significant (p < 0.001). There were differences in factor scores among groups with different menopausal status. Factor scores were significantly correlated with scores of a global quality-of-life instrument (p < or = 0.001). CONCLUSIONS: Our study suggests that the scale is appropriate for use for Hispanic and White non-Hispanic women and is capable of detecting differences among women with different menopause-related symptoms. Menopause-related symptoms measured by the scale were correlated with global quality of life.

Insulin resistance but not visceral adipose tissue is associated with plasminogen activator inhibitor type 1 levels in overweight and obese premenopausal African-American women.

OBJECTIVE: To compare plasma plasminogen activator inhibitor type 1 (PAI-1) levels and to examine the association of PAI-1 with visceral adiposity and other components of the metabolic syndrome in overweight and obese premenopausal African-American (AA) and Caucasian (CC) women. DESIGN: Cross-sectional study. SUBJECTS: 33 CC and 23 AA healthy, overweight and obese, premenopausal women (age 19-53 y, body mass index 28.1-48.9 kg/m(2)). MEASUREMENTS: Body mass index, sagittal diameter, waist circumference, percentage body fat, visceral and subcutaneous adipose tissue (by anthropometry, magnetic resonance imaging (MRI), and bioelectric impedance techniques), PAI-1, leptin, lipids, glucose, insulin, and insulin resistance (by HOMA IR). RESULTS: AA women had lower triglyceride levels and less visceral adipose tissue (VAT) volume than CC despite similar BMI. PAI-1 levels were not significantly different in the two groups. Insulin resistance was associated with PAI-1 in both groups but only in CC women were VAT, triglyceride, HDL cholesterol and blood pressure related to plasma PAI-1 levels. Multiple regression analysis showed that VAT in CC and insulin resistance in AA were independent predictors of PAI-1. CONCLUSION: VAT is significantly associated with circulating PAI-1 levels in overweight and obese CC but not AA premenopausal women.

Dose-response effect of walking exercise on weight loss. How much is enough?

OBJECTIVE: Exercise is the cornerstone of behavioral weight loss programs. The total volume of exercise needed to both promote weight loss and elicit health benefits has not been sufficiently investigated. The aim of this study was to examine the effects of two different volumes of walking 'metabolic fitness' exercise prescriptions, in combination with a low-fat, ad libitum diet (LFAL) on weight loss and additional modifiable health-related variables (HRV) in an ethnically diverse sample of overweight premenopausal women. DESIGN: Clinical 12 week weight loss intervention study with a 5.0-5.8 MJ diet daily with (a). participants walking 30 min, 5 days per week (DEX1), (b). participants walking 60 min, five times per week (DEX2) or (c). a diet only control group (DO). SUBJECTS: A mixed racial sample (predominantly Hispanic) of 56 subjects (mean BMI=34.26+/-6.61, mean age= 39.45+/-7.34) completed the 12 week program. MEASUREMENTS: Various body weight, body composition and fat distribution variables, dietary intake and additional HRV such as blood lipids, blood pressure and an estimate of cardiorespiratory fitness at baseline and after 3 months. RESULTS: All groups showed similar and significant (P<0.001) declines in body weight, percentage body fat, BMI, WHR, fat mass, fat-free mass and diastolic blood pressure following the program. In addition, total cholesterol, triacylglycerol and the TC:HDL ratio displayed a significant time effect (P<0.05). Significant interactions (P<0.05) were found for waist circumference, sagittal diameter, estimated VO(2max) and LDL-C, with both exercise groups showing similar and significantly greater (P<0.05) improvements than DO. Significant interactions (P<0.05) were also observed for several dietary variables. CONCLUSION: Our study showed no dose-response effect of walking exercise on weight loss over diet alone. Both lower and higher volume metabolic fitness prescriptions resulted in similar and significant beneficial changes in several HRV. This data suggests that 30 min of walking on most days of the week may be as beneficial as 60 min (in combination with diet) in promoting numerous additional healthful outcomes over diet alone following a 12 week weight loss program.

Efficient metaphase II transgenesis with different transgene archetypes.

Mammalian genome characterization and biotechnology each require the mobilization of large DNA segments to produce transgenic animals. We recently showed that mouse metaphase II (mII) oocytes could efficiently promote transgenesis (mII transgenesis) when coinjected with sperm and small (<5 kilobases) ubiquitously expressed transgenes (tgs). We have extended this work and now report that mII transgenesis can readily be applied to a range of larger tgs (11.9-170 kilobases), including bacterial and mammalian artificial chromosome (BAC and MAC) constructs. The efficiency of large-construct mII transgenesis was at least as high as that with small constructs; 11-47% of offspring carried the large tgs. More than 95% of these transgenic founders transmitted the tg to offspring. These data demonstrate the ability of mII transgenesis to deliver large tgs efficiently.

Behavioral economic influences on treatments designed to decrease destructive behavior.

In behavioral economics terms, response allocation is viewed as an exchange between the price of and the demand for reinforcers associated with various responses. In this study, behavioral economics principles were used to develop and evaluate a treatment package that reduced destructive behavior to zero while communication and compliance were increased.

Differentiation of embryonic stem cell lines generated from adult somatic cells by nuclear transfer.

Embryonic stem (ES) cells are fully pluripotent in that they can differentiate into all cell types, including gametes. We have derived 35 ES cell lines via nuclear transfer (ntES cell lines) from adult mouse somatic cells of inbred, hybrid, and mutant strains. ntES cells contributed to an extensive variety of cell types, including dopaminergic and serotonergic neurons in vitro and germ cells in vivo. Cloning by transfer of ntES cell nuclei could result in normal development of fertile adults. These studies demonstrate the full pluripotency of ntES cells.

Molecular cloning and phylogenetic analysis of a cDNA encoding the cat (Felis domesticus) Ig epsilon constant region.

A feline splenic cDNA library was screened with a (32)P-labelled cDNA probe encoding the canine IgE epsilon heavy chain subunit. A cDNA sequence of 1614 nucleotides encoding the complete feline IgE heavy chain, as well as a portion of a variable region, was identified. A search of the GenBank database revealed an identity of 82% at the nucleotide level and 76% at the amino acid level between the feline epsilon heavy chain sequence and the canine homologue. In a separate study, feline genomic DNA, isolated from whole feline embryo cells, was subjected to PCR amplification using primers based on known partial genomic DNA sequences for the feline C epsilon gene. Following removal of an intron from the 683 bp PCR product, the coding sequence yielded an ORF of 506 bp. The DNA sequence of this PCR clone differed by a single nucleotide from the cDNA clone. This difference is silent, and therefore the proteins encoded by the two sequences are identical over the regions cloned and sequenced. Phylogenetic analysis of the constant regions of nine immunoglobulin epsilon genes revealed that the feline cDNA is most similar to the canine homologue.

Pig cloning by microinjection of fetal fibroblast nuclei.

Pig cloning will have a marked impact on the optimization of meat production and xenotransplantation. To clone pigs from differentiated cells, we microinjected the nuclei of porcine (Sus scrofa) fetal fibroblasts into enucleated oocytes, and development was induced by electroactivation. The transfer of 110 cloned embryos to four surrogate mothers produced an apparently normal female piglet. The clonal provenance of the piglet was indicated by her coat color and confirmed by DNA microsatellite analysis.

Racial differences in visceral adipose tissue but not anthropometric markers of health-related variables.

This study sought to determine whether visceral adipose tissue (VAT) and/or its anthropometric surrogates could significantly predict health-related variables (HRV) in overweight Caucasian (CC) (n = 36) and African-American (AA) (n = 30) women. With the use of magnetic resonance imaging, findings showed significantly higher volume and area of VAT (P < 0.0001 for both) as well as higher triacylglycerol (P = 0.009) in CC compared with AA women. Furthermore, VAT volume, race, and VAT volume x race interaction could significantly predict triacylglycerol (P = 0.0094), high-density lipoprotein cholesterol (P = 0.0057), insulin (P = 0.0002), and insulin resistance (P < 0. 0001). Additionally, the VAT volume x race interaction for insulin (P = 0.040) and insulin resistance (P = 0.003) was significant. In a separate analysis, waist circumference and race predicted the identical variables. Our results support the use of volume or area of VAT in predicting HRV in CC women; however, its use in AA women appears limited. In contrast, waist circumference can provide a suitable VAT alternative for both CC and AA women; however, VAT clearly represents the more powerful predictor.

Hijacking oocyte DNA repair machinery in transgenesis?

Transgenesis refers to the modification of a genome to carry specified exogenous DNA sequences (transgenes, tgs) in a plant or animal; tgs are ideally transmissible through the germline. It has recently been shown that a membrane-disrupted spermatozoon mixed with exogenous DNA can be microinjected into an unfertilized, metaphase II (mII) oocyte to generate transgenic mouse embryos and offspring. This is here referred to as metaphase II (mII) transgenesis. Exogenous DNA clearly becomes genomically integrated in this process, but how? Presumably, the integration process utilizes DNA repair and recombinational machinery resident within mII oocytes. With recent advances in the description of DNA recombination and repair per se and in relation to meiotic and mitotic cell cycles, we are now poised to explain features of mIl transgenesis. Conversely, the method may of itself provide a new tool to probe these aspects of DNA metabolism. This article describes mil transgenesis in the context of DNA recombination and homeostasis in mII oocytes. A fuller understanding of the underlying recombinational mechanisms may enable improved methods of manipulating mammalian genomes and lead to gene targeting and genetic surgery in mII oocytes.

Mammalian oocyte activation by the synergistic action of discrete sperm head components: induction of calcium transients and involvement of proteolysis.

Sperm-borne oocyte-activating factor (SOAF) elicits activation sufficient for full development and originates from sperm head submembrane matrices. SOAF comprises discrete, heat-sensitive and -stable components (referred to here respectively as SOAF-I and -II) which are each necessary but not sufficient to activate oocytes. The heat-sensitive SOAF component, SOAF-I(m), becomes solubilized from the perinuclear matrix under reducing conditions (the SOAF transition) to generate SOAF-I(s). Although calcium transients likely play an important role in oocyte activation at fertilization, the question is open as to whether demembranated heads or SOAF-I(s) and/or SOAF-II can induce calcium transients. We now report that injection of demembranated sperm heads into mouse oocytes efficiently induced Ca(2+) oscillations. When injected independently, SOAF-I(s) and demembranated heads heated to 48 degrees C failed to generate Ca(2+) oscillations. However, co-injection of SOAF-I(s) and 48 degrees C-heated heads induced oscillations, mirroring their synergistic ability to activate oocytes. This suggests that SOAF-mediated activation proceeds via pathways resembling those at fertilization and provides the first direct evidence that multiple sperm components are required to induce Ca(2+) oscillations. We probed the SOAF-I(s) liberation at the center of this activation and show that in vitro it was sensitive to a profile of serine protease inhibitors. These findings support a model in which mammalian oocyte activation, including the induction of calcium transients, involves proteolytic processing of SOAF from sperm head submembrane compartments.

Physical and psychological changes with vigorous exercise in sedentary primigravidae.

PURPOSE: The present study examined the effects of exercise on physical and psychological variables in sedentary primigravidae (PRA). METHODS: A total of nine women randomly assigned to an exercise (E) (mean age = 31.3 +/- 3.1 yr) and six subjects randomly assigned to a control (C) group (mean age = 27.8 +/- 3.1 yr) fulfilled all requirements for the study. Exercise included a variety of exercise activities performed to a target heart rate of 150-156 beats x min(-1), three times per week for 15 wk. RESULTS: Results showed no significant differences between E and C groups in physical characteristics initially. A repeated measures ANOVA showed a significant group effect (P < 0.05) and a significant group by time interaction (P = 0.001) with the E group showing a significantly longer amount of time on the PWC150 test than the C group. There was no significant group, time, or group by time interaction for lactate accumulation. This occurred, despite the fact that the E group spent 56% longer on the PWC150 test and the C group spent 30% less time on the same test at the conclusion of the study. Finally, the E group in comparison to the C group showed more favorable improvements in several items related to health and well-being on the Body Cathexis Scale. There were no significant differences between E and C groups in any pregnancy outcome measures. All babies were delivered healthy at term. CONCLUSIONS: These data suggest that a vigorous exercise program can lead to significant improvements in aerobic fitness at similar lactate concentrations compared to a control group and can be well tolerated by low risk sedentary PRA without any deleterious effects occurring to herself or unborn child.

Mice cloned from embryonic stem cells.

Cloning allows the asexual reproduction of selected individuals such that the offspring have an essentially identical nuclear genome. Cloning by nuclear transfer thus far has been reported only with freshly isolated cells and cells from primary cultures. We previously reported a method of cloning mice from adult somatic cells after nuclear transfer by microinjection. Here, we apply this method to clone mice from widely available, established embryonic stem (ES) cell lines at late passage. With the ES cell line R1, 29% of reconstructed oocytes developed in vitro to the morula/blastocyst stage, and 8% of these embryos developed to live-born pups when transferred to surrogate mothers. We thus cloned 26 mice from R1 cells. Nuclei from the ES cell line E14 also were shown to direct development to term. We present evidence that the nuclei of ES cells at G(1)- or G(2)/M-phases are efficiently able to support full development. Our findings demonstrate that late-passage ES cells can be used to produce viable cloned mice and provide a link between the technologies of ES cells and animal cloning. It thus may be possible to clone from a single cell a large number of individuals over an extended period.

Identification, sequence analysis and expression of transcripts encoding a putative metalloproteinase, eMDC II, in human and macaque epididymis.

The metalloproteinase-like, disintegrin-like, cysteine-rich (MDC) family is a large group of sequence-related proteins, first characterized in the male reproductive tract, but subsequently also identified in non-reproductive tissues. Their primary translation products are of approximately 90 kDa and each can be divided into distinct domains which show remarkable homology to reprolysins; snake venom haemorrhagic components possessing metalloproteinase and/or disintegrin domains. Several MDC proteins are abundantly-expressed in the male reproductive tract, suggesting functions in fertility. We now describe the cloning, sequence determination and characterization of transcripts encoding the human and macaque (Macaca fascicularis) orthologues of a novel member of the MDC family (eMDC II) which is abundantly-expressed in the epididymis. Unlike many MDC proteins expressed in the reproductive tract, eMDC II possesses the extended 'catalytic centre' consensus sequence characteristic of a reprolysin-like metalloproteinase. This suggests that eMDC II has proteolytic activity.

Fibrovascular ingrowth in porous ocular implants: the effect of material composition, porosity, growth factors, and coatings.

PURPOSE: Fibrovascular ingrowth into various porous ocular implants as a function of implant material composition, porosity, growth factors, and coatings was investigated in a pilot study in an animal model. METHODS: Eighty-one New Zealand white rabbits underwent unilateral enucleation and implantation with ocular implants composed of the following materials: coralline hydroxyapatite (HA) with 200-microm pores (HA200) or 500-microm pores (HA500), synthetic HA (synHA), and high-density porous polyethylene (PP). The HA200, HA500, and PP implants were implanted untreated or after treatment with recombinant human basic fibroblast growth factor (Rh-bFGF). Nine HA500 implants were implanted after coating with calcium sulfate (plaster of Paris) to provide a smooth outer surface. Implants were harvested at 1-, 2-, 4-, or 8-week intervals and were examined histologically. RESULTS: A significant difference was found between untreated HA500 and PP, with PP showing better ingrowth. There was no significant difference between untreated HA and PP, nor between untreated HA500 and synHA. Significant increases in ingrowth were found in HA200 compared with HA500, and in Rh-bFGF-treated implants compared with untreated controls. The calcium sulfate-coated implants showed less vascularization compared with the uncoated implants, although the difference was not significant. CONCLUSIONS: Fibrovascular ingrowth occurred earlier in HA200 implants than in HA500 implants, and was enhanced when implants were treated with Rh-bFGF.