Examination of X chromosome markers in Rett syndrome: exclusion mapping with a novel variation on multilocus linkage analysis.

Rett syndrome is a neurologic disorder characterized by early normal development followed by regression, acquired deceleration of head growth, autism, ataxia, and stereotypic hand movements. The exclusive occurrence of the syndrome in females and the occurrence of a few familial cases with inheritance through maternal lines suggest that this disorder is most likely secondary to a mutation on the X chromosome. To address this hypothesis and to identify candidate regions for the Rett syndrome gene locus, genotypic analysis was performed in two families with maternally related affected half-sisters by using 63 DNA markers from the X chromosome. Maternal and paternal X chromosomes from the affected sisters were separated in somatic cell hybrids and were examined for concordance/discordance of maternal alleles at the tested loci. Thirty-six markers were informative in at least one of the two families, and 25 markers were informative in both families. Twenty loci were excluded as candidates for the Rett syndrome gene, on the basis of discordance for maternal alleles in the half-sisters. Nineteen of the loci studied were chosen for multipoint linkage analysis because they have been previously genetically mapped using a large number of meioses from reference families. Using the exclusion criterion of a lod score less than -2, we were able to exclude the region between the Duchenne muscular dystrophy locus and the DXS456 locus. This region extends from Xp21.2 to Xq21-q23. The use of the multipoint linkage analysis approach outlined in this study should allow the exclusion of additional regions of the X chromosome as new markers are analyzed. This in turn will result in a defined region of the X chromosome that should be searched for candidate sequences for the Rett syndrome gene in both familial and sporadic cases.     

Generating social network data using partially described networks: an example informing avian influenza control in the British poultry industry

Background

The United Kingdom (UK) government has been recording the births, deaths, and movements of cattle for the last decade. Despite reservations about the accuracy of these data, they represent a large and valuable body of information about the demographics of the UK cattle herd and its contact structure. In this article, a range of demographic data about UK cattle, and particularly their movements, are presented, as well as yearly trends in the patterns of movements.

Results

A clear seasonal pattern is evident in the number of movements of cattle, as are the reductions in movement volume due to foot and mouth disease outbreaks in 2001 and 2007. The distribution of ages of cattle at their time of death is multimodal, and the impact of the over thirty months rule is marked. Most movements occur between agricultural holdings, markets, and slaughterhouses, and there is a non-random pattern to the types of holdings movements occur between. Most animals move only a short distance and a few times in their life. Most movements between any given pair of holdings only occurred once in the last 10 years, but about a third occurred between 2 and 10 times in that period. There is no clear trend to movement patterns in the UK since 2002.

Conclusions

Despite a substantial number of regulatory interventions during the last decade, movement patterns show no clear trend since 2002. The observed patterns in the repeatability of movements, the types of holdings involved in movements, the distances and frequencies of cattle movements, and the batch sizes involved give an insight into the structure of the UK cattle industry, and could act as the basis for a predictive model of livestock movements in the UK.     

Characterization of I-Ppo, an intron-encoded endonuclease that mediates homing of a group I intron in the ribosomal DNA of Physarum polycephalum.

A novel and only recently recognized class of enzymes is composed of the site-specific endonucleases encoded by some group I introns. We have characterized several aspects of I-Ppo, the endonuclease that mediates the mobility of intron 3 in the ribosomal DNA of Physarum polycephalum. This intron is unique among mobile group I introns in that it is located in nuclear DNA. We found that I-Ppo is encoded by an open reading frame in the 5' half of intron 3, upstream of the sequences required for self-splicing of group I introns. Either of two AUG initiation codons could start this reading frame, one near the beginning of the intron and the other in the upstream exon, leading to predicted polypeptides of 138 and 160 amino acid residues. The longer polypeptide was the major form translated in vitro in a reticulocyte extract. From nuclease assays of proteins synthesized in vitro with partially deleted DNAs, we conclude that both polypeptides possess endonuclease activity. We also have expressed I-Ppo in Escherichia coli, using a bacteriophage T7 RNA polymerase expression system. The longer polypeptide also was the predominant form made in this system. It showed enzymatic activity in bacteria in vivo, as demonstrated by the cleavage of a plasmid carrying the target site. Like several other intron-encoded endonucleases, I-Ppo makes a four-base staggered cut in its ribosomal DNA target sequence, very near the site where intron 3 becomes integrated in crosses of intron 3-containing and intron 3-lacking Physarum strains.     

The SARS-CoV2 envelope differs from host cells,exposes procoagulant lipids,and is disrupted in vivo by oral rinses

The lipid envelope of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an essential component of the virus; however, its molecular composition is undetermined. Addressing this knowledge gap could support the design of antiviral agents as well as further our understanding of viral-host protein interactions, infectivity, pathogenicity, and innate immune system clearance. Lipidomics revealed that the virus envelope comprised mainly phospholipids (PLs), with some cholesterol and sphingolipids, and with cholesterol/phospholipid ratio similar to lysosomes. Unlike cellular membranes, procoagulant amino-PLs were present on the external side of the viral envelope at levels exceeding those on activated platelets. Accordingly, virions directly promoted blood coagulation. To investigate whether these differences could enable selective targeting of the viral envelope in vivo, we tested whether oral rinses containing lipid-disrupting chemicals could reduce infectivity. Products containing PL-disrupting surfactants (such as cetylpyridinium chloride) met European virucidal standards in vitro; however, components that altered the critical micelle concentration reduced efficacy, and products containing essential oils, povidone-iodine, or chlorhexidine were ineffective. This result was recapitulated in vivo, where a 30-s oral rinse with cetylpyridinium chloride mouthwash eliminated live virus in the oral cavity of patients with coronavirus disease 19 for at least 1 h, whereas povidone-iodine and saline mouthwashes were ineffective. We conclude that the SARS-CoV-2 lipid envelope i) is distinct from the host plasma membrane, which may enable design of selective antiviral approaches; ii) contains exposed phosphatidylethanolamine and phosphatidylserine, which may influence thrombosis, pathogenicity, and inflammation; and iii) can be selectively targeted in vivo by specific oral rinses.     

Girl helpers and time allocation of nursing women among the Toba of Argentina

In this paper we outline the activities of young girls in a Toba community of northern Argentina and examine the effect of girl helpers on time allocation of nursing women. Activity budgets were obtained for 41 girls aged 3 to 15 using spot observations. Girls spent substantial portions of observations engaged in helping behaviors. Individual values varied with age, anthropometric characteristics, and birth order. Activity budgets of 21 nursing women were obtained through focal observation sessions. Women living in households with girls aged 7 to 15 allocated 17% less time to domestic work and 9% more time to socializing during afternoon observation sessions. For nursing women in this community, direct childcare (provided by the infant’s own mother) seemed to be a priority. Living with a girl helper did not have any measurable effect on the frequency or duration of nursing, or on the time that women spent caring for their infants. Based on these findings, hypotheses are outlined for future work on the effect of girl helpers on women’s fertility. Research for this project was funded by the Nestlé Foundation, the Harvard Anthropology Goelet Fund, and the Harvard College Research Program. Eduardo Fernandez-Duque commented on an early draft of this paper. Riley Bove (A.B., Harvard, 1998) is a research associate in the Department of Anthropology at Harvard University with research interests in intergenerational female networks and polygyny as a social mediator of women’s health (esp. urban, African). Claudia Valeggia (Ph.D., U.C. Davis, 1996) is a postdoctoral fellow in the Department of Anthropology at Harvard University with research interests in reproductive ecology and ethnopediatry. Peter Ellison (Ph.D., Harvard, 1983) is a professor of anthropology and director of the Reproductive Ecology Laboratory at Harvard University. He is the author of On Fertile Ground: Ecology, Evolution and Human Reproduction (Harvard University Press, 2001).     

The Cultural Evolution of Human Communication Systems in Different Sized Populations: Usability Trumps Learnability

This study examines the intergenerational transfer of human communication systems. It tests if human communication systems evolve to be easy to learn or easy to use (or both), and how population size affects learnability and usability. Using an experimental-semiotic task, we find that human communication systems evolve to be easier to use (production efficiency and reproduction fidelity), but harder to learn (identification accuracy) for a second generation of naïve participants. Thus, usability trumps learnability. In addition, the communication systems that evolve in larger populations exhibit distinct advantages over those that evolve in smaller populations: the learnability loss (from the Initial signs) is more muted and the usability benefits are more pronounced. The usability benefits for human communication systems that evolve in a small and large population is explained through guided variation reducing sign complexity. The enhanced performance of the communication systems that evolve in larger populations is explained by the operation of a content bias acting on the larger pool of competing signs. The content bias selects for information-efficient iconic signs that aid learnability and enhance usability.     

An international comparability study on quantification of total methyl cytosine content

A novel biocomposite film (MWCNTs-PNDGAChi), which contains multiwalled carbon nanotubes (MWCNTs) along with the incorporation of poly(nordihydroguaiaretic acid) and chitosan copolymer (PNDGAChi), has been synthesized on gold electrode by potentiostatic methods. The presence of MWCNTs in the biocomposite film enhances PNDGAChi’s surface coverage concentration (Γ) on the electrode and decreases degradation of PNDGAChi during cycling. The biocomposite film also exhibits promising enhanced electrocatalytic activity toward the oxidation of biochemical compounds such as epinephrine (EP) and norepinephrine (NEP). Cyclic voltammetry was used for the measurement of electroanalytical properties of analytes by means of MWCNTs-PNDGAChi biocomposite film modified gold electrode. The sensitivity values of MWCNTs-PNDGAChi biocomposite film modified gold electrode are higher than the values obtained for PNDGAChi film modified gold electrode. Electrochemical quartz crystal microbalance studies reveal the enhancements in the functional properties of MWCNTs and PNDGAChi present in MWCNTs-PNDGAChi biocomposite film. Surface morphology of the biocomposite films was studied using scanning electron microscopy, atomic force microscopy, and scanning tunneling microscopy. The surface morphology results reveal that PNDGAChi incorporated on MWCNTs. Finally, flow injection analysis was used for the amperometric detection of EP and NEP at MWCNTs-PNDGAChi film modified screen printed carbon electrode.     

Structure of a conjugating enzyme-ubiquitin thiolester intermediate reveals a novel role for the ubiquitin tail

BACKGROUND: Ubiquitin-conjugating enzymes (E2s) are central enzymes involved in ubiquitin-mediated protein degradation. During this process, ubiquitin (Ub) and the E2 protein form an unstable E2-Ub thiolester intermediate prior to the transfer of ubiquitin to an E3-ligase protein and the labeling of a substrate for degradation. A series of complex interactions occur among the target substrate, ubiquitin, E2, and E3 in order to efficiently facilitate the transfer of the ubiquitin molecule. However, due to the inherent instability of the E2-Ub thiolester, the structural details of this complex intermediate are not known. RESULTS: A three-dimensional model of the E2-Ub thiolester intermediate has been determined for the catalytic domain of the E2 protein Ubc1 (Ubc1(Delta450)) and ubiquitin from S. cerevisiae. The interface of the E2-Ub intermediate was determined by kinetically monitoring thiolester formation by 1H-(15)N HSQC spectra by using combinations of 15N-labeled and unlabeled Ubc1(Delta450) and Ub proteins. By using the surface interface as a guide and the X-ray structures of Ub and the 1.9 A structure of Ubc1(Delta450) determined here, docking simulations followed by energy minimization were used to produce the first model of a E2-Ub thiolester intermediate. CONCLUSIONS: Complementary surfaces were found on the E2 and Ub proteins whereby the C terminus of Ub wraps around the E2 protein terminating in the thiolester between C88 (Ubc1(Delta450)) and G76 (Ub). The model supports in vivo and in vitro experiments of E2 derivatives carrying surface residue substitutions. Furthermore, the model provides insights into the arrangement of Ub, E2, and E3 within a ternary targeting complex.