The admixed brushtail possum genome reveals invasion history in New Zealand and novel imprinted genes.

Combining genome assembly with population and functional genomics can provide valuable insights to development and evolution, as well as tools for species management. Here, we present a chromosome-level genome assembly of the common brushtail possum (Trichosurus vulpecula), a model marsupial threatened in parts of their native range in Australia, but also a major introduced pest in New Zealand. Functional genomics reveals post-natal activation of chemosensory and metabolic genes, reflecting unique adaptations to altricial birth and delayed weaning, a hallmark of marsupial development. Nuclear and mitochondrial analyses trace New Zealand possums to distinct Australian subspecies, which have subsequently hybridised. This admixture allowed phasing of parental alleles genome-wide, ultimately revealing at least four genes with imprinted, parent-specific expression not yet detected in other species (MLH1, EPM2AIP1, UBP1 and GPX7). We find that reprogramming of possum germline imprints, and the wider epigenome, is similar to eutherian mammals except onset occurs after birth. Together, this work is useful for genetic-based control and conservation of possums, and contributes to understanding of the evolution of novel mammalian epigenetic traits.

Core Surface Flow Changes Associated With the 2017 Pacific Geomagnetic Jerk.

A geomagnetic jerk was seen in Swarm satellite data in 2017 over the Pacific region. We invert time series of spatial gradient secular variation data between 2014 and 2020, reduced to a grid of points at satellite altitude, for spatially- and temporally-regularized core surface flow. Pacific region flow acceleration was almost constant before and after the jerk, with a sharp change, especially in the azimuthal component, at the jerk epoch, despite the temporal regularization. Azimuthal acceleration is oppositely signed either side of 160°W, where it effectively vanishes, and also reverses sign at the jerk epoch. Acceleration features drift westward at about 900 km year-1. Unlike previous studies, the evidence presented here for low latitude waves does not depend on imposing flow equatorial symmetry, quasi- or tangential geostrophy, or band-pass filtering, and has no reliance on stochastic models or numerical simulations.

Scalable, Green Fabrication of Single-Crystal Noble Metal Films and Nanostructures for Low-Loss Nanotechnology Applications.

The confinement of spatially extended electromagnetic waves to nanometer-scale metal structures can be harnessed for application in information processing, energy harvesting, sensing, and catalysis. Metal nanostructures enable negative refractive index, subwavelength resolution imaging, and patterning through engineered metamaterials and promise technologies that will operate in the quantum plasmonics regime. However, the controlled fabrication of high-definition single-crystal subwavelength metal nanostructures has remained a significant hurdle due to the tendency for polycrystalline metal growth using conventional physical vapor deposition methods and the challenges associated with placing solution-grown nanocrystals in desired orientations and locations on a surface to manufacture functional devices. Here, we introduce a scalable and green wet chemical approach to monocrystalline noble metal thin films and nanostructures. The method enables the fabrication of ultrasmooth, epitaxial, single-crystal films of controllable thickness that are ideal for the subtractive manufacture of nanostructures through ion beam milling and additive crystalline nanostructure via lithographic patterning for large-area, single-crystal metasurfaces and high aspect ratio nanowires. Our single-crystal nanostructures demonstrate improved feature quality, pattern transfer yield, reduced optical and resistive losses, and tailored local fields to yield greater optical response and improved stability compared to those of polycrystalline structures-supporting greater local field enhancements and enabling practical advances at the nanoscale.

Secreted products of Fasciola hepatica inhibit the induction of T cell responses that mediate allergy.

There is evidence from epidemiology studies of a negative association between infection with helminth parasites and the development of allergy and asthma. Here, we demonstrate that the excretory/secretory products of the helminth Fasciola hepatica (FHES) protected mice against ovalbumin (OVA)-induced allergic asthma when administered at time of allergen sensitization. FHES reduced the accumulation of mucus, eosinophils and lymphocytes into the airways of allergen-challenged mice. Furthermore, FHES treatment suppressed Th2 responses in the airways. Interestingly, systemic administration of FHES at allergen challenge had no effect on airway inflammation, demonstrating that alum-induced Th2 response is set following initial allergen sensitization. Our findings highlight the immunomodulatory potential of molecules secreted by F. hepatica.

A critical role for the TLR signaling adapter Mal in alveolar macrophage-mediated protection against Bordetella pertussis.

Bordetella pertussis causes whooping cough, an infectious disease of the respiratory tract that is re-emerging despite high vaccine coverage. Here we examined the role of Toll-like receptor (TLR) adapter protein Mal in the control of B. pertussis infection in the lungs. We found that B. pertussis bacterial load in the lungs of Mal-defective (Mal(-/-)) mice exceeded that of wild-type (WT) mice by up to 100-fold and bacteria disseminated to the liver in Mal(-/-) mice and 50% of these mice died from the infection. Macrophages from Mal(-/-) mice were defective in an early burst of pro-inflammatory cytokine production and in their ability to kill or constrain intracellular growth of B. pertussis. Importantly, the B. pertussis bacterial load in the lungs inversely correlated with the number of alveolar macrophages. Despite the maintenance and expansion of other cell populations, alveolar macrophages were completely depleted from the lungs of infected Mal(-/-) mice, but not from infected WT mice. Our findings define for the first time a role for a microbial pattern-recognition pathway in the survival of alveolar macrophages and uncover a mechanism of macrophage-mediated immunity to B. pertussis in which Mal controls intracellular survival and dissemination of bacteria from the lungs.

Effects of dynamic upper-body exercise on lower-limb isometric endurance.

Following preliminary indications that in some individuals arm exercise enhanced rather than reduced simultaneous leg endurance, ten young men and women performed three forms of intermittent work to volitional exhaustion, under duty cycles of 45 s work, 15 s rest. The protocols were as follows: (A) knee extensions at 30% maximum voluntary contraction (MVC); (B) 30% MVC knee extensions combined with arm cranking at 130% of their own lactate threshold; (C) combined 30% MVC knee extensions and arm cranking at 20% of their own lactate threshold. Heart rate, oxygen uptake (VO(2)), and blood lactate concentration were among the variables recorded throughout. All physiological indicators of demand were substantially higher in protocol B than in protocols A or C [heart rate: (A) 154 beats . min(-1), (B) 171 beats . min(-1), (C) 150 beats . min(-1); VO(2): (A) 11.9 ml . kg(-1) . min(-1), (B) 21.7 ml . kg(-1) . min(-1), (C) 14.2 ml . kg(-1) . min(-1); blood lactate concentration: (A) 3.3 mmol . l(-1), (B) 5.1 mmol . l(-1), (C) 2.8 mmol . l(-1)], yet there were no significant differences (P > 0.05) in the endurance times between the three conditions [(A) 11.43 min, (B) 11.1 min, (C) 10.57 min] and seven participants endured longest in protocol B. Results from protocol (C) cast doubt on explanations in terms of psychological distraction. We suggest that lactic acid produced by the arms is shuttled to the legs and acts there either as a supplementary fuel source or as an antagonist to the depressing effects of increased potassium concentration.

[Training for diagnosis of intestinal parasitic diseases in the national laboratory system of Cuba]. / Adiestramiento en el diagnóstico de las parasitosis intestinales en la red de laboratorios de Cuba.

A national training project in the diagnosis of intestinal parasites was conducted in 1997. An initial national course was followed respectively by courses in the Central, Eastern, and Western Provinces. Our results showed that Cryptosporidium parvum, Cyclospora cayetanensis, and leukocytes showed a significantly lower percentage of errors after the training than before (p < 0.01). The same occurred with Entamoeba histolytica/E. dispar and Chilomastix mesnilii (p < 0.05). Among the helminths, Taenia spp., Fasciola hepatica, and hookworm showed significantly fewer errors after the training (p < 0.01). In the other specimens, few mistakes were found both at the beginning and after the training, and the percentage of errors did not change (p > 0.05). Furthermore, when comparing scores before and after training, a significant increase in median scores appeared in the Central Provinces (p < 0.05), Western Provinces (p < 0.01), and in the entire series (p < 0.01). The results showed the effectiveness of this intervention; these periodic mandatory training courses should be developed together with national programs for quality assessment in Parasitology.

Absorption of dimethylacetamide (DMAC) following application of a polymer film to the skin of rabbits.

Kapton Film is a polymer with high strength and thermal resistance finding use in a wide variety of applications. In the preparation of this film, the uncured material could contain up to 30% dimethylacetamide (DMAC). Note that the final product Kapton Film has been thermally cured with the DMAC removed. During processing, dermal contact with the film is anticipated, and the possibility exists of DMAC transfer from the uncured film, to and through the skin. In this study, 2 x 2-inch pieces of film were applied to the skin of a group of rabbits and secured in place for a single 4-hour contact time. An amount of liquid DMAC corresponding to the amount contained in the 2-inch square was applied to the skin of a separate group of rabbits for 4 hours. Urine samples were collected over the intervals of application to 4 hours post application (8 hours total) or 4 to 20 hours post application (16 hours). The amount of the urinary metabolite monomethylacetamide (MMAC) was determined analytically in these samples. Urine from 2 untreated rabbits was collected at the same time to serve as controls. The amount of urinary MMAC found in the rabbit rine from animals exposed to uncured Kapton Film at both ollection intervals was similar to the amount seen in the controls (background). Rabbits treated with DMAC liquid had measurable urinary MMAC levels that were approximately 100 times background. It is concluded that, under the conditions of this study, very little, if any, DMAC from the uncured Kapton Film was absorbed through the skin (and excreted in the urine as MMAC).

Anxiogenic effects of repeated administrations of 20% CO2-enriched air: stability within sessions and habituation across time.

Increasingly carbon dioxide-enriched air is being used as an aversive unconditioned stimulus in laboratory examinations of anxiety. Yet, little is known about the stability of the autonomic and subjective effects of this stimulus across repeated inhalations and sessions. We examined whether repeated administrations of high concentrations of CO2-enriched air produced either habituation, stability, or sensitization across several autonomic and self-report indices within one session (Experiment 1) and then several sessions (Experiment 2) of exposure. Results suggest that non-clinical participants do not habituate to CO2 within sessions, but do show habituation on cardiac and subjective report of anxiety across sessions. Individual difference factors such as anxiety sensitivity and suffocation fear seem to moderate some of these effects, including self-reported distress and anxiety in response to the challenge. These results support the use of CO2 as a panicogenic aversive stimulus in laboratory models of fear onset and in clinical settings for interoceptive exposure treatments of panic.

Kinetics of antibody-based antigen detection in serum and faeces of sheep experimentally infected with Fasciola hepatica.

The monoclonal antibody ES78 was used in a sandwich immunosorbent assay (Sandwich ELISA) for the detection of antigens in sera and faeces in the course of Fasciola hepatica infection in 10 experimentally infected sheep. All infected sheep had circulating antigens in the first week post-infection (WPI). Antigenemia was detectable until WPI 3 in four infected sheep, WPI 4 in five infected sheep and in only one sheep by WPI 5. The detection of coproantigens (Fa(g)) was possible in five infected sheep at WPI-4, in four sheep at WPI-5 and in one sheep only at WPI-6. This technique was compared to an indirect ELISA for the detection of antibodies using excretory secretory antigens of F. hepatica. A significant correlation was found between Fa(g) and egg output and also with adult worm numbers. Our method demonstrated that the diagnosis of active fasciolosis in sheep is possible during all periods of infection.

Longitudinal study of giardiasis in three day care centres of Havana City.

The prevalence, incidence and reinfection of giardiasis were studied in 365 children attending three day care centres (DCCs) in Havana City. Three stool samples were obtained from each child every 6 months during an 18-month period. We identified three distinct groups of children according to their patterns of infection. In the largest group (51%) children were never found infected. In the second group, they were found infected once or twice during the study period, and in the third and the smallest group (9%) they were found infected in most or all the study periods. This last group seems to be children 'predisposed' to Giardia lamblia infection. The prevalence of giardiasis (20%) remained almost constant throughout the study period. The incidence declined from 16 to 11%, and reinfection increased from 36 to 49%. All the children had normal nutritional status and the only clinical manifestation that correlated strongly with the frequency of cross-sectional surveys positive to Giardia was the number of diarrhoeal episodes recorded during the last 6 months of the study period. Further studies will be necessary to ascertain the causes that determine the 'predisposition' to giardiasis in children.

Kinetics of antibody-based antigen detection in serum and faeces of sheep experimentally infected with Fasciola hepatica.

The monoclonal antibody ES78 was used in a sandwich immunosorbent assay (Sandwich ELISA) for the detection of antigens in sera and faeces in the course of Fasciola hepatica infection in 10 experimentally infected sheep. All infected sheep had circulating antigens in the first week post-infection (WPI). Antigenemia was detectable until WPI 3 in four infected sheep, WPI 4 in five infected sheep and in only one sheep by WPI 5. The detection of coproantigens (Fag) was possible in five infected sheep at WPI-4, in four sheep at WPI-5 and in one sheep only at WPI-6. This technique was compared to an indirect ELISA for the detection of antibodies using excretory secretory antigens of F. hepatica. A significant correlation was found between Fag and egg output and also with adult worm numbers. Our method demonstrated that the diagnosis of active fasciolosis in sheep is possible during all periods of infection.

Overproduction of MDM2 in vivo disrupts S phase independent of E2F1.

Expression of a beta-lactoglobulin (BLG)/mdm2 transgene (BLGmdm2) in the epithelial cells of the mouse mammary gland causes an uncoupling of S phase from M phase, resulting in polyploidy and tumor formation. The cell cycle defects are independent of interactions with p53. Because MDM2 also binds and activates the S phase-specific transcription factor E2F1, we hypothesized that increased E2F1 activity causes the development of the BLGmdm2 phenotype. We, therefore, generated BLGmdm2 mice that were null for E2F1. We observed no notable differences in histology or cyclin gene expression between BLGmdm2 and BLGmdm2/E2F1-/- mice, indicating that endogenous E2F1 activity was not required for the BLGmdm2 phenotype. Because, depending on the experimental system, either loss of E2F1 function or overexpression of E2F1 results in transformation, we also tested whether overexpression of E2F1 augmented the severity of the BLGmdm2 phenotype by generating mice that were bitransgenic for BLGmdm2 and BLGE2F1. We observed a unique mixture of the two single transgenic phenotypes histologically and found no significant changes in cyclin levels, indicating that overexpression of E2F1 had no effect on the BLGmdm2 transgenic phenotype. Thus, increased expression or absence of E2F1 does not affect the ability of MDM2 to disrupt the cell cycle.

[Knowledge, opinions, and practices regarding Aedes aegypti]. / Conocimientos, opiniones y prácticas sobre Aedes aegypti.

The active participation of the community is indispensable for reducing Aedes aegypti mosquito populations so causes leading to their growth, actions taken, effects on the health and the people in charge of controlling such mosquitoes are all dealt with in this paper. 210 persons were interviewed from June 25 to 27, 1997. The causes of the growth of the mosquito population were: excessive garbage (59%), dirty waters (58%), open water reservoirs (50%) and lack of fumigation (21%). 40% of interviewed people did not know that these vectors can breed even in clean waters. They mentioned some effects on the health due to the mosquito bites. 51% said that mosquitoes should be jointly controlled by the State and the community but 35% believed that the State should be the sole responsible for this task. These opinions may determine that steps are not properly taken to avoid proliferation of vector and that the activities of the anti-vector program are not really understood since they do not meet the expectations.

Dynamics of antigenemia and coproantigens during a human Fasciola hepatica outbreak.

In the present study the dynamics of antigenemia and coproantigens were studied in patients with Fasciola hepatica infection during an outbreak occurring in La Palma, Pinar del Río, in the West Province of Cuba. Stool and serum samples were collected from 67 patients and 40 healthy subjects. Stool samples were studied by a simple gravity sedimentation technique and an ES78 sandwich enzyme-linked immunosorbent assay (ELISA) for observation of eggs and detection of parasite coproantigens, respectively. Serum samples were also studied by the ES78 sandwich ELISA and an indirect ELISA to detect circulating antigens and antibodies, respectively. At the beginning of the study, 8 of 67 patients had patent infections and 59 had prepatent infections, which was determined by the recent consumption of lettuce contaminated with metacercariae of F. hepatica, the presence of clinical symptoms, and the absence of Fasciola eggs in their stools. Patients with prepatent infections were monitored by all techniques until patency. Circulating antigens were not detected in patients with patent infections. However, coproantigens were clearly detected in all patients with patent infections. On the other hand, 28.8% of patients with prepatent infections tested positive for circulating antigens and 81.4% tested positive for coproantigens in the first stool sample studied. Only two other coproantigen determinations were necessary to diagnose 93.2% of the patients. While circulating antigen levels diminished in all patients during the infection, coproantigen levels increased. The present study demonstrates that the ES78 sandwich ELISA is a better tool than parasitological examination for diagnosis of active early infection, since by the combination of the circulating-antigen detection assay and the coproantigen detection assay 91% of patients were able to be diagnosed at the beginning of the study. In contrast, a coprologic analysis repeated over several weeks was necessary to diagnose 100% of the patients.

Specific cellular and humoral immune response in Balb/c mice immunised with an expression genomic library of Trypanosoma cruzi.

An expression genomic library of Trypanosoma cruzi (T. cruzi) constructed using pcDNA3 plasmid was used for the immunisation (25 micrograms) of Balb/c mice. Expression of T. cruzi antigens in the muscle of inoculated mice was detected by indirect immunofluorescence 7 days after immunisation. Specific IgG antibodies were significatively increased (P < 0.05) in animals that were reimmunized with 50 micrograms of the genomic library. An antigen specific lymphoproliferative response was detected in one animal of the group inoculated with one dose of the library.

Partial characterization of the epitope on excretory-secretory products of Fasciola hepatica recognized by monoclonal antibody ES78.

This report contains a partial characterization of the epitope recognized by monoclonal antibody (MAb) ES78 produced against excretory-secretory (ES) antigens of Fasciola hepatica. ES78 is currently used for the detection of ES antigens in serum and stool samples of cattle and humans with fasciolosis, using a highly sensitive and specific sandwich enzyme-linked immunosorbent assay (ELISA). The epitope was characterized by periodate oxidation, alkaline borohydride reduction, trichloroacetic acid precipitation, beta-mercaptoethanol treatment, and enzymatic proteolysis. These results, together with those of the 2-site ELISA, lectin immunoassays, and beta-galactosidase digestion, showed that MAb ES78 reacts with a partly protein/partly carbohydrate antigenic determinant that is found on several ES molecules of adult specimens of F. hepatica and contains at least 1 disulfide bond and beta-galactose probably as galactose-beta(1-3)-N-acetylgalactosamine disaccharide.

Targeted expression of MDM2 uncouples S phase from mitosis and inhibits mammary gland development independent of p53.

MDM2 is a cellular protein that binds to and inactivates the p53 tumor suppressor protein. Although mdm2 has been shown to function as an oncogene in vitro, all studies to date have assessed MDM2 activities in the presence of p53, implicating p53 inactivation in MDM2-directed transformation. To determine the role of MDM2 in the cell cycle and in tumorigenesis and whether or not this role is dependent on p53, an MDM2 minigene was expressed during gestation and lactation in the mammary gland of both wild-type p53 (p53+/+) and p53 knockout (p53-/-) mice using the bovine beta-lactoglobulin promoter. In six different transgenic mouse lines, deregulated expression of MDM2 inhibited normal development and morphogenesis of the mammary gland, and caused cellular hypertrophy and nuclear abnormalities. These abnormalities included both multinucleated cells and enlarged cells with giant nuclei. Although there were fewer epithelial cells present in the transgenic mammary gland, no apoptosis was observed. Instead, BrdU incorporation and PCNA staining showed that 12%-27% of the transgenic mammary epithelial cells were in S phase at a time when normal cells were terminally differentiated. Analysis of DNA content showed that 30%-45% of the cells were polyploid, with DNA contents up to 16N, indicating that overexpression of MDM2 caused mammary epithelial cells to undergo multiple rounds of S phase without cell division. This phenotype was similar in the p53+/+ and p53-/- background, demonstrating a role for MDM2 in the regulation of DNA synthesis that is independent of the ability of MDM2 to inhibit p53 activity. Additionally, multiple lines of BLGMDM2 transgenic mice developed mammary tumors, confirming that overproduction of MDM2 contributes to tumorigenesis in epithelial cells in vivo.