Comparative cytogenetics of three economically important Piper L. species from the Brazilian Amazon

The species Piper hispidinervum, Piper aduncum, and Piper affinis hispidinervum have essential oils with high levels of safrole, dillapiole, and sarisan, respectively. Safrole is important for pharmaceutical and chemical industries, while dillapiole and sarisan are promising compounds to control insects and fungi. These species are very similar morphologically and their taxonomy is controversial. Divergent hypotheses consider P. aduncum and P. hispidinervum either as a single species or as distinct taxa, while P. affinis hispidinervum is inferred to be a natural hybrid or a chemotype of P. hispidinervum. Delimiting the taxonomic boundaries would be helpful for germplasm conservation and breeding programs. This study aimed to undertake a detailed analysis of P. aduncum, P. hispidinervum, and P. affinis hispidinervum karyotype and rDNA sites. Genomic in situ hybridization (GISH) was used to establish genomic homology among species and to test the natural hybridization hypothesis for origin of P. affinis hispidinervum. Karyotype traits were similar for all three species: 2n = 26 small chromosomes, predominantly metacentric. All three species exhibited CMA+ bands on the secondary constriction of chromosome pair 4. A size-heteromorphic 35S rDNA site was co-localized with the CMA+ band. A 5S rDNA site was located in the proximal region of chromosome pair 7. The patterns of genomic hybridization revealed that the repetitive DNA fraction of the species is highly similar in terms of proportion of genome, sequence type, and distribution. Our findings did not allow us to differentiate the three species and point to the importance of deeper genomic studies to elucidate the taxonomic controversy.

     

Análisis del impacto presupuestal en Colombia de la prueba de HPV con genotipificación comparada con la citología

Introduction:

The detection of the human papillomavirus (HPV) through the combination of the HPV test and other techniques such as cytology has impacted the detection and timely treatment of lesions associated with cervical cancer.

Objective:

To estimate the budgetary impact of the strategy of early detection of HPV with DNA test genotyping with reflex cytology versus conventional cytology in women aged 30 to 65 years attending the cervical cancer screening program at a health benefit managing entity in Colombia.

Materials and methods:

Using a decision tree and a Markov model, the clinical implications and direct costs of screening, diagnosis, and treatment were estimated in a cohort of women. The analysis considered two screening cycles and their annual costs. The data on the prevalence of clinical results and the costs were taken from the health managing entity. The information on the progression, persistence, and regression of the health states were taken from the ATHENA study.

Results:

The screening scheme with the HPV test, genotyping, and reflex cytology compared to conventional cytology was cost-saving. The average cost per screening cycle with the HPV test was estimated at COP $ 129,201,363 and with cytology at COP $ 186,309,952, i.e., a saving of COP $ 57,108,589 (30.7%).

Conclusion:

The implementation of the screening strategy under evaluation suggests prospective savings derived from the early detection of health states associated with the development of cervical cancer.     

The cloning, expression, purification, characterization and modeled structure of Caulobacter crescentus β-Xylosidase I

The xynB1 gene (CCNA 01040) of Caulobacter crescentus that encodes a bifunctional enzyme containing the conserved β-Xylosidase and α-L: -Arabinofuranosidase (β-Xyl I-α-L: -Ara) domains was amplified by PCR and cloned into the vector pJet1.2Blunt. The xynB1 gene was subcloned into the vector pPROEX-hta that produces a histidine-fused translation product. The overexpression of recombinant β-Xyl I-α-L: -Ara was induced with IPTG in BL21 (DE3) and the resulting intracellular protein was purified with pre-packaged nickel-Sepharose columns. The recombinant β-Xyl I-α-L: -Ara exhibited a specific β-Xylosidase I activity of 1.25?U?mg(-1) to oNPX and a specific α-L: -Arabinofuranosidase activity of 0.47?U?mg(-1) to pNPA. The predominant activity of the recombinant enzyme was its β-Xylosidase I activity, and the enzymatic characterization was focused on it. The β-Xylosidase I activity was high over the pH range 3-10, with maximal activity at pH 6. The enzyme activity was optimal at 45?°C, and a high degree of stability was verified over 240?min at this temperature. Moreover, β-Xylosidase activity was inhibited in the presence of the metals Zn(2+) and Cu(2+), and the enzyme exhibited K(M) and V(Max) values of 2.89?±?0.13?mM and 1.4?±?0.04?μM?min(-1) to oNPX, respectively. The modeled structure of β-xylosidase I showed that its active site is highly conserved compared with other structures of the GH43 family. The increase in the number of contact residues responsible for maintaining the dimeric structure indicates that this dimer is more stable than the tetramer form.     

Using the Hard, Randy, and Rittler Test to Evaluate Color Vision in Capuchins (Cebus libidinosus)

The identification of color vision types in primates is fundamental to understanding the evolution and biological function of color perception. The Hard, Randy, and Rittler (HRR) pseudoisochromatic test categorizes human color vision types successfully. Here we provide an experimental setup to employ HRR in a nonhuman primate, the capuchin (Cebus libidinosus), a platyrrhine with polymorphic color vision. The HRR test consists of plates with a matrix composed of gray circles that vary in size and brightness. Differently colored circles form a geometric shape (X, O, or Δ) that is discriminated visually from the gray background pattern. The ability to identify these shapes determines the type of dyschromatopsy (deficiency in color vision). We tested six capuchins in their own cages under natural sunlight. The subjects chose between two HRR plates in each trial: one with the gray pattern only and the other with a colored shape, presented on the left or right side at random. We presented the test 40 times and calculated the 95?% confidence limits for chance performance based on the binomial test. We also genotyped all subjects for exons 3 and 5 of the X-linked opsin genes. The HRR test diagnosed two subjects as protan dichromats (missing or defective L-cone), three as deutan dichromats (missing or defective M-cone), and one female as trichromat. Genetic analysis supported the behavioral data for all subjects. These findings show that the HRR test can be applied to diagnose color vision in nonhuman primates.     

Brain Pharmacokinetics of Non‐Imidazole Biphenyl H3 Receptor Antagonists: a Liquid Chromatography/Electrospray‐Mass Spectrometry and ex vivo Binding Study in Rats

In the present article, we report on the kinetics of brain penetration in rats of the H3R antagonist 1,1′‐[1,1′‐biphenyl‐4,4′‐diylbis(methylene)]bis‐[piperidine] ( 1 ), which had shown a favorable in vitro pharmacological profile and in vivo potency in preventing scopolamine‐induced amnesia. Two different approaches were employed: high‐performance liquid chromatography/electrospray‐mass spectrometry (HPLC/ESI‐MS) and ex vivo binding against the labeled agonist [³H]‐(R)‐α‐methylhistamine ([³H]RAMHA). Starting from the structure of 1 , the rigid piperidine ring was replaced by a flexible dipropylamino group (see 2 ) or by a morpholino ring (see 3 ), endowed with lower basicity. The effect of replacement on rat plasma and brain disposition in the 24 h after administration was analyzed. High (μM ) and persistent concentrations of 1 were found in rat plasma, while plasma levels were significantly lower (range: 0–200 nM ) for the other two derivatives. This could be explained, among other factors, by the higher stability, observed for 1 , to liver metabolic cleavage. The applied chemical modulation had an important effect on in vivo brain disposition, as, despite the comparable physico‐chemical properties, 2 did not show the tendency to accumulate within the brain, as stated by its brain vs. plasma concentration ratios, if compared to 1 . These structure? property relationships should be taken into account in the pharmacokinetic optimization of new series of H3 receptor antagonists.     

The Jurassic succession in west-central Argentina: stratal patterns, sequences and paleogeographic evolution

Jurassic rocks in west-central Argentina are predominantly marine and marginal-marine siliciclastics, associated with prominent but volumetrically subordinate carbonates and evaporites. Facies developments were ruled by paleogeographic persistence within the southern-infratropical latitude belt (within 40–50°S) and by siliciclastic derivation from the Patagonian hinterland to the southwest. Minor volcanic and volcaniclastic supply arrived from the west, out of a magmatic belt related to Circum-Pacific convergence. Timing of the main marine-flooding events and general correspondence with the high-rank stratal packaging recorded in western North America, suggest that global eustasy was also a factor in controlling the local stratigraphic record. Early Jurassic sedimentation occurred within a series of semi-isolated depocentres linked to fault-bounded Triassic troughs. The Sinemurian-Toarcian deposits record depocentre expansion and coalescence. These trends were coeval with progressively more widespread marine invasions from the northwest and west, leading to an elongate marine seaway which connected central Patagonia with the Pacific domain. During the Aalenian-Bajocian the region was subject to a more subdued tectonism and the foreland side of the basin became fringed by an extensive clastic embankment. Bathonian and Early Callovian were times when coarse clastics prograded into the basin, while the marine embayment shrank as a result of stepwise forced regressions. During Late Callovian to Oxfordian globally rising sea-level, the depocentre witnessed the appearance of cosmopolitan invertebrates and a stratal pattern of basin widening and depositional underbalance, that promoted cratonward onlap and inception of widespread carbonate deposition. Ooidal-coralline carbonate development was terminated after a relatively sudden (Messinian-style) event that desiccated a large tract of the Andean basin, and favored massive precipitation of anhydrite. In the course of the Kimmeridgian the evaporite basin was largely flooded by siliciclastics and turned into a broad and featureless mudflat-salina complex, linked to a widespread erg and to an ephemeral drainage system. By the Tithonian, at a period of peak oceanic stand, marine connection was reinstated and recorded as the most widespread Jurassic transgression across the Neuquén-Aconcagua embayment. Shelfal deposition consisted of molluscs and ooid-dominated carbonate terraces that grew in pulses tuned to eustatic fluctuations. Like in other prolific petroleum provinces around the world the Tithonian basinal strata involve widespread euxinic deposits featuring unusually high organic content.     

Physical activity in osteopenia treatment improved the mass of bones directly and indirectly submitted to mechanical impact

The effect of physical activity in the treatment of osteopenia induced by ovariectomy was studied in 34 two-month-old Wistar female rats. Animals were divided into three groups in which two were formed by ovariectomized (OVX) animals and the other one had sham-operated animals. Group 1, active OVX'd rats; group 2, sedentary OVX'd rats and group 3, sham-operated ones (control). After three months of daily physical activity in a motor-driven treadmill all rats were sacrificed. In order to perform a histomorphometric analysis, long bones, vertebrae, and nasal bone were selected at necropsy. Ovariectomized rats which exercised showed an increased trabecular bone volume, cortical thickness in the long bones and vertebrae and also an increased nasal bone thickness. Physical activity also increased the connection of osteocytes. It was concluded that physical activity in osteopenia treatment increases and restores the mass of bones directly and indirectly submitted to physical impact.     

Fire ants: What do rural and urban areas show us about occurrence, diversity,and ancestral state reconstruction?

In South America, Solenopsis saevissima and S. invicta are the most common fire ants. Nests are founded in areas under anthropic interference like urban or rural areas, but S. invicta is found preferentially in those with the greatest anthropic interference. However, we do not know the rates at which they exist in anthropized areas next to high density of native vegetation. Areas with 60 to 90% of native Atlantic Forest were selected to verify the occurrence of both species in rural and urban areas. We investigated the molecular diversity and applied the reconstruction of the ancestral state analysis for each species. A total of 186 nests were analyzed and we found that the two species had the same proportion in the urban area. However, S. saevissima had a higher rate of prevalence in the rural area, in addition to having a greater number of haplotypes and ancestry associated with this type of habitat for the region. S. invicta had the same number of haplotypes in both rural and urban regions, and less haplotypic diversity. We conclude that S. saevissima is a species typically associated with rural areas and S. invicta, although present, is not dominant in urban areas. Keywords: Biodiversity, haplotype diversity, DNA barcoding, mtDNA     

Solid�CLiquid Transitions and Stability of HPKO-in-Water Systems Emulsified by Dairy Proteins

Stability of oil-in-water emulsions during freezing and thawing is regulated by the phase transitions occurring in the continuous and dispersed phases upon thermal treatments and by the composition of the interfacial membrane. In the present study, the impact of the water phase formulation (0–2.5–5–10–20–30–40% w/w sucrose), the interfacial composition [whey protein isolates (WPI) or sodium caseinate (NaCas) used at different concentrations], and the particle size on the stability of hydrogenated palm kernel oil (30% w/w)-in-water systems was investigated. Phase/state behaviour of the continuous and dispersed phases and emulsion destabilisation were studied by differential scanning calorimetry. System morphology was observed by particle size analysis and optical microscopy. The presence of sucrose in the aqueous phase and reduced particle size distribution significantly improved emulsion stability. WPI showed better stabilising properties than NaCas at lipid to protein ratios of 10:1, 7.5:1, 5:1 and 4:1. Increased WPI concentration significantly improved emulsion resistance to breakdown during freeze–thaw cycling. NaCas showed poor stabilising properties and was ineffective in reducing emulsion destabilisation at 0% sucrose at all the lipid to protein ratios.