Supporting clinical decision making in advanced melanoma by preclinical testing in personalized immune-humanized xenograft mouse models.

BACKGROUND: The mouse strains usually used to generate patient-derived xenografts (PDXs) are immunocompromised, rendering them unsuitable for immunotherapy studies. Here we assessed the value of immune-PDX mouse models for predicting responses to anti-PD-1 checkpoint inhibitor therapy in patients. PATIENTS AND METHODS: Melanoma biopsies contained in a retrospective biobank were transplanted into NOG mice or NOG mice expressing interleukin 2 (hIL2-NOG mice). Tumor growth was monitored, and comparisons were made with clinical data, sequencing data, and current in silico predictive tools. RESULTS: Biopsies grew readily in NOG mice but growth was heterogeneous in hIL2-NOG mice. IL2 appears to activate T-cell immunity in the biopsies to block tumor growth. Biopsy growth in hIL2-NOG mice was negatively associated with survival in patients previously treated with PD-1 checkpoint blockade. In two cases, the prospective clinical decisions of anti-PD-1 therapy or targeted BRAF/MEK inhibitors were supported by the observed responses in mice. CONCLUSIONS: Immune-PDX models represent a promising addition to future biomarker discovery studies and for clinical decision making in patients receiving immunotherapy.

One-year follow-up on liraglutide treatment for prediabetes and overweight/obesity in clozapine- or olanzapine-treated patients.

OBJECTIVE: Treatment with most antipsychotics is associated with an increased risk of weight gain and metabolic disturbances. In a randomized trial, we previously demonstrated that 16 weeks of glucagon-like peptide-1 receptor agonist liraglutide treatment vs. placebo significantly reduced glucometabolic disturbances and body weight in prediabetic, overweight/obese schizophrenia-spectrum disorder patients treated with clozapine or olanzapine. The aim of this study was to investigate whether the beneficial effects of the 16-week intervention were sustained beyond the intervention period. METHOD: One year after completion of the intervention, we investigated changes in body weight, fasting glucose, glycated hemoglobin, C-peptide, and lipids comparing 1-year follow-up levels to end of treatment (week 16) and baseline (week 0) levels. RESULTS: From end of treatment to the 1-year follow-up, body weight had increased in the liraglutide-treated group. However, compared to baseline levels, the placebo-subtracted body weight loss remained significantly reduced (-3.8 kg, 95% CI: -7.3 to -0.2, P = 0.04). Fasting glucose, glycated hemoglobin, C-peptide, and lipids had each returned to baseline levels 1 year after stopping liraglutide. CONCLUSION: The body weight reduction during 16 weeks of liraglutide treatment was partially sustained 1 year after the intervention was completed. However, the improvements in other metabolic parameters returned to baseline levels.

Genetic differentiation of southeast Baltic populations of sea trout inferred from single nucleotide polymorphisms.

Sea trout (Salmo trutta m. trutta) is a migratory form of brown trout common in the Baltic Sea. Nine populations from the southeast Baltic (Poland; Lithuania; Denmark, Bornholm; Estonia and Russia) were genotyped using iPLEX Gold technology (Sequenom) with 62 informative SNPs. A diagnostic panel of 23 SNPs was applied to estimate genetic differentiation and assess the population structure of Baltic sea trout. The highest level of pairwise FST differences was observed between the Russian (East Gulf of Finland) and Polish (Baltic main basin) populations. The lowest differences were between the two Polish and the Polish and Lithuanian populations. A genetic similarity was noted between the Estonian Riguldi River and Danish Bornholm populations, and this finding was supported by a Bayesian and factorial correspondence analysis. Diversity within populations was highest for populations from Estonia and lowest for the Lithuanian population. Genetic structure analysis indicated that individuals from the nine populations were clustered into four groups.

Long-term leaching through clayey till of N,N-dimethylsulfamide, a Persistent and Mobile Organic Compound (PMOC).

Environmental pollution with Persistent and Mobile Organic Compounds (PMOC) from anthropogenic activities is an increasing cause for concern. These compounds are readily leached to groundwater aquifers and are likely to resist degradation, putting pressure on groundwater resources. Pesticides can form PMOCs upon degradation in the environment. The PMOC N,N-dimethylsulfamide (DMS) was the most frequently detected pesticide metabolite in Danish drinking water wells in 2020, although the pesticidal use of the last parent compound (tolylfluanid) ended in 2007. This study aimed to improve the understanding of the leaching of the PMOC DMS from clayey tills by combining a review of compound properties, sources and use, comprehensive field observations and numerical flow and solute transport modeling. The modeling explored the mechanisms of DMS retention during vertical transport in clayey till and the fingerprint in the underlying aquifer. The results were supported by detailed field observations at an agricultural site with strawberry production. Porewater samples were collected from clayey till to a depth of 12 m bgs by a custom designed installation method of suction cups. Groundwater sampling (249 samples) was designed to provide vertical concentration profiles at various distances from the presumed sources. The review of properties showed that the parent compounds and intermediates degrade quickly in topsoil, releasing the highly persistent and mobile DMS. We tested the effect of fractures on transport with different hydraulic apertures and a scenario without fractures by numerical modeling. The results showed that the presence of fractures can smooth the breakthrough curve below the clayey till, leading to faster breakthrough, lower maximum concentration, and several decades of prolonged leaching in simulations with the largest aperture (20 µm). The fracture-matrix interaction is a possible explanation for the observed delay of leaching from clayey till. The vertical concentration profiles in groundwater were used for identifying the sources at the field site and testing source strengths. Assigning one point source (200 µg/L) and two diffuse sources (40-50 µg/L) to the model produced vertical concentration profiles that compared well with observed field data in clayey till and the aquifer. All results were integrated into a conceptual model for the environmental fate of PMOCs in soil and groundwater. The findings of this study imply that the presence of fractures in clayey till should be considered in conceptual site models, since they can substantially prolong the leaching of PMOCs to groundwater. The integration of comprehensive field investigations and numerical modeling is key to understand the fate of PMOCs in complex field systems with different source types. Together with widespread occurrences of PMOCs in groundwater systems, the results highlight the need for improved approval procedures for pesticides and biocides which considers their persistent and mobile metabolites.

Analysis of two incompletely spliced Arabidopsis cDNAs encoding novel types of peroxidase.

Most known class III peroxidase genes contain three introns at conserved positions. Two Arabidopsis cDNAs (ESTs), encoding novel type peroxidases ATP9a and ATP15a were sequenced, and found to contain inserts for intron 2. PCR and sequence analysis of genomic DNA revealed that the atp9a gene contains all three introns, whereas atp15a contains only introns 2 and 3. The ATP15a cDNA intron contained a single base substitution reducing the splicing potential significantly as compared with the genomic sequence. The putative enzymes share essential catalytic and structural features with horseradish peroxidase, despite a pair-wise sequence identity of only 40-45% among the three.

Amino acid sequences and structures of chicken and turkey beta 2-microglobulin.

The complete amino acid sequences of chicken and turkey beta 2-microglobulins have been determined by analyses of tryptic, V8-proteolytic and cyanogen bromide fragments, and by N-terminal sequencing. Mass spectrometric analysis of chicken beta 2-microglobulin supports the sequence-derived Mr of 11,048. The higher apparent Mr obtained for the avian beta 2-microglobulins as compared to human beta 2-microglobulin by SDS-PAGE is not understood. Chicken and turkey beta 2-microglobulin consist of 98 residues and deviate at seven positions: 60, 66, 74-76, 78 and 82. The chicken and turkey sequences are identical to human beta 2-microglobulin at 46 and 47 positions, respectively, and to bovine beta 2-microglobulin at 47 positions, i.e. there is about 47% identity between avian and mammalian beta 2-microglobulins. The known X-ray crystallographic structures of bovine beta 2-microglobulin and human HLA-A2 complex suggest that the seven chicken to turkey differences are exposed to solvent in the avian MHC class I complex. The key residues of beta 2-microglobulin involved in alpha chain contacts within the MHC class I molecule are highly conserved between chicken and man. This explains that heterologous human beta 2-microglobulin can substitute the chicken beta 2-microglobulin in exchange studies with B-F (chicken MHC class I molecule), and suggests that the MHC class I structure is conserved over long evolutionary distances.

A circularly permuted alpha-amylase-type alpha/beta-barrel structure in glucan-synthesizing glucosyltransferases.

A motif of amino acid residues, located at the active site and specific beta-strands in alpha-amylases, is recognized in alpha-1,3- and alpha-1,6-glucan-synthesizing glucosyltransferases, leading to the conclusion that these enzymes contain an alpha/beta-barrel closely related to the (beta/alpha)8-fold of the alpha-amylase superfamily. The secondary structure elements of the transferase barrel, however, are circularly permuted to start with an alpha-helix equivalent to helix 3 in the alpha-amylases. Thus, the transferase counterpart of the long third beta-->alpha connection--constituting a domain in the alpha-amylases--is divided to precede and succeed the barrel. This architectural arrangement may be coupled to sucrose scission and glucosyl transfer. The involvement in the mechanism in glucosyltransferases of active site residues recurring in amylolytic enzymes is discussed.

Widespread occurrence of a highly conserved RING-H2 zinc finger motif in the model plant Arabidopsis thaliana.

Several novel Arabidopsis thaliana proteins containing a RING-H2 zinc finger motif were predicted after database searches. Alignment of 29 RING-H2 finger sequences shows that the motif is strikingly conserved in otherwise unrelated proteins. Only short, non-conserved polar/charged sequences distinguish these domains. The RING-H2 domain is most often present in multi-domain structures, a number of which are likely to contain a membrane-spanning region or an additional zinc finger. However, there are several small (126-200 residues) proteins consisting of an N-terminal domain, rich in aliphatic residues, and a C-terminal RING-H2 domain. Reverse-transcription PCR suggests that the RING-H2 genes are widely expressed at low levels.

Computational analyses and annotations of the Arabidopsis peroxidase gene family.

Classical heme-containing plant peroxidases have been ascribed a wide variety of functional roles related to development, defense, lignification, and hormonal signaling. More than 40 peroxidase genes are now known in Arabidopsis thaliana for which functional association is complicated by a general lack of peroxidase substrate specificity. Computational analysis was performed on 30 near full-length Arabidopsis peroxidase cDNAs for annotation of start codons and signal peptide cleavage sites. A compositional analysis revealed that 23 of the 30 peroxidase cDNAs have 5' untranslated regions containing 40-71% adenine, a rare feature observed also in cDNAs which predominantly encode stress-induced proteins, and which may indicate translational regulation.

Headache in stroke: the Copenhagen Stroke Study.

BACKGROUND: Many aspects of stroke-related headache, such as its incidence, pathophysiology, risk factors, and relation to stroke severity and outcome, are uncertain. DESIGN: A prospective, community-based study including 1,128 consecutive acute stroke patients; 867 (77%) were able to communicate. Stroke severity and the clinical course were assessed using the Scandinavian Stroke Scale (SSS). CT was performed in 84% of patients. In the data analysis, multivariate regression analyses were used when appropriate. RESULTS: Two hundred forty-one (28%) of the 867 patients had headache in relation to stroke onset. Independent risk factors were age (odds ratio [OR] per 20 years, 0.57; 95% confidence interval [CI], 0.43 to 0.86; p = 0.0001), female sex (OR, 1.6; CI, 1.2 to 2.2; p = 0.003), intracerebral hemorrhage (OR, 2.8; CI, 1.5 to 5.4; p = 0.002), stroke in the vertebrobasilar territory (OR, 1.7; CI, 1.1 to 2.3; p = 0.02), and ischemic heart disease (OR, 1.5; CI, 1.1 to 2.2; p = 0.01). In those patients in whom headache was lateralized (46%), it was ipsilateral to the lesion in 68% of cases (p = 0.01). There was no correlation between headache and (1) initial stroke severity as measured with the SSS (p = 0.75), (2) lesion size (p = 0.77), (3) whether or not CT showed cortical involvement (p = 0.29), (4) stroke outcome as measured with the SSS (p = 0.9), or (5) mortality (p = 0.75). CONCLUSION: Stroke-related headache is associated with factors such as age and sex. In patients capable of providing reliable headache information, headache is more frequent in strokes confined to the vertebrobasilar than to the carotid territory and is not associated with stroke severity or whether or not the cortex was involved. Stroke-related headache is not related to stroke outcome.

[Intracranial venous thrombosis]. / Intrakranial venøs trombose.

Thrombosis of intracranial sinuses and veins is a rare but potentially lethal syndrome. It often affects young adults, but occurs in all ages. It is frequently seen in association with hormonal changes (puerperium, pregnancy and the use of oral contraceptives), inflammatory bowel disease, cancer, and various states of hypercoagulability, or as a consequence of infection. The clinical picture is extremely variable and the diagnosis can be difficult unless one specifically searches for it. The combination of magnetic resonance imaging and magnetic resonance angiography is a powerful and non-invasive tool for diagnosing the syndrome. Treatment with heparin (and for the septic form antibiotics) improves the outcome.

[Whiplash injury syndrome. A prospective study of 39 patients with whiplash injury]. / Piskesmaeldsyndromet. En prospektiv undersøgelse af 39 patienter med piskesmaeldlaesioner.

The acute symptoms after whiplash trauma can be explained by the neck sprain, but the pathogenesis of the "late whiplash syndrome" and the reasons why only some people have persistent symptoms more than six months are still unknown. Thirty-four consecutive cases of piskesmaeld injury were examined clinically three times; respectively within 14 days, after one month and finally seven months post-injury. In addition, MRI of the brain and the cervical spine, neuropsychological tests and motor evoked potentials (MEP) were done one month post-injury and repeated after six months, if abnormalities were found. We found the total recovery rate (asymptomatic patients) was 29% after seven months. All MEP examinations were normal. The correlation between MRI and the clinical findings was poor. Cognitive dysfunction as a symptom of brain injury was not found. Stress at the same time as the accident predicted more symptoms at follow-up. We conclude that long-lasting distress and poor outcome were more related to the occurrence of stressful life events than to clinical and paraclinical findings.

Starch- and glycogen-debranching and branching enzymes: prediction of structural features of the catalytic (beta/alpha)8-barrel domain and evolutionary relationship to other amylolytic enzymes.

Sequence alignment and structure prediction are used to locate catalytic alpha-amylase-type (beta/alpha)8-barrel domains and the positions of their beta-strands and alpha-helices in isoamylase, pullulanase, neopullulanase, alpha-amylase-pullulanase, dextran glucosidase, branching enzyme, and glycogen branching enzymes--all enzymes involved in hydrolysis or synthesis of alpha-1,6-glucosidic linkages in starch and related polysaccharides. This has allowed identification of the transferase active site of the glycogen debranching enzyme and the locations of beta-->alpha loops making up the active sites of all enzymes studied. Activity and specificity of the enzymes are discussed in terms of conserved amino acid residues and loop variations. An evolutionary distance tree of 47 amylolytic and related enzymes is built on 37 residues representing the four best conserved beta-strands of the barrel. It exhibits clusters of enzymes close in specificity, with the branching and glycogen debranching enzymes being the most distantly related.

Comparison of the domain-level organization of starch hydrolases and related enzymes.

Structure-prediction and hydrophobic-cluster analysis of several starch hydrolases and related enzymes indicated the organization of eleven domain types. Most enzymes possess a catalytic (beta/alpha)8-barrel and a smaller C-terminal domain as seen in crystal structures of alpha-amylase and cyclodextrin glucanotransferase. Some also have a starch-granule-binding domain. Enzymes breaking or forming endo-alpha-1,6 linkages contain domains N-terminal to the (beta/alpha)8-barrel.