Seven cases of probable endotoxin poisoning related to contaminated glutathione infusions.

We report seven cases of probable endotoxin poisoning linked to contaminated compounded glutathione. Five of the cases were using the infusions for treatment of Lyme disease highlighting the risks of using compounded sterile preparations for unapproved indications, especially if the quality of source products cannot be assured.

A genome-based model for adjusting radiotherapy dose (GARD): a retrospective, cohort-based study.

BACKGROUND: Despite its common use in cancer treatment, radiotherapy has not yet entered the era of precision medicine, and there have been no approaches to adjust dose based on biological differences between or within tumours. We aimed to assess whether a patient-specific molecular signature of radiation sensitivity could be used to identify the optimum radiotherapy dose. METHODS: We used the gene-expression-based radiation-sensitivity index and the linear quadratic model to derive the genomic-adjusted radiation dose (GARD). A high GARD value predicts for high therapeutic effect for radiotherapy; which we postulate would relate to clinical outcome. Using data from the prospective, observational Total Cancer Care (TCC) protocol, we calculated GARD for primary tumours from 20 disease sites treated using standard radiotherapy doses for each disease type. We also used multivariable Cox modelling to assess whether GARD was independently associated with clinical outcome in five clinical cohorts: Erasmus Breast Cancer Cohort (n=263); Karolinska Breast Cancer Cohort (n=77); Moffitt Lung Cancer Cohort (n=60); Moffitt Pancreas Cancer Cohort (n=40); and The Cancer Genome Atlas Glioblastoma Patient Cohort (n=98). FINDINGS: We calculated GARD for 8271 tissue samples from the TCC cohort. There was a wide range of GARD values (range 1·66-172·4) across the TCC cohort despite assignment of uniform radiotherapy doses within disease types. Median GARD values were lowest for gliomas and sarcomas and highest for cervical cancer and oropharyngeal head and neck cancer. There was a wide range of GARD values within tumour type groups. GARD independently predicted clinical outcome in breast cancer, lung cancer, glioblastoma, and pancreatic cancer. In the Erasmus Breast Cancer Cohort, 5-year distant-metastasis-free survival was longer in patients with high GARD values than in those with low GARD values (hazard ratio 2·11, 95% 1·13-3·94, p=0·018). INTERPRETATION: A GARD-based clinical model could allow the individualisation of radiotherapy dose to tumour radiosensitivity and could provide a framework to design genomically-guided clinical trials in radiation oncology. FUNDING: None.

Shrubline but not treeline advance matches climate velocity in montane ecosystems of south-central Alaska.

Tall shrubs and trees are advancing into many tundra and wetland ecosystems but at a rate that often falls short of that predicted due to climate change. For forest, tall shrub, and tundra ecosystems in two pristine mountain ranges of Alaska, we apply a Bayesian, error-propagated calculation of expected elevational rise (climate velocity), observed rise (biotic velocity), and their difference (biotic inertia). We show a sensitive dependence of climate velocity on lapse rate and derive biotic velocity as a rigid elevational shift. Ecosystem presence identified from recent and historic orthophotos ~50 years apart was regressed on elevation. Biotic velocity was estimated as the difference between critical point elevations of recent and historic logistic fits divided by time between imagery. For both mountain ranges, the 95% highest posterior density of climate velocity enclosed the posterior distributions of all biotic velocities. In the Kenai Mountains, mean tall shrub and climate velocities were both 2.8 m y(-1). In the better sampled Chugach Mountains, mean tundra retreat was 1.2 m y(-1) and climate velocity 1.3 m y(-1). In each mountain range, the posterior mode of tall woody vegetation velocity (the complement of tundra) matched climate velocity better than either forest or tall shrub alone, suggesting competitive compensation can be important. Forest velocity was consistently low at 0.1-1.1 m y(-1), indicating treeline is advancing slowly. We hypothesize that the high biotic inertia of forest ecosystems in south-central Alaska may be due to competition with tall shrubs and/or more complex climate controls on the elevational limits of trees than tall shrubs. Among tall shrubs, those that disperse farthest had lowest inertia. Finally, the rapid upward advance of woody vegetation may be contributing to regional declines in Dall's sheep (Ovis dalli), a poorly dispersing alpine specialist herbivore with substantial biotic inertia due to dispersal reluctance.

Analysis of liquid metal foams through X-ray radioscopy and microgravity experiments.

Complex liquid structures such as metallic foams were produced in a furnace that allowed in situ X-ray monitoring of the evolution of the structure and distribution of the liquid in the foam. The experiments were carried out during parabolic flights which provided varying levels of gravity. The evolution of the characteristic liquid fraction profiles due to gravity induced drainage was measured and analysed in terms of the foam drainage equation, obtaining viscosity and surface tension by fitting solutions of the equation to the experimental data. The surface tension of the melt in the foam was decreased up to 40%. Effective viscosities of up to 139 times the viscosity of a pure bulk melt were observed. These effects could be attributed to the smaller influence of solid particles dispersed in the melt and the larger influence of the complex foam structure.

Partial antagonism of propofol anaesthesia by physostigmine in rats is associated with potentiation of fast (80-200 Hz) oscillations in the thalamus.

BACKGROUND: Positron emission tomography studies in human subjects show that propofol-induced unconsciousness in humans is associated with a reduction in thalamic blood flow, suggesting that anaesthesia is associated with impairment of thalamic function. A recent study showed that antagonism of propofol-induced unconsciousness by the anticholinesterase physostigmine is associated with a marked increase in thalamic blood flow, supporting the implication of the thalamus. The aim of the present study was to assess the role of the thalamus in the antagonistic effects of physostigmine during propofol anaesthesia using electrophysiological recordings in a rat model. METHODS: Local field potentials were recorded from the barrel cortex and ventroposteromedial thalamic nucleus in 10 chronically instrumented rats to measure spectral power in the gamma/high-gamma range (50-200 Hz). Propofol was given i.v. by target-controlled infusion at the lowest concentration required to abolish righting attempts. Physostigmine was given during anaesthesia to produce behavioural arousal without changing anaesthetic concentration. RESULTS: Compared with baseline, gamma/high-gamma power during anaesthesia was reduced by 31% in the cortex (P=0.006) and by 65% in the thalamus (P=0.006). Physostigmine given during anaesthesia increased gamma/high-gamma power in the thalamus by 60% (P=0.048) and caused behavioural arousal that correlated (P=0.0087) with the increase in power. Physostigmine caused no significant power change in the cortex. CONCLUSIONS: We conclude that partial antagonism of propofol anaesthesia by physostigmine is associated with an increase in thalamic activity reflected in gamma/high-gamma (50-200 Hz) power. These findings are consistent with the view that anaesthetic-induced unconsciousness is associated with impairment of thalamic function.

Neonatal pulse oximetry screening: a national survey.

AIM: Studies suggest that universal pulse oximetry (PO) screening of all neonates before hospital discharge improves the detection rate of congenital heart disease (CHD). The aim is to survey the use of Pulse Oximetry (PO) screening in UK. METHODS: A telephone interview was conducted between late 2009 to mid-2010 of all maternity units using a standardised questionnaire. RESULTS: 209(93%) of 224 responding units did not routinely use PO. Among the 15 that performed PO, 5 measured pre and post ductal saturations, 9 measured only post-ductal saturations, and 1 measured only pre-ductal saturations. There were differences in the values used to trigger further investigation, ranging from <94% to <96% and/or difference of > 2-3% between pre and post-ductal saturations. When saturations were abnormal, 13 units performed echocardiography locally. In addition to an echocardiogram, 2 units performed chest x-ray (CXR); 2 units performed electrocardiogram (ECG) and 2 units performed both CXR and ECG. CONCLUSION: Only a minority of hospitals across the UK use PO to supplement the postnatal examination with inconsistent practice. National guidelines should be developed if PO screening is implemented with an agreed management plan if abnormal results are obtained.

Primary standardisation of technetium-99m by liquid scintillation coincidence counting.

Technetium-99m was standardised by the 4π(LS)ce-γ coincidence extrapolation method. Sensitivity of the 4π(LS) channel to two types of radiation, namely conversion electrons and γ-rays, resulted in incorrect activity values being obtained when this was not adequately accounted for. Measurements were more robust when the LS detection efficiency was optimised, and when a γ-window setting was used that monitored the combined LS efficiency for conversion electrons and γ-rays. The primary standard was internationally compared through participation in the BIPM.RI(II)-K4.Tc99m key comparison.

Alteration by heat shock and immunological characterization of Drosophila small nuclear ribonucleoproteins.

Sera from human patients with systemic lupus erythematosus (SLE) have been shown to react with snRNP particles of both mammals and Drosophila (Mount, S. M. and J. A. Steitz. 1981. Nucleic Acids Res. 9:6351-6368). We have utilized fully characterized monospecific sera and specifically purified antibodies to carry out indirect immunofluorescence experiments with frozen sections of Drosophila embryos. Embryos subjected to severe heat shock before sectioning showed reduced binding of anti-Sm sera. Anti-nRNP sera reacted identically with antigens of heat shocked and non-heat-shocked sections. The reduction in anti-Sm fluorescence was restored by a brief salt wash. These results imply a noncovalent alteration in the conformation of Sm antigens with the administration of heat shock that can revert with exposure to salt. Drosophila antigens have been compared to mammalian standards, showing partial identity with bovine spleen extract (BSE) antigens when reacted with anti-Sm sera. The antigenic relatedness between affinity-purified heat-shocked and non-heat-shocked Drosophila antigens and their mammalian homologues was examined by quantitative ELISA methodology. In all cases, the Drosophila antigens from heat-shocked and non-heat-shocked embryos were identical. We theorize that the heat shock-induced alteration of Sm antigen reverst during extraction. Because the snRNP antigens have been shown to be involved in splicing, and because splicing is inhibited during heat shock (Yost, H. J., and S. Lindquist. 1986. Cell. 45:185-193), our results provide information on the nature and stability of a change in these antigens which may be a central element in control of the heat shock response.

Development of the Australian Standard for Germanium-68 by two Liquid Scintillation Counting methods.

In response to the increasing application of 68Ge/68Ga and 68Ga in nuclear medicine, an international comparison of activity measurement of 68Ge in equilibrium with 68Ga was organised. ANSTO standardised the comparison solution by the 4π(LS)ß+-γ coincidence extrapolation and TDCR efficiency calculation methods, with excellent agreement between the two results. The primary standard was transferred to the ANSTO Secondary Standard Ionisation Chamber. Internationally traceable Australian Certified Reference Materials (ACRMs) of 68Ge/68Ga can now be prepared in various measurement geometries applied in nuclear medicine.

The contribution of GJB2 mutations to slight or mild hearing loss in Australian elementary school children.

BACKGROUND: There is a lack of information on prevalence, cause and consequences of slight/mild bilateral sensorineural hearing loss (SNHL) in children. We report the first systematic genetic analysis of the GJB2 gene in a population-derived sample of children with slight/mild bilateral SNHL. METHODS: Hearing tests were conducted in 6240 Australian elementary school children in Grades 1 and 5. 55 children (0.88%) were found to have a slight/mild sensorineural hearing loss. 48 children with slight/mild sensorineural hearing loss and a matched group of 90 children with normal hearing participated in a genetic study investigating mutations in the GJB2 gene, coding for connexin 26, and the presence of the del(GJB6-D13S1830) and del(GJB6-D13S1854) deletions in the GJB6 gene, coding for connexin 30. RESULTS: Four of 48 children with slight/mild sensorineural hearing loss were homozygous for the GJB2 V37I change. The four children with homozygous V37I mutations were all of Asian background and analysis of SNPs in or near the GJB2 gene suggests that the V37I mutation arose from a single mutational event in the Asian population. DISCUSSION: Based on the prevalence of carriers of this change we conclude that V37I can be a causative mutation that is often associated with slight/mild sensorineural hearing loss. No other children in the slight/mild hearing loss group had a hearing loss related to a GJB2 mutation. One child with normal hearing was homozygous for the R127H change and we conclude that this change does not cause hearing loss. Two children of Asian background were carriers of the V37I mutation. Our data indicate that slight/mild sensorineural hearing loss due to the GJB2 V37I mutation is common in people of Asian background.

Heart Failure and MEF2 Transcriptome Dynamics in Response to ß-Blockers.

Myocyte Enhancer Factor 2 (MEF2) mediates cardiac remodelling in heart failure (HF) and is also a target of ß-adrenergic signalling, a front-line treatment for HF. We identified global gene transcription networks involved in HF with and without ß-blocker treatment. Experimental HF by transverse aortic constriction (TAC) in a MEF2 "sensor" mouse model (6 weeks) was followed by four weeks of ß-blockade with Atenolol (AT) or Solvent (Sol) treatment. Transcriptome analysis (RNA-seq) from left ventricular RNA samples and MEF2A depleted cardiomyocytes was performed. AT treatment resulted in an overall improvement in cardiac function of TAC mice and repression of MEF2 activity. RNA-seq identified 65 differentially expressed genes (DEGs) due to TAC treatment with enriched GO clusters including the inflammatory system, cell migration and apoptosis. These genes were mapped against DEGs in cardiomyocytes in which MEF2A expression was suppressed. Of the 65 TAC mediated DEGs, AT reversed the expression of 28 mRNAs. Rarres2 was identified as a novel MEF2 target gene that is upregulated with TAC in vivo and isoproterenol treatment in vitro which may have implications in cardiomyocyte apoptosis and hypertrophy. These studies identify a cohort of genes with vast potential for disease diagnosis and therapeutic intervention in heart failure.

Two Drosophila actin genes in detail. Gene structure, protein structure and transcription during development.

DNA fragments representing the six Drosophila actin genes have been isolated by recombinant DNA techniques. We have compared the transcriptional characteristics of the actin genes at the cytological loci 79B and 88F. The activity of each gene in vivo was examined using gene-specific probes from transcribed, but non-translated 3' regions of each gene. The genes show similar patterns of transcriptional activity during development until the pupal stage, with two periods showing RNA accumulation at two to three hours and 12 to 15 hours during embryonic development, followed by large increases in the proportion of message from each gene in first and second instar larvae. During pupal development, the 88F gene apparently produces a larger proportion of transcripts than at any other developmental stage, while the transcripts of the 79B gene are reduced to a level lower than in first and second instar larvae. The 5' end of each messenger RNA in larvae has been mapped by nuclease S1 digestion of hybrids between restriction fragments of genes and homologous mRNAs. The two genes display widely differing capacities to serve as templates for transcription in vitro in HeLa cell extracts. The complete DNA sequences of both genes including the flanking regions immediately 3' and 5' to the gene are presented. These data permit comparison of the DNA sequences of these Drosophila actin genes with each other and with the DNA sequence and protein sequence information available for the actins of Drosophila and other organisms. These two genes share the common structural feature of an intervening sequence at amino acid 307, though the sequences within each intron differ greatly. This may be a reflection of a duplication event, followed by divergence of the intervening sequences. We discuss possible correlations between the DNA sequences of each 5' flanking region and the differences in transcriptional characteristics of these two distinct but closely related genes.

Developmental expression of the Drosophila melanogaster calmodulin gene.

The highly conserved, intracellular calcium binding protein calmodulin is present in all cells at all times. In addition to this constitutive level, the amount of calmodulin is highly regulated according to the tissue or stage of development. Since there are only a few genes or a single gene for this protein in most species, intricate regulatory elements may be necessary to effect its complex regulation. This report adds new information concerning the gene structure and outlines the developmental and spatial regulation of Drosophila melanogaster calmodulin transcripts. The gene contains five exons, including a 49 bp exon in the 5' untranslated region, and spans over 16 kb. Homologues to this small, 5' noncoding exon have not been found in other calmodulin genes. The combined level of the transcripts is developmentally regulated, and the relative amounts of the two transcript size classes (1.65 kb and 1.9 kb) are differentially regulated during development. Primer extension experiments and RNase protection mapping show that both size classes of Drosophila calmodulin transcripts initiate at the same site but undergo alternative termination within the final exon. The spatial distribution of calmodulin transcripts was examined by in situ hybridization to sections of adults and to developmentally staged whole mount embryos. Calmodulin transcripts are evenly distributed early in embryogenesis. In later stages of embryogenesis, higher levels accumulate in the developing nerve cord and other tissues. Elevated levels of calmodulin transcripts are seen quite distinctly in the adult neural tissues and in the photoreceptor region of the compound eye.

Multiple antibody formation in a primigravida: anti-D, anti-C, anti-Kpb.

A case of primigravida in whom anti-C, anti-D, and anti-Kpb developed during the third trimester is described. Her first-born infant, delivered at 40 weeks' gestation, was mildly affected with hemolytic disease of the newborn and responded well to phototherapy. Her second-born infant, delivered at 36 weeks' gestation, had a positive direct antiglobulin test, but required no treatment. Clinical and laboratory aspects of both pregnancies are discussed.

Organ culture model for the study of HVH-II infections in carcinoma of the cervix.

An experimental model is described whereby human and monkey cervical tissues may be maintained as organ cultures for 21 and 40 days, respectively. Inclusion of sodium carboxymethyl cellulose in the culture medium prolongs the survival time of tissues considerably. The sequential cytologic changes associated with herpesvirus hominis type II (HVH-II) infection are reported. These changes are considered in relation to the possible causal role of HVH-II infection in cervical carcinogenesis.

Relation of HVH-II to carcinoma of the cervix.

An animal model is described that demonstrates the development of varying degrees of basal cell hyperplasia, metaplasia, and dysplasia following cervicovaginal herpesvirus hominis type II (HVH-II) infection. Although the study spanned a period of 30 months, the equivalent of 60 human years, there was no evidence of anaplasia. These results are considered in relation to the role of HVH-II as an initiator in the multistep process leading to cervical carcinoma.

Ascending infection following cervicovaginal exposure to herpesvirus hominis type II.

An animal model is presented that demonstrates that following cervicovaginal inoculation, infective doses of herpesvirus hominis type II induce a form of herpetic encephalitis. Immunofluorescent results indicate that there is both neural and hematogenous spread of the herpes infection. The possible role of a latent viral infection in the genesis of an acute disease of the central nervous system is investigated in this experimental system.