Uniparental disomy screen of Irish rare disorder cohort unmasks homozygous variants of clinical significance in the TMCO1 and PRKRA genes.

A uniparental disomy (UPD) screen using whole genome sequencing (WGS) data from 164 trios with rare disorders in the Irish population was performed to identify large runs of homozygosity of uniparental origin that may harbour deleterious recessive variants. Three instances of whole chromosome uniparental isodisomy (UPiD) were identified: one case of maternal isodisomy of chromosome 1 and two cases of paternal isodisomy of chromosome 2. We identified deleterious homozygous variants on isodisomic chromosomes in two probands: a novel p (Glu59ValfsTer20) variant in TMCO1, and a p (Pro222Leu) variant in PRKRA, respectively. The overall prevalence of whole chromosome UPiD in our cohort was 1 in 55 births, compared to 1 in ∼7,500 births in the general population, suggesting a higher frequency of UPiD in rare disease cohorts. As a distinct mechanism underlying homozygosity compared to biallelic inheritance, the identification of UPiD has important implications for family planning and cascade testing. Our study demonstrates that UPD screening may improve diagnostic yields by prioritising UPiD chromosomes during WGS analysis.

Adsorption of Small Cationic Nanoparticles onto Large Anionic Particles from Aqueous Solution: A Model System for Understanding Pigment Dispersion and the Problem of Effective Particle Density.

The present study focuses on the use of copolymer nanoparticles as a dispersant for a model pigment (silica). Reversible addition-fragmentation chain transfer (RAFT) alcoholic dispersion polymerization was used to synthesize sterically stabilized diblock copolymer nanoparticles. The steric stabilizer block was poly(2-(dimethylamino)ethyl methacrylate) (PDMA) and the core-forming block was poly(benzyl methacrylate) (PBzMA). The mean degrees of polymerization for the PDMA and PBzMA blocks were 71 and 100, respectively. Transmission electron microscopy (TEM) studies confirmed a near-monodisperse spherical morphology, while dynamic light scattering (DLS) studies indicated an intensity-average diameter of 30 nm. Small-angle X-ray scattering (SAXS) reported a volume-average diameter of 29 ± 0.5 nm and a mean aggregation number of 154. Aqueous electrophoresis measurements confirmed that these PDMA71-PBzMA100 nanoparticles acquired cationic character when transferred from ethanol to water as a result of protonation of the weakly basic PDMA chains. Electrostatic adsorption of these nanoparticles from aqueous solution onto 470 nm silica particles led to either flocculation at submonolayer coverage or steric stabilization at or above monolayer coverage, as judged by DLS. This technique indicated that saturation coverage was achieved on addition of approximately 465 copolymer nanoparticles per silica particle, which corresponds to a fractional surface coverage of around 0.42. These adsorption data were corroborated using thermogravimetry, UV spectroscopy and X-ray photoelectron spectroscopy. TEM studies indicated that the cationic nanoparticles remained intact on the silica surface after electrostatic adsorption, while aqueous electrophoresis confirmed that surface charge reversal occurred below pH 7. The relatively thick layer of adsorbed nanoparticles led to a significant reduction in the effective particle density of the silica particles from 1.99 g cm-3 to approximately 1.74 g cm-3, as judged by disk centrifuge photosedimentometry (DCP). Combining the DCP and SAXS data suggests that essentially no deformation of the PBzMA cores occurs during nanoparticle adsorption onto the silica particles.

The genetics of an early Neolithic pastoralist from the Zagros, Iran.

The agricultural transition profoundly changed human societies. We sequenced and analysed the first genome (1.39x) of an early Neolithic woman from Ganj Dareh, in the Zagros Mountains of Iran, a site with early evidence for an economy based on goat herding, ca. 10,000 BP. We show that Western Iran was inhabited by a population genetically most similar to hunter-gatherers from the Caucasus, but distinct from the Neolithic Anatolian people who later brought food production into Europe. The inhabitants of Ganj Dareh made little direct genetic contribution to modern European populations, suggesting those of the Central Zagros were somewhat isolated from other populations of the Fertile Crescent. Runs of homozygosity are of a similar length to those from Neolithic farmers, and shorter than those of Caucasus and Western Hunter-Gatherers, suggesting that the inhabitants of Ganj Dareh did not undergo the large population bottleneck suffered by their northern neighbours. While some degree of cultural diffusion between Anatolia, Western Iran and other neighbouring regions is possible, the genetic dissimilarity between early Anatolian farmers and the inhabitants of Ganj Dareh supports a model in which Neolithic societies in these areas were distinct.

How Do Spherical Diblock Copolymer Nanoparticles Grow during RAFT Alcoholic Dispersion Polymerization?

A poly(2-(dimethylamino)ethyl methacrylate) (PDMA) chain transfer agent (CTA) is used for the reversible addition-fragmentation chain transfer (RAFT) alcoholic dispersion polymerization of benzyl methacrylate (BzMA) in ethanol at 70 °C. THF GPC analysis indicated a well-controlled polymerization with molecular weight increasing linearly with conversion. GPC traces also showed high blocking efficiency with no homopolymer contamination apparent and Mw/Mn values below 1.35 in all cases. 1H NMR studies confirmed greater than 98% BzMA conversion for a target PBzMA degree of polymerization (DP) of up to 600. The PBzMA block becomes insoluble as it grows, leading to the in situ formation of sterically stabilized diblock copolymer nanoparticles via polymerization-induced self-assembly (PISA). Fixing the mean DP of the PDMA stabilizer block at 94 units and systematically varying the DP of the PBzMA block enabled a series of spherical nanoparticles of tunable diameter to be obtained. These nanoparticles were characterized by TEM, DLS, MALLS, and SAXS, with mean diameters ranging from 35 to 100 nm. The latter technique was particularly informative: data fits to a spherical micelle model enabled calculation of the core diameter, surface area occupied per copolymer chain, and the mean aggregation number (Nagg). The scaling exponent derived from a double-logarithmic plot of core diameter vs PBzMA DP suggests that the conformation of the PBzMA chains is intermediate between the collapsed and fully extended state. This is in good agreement with 1H NMR studies, which suggest that only 5-13% of the BzMA residues of the core-forming chains are solvated. The Nagg values calculated from SAXS and MALLS are in good agreement and scale approximately linearly with PBzMA DP. This suggests that spherical micelles grow in size not only as a result of the increase in copolymer molecular weight during the PISA synthesis but also by exchange of individual copolymer chains between micelles and/or by sphere-sphere fusion events.

Ancient Ethiopian genome reveals extensive Eurasian admixture throughout the African continent.

Characterizing genetic diversity in Africa is a crucial step for most analyses reconstructing the evolutionary history of anatomically modern humans. However, historic migrations from Eurasia into Africa have affected many contemporary populations, confounding inferences. Here, we present a 12.5× coverage ancient genome of an Ethiopian male ("Mota") who lived approximately 4500 years ago. We use this genome to demonstrate that the Eurasian backflow into Africa came from a population closely related to Early Neolithic farmers, who had colonized Europe 4000 years earlier. The extent of this backflow was much greater than previously reported, reaching all the way to Central, West, and Southern Africa, affecting even populations such as Yoruba and Mbuti, previously thought to be relatively unadmixed, who harbor 6 to 7% Eurasian ancestry.

MMP and TIMP temporal gene expression during osteocytogenesis.

Osteocytes within bone differentiate from osteoblast precursors which reside in a mineralised extracellular matrix (ECM). Fully differentiated osteocytes are critical for bone development and function but the factors that regulate this differentiation process are unknown. The enzymes primarily responsible for ECM remodelling are matrix metalloproteinases (MMP); however, the expression and role of MMPs during osteocytogenesis is undefined. Here we used MLO-A5 cells to determine the temporal gene expressions of the MMP family and their endogenous inhibitors--tissue inhibitors of metalloproteinases (TIMPs) during osteocytogenesis. RT-qPCR revealed expression of 14 Mmps and 3 Timps in MLO-A5 cells. Mmp2, Mmp23 and Mmp28 were decreased concurrent with mineralisation onset (P < 0.05*). Mmp14 and Mmp19 mRNAs were also significantly increased at day 3 (P < 0.05*) before returning to baseline levels at day 6. Decreased expressions of Timp1, Timp2 and Timp3 mRNA were observed by day 6 compared to day 0 (P < 0.05*). To examine whether these changes are linked to osteocytogenesis, we determined Mmp/Timp mRNA expressions in mineralisation-limited conditions. RT-qPCR revealed that the previously observed decreases in Mmp2, Mmp23 and Mmp28 were not observed in these mineralisation-limited cultures, therefore closely linking these MMPs with osteocyte differentiation. Similarly, we found differential expression of Timp1, Timp2 and Timp3 mRNA in mineralisation-restricted cultures (P < 0.05*). In conclusion, we have identified several members of the MMP/TIMP families as regulators of ECM remodelling necessary for the acquisition of the osteocyte phenotype.

Are block copolymer worms more effective Pickering emulsifiers than block copolymer spheres?

RAFT-mediated polymerisation-induced self-assembly (PISA) is used to prepare six types of amphiphilic block copolymer nanoparticles which were subsequently evaluated as putative Pickering emulsifiers for the stabilisation of n-dodecane-in-water emulsions. It was found that linear poly(glycerol monomethacrylate)-poly(2-hydroxypropyl methacrylate) (PGMA-PHPMA) diblock copolymer spheres and worms do not survive the high shear homogenisation conditions used for emulsification. Stable emulsions are obtained, but the copolymer acts as a polymeric surfactant; individual chains rather than particles are adsorbed at the oil-water interface. Particle dissociation during emulsification is attributed to the weakly hydrophobic character of the PHPMA block. Covalent stabilisation of these copolymer spheres or worms can be readily achieved by addition of ethylene glycol dimethacrylate (EGDMA) during the PISA synthesis. TEM studies confirm that the resulting cross-linked spherical or worm-like nanoparticles survive emulsification and produce genuine Pickering emulsions. Alternatively, stabilisation can be achieved by either replacing or supplementing the PHPMA block with the more hydrophobic poly(benzyl methacrylate) (PBzMA). The resulting linear spheres or worms also survive emulsification and produce stable n-dodecane-in-water Pickering emulsions. The intrinsic advantages of anisotropic worms over isotropic spheres for the preparation of Pickering emulsions are highlighted. The former particles are more strongly adsorbed at similar efficiencies compared to spheres and also enable smaller oil droplets to be produced for a given copolymer concentration. The scalable nature of PISA formulations augurs well for potential applications of anisotropic block copolymer nanoparticles as Pickering emulsifiers.

A ten-year review of the validity and clinical utility of depression screening.

OBJECTIVE: Use of depression screening instruments in primary care is controversial. The authors reviewed research studies published since the development of national practice guidelines to determine whether new evidence might favor screening. The review focused on evidence-related validity and clinical utility of depression screening instruments. METHODS: Silver Platter MEDLINE was searched for English-language studies of depression screening instruments published between 1986 and 1995. Studies were classified by type--reviews of studies, outcome studies, validation studies. RESULTS AND CONCLUSIONS: Fifty-nine studies met criteria for review. Validation studies were the most frequent type (39 studies) and were subclassified according to population, type of comparison, and analytical method. These studies documented the validity of screening instruments compared with formal criteria and demonstrated consistently better performance for systematic approaches compared with clinical impressions. Thirteen studies were reviews; those reviewing evidence for effectiveness disagreed in their conclusions. Only seven outcome studies related to depression screening instruments were found, and none showed measurable benefit in a screened population. Several studies showed that very brief instruments performed about as well as longer, well-validated questionnaires for screening in general populations.

The magnitude of metabolic acidosis is dependent on differences in bicarbonate assays.

Metabolic acidosis has been recently recognized as an important comorbid event in the high mortality rates seen in patients with end-stage renal disease. The recognition of hypobicarbonatemia is dependent on a reliable assay for total carbon dioxide (TCO2). It is common practice for dialysis facilities to send blood samples for testing to remote laboratories, which may assay bicarbonate differently than the local hospital. We noted that serum bicarbonate concentrations from blood samples sent to our reference laboratory were significantly lower (4 mEq/L) compared with blood samples sent to our local laboratory. Blood samples were assayed for TCO2 using an enzymatic technique (in the reference laboratory) and direct measurement using an electrode (in the local laboratory). The blood test results for TCO2 sent to the reference laboratory (18.7 +/- 0.8 mEq/L) were significantly lower than samples assayed in our local laboratory (22.2 +/- 0.7 mEq/L). In conclusion, recognition of the differences in assays used in the laboratory for routine bicarbonate measurements is important in defining the magnitude of metabolic acidosis and in helping to dictate appropriate therapy.

A pragmatist's approach to pathology costing: the Welsh Datatree project.

The Datatree costing project in Wales has provided the Welsh pathology laboratories with a standard costing package that allows pathologists to understand how their own laboratory's test costs are compiled. The software provides answers to the question "what if? ..." and shows instantly the effect of salary or consumable cost alterations. Resource management at a laboratory level is enhanced by a greater knowledge of costs, particularly in relation to volumes of work. Perhaps this is one of the stepping-stones across the river to the "open market." In the United Kingdom NHS any information of this kind must be regarded as invaluable.

Early English steroid history.

Following the revelation of the presence of vitamin D in fish liver oils and of estrogenic hormones in pregnancy urine in the 1920s, active interest in the steroids began in England. Most of this interest originated from the studies of Ian Heilbron at Liverpool and of Otto Rosenheim at the National Institute for Medical Research in London.

Case study. Erythropoietin for anaemia in a preterm Jehovah's Witness baby.

An infant born at 24 weeks gestation to Jehovah's Witness parents was made a Ward of Court and treated against their wishes with blood products. Erythropoietin was used without obvious benefit, but the child did well. The parents did not reject the child and maintained a good relationship with medical and nursing staff. We present this case in the light of current discussions on child welfare and recent reform of the law relating to child protection and highlight the many difficult dilemmas faced by the medical team.

Attitudes of paediatricians to HIV and hepatitis B virus infection.

There is a recent upsurge of interest among health care professionals regarding the risk of accidental occupational exposure to HIV virus. We evaluated knowledge of virus carriage prevalence, needlestick injuries, venepuncture practices, and glove use among paediatricians in Wales and South West Regional Health Authorities. We also attempted to evaluate hepatitis B immunisation uptake in this group. Paediatricians have traditionally been considered a low risk group in the context of accidental occupational exposure to these viruses. We targeted a four point questionnaire at 221 paediatricians. Results suggested that despite recent increasing concern about these viruses, that is reflected in the amount of medical literature recently published, and the issuing of Department of Health guidelines on venepuncture, knowledge of prevalence of HIV and hepatitis B carriage rates, and hence assessment of risk magnitude, was surprisingly poor. Safe venepuncture practices were not widely used. In the 12 months before receiving the questionnaire 55% had suffered a needlestick injury with only 10% reporting the fact. Hepatitis B immunisation uptake was highest in the junior grades (but this does not necessarily mean those at greatest risk). There were many inconsistencies between the clinicians' perceptions of risk and their practices. As the virus attains a firm hold in the heterosexual population paediatricians by virtue of the nature of venepuncture in children will almost certainly see their risk of acquiring HIV/hepatis B viruses secondary to accidental occupational exposure increase over the next decade. Without an improvement in current knowledge of carriage prevalence in high risk areas and alteration in venepuncture practices/hepatitis B immunisation uptake some will unfortunately, though avoidably, contract these bloodborne viral infections.