Intravitreal antisense oligonucleotide sepofarsen in Leber congenital amaurosis type 10: a phase 1b/2 trial

CEP290-associated Leber congenital amaurosis type 10 (LCA10) is a retinal disease resulting in childhood blindness. Sepofarsen is an RNA antisense oligonucleotide targeting the c.2991+1655A>G variant in the CEP290 gene to treat LCA10. In this open-label, phase 1b/2 (NCT03140969), 12-month, multicenter, multiple-dose, dose-escalation trial, six adult patients and five pediatric patients received ≤4 doses of intravitreal sepofarsen into the worse-seeing eye. The primary objective was to evaluate sepofarsen safety and tolerability via the frequency and severity of ocular adverse events (AEs); secondary objectives were to evaluate pharmacokinetics and efficacy via changes in functional outcomes. Six patients received sepofarsen 160 µg/80 µg, and five patients received sepofarsen 320 µg/160 µg. Ten of 11 (90.9%) patients developed ocular AEs in the treated eye (5/6 with 160 µg/80 µg; 5/5 with 320 µg/160 µg) versus one of 11 (9.1%) in the untreated eye; most were mild in severity and dose dependent. Eight patients developed cataracts, of which six (75.0%) were categorized as serious (2/3 with 160 µg/80 µg; 4/5 with 320 µg/160 µg), as lens replacement was required. As the 160-µg/80-µg group showed a better benefit–risk profile, higher doses were discontinued or not initiated. Statistically significant improvements in visual acuity and retinal sensitivity were reported (post hoc analysis). The manageable safety profile and improvements reported in this trial support the continuation of sepofarsen development

Antisense oligonucleotide-based treatment of retinitis pigmentosa caused by USH2A exon 13 mutations

Mutations in USH2A are among the most common causes of syndromic and non-syndromic retinitis pigmentosa (RP). The two most recurrent mutations in USH2A, c.2299delG and c.2276G > T, both reside in exon 13. Skipping exon 13 from the USH2A transcript presents a potential treatment modality in which the resulting transcript is predicted to encode a slightly shortened usherin protein. Morpholino-induced skipping of ush2a exon 13 in zebrafish ush2(armc1) mutants resulted in the production of usherin Delta exon 13 protein and a completely restored retinal function. Antisense oligonucleotides were investigated for their potential to selectively induce human USH2A exon 13 skipping. Lead candidate QR-421a induced a concentration-dependent exon 13 skipping in induced pluripotent stem cell (iPSC)-derived photoreceptor precursors from an Usher syndrome patient homozygous for the c.2299delG mutation. Mouse surrogate mQR-421a reached the retinal outer nuclear layer after a single intravitreal injection and induced a detectable level of exon skipping until at least 6 months post-injection. In conclusion, QR-421a-induced exon skipping proves to be a highly promising treatment option for RP caused by mutations in USH2A exon 13

Cattle Mammary Bioreactor Generated by a Novel Procedure of Transgenic Cloning for Large-Scale Production of Functional Human Lactoferrin

Large-scale production of biopharmaceuticals by current bioreactor techniques is limited by low transgenic efficiency and low expression of foreign proteins. In general, a bacterial artificial chromosome (BAC) harboring most regulatory elements is capable of overcoming the limitations, but transferring BAC into donor cells is difficult. We describe here the use of cattle mammary bioreactor to produce functional recombinant human lactoferrin (rhLF) by a novel procedure of transgenic cloning, which employs microinjection to generate transgenic somatic cells as donor cells. Bovine fibroblast cells were co-microinjected for the first time with a 150-kb BAC carrying the human lactoferrin gene and a marker gene. The resulting transfection efficiency of up to 15.79×10−2 percent was notably higher than that of electroporation and lipofection. Following somatic cell nuclear transfer, we obtained two transgenic cows that secreted rhLF at high levels, 2.5 g/l and 3.4 g/l, respectively. The rhLF had a similar pattern of glycosylation and proteolytic susceptibility as the natural human counterpart. Biochemical analysis revealed that the iron-binding and releasing properties of rhLF were identical to that of native hLF. Importantly, an antibacterial experiment further demonstrated that rhLF was functional. Our results indicate that co-microinjection with a BAC and a marker gene into donor cells for somatic cell cloning indeed improves transgenic efficiency. Moreover, the cattle mammary bioreactors generated with this novel procedure produce functional rhLF on an industrial scale

Criminaliteit en rechtshandhaving 2018: ontwikkelingen en samenhangen

De publicatie Criminaliteit en rechtshandhaving brengt ontwikkelingen in en samenhangen tussen criminaliteit en rechtshandhaving in kaart. Deze 17e editie richt zich daarbij met name op de periode 2008 tot en met 2018. Deze editie staat weer boordevol informatie over de aard en omvang van de criminaliteit in Nederland, welk deel hiervan wordt opgespoord, de strafrechtelijke reactie die justitie hierop heeft, de tenuitvoerlegging van opgelegde sancties en ontwikkelingen die zich hierin hebben voorgedaan

Criminaliteit en rechtshandhaving 2018

De publicatie Criminaliteit en rechtshandhaving brengt ontwikkelingen in en samenhangen tussen criminaliteit en rechtshandhaving in kaart. Deze 17e editie richt zich daarbij met name op de periode 2008 tot en met 2018. Deze editie staat weer boordevol informatie over de aard en omvang van de criminaliteit in Nederland, welk deel hiervan wordt opgespoord, de strafrechtelijke reactie die justitie hierop heeft, de tenuitvoerlegging van opgelegde sancties en ontwikkelingen die zich hierin hebben voorgedaan. De publicatie 'Criminaliteit en rechtshandhaving 2018' is ook terug te vinden op de websites van het CBS en de Raad voor de rechtspraak. Daarnaast is de publicatie beschikbaar op het nieuwe digitale platform www.criminaliteitinbeeld.nl, een initiatief gedragen door het WODC, het CBS, de Raad voor de rechtspraak, het Openbaar Ministerie en de Nationale Politie. Een overzicht van vorige edities van C&R is te vinden op de speciale webpagina 'Criminaliteit en rechtshandhaving'. iNHOUD: 1. Inleiding 2. Het Nederlandse strafrechtssysteem 3. Criminaliteit en slachtofferschap 4. Misdrijven en opsporing 5. Vervolging 6. Berechting 7. Tenuitvoerlegging van sancties 8. De strafrechtsketen in samenhang 9. Overtredingen 10. Kosten van criminaliteit 11. Nederland in internationaal perspectie

Vaccine-induced effector-memory CD8+ T cell responses predict therapeutic efficacy against tumors

CD8(+) T cells have the potential to attack and eradicate cancer cells. The efficacy of therapeutic vaccines against cancer, however, lacks defined immune correlates of tumor eradication after (therapeutic) vaccination based on features of Ag-specific T cell responses. In this study, we examined CD8(+) T cell responses elicited by various peptide and TLR agonist-based vaccine formulations in nontumor settings and show that the formation of CD62L(-)KLRG1(+) effector-memory CD8(+) T cells producing the effector cytokines IFN-γ and TNF predicts the degree of therapeutic efficacy of these vaccines against established s.c. tumors. Thus, characteristics of vaccine-induced CD8(+) T cell responses instill a predictive determinant for the efficacy of vaccines during tumor therapy.Tumorimmunolog

Household drinking water in developing countries: a systematic review of microbiological contamination between source and point-of-use

Summary Objective To assess the extent and causes of microbiological contamination of household drinking water between source and point-of-use in developing countries. Methods A systematic meta-analysis of 57 studies measuring bacteria counts for source water and stored water in the home to assess how contamination varied between settings. Results The bacteriological quality of drinking water significantly declines after collection in many settings. The extent of contamination after water collection varies considerably between settings, but is proportionately greater where faecal and total coliform counts in source water are low. Conclusions Policies that aim to improve water quality through source improvements may be compromised by post-collection contamination. Safer household water storage and treatment is recommended to prevent this, together with point-of-use water quality monitoring

Effect of an intravitreal antisense oligonucleotide on vision in Leber congenital amaurosis due to a photoreceptor cilium defect

Photoreceptor ciliopathies constitute the most common molecular mechanism of the childhood blindness Leber congenital amaurosis. Ten patients with Leber congenital amaurosis carrying the c.2991+1655A>G allele in the ciliopathy gene centrosomal protein 290 (CEP290) were treated (ClinicalTrials.gov no. NCT03140969) with intravitreal injections of an antisense oligonucleotide to restore correct splicing. There were no serious adverse events, and vision improved at 3 months. The visual acuity of one exceptional responder improved from light perception to 20/400