Retrospective harm benefit analysis of pre-clinical animal research for six treatment interventions

The harm benefit analysis (HBA) is the cornerstone of animal research regulation and is considered to be a key ethical safeguard for animals. The HBA involves weighing the anticipated benefits of animal research against its predicted harms to animals but there are doubts about how objective and accountable this process is.i. To explore the harms to animals involved in pre-clinical animal studies and to assess these against the benefits for humans accruing from these studies; ii. To test the feasibility of conducting this type of retrospective HBA.Data on harms were systematically extracted from a sample of pre-clinical animal studies whose clinical relevance had already been investigated by comparing systematic reviews of the animal studies with systematic reviews of human studies for the same interventions (antifibrinolytics for haemorrhage, bisphosphonates for osteoporosis, corticosteroids for brain injury, Tirilazad for stroke, antenatal corticosteroids for neonatal respiratory distress and thrombolytics for stroke). Clinical relevance was also explored in terms of current clinical practice. Harms were categorised for severity using an expert panel. The quality of the research and its impact were considered. Bateson's Cube was used to conduct the HBA.The most common assessment of animal harms by the expert panel was 'severe'. Reported use of analgesia was rare and some animals (including most neonates) endured significant procedures with no, or only light, anaesthesia reported. Some animals suffered iatrogenic harms. Many were kept alive for long periods post-experimentally but only 1% of studies reported post-operative care. A third of studies reported that some animals died prior to endpoints. All the studies were of poor quality. Having weighed the actual harms to animals against the actual clinical benefits accruing from these studies, and taking into account the quality of the research and its impact, less than 7% of the studies were permissible according to Bateson's Cube: only the moderate bisphosphonate studies appeared to minimise harms to animals whilst being associated with benefit for humans.This is the first time the accountability of the HBA has been systematically explored across a range of pre-clinical animal studies. The regulatory systems in place when these studies were conducted failed to safeguard animals from severe suffering or to ensure that only beneficial, scientifically rigorous research was conducted. Our findings indicate a pressing need to: i. review regulations, particularly those that permit animals to suffer severe harms; ii. reform the processes of prospectively assessing pre-clinical animal studies to make them fit for purpose; and iii. systematically evaluate the benefits of pre-clinical animal research to permit a more realistic assessment of its likely future benefits

Genome-wide copy number variation (CNV) in patients with autoimmune Addison's disease

<p>Abstract</p> <p>Background</p> <p>Addison's disease (AD) is caused by an autoimmune destruction of the adrenal cortex. The pathogenesis is multi-factorial, involving genetic components and hitherto unknown environmental factors. The aim of the present study was to investigate if gene dosage in the form of copy number variation (CNV) could add to the repertoire of genetic susceptibility to autoimmune AD.</p> <p>Methods</p> <p>A genome-wide study using the Affymetrix GeneChip^®Genome-Wide Human SNP Array 6.0 was conducted in 26 patients with AD. CNVs in selected genes were further investigated in a larger material of patients with autoimmune AD (n = 352) and healthy controls (n = 353) by duplex Taqman real-time polymerase chain reaction assays.</p> <p>Results</p> <p>We found that low copy number of <it>UGT2B28 </it>was significantly more frequent in AD patients compared to controls; conversely high copy number of <it>ADAM3A </it>was associated with AD.</p> <p>Conclusions</p> <p>We have identified two novel CNV associations to <it>ADAM3A </it>and <it>UGT2B28 </it>in AD. The mechanism by which this susceptibility is conferred is at present unclear, but may involve steroid inactivation (<it>UGT2B28</it>) and T cell maturation (<it>ADAM3A</it>). Characterization of these proteins may unravel novel information on the pathogenesis of autoimmunity.</p

Biallelic NAA60 variants with impaired n-terminal acetylation capacity cause autosomal recessive primary familial brain calcifications

Primary familial brain calcification (PFBC) is characterized by calcium deposition in the brain, causing progressive movement disorders, psychiatric symptoms, and cognitive decline. PFBC is a heterogeneous disorder currently linked to variants in six different genes, but most patients remain genetically undiagnosed. Here, we identify biallelic NAA60 variants in ten individuals from seven families with autosomal recessive PFBC. The NAA60 variants lead to loss-of-function with lack of protein N-terminal (Nt)-acetylation activity. We show that the phosphate importer SLC20A2 is a substrate of NAA60 in vitro. In cells, loss of NAA60 caused reduced surface levels of SLC20A2 and a reduction in extracellular phosphate uptake. This study establishes NAA60 as a causal gene for PFBC, provides a possible biochemical explanation of its disease-causing mechanisms and underscores NAA60-mediated Nt-acetylation of transmembrane proteins as a fundamental process for healthy neurobiological functioning

Biallelic NAA60 variants with impaired n-terminal acetylation capacity cause autosomal recessive primary familial brain calcifications.

Primary familial brain calcification (PFBC) is characterized by calcium deposition in the brain, causing progressive movement disorders, psychiatric symptoms, and cognitive decline. PFBC is a heterogeneous disorder currently linked to variants in six different genes, but most patients remain genetically undiagnosed. Here, we identify biallelic NAA60 variants in ten individuals from seven families with autosomal recessive PFBC. The NAA60 variants lead to loss-of-function with lack of protein N-terminal (Nt)-acetylation activity. We show that the phosphate importer SLC20A2 is a substrate of NAA60 in vitro. In cells, loss of NAA60 caused reduced surface levels of SLC20A2 and a reduction in extracellular phosphate uptake. This study establishes NAA60 as a causal gene for PFBC, provides a possible biochemical explanation of its disease-causing mechanisms and underscores NAA60-mediated Nt-acetylation of transmembrane proteins as a fundamental process for healthy neurobiological functioning

Gauging the epidemic potential of a widely circulating non-invasive meningococcal strain in Africa

Neisseria meningitidis colonizes the human oropharynx and transmits mainly via asymptomatic carriage. Actual outbreaks of meningococcal meningitis are comparatively rare and occur when susceptible populations are exposed to hypervirulent clones, genetically distinct from the main carriage isolates. However, carriage isolates can evolve into pathogens through a limited number of recombination events. The present study examines the potential for the sequence type (ST)-192, by far the dominant clone recovered in recent meningococcal carriage studies in sub-Saharan Africa, to evolve into a pathogen. We used wholegenome sequencing on a collection of 478 meningococcal isolates sampled from 1- to 29- year-old healthy individuals in Arba Minch, southern Ethiopia in 2014. The ST-192 clone was identified in nearly 60% of the carriers. Using complementary shortand long-read techniques for whole-genome sequencing, we were able to completely resolve genomes and thereby identify genomic differences between the ST-192 carriage strain and known pathogenic clones with the highest possible resolution. We conclude that it is possible, but unlikely, that ST-192 could evolve into a significant pathogen, thus, becoming the major invasive meningococcus clone in the meningitis belt of Africa following upcoming mass vaccination with a polyvalent conjugate vaccine that targets the A, C, W, Y and X capsules.publishedVersio

Kartlegging av miljøgifter i sedimenter i Indre Drammensfjorden 1993

Totalt 18 sedimentstasjoner i indre fjordområde og 2 løsmasseprøver fra Holmen i utløpet av Drammenselva ble undersøkt for innholdet av miljøgifter. I tillegg er det foretatt evaluering av en rekke mulige forurensningskilder, samt nærmere karakterisering av ren tjæreprøve fra Gilhusbukta. Hovedresultatene viser at belastningen mht. tungmetaller er liten til moderat i sedimentene og samlet viser metallene maksimalt tilstandsklasse III. Flere ulike kilder til tungmetallforekomstene kan antydes. Av de organiske miljøgiftene er forekomstene av PCB mest markert, og antyder tilstandsklasse III-IV for store deler av fjorden. PAH forekommer i svært høye konsentrasjoner, men har hovedsakelig en begrenset og lokal utbredelse. Både PCB og PAH forekomstene kan stamme fra flere ulike kilder i indre del av fjorden. Av andre klororganiske forbindelser ble kun DDT registrert i forhøyde konsentrasjoner og synes å være knyttet til Lierelva. Oljeforurensningen (THC) var mer moderat og var mest framtredende i elva utenfor Holmen. Referansestasjonen utenfor Sandtangen, synes å ligge i et akkumulasjonsområde for forurensninger i fjorden, da det gjennomgående ble registrert høye konsentrasjoner her for alle miljøgiftene. Løsmasseprøvene fra Holmen var ikke forurenset

Kartlegging av miljøgifter i sedimenter i Indre Drammensfjorden 1993

Totalt 18 sedimentstasjoner i indre fjordområde og 2 løsmasseprøver fra Holmen i utløpet av Drammenselva ble undersøkt for innholdet av miljøgifter. I tillegg er det foretatt evaluering av en rekke mulige forurensningskilder, samt nærmere karakterisering av ren tjæreprøve fra Gilhusbukta. Hovedresultatene viser at belastningen mht. tungmetaller er liten til moderat i sedimentene og samlet viser metallene maksimalt tilstandsklasse III. Flere ulike kilder til tungmetallforekomstene kan antydes. Av de organiske miljøgiftene er forekomstene av PCB mest markert, og antyder tilstandsklasse III-IV for store deler av fjorden. PAH forekommer i svært høye konsentrasjoner, men har hovedsakelig en begrenset og lokal utbredelse. Både PCB og PAH forekomstene kan stamme fra flere ulike kilder i indre del av fjorden. Av andre klororganiske forbindelser ble kun DDT registrert i forhøyde konsentrasjoner og synes å være knyttet til Lierelva. Oljeforurensningen (THC) var mer moderat og var mest framtredende i elva utenfor Holmen. Referansestasjonen utenfor Sandtangen, synes å ligge i et akkumulasjonsområde for forurensninger i fjorden, da det gjennomgående ble registrert høye konsentrasjoner her for alle miljøgiftene. Løsmasseprøvene fra Holmen var ikke forurenset.Fylkesmannen i Buskerud, Miljøvernavdelinge