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.

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.

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.

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.

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.

Mammalian transgenesis by intracytoplasmic sperm injection.

Coinjection of unfertilized mouse oocytes with sperm heads and exogenous DNA encoding either a green fluorescent protein (GFP) or beta-galactosidase reporter produced 64 to 94 percent transgene-expressing embryos, reflecting DNA-sperm head association before coinjection. Nonselective transfer to surrogate mothers of embryos in the GFP series generated about 20 percent offspring expressing the integrated transgene. These data indicate that exogenous DNA can reproducibly be delivered into an oocyte by microinjected spermatozoa and suggest an adaptable method of transgenesis.

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.

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.

Function from structure? The crystal structure of human phosphatidylethanolamine-binding protein suggests a role in membrane signal transduction.

BACKGROUND: Proteins belonging to the phosphatidylethanolamine-binding protein (PEBP) family are highly conserved throughout nature and have no significant sequence homology with other proteins of known structure or function. A variety of biological roles have previously been described for members of this family, including lipid binding, roles as odorant effector molecules or opioids, interaction with the cell-signalling machinery, regulation of flowering plant stem architecture, and a function as a precursor protein of a bioactive brain neuropeptide. To date, no experimentally derived structural information has been available for this protein family. In this study we have used X-ray crystallography to determine the three-dimensional structure of human PEBP (hPEBP), in an attempt to clarify the biological role of this unique protein family. RESULTS: The crystal structures of two forms of hPEBP have been determined: one in the native state (at 2.05 A resolution) and one in complex with cacodylate (at 1.75 A resolution). The crystal structures reveal that hPEBP adopts a novel protein topology, dominated by the presence of a large central beta sheet, and is expected to represent the archaetypal fold for this family of proteins. Two potential functional sites have been identified from the structure: a putative ligand-binding site and a coupled cleavage site. hPEBP forms a dimer in the crystal with a distinctive dipole moment that may orient the oligomer for membrane binding. CONCLUSIONS: The crystal structure of hPEBP suggests that the ligand-binding site could accommodate the phosphate head groups of membrane lipids, therefore allowing the protein to adhere to the inner leaf of bilipid membranes where it would be ideally positioned to relay signals from the membrane to the cytoplasm. The structure also suggests that ligand binding may lead to coordinated release of the N-terminal region of the protein to form the hippocampal neurostimulatory peptide, which is known to be active in the development of the hippocampus. These studies are consistent with a primary biological role for hPEBP as a transducer of signals from the interior membrane surface.

Sequence analysis of monkey acrosin-trypsin inhibitor transcripts and their abundant expression in the epididymis.

Seven independent cross-hybridizing clones have been isolated from a Macaca fascicularis epididymal cDNA library and their inserts shown to correspond to a secreted inhibitor of the sperm serine proteinase, acrosin. Hybridizing transcripts of approx. 0.6 kb have been shown by Northern blot analysis to be considerably more abundant in the epididymis than the testis. This finding is in surprising contrast to a previous report that human acrosin-trypsin inhibitor mRNA predominates in the testis.

Identification of an abundant monkey epididymal transcript encoding a homologue of human CAMPATH-1 antigen precursor.

A number of cDNA clones encoding a small (0.5-0.6 kb) transcript have been isolated from a monkey (Macaca fascicularis) epididymal cDNA library. DNA sequence analysis indicates that this abundant epididymal transcript is homologous to the human CAMPATH-1 (CDw52) antigen precursor, a GPI-anchored membrane glycoprotein previously described on lymphocytes and monocytes.

Genetic evidence for an additional member of the metalloproteinase-like, disintegrin-like, cysteine-rich (MDC) family of mammalian proteins and its abundant expression in the testis.

Full-length clones have been isolated from a Macaca fascicularis cDNA library corresponding to a 2.65 kb mRNA present at high steady state levels in the testis. Sequence analysis suggests that the predicted translation product, tMDC II protein, represents a novel member of the metalloproteinase-like, disintegrin-like, cysteine-rich (MDC) family of mammalian proteins.

Sequence and expression of a monkey testicular transcript encoding tMDC I, a novel member of the metalloproteinase-like, disintegrin-like, cysteine-rich (MDC) protein family.

Full-length clones encoding a novel member of the metalloproteinase-like, disintegrin-like, cysteine-rich (MDC) family of proteins have been isolated from a monkey (Macaca fascicularis) testicular cDNA library. The encoded putative 82 kDa transmembrane protein (tMDC I) shows striking sequence similarity to other members of the MDC family including rat and monkey EAP I, guinea-pig PH-30 and human MDC protein, as well as a number of snake venom components.

Nucleotide and deduced amino acid sequence of porcine interleukin 4 cDNA derived from lamina propria lymphocytes.

Total RNA was isolated from in vitro activated lamina propria lymphocytes and used to direct the synthesis of cDNA. Interleukin 4 transcripts were then specifically amplified by PCR. Comparison of the nucleotide sequence with its human homologue demonstrates deletion within the coding region of pig interleukin 4 centred around amino acid residue 70 in the mature human protein.

The monkey ESP14.6 mRNA, a novel transcript expressed at high levels in the epididymis.

A screening strategy designed to identify cloned cDNAs encoding abundant Macaca fascicularis epididymal transcripts has yielded a clone corresponding to a 1.03-kb transcript present at high levels in the epididymis. Following library rescreening, DNA sequence analysis of several near full-length clones predicts a novel, 151-amino-acid protein, epididymal secretory protein 14.6 (ESP14.6), which contains a strong candidate signal peptide characteristic of secretory proteins.

Structural analysis of the pilE region of Neisseria gonorrhoeae P9.

We have determined the nucleotide sequence of an expressed structural pilus gene (pilE) derived from Neisseria gonorrhoeae strain P9-2. Detailed analysis of nucleotide sequences upstream from pilE revealed a silent, truncated pilin gene segment that was linked to families of DNA elements (RS1 and RS3) that have previously been identified at the major silent pilin gene locus (pilS1) and at pilE of the independently isolated N. gonorrhoeae strain MS11ms. A nucleotide sequence downstream from pilE was reminiscent of the recognition sequences of several recombinases, including Tn3 tnpR product (resolvase), suggesting a possible role for site-specific events in the recombinational modulation of pilus expression.

Relation between anthropometric measures of fat distribution and cardiovascular risk factors in overweight pre- and postmenopausal women.

This study compared three different measures of central adiposity: waist-to-hip ratio (WHR), waist-to-height ratio (WSHT), and waist circumference with cardiovascular risk factors, including serum lipoproteins and blood pressure in overweight pre- (n = 115) and postmenopausal (n = 46) women. Premenopausal women had a mean age of 35.6 +/- 6.79 y and a mean body mass index (BMI; in kg/m2) of 37.08 +/- 6.01. Postmenopausal women had a mean age of 52.5 +/- 8.19 y and a mean BMI of 38.75 +/- 6.9. Although several correlations between central adiposity and serum lipoproteins and blood pressure were significant, they were unaffected by menopausal status. There were also no significant differences among the three measures of central adiposity in relation to cardiovascular risk factors within premenopausal and postmenopausal groups. An analysis of covariance controlling for BMI showed that after stratifying WSHT into tertiles, a significant interaction of WSHT group by menopausal status was found for systolic blood pressure (SBP) (P = 0.019). Postmenopausal women had a significantly greater SBP than premenopausal women in the lowest and highest tertiles (P = 0.001); however, this pattern was not shown in the middle WSHT tertile. The relation between central adiposity and cardiovascular risk factors appears to be unchanged after menopause, except when WSHT is used to indicate SBP. Because increased central adiposity may also indicate an increase in cardiovascular risk factors, measurements of central adiposity can be used to supplement the routine clinical evaluation of cardiovascular risk factors in both pre- and postmenopausal overweight women.

Clinical predictability of the waist-to-hip ratio in assessment of cardiovascular disease risk factors in overweight, premenopausal women.

The waist-to-hip ratio (WHR) is one of the most commonly used anthropometric measures to indicate a central obesity pattern and an increased risk of cardiovascular disease in normal-weight women. Although the American Heart Association has reported that a WHR >0.80 be used to indicate increased risk of cardiovascular disease in women, the present study assessed the WHR above which is seen elevations in cardiovascular disease risk factors in a sample of overweight women. Using data from 240 women aged 27.5-47.5 y enrolled in a university weight-loss program, we determined WHR quartiles: <0.80, 0.80 to <0.84, 0.84 to <0.90, and > or =0.90. Subjects were placed into high-risk categories for cardiovascular disease on the basis of age- and population-defined norms. Women had an increased likelihood of elevated VLDL cholesterol, triacylglycerol, diastolic blood pressure, and composite risk (ie, having > or =4 cardiovascular disease risk factors) and an increased risk of having low concentrations of HDL at a WHR > or =0.90. All aforementioned variables had a significant odds ratio at a WHR > or =20.90 after adjustment for smoking, whereas elevated VLDL, triacylglycerol, and diastolic blood pressure were observed at this WHR after adjustment for a body mass index (in kg/m2) < or > or =35. Only 2 variables, VLDL and triacylglycerol, had a significant odds ratio at a WHR <0.90 before and after adjustment for BMI and smoking. These data suggest an upward shift in the critical threshold for WHR to > or =0.90, at which point there was an elevation in cardiovascular disease risk factors in already overweight women. This trend persisted regardless of whether the women smoked or whether their body mass index was < or > or =35.

Sequence analysis and immunolocalisation of phosphatidylethanolamine binding protein (PBP) in human brain tissue.

We have cloned and sequenced cDNA corresponding to human brain phosphatidylethanolamine binding protein (PBP) and have used the resultant data to generate PBP-specific anti-peptide antisera for immunohistochemical studies. The distribution of PBP was assessed immunohistochemically in sections from multiple regions of neonatal brain and spinal cord, including spinal nerve roots. Strong PBP immunoreactivity was present in the cytoplasm of oligodendrocytes in the white matter and Schwann cells in the nerve roots. There was only weak immunostaining within the neurones and neuropil. The findings are in keeping with a role for PBP in the organisation of phospholipids in the myelin sheath.