Detection of Crosslinks within and between Proteins by LC-MALDI-TOFTOF and the Software FINDX to Reduce the MSMS-Data to Acquire for Validation

Lysine-specific chemical crosslinking in combination with mass spectrometry is emerging as a tool for the structural characterization of protein complexes and protein-protein interactions. After tryptic digestion of crosslinked proteins there are thousands of peptides amenable to MSMS, of which only very few are crosslinked peptides of interest. Here we describe how the advantage offered by off-line LC-MALDI-TOF/TOF mass spectrometry is exploited in a two-step workflow to focus the MSMS-acquisition on crosslinks mainly. In a first step, MS-data are acquired and all the peak list files from the LC-separated fractions are merged by the FINDX software and screened for presence of crosslinks which are recognized as isotope-labeled doublet peaks. Information on the isotope doublet peak mass and intensity can be used as search constraints to reduce the number of false positives that match randomly to the observed peak masses. Based on the MS-data a precursor ion inclusion list is generated and used in a second step, where a restricted number of MSMS-spectra are acquired for crosslink validation. The decoupling of MS and MSMS and the peptide sorting with FINDX based on MS-data has the advantage that MSMS can be restricted to and focused on crosslinks of Type 2, which are of highest biological interest but often lowest in abundance. The LC-MALDI TOF/TOF workflow here described is applicable to protein multisubunit complexes and using 14N/15N mixed isotope strategy for the detection of inter-protein crosslinks within protein oligomers

Changes in life expectancy and disease burden in Norway, 1990–2019: an analysis of the Global Burden of Disease Study 2019

Bill & Melinda Gates FoundationpublishedVersio

Local host-dependent persistence of the entomopathogenic nematode Steinernema carpocapsae used to control the large pine weevil Hylobius abietis

Entomopathogenic nematodes (EPN) applied inundatively to suppress insect pests are more likely to persist and establish in stable agroecosystems than in annual crops. We investigated a system of intermediate stability: three stumps harbouring the large pine weevil (Hylobius abietis L.; Coleoptera: Curculionidae), a major European forestry pest. We tested whether persistence of EPN Steinernema carpocapsae Weiser (Rhabditida: Steinernematidae) applied around stumps is maintained by recycling of EPN through pine weevils developing within stumps. Steinernema carpocapsae was detected in soil around and under the bark of treated tree stumps up to two years, but not 4–5 years after application. Differences in nematode presence between sites were better explained by tree species (pine or spruce) than soil type (mineral or peat). Presence of S. carpocapsae in soil was positively correlated with the number of H. abietis emerging from untreated stumps the previous year, which was greater for pine stumps than spruce stumps

Vitamin D Induction of the Human Antimicrobial Peptide Cathelicidin in the Urinary Bladder

The urinary tract is frequently being exposed to potential pathogens and rapid defence mechanisms are therefore needed. Cathelicidin, a human antimicrobial peptide is expressed and secreted by bladder epithelial cells and protects the urinary tract from infection. Here we show that vitamin D can induce cathelicidin in the urinary bladder. We analyzed bladder tissue from postmenopausal women for expression of cathelicidin, before and after a three-month period of supplementation with 25-hydroxyvitamin D3 (25D3). Cell culture experiments were performed to elucidate the mechanisms for cathelicidin induction. We observed that, vitamin D per se did not up-regulate cathelicidin in serum or in bladder tissue of the women in this study. However, when the bladder biopsies were infected with uropathogenic E. coli (UPEC), a significant increase in cathelicidin expression was observed after 25D3 supplementation. This observation was confirmed in human bladder cell lines, even though here, cathelicidin induction occurred irrespectively of infection. Vitamin D treated bladder cells exerted an increased antibacterial effect against UPEC and colocalization to cathelicidin indicated the relevance of this peptide. In the light of the rapidly growing problem of resistance to common urinary tract antibiotics, we suggest that vitamin D may be a potential complement in the prevention of UTI

Surveillance after surgery for pancreatic cancer : a global scoping review of guidelines and a nordic Survey of contemporary practice

Objectives: Most patients with pancreatic cancer who have undergone surgical resection eventually develop disease recurrence. ‍This study aimed to investigate whether there is evidence to support routine surveillance after pancreatic cancer surgery, with a secondary aim of analyzing the implementation of surveillance strategies in the Nordic countries. Materials and Methods: A scoping review was conducted to identify clinical practice guidelines globally and research studies relating to surveillance after pancreatic cancer resection. This was followed by a survey among 20 pancreatic units from four Nordic countries to assess their current practice of follow-up for operated patients. Results: Altogether 16 clinical practice guidelines and 17 research studies were included. The guidelines provided inconsistent recommendations regarding postoperative surveillance of pancreatic cancer. The clinical research data were mainly based on retrospective cohort studies with low level of evidence and lead-time bias was not addressed. Active surveillance was recommended in Sweden and Denmark, but not in Norway beyond the post-operative/adjuvant period. Finland had no national recommendations for surveillance. The Nordic survey revealed a wide variation in reported practice among the different units. About 75% (15 of 20 units) performed routine postoperative surveillance. Routine CA 19-9 testing was used by 80% and routine CT by 67% as part of surveillance. About 73% of centers continued follow-up until 5 years postoperatively. Conclusion: Evidence for routine long-term (i.e. 5 years) surveillance after pancreatic cancer surgery remains limited. Most pancreatic units in the Nordic countries conduct regular follow-up, but protocols vary.Peer reviewe

Oral rehydration therapies in Senegal, Mali, and Sierra Leone: a spatial analysis of changes over time and implications for policy.

BACKGROUND: Oral rehydration solution (ORS) is a simple intervention that can prevent childhood deaths from severe diarrhea and dehydration. In a previous study, we mapped the use of ORS treatment subnationally and found that ORS coverage increased over time, while the use of home-made alternatives or recommended home fluids (RHF) decreased, in many countries. These patterns were particularly striking within Senegal, Mali, and Sierra Leone. It was unclear, however, whether ORS replaced RHF in these locations or if children were left untreated, and if these patterns were associated with health policy changes. METHODS: We used a Bayesian geostatistical model and data from household surveys to map the percentage of children with diarrhea that received (1) any ORS, (2) only RHF, or (3) no oral rehydration treatment between 2000 and 2018. This approach allowed examination of whether RHF was replaced with ORS before and after interventions, policies, and external events that may have impacted healthcare access. RESULTS: We found that RHF was replaced with ORS in most Sierra Leone districts, except those most impacted by the Ebola outbreak. In addition, RHF was replaced in northern but not in southern Mali, and RHF was not replaced anywhere in Senegal. In Senegal, there was no statistical evidence that a national policy promoting ORS use was associated with increases in coverage. In Sierra Leone, ORS coverage increased following a national policy change that abolished health costs for children. CONCLUSIONS: Children in parts of Mali and Senegal have been left behind during ORS scale-up. Improved messaging on effective diarrhea treatment and/or increased ORS access such as through reducing treatment costs may be needed to prevent child deaths in these areas

Targeting the RNA m⁶A reader YTHDF2 selectively compromises cancer stem cells in acute myeloid leukemia

International audienc

Irreversible electroporation of pancreatic adenocarcinoma

Pancreatic cancer (PC) is a severe diagnosis with poor prognosis. Radical surgery is the only treatment that can possibly lead to a cure, and even with surgery, the 5-year survival is only 20%–25%. The majority of patients cannot be resected due to metastases or having a tumour that is too advanced locally (LAPC) with encasement of blood-vessels. Short electrical pulses can change the cell membrane, creating reversible pores in it. With a higher current, the pores become permanent, resulting in irreversible electroporation (IRE). This leads to specific cell death, with the chance to save surrounding scaffold material, such as the walls of blood vessels and bile ducts. This led to the theory that IRE might be suitable for treating LAPC. In Paper I, we found that IRE can be safely performed percutaneously with ultrasound guidance in humans with PC, with promising efficacy, since one of the five patients included was downstaged due to the IRE and could be surgically resected. In Paper II, which is an extension of Paper I, we treated 24 patients with LAPC (3 were also included in Paper I) who had received chemotherapy and, after IRE, stable disease was seen. Median overall survival was 17.9 months. Eleven patients had some form of complication, but we still concluded that IRE is reasonably safe in LAPC patients, with promising efficacy. In Paper III, we chose to treat LAPC with IRE followed by adjuvant chemotherapy. We compared the overall survival of our patients with those with LAPC in the National Quality Registry for Pancreatic and Periampullary Cancer. No significant survival gain could be seen in the group that received IRE compared to the registry group (13.3 months versus 9.9 months, p=0.511). In the IRE group, there were six major complications and we found no support for using IRE in this setting. Paper IV examines the response on the tumour marker CA19-9 in PC treated with IRE. We found 35 patients suitable for this analysis. The hypothesis that IRE would lower the CA19-9 value could not be proven. In fact, the CA19-9 was slightly higher one month after IRE (282 U/ml versus 315 U/ml). However, the 25th percentile of patients with the best CA19-9 response had a better survival (p=0.01) compared to the 25th percentile with the worst response, indicating that CA19-9 can be used as a prognostic marker after IRE in PC

Irreversible electroporation of pancreatic adenocarcinoma

Pancreatic cancer (PC) is a severe diagnosis with poor prognosis. Radical surgery is the only treatment that can possibly lead to a cure, and even with surgery, the 5-year survival is only 20%–25%. The majority of patients cannot be resected due to metastases or having a tumour that is too advanced locally (LAPC) with encasement of blood-vessels. Short electrical pulses can change the cell membrane, creating reversible pores in it. With a higher current, the pores become permanent, resulting in irreversible electroporation (IRE). This leads to specific cell death, with the chance to save surrounding scaffold material, such as the walls of blood vessels and bile ducts. This led to the theory that IRE might be suitable for treating LAPC. In Paper I, we found that IRE can be safely performed percutaneously with ultrasound guidance in humans with PC, with promising efficacy, since one of the five patients included was downstaged due to the IRE and could be surgically resected. In Paper II, which is an extension of Paper I, we treated 24 patients with LAPC (3 were also included in Paper I) who had received chemotherapy and, after IRE, stable disease was seen. Median overall survival was 17.9 months. Eleven patients had some form of complication, but we still concluded that IRE is reasonably safe in LAPC patients, with promising efficacy. In Paper III, we chose to treat LAPC with IRE followed by adjuvant chemotherapy. We compared the overall survival of our patients with those with LAPC in the National Quality Registry for Pancreatic and Periampullary Cancer. No significant survival gain could be seen in the group that received IRE compared to the registry group (13.3 months versus 9.9 months, p=0.511). In the IRE group, there were six major complications and we found no support for using IRE in this setting. Paper IV examines the response on the tumour marker CA19-9 in PC treated with IRE. We found 35 patients suitable for this analysis. The hypothesis that IRE would lower the CA19-9 value could not be proven. In fact, the CA19-9 was slightly higher one month after IRE (282 U/ml versus 315 U/ml). However, the 25th percentile of patients with the best CA19-9 response had a better survival (p=0.01) compared to the 25th percentile with the worst response, indicating that CA19-9 can be used as a prognostic marker after IRE in PC