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Acquired aplastic anaemia is a rare disease, and occurrence in more than one member of the same family is uncommon. With this case report, we wish to highlight the importance of searching for an underlying genetic cause when this occurs. It may have consequences for future generations in affected families. CTLA4 haploinsufficiency is a heterogeneous disease entity with severe systemic immune dysregulation associated with several autoimmune diseases including aplastic anaemia.

Introduction: The established method for assessment of mediastinal and hilar lymph nodes is endobronchial ultrasound bronchoscopy (EBUS) with needle aspirations. Previously, we presented an electromagnetic navigation platform for this purpose. There were several issues with the permanent electromagnetic tracking (EMT) sensor attachment on the tip of the experimental EBUS bronchoscope. The purpose was to develop a device for on-site attachment of the EMT sensor. Material and methods: A clip-on EMT sensor attachment device was 3D-printed in Ultem™ and attached to an EBUS bronchoscope. A specially designed ultrasound probe calibration adapter was developed for on-site and quick probe calibration. Navigation accuracy was studied using a wire cross water phantom and clinical feasibility was tested in a healthy volunteer. Results: The device attached to the EBUS bronchoscope increased its diameter from 6.9 mm to 9.5 mm. Average preclinical navigation accuracy was 3.9 mm after adapter calibration. The maneuvering of the bronchoscope examining a healthy volunteer was adequate without harming the respiratory epithelium, and the device stayed firmly attached. Conclusion: Development, calibration and testing of a clip-on EMT sensor attachment device for EBUS bronchoscopy was successfully demonstrated. Acceptable accuracy results were obtained, and the device is ready to be tested in patient studies.

Carbapenem-resistant Gram-negative pathogens are a critical public health threat and there is an urgent need for new treatments. Carbapenemases (β-lactamases able to inactivate carbapenems) have been identified in both serine β-lactamase (SBL) and metallo-β-lactamase (MBL) families. The recent introduction of SBL carbapenemase inhibitors has provided alternative therapeutic options. Unfortunately, there are no approved inhibitors of MBL-mediated carbapenem-resistance and treatment options for infections caused by MBL-producing Gram-negatives are limited., Carbapenem-resistant Gram-negative pathogens are a critical public health threat and there is an urgent need for new treatments. Carbapenemases (β-lactamases able to inactivate carbapenems) have been identified in both serine β-lactamase (SBL) and metallo-β-lactamase (MBL) families. The recent introduction of SBL carbapenemase inhibitors has provided alternative therapeutic options. Unfortunately, there are no approved inhibitors of MBL-mediated carbapenem-resistance and treatment options for infections caused by MBL-producing Gram-negatives are limited. Here, we present ZN148, a zinc-chelating MBL-inhibitor capable of restoring the bactericidal effect of meropenem and in vitro clinical susceptibility to carbapenems in >98% of a large international collection of MBL-producing clinical Enterobacterales strains (n = 234). Moreover, ZN148 was able to potentiate the effect of meropenem against NDM-1-producing Klebsiella pneumoniae in a murine neutropenic peritonitis model. ZN148 showed no inhibition of the human zinc-containing enzyme glyoxylase II at 500 μM, and no acute toxicity was observed in an in vivo mouse model with cumulative dosages up to 128 mg/kg. Biochemical analysis showed a time-dependent inhibition of MBLs by ZN148 and removal of zinc ions from the active site. Addition of exogenous zinc after ZN148 exposure only restored MBL activity by ∼30%, suggesting an irreversible mechanism of inhibition. Mass-spectrometry and molecular modeling indicated potential oxidation of the active site Cys221 residue. Overall, these results demonstrate the therapeutic potential of a ZN148-carbapenem combination against MBL-producing Gram-negative pathogens and that ZN148 is a highly promising MBL inhibitor that is capable of operating in a functional space not presently filled by any clinically approved compound.

The rise of antimicrobial resistance (AMR) worldwide and the increasing spread of multi-drug-resistant organisms expressing metallo-β-lactamases (MBL) require the development of efficient and clinically available MBL inhibitors. At present, no such inhibitor is available, and research is urgently needed to advance this field. We report herein the development, synthesis, and biological evaluation of chemical compounds based on the selective zinc chelator tris-picolylamine (TPA) that can restore the bactericidal activity of Meropenem (MEM) against Pseudomonas aeruginosa and Klebsiella pneumoniae expressing carbapenemases Verona integron-encoded metallo-β-lactamase (VIM-2) and New Delhi metallo-β-lactamase 1 (NDM-1), respectively. These adjuvants were prepared via standard chemical methods and evaluated in biological assays for potentiation of MEM against bacteria and toxicity (IC50) against HepG2 human liver carcinoma cells. One of the best compounds, 15, lowered the minimum inhibitory concentration (MIC) of MEM by a factor of 32-256 at 50 μM within all tested MBL-expressing clinical isolates and showed no activity toward serine carbapenemase expressing isolates. Biochemical assays with purified VIM-2 and NDM-1 and 15 resulted in inhibition kinetics with kinact/ KI of 12.5 min-1 mM-1 and 0.500 min-1 mM-1, respectively. The resistance frequency of 15 at 50 μM was in the range of 10-7 to 10-9. 15 showed good tolerance in HepG2 cells with an IC50 well above 100 μM, and an in vivo study in mice showed no acute toxic effects even at a dose of 128 mg/kg.

The uptake of specific fatty acids in humans is dependent on their position on the glycerol backbone. There is a great interest in methods that can access this information fast and accurately. By way of high-resolution NMR, we have analyzed TAG extracted from pig and beef tissues and obtained quantitative data for the composition and regioisomeric distribution of all major unsaturated fatty acids usually found in these source materials, using a combination of manual integration and deconvolution of

The uptake of specific fatty acids in humans is dependent on their position on the glycerol backbone. There is a great interest in methods that can access this information fast and accurately. By way of high-resolution NMR, we have analyzed TAG extracted from pig and beef tissues and obtained quantitative data for the composition and regioisomeric distribution of all major unsaturated fatty acids usually found in these source materials, using a combination of manual integration and deconvolution of 13 C NMR spectra. In addition, we have developed a method for determining composition and regioisomeric distribution of the two main saturated fatty acids found in pork (16:0, 18:0). The results are discussed in relation to species-specific genetic characteristics of fatty acid and TAG biosynthesis. The developed method could support decisions related to breeding for desired fatty acid profiles, and stimulate further methodology developments using high field NMR.

International audience; The syntheses of metallo-β-lactamase inhibitors comprising chelating moieties, with varying zinc affinities,and peptides partly inspired from bacterial peptide sequences, have been undertaken. The zinc chelatorstrength was varied using the following chelators, arranged in order of ascending binding affinity:dipicolylamine (DPA, tridentate), dipicolyl-1,2,3-triazolylmethylamine (DPTA, tetradentate) dipicolyl ethylenediamine(DPED, tetradentate) and trispicolyl ethylenediamine (TPED, pentadentate). The chosen peptideswere mainly based on the known sequence of the C-terminus of the bacterial peptidoglycan precursors.Biological evaluation on clinical bacterial isolates, harbouring either the NDM-1 or VIM-2 metallo-β-lactamase, showed a clear relationship between the zinc chelator strength and restoration of meropenemactivity. However, evaluation of toxicity on different cancer cell lines demonstrated a similar trend, and thusinclusion of the bacterial peptides did possess rather high toxicity towards eukaryotic cells.

Four hexacyclic oleananes having characteristic mass spectra with an intense fragment at m / z 325 were found in Late Cretaceous/Tertiary terrigenous oils and immature source rocks containing 18α-oleanane and other saturated pentacyclic compounds of higher plant origin. They are sometimes as abundant as 18α-oleanane itself. They were not found in oils without oleanane. The major hexacyclic oleanane, comprising 50–80% of the hexacyclic oleananes, was isolated using high performance liquid chromatography (HPLC) and characterized using nuclear magnetic resonance spectroscopy (NMR) as 18α-oleanane with the Me group at C-10 shifted to C-9 and with the Me at C-14 creating a new methylene bridge to C-10. Hexacyclic oleananes were not visible in m / z 191 chromatograms. They are most effectively detected by way of GC-MS-MS using the m / z 410 → 325 transition. Since few other compounds produce any m / z 410 → 325 signal, they may sometimes be easier to detect than regular and rearranged oleananes in samples with a low concentration of angiosperm markers. Precursor natural products have not been identified among plant triterpenoids, but might be related to (poly)functionalized oleanoids. Oleanoids hydroxylated at C-27 are obvious precursors. Oxic/clay-rich depositional environments seem to favour the formation of hexacyclic oleananes. A series of 2-alkylated oleananes had previously been characterized using NMR. Their hexacyclic counterparts were tentatively assigned in this study.

series of novel C-33-C-35 hexacyclic benzohopanes (C(33)b-C(35)b) were identified in 39 samples of coal extracts and 39 crude oils of different ages from all over the world. C(33)b and C(34)b homologues were isolated, and their structures were determined by nuclear magnetic resonance. The structure of C(35)b benzohopane was proposed based on the mass spectrum and its similarity with the mass spectra of structurally defined C(33)b and C(34)b homologues. The structures of the C(33)b-C(35)b hexacyclic benzohopanes are closely related to isohopanes; both groups are typical for terrestrial organic matter and can be useful in the correlation analysis. A possible pathway of formation of these novel benzohopanes and their hopanoid precursors with an additional branch in the aliphatic side chain is proposed. C(33)b-C(35)b hexacyclic benzohopanes are stable up to the maturity level corresponding to random vitrinite reflectance (Rr) of similar to 0.80%, which was demonstrated by analyzing the samples of different maturity and by the maturation simulation experiments: hydrous pyrolysis of two bituminous coals (Rr = 0.55 and 0.59%) and pyrolysis of an extracted bituminous coal (Rr = 0.56%) and its asphaltenes. This represents a confirmation that the formation of these novel benzohopanes is related to specific depositional conditions and microbial activity during diagenesis. Mature samples (Rr = 0.8%) and hydrous pyrolysate of the bituminous coals (Rr lt 0.60%) obtained at 330 degrees C show a distinct distribution of benzohopanes in comparison to immature and moderately mature samples, which is characterized by a low abundance of the b series benzohopanes and the presence of regular and numerous other benzohopane isomers. The latter most likely represent isomers of regular and novel benzohopanes with different substitution patterns on the aromatic ring. This isomerization of alkyl groups attached to the aromatic ring, leading to the formation of thermodynamically more stable isomers, is a well-known maturation scenario so far reported in the series of alkylated naphthalenes, phenanthrenes, and dibenzothiophenes. Therefore, in the same way, a distribution of benzohopanes can indicate thermal maturity. In addition to the novel benzohopanes, three series (2 alpha, 2 beta, and 3 beta) of their methylated derivatives were identified in numerous samples. Finally, a novel C-35 heptacyclic benzohopane with an additional cyclopentane ring was also observed in the studied samples, and its structure was tentatively identified based on the mass spectrum. Opposite to the hexacyclic C(33)b-C(35)b benzohopanes, the formation of the C-35 heptacyclic benzohopane does not require a specific hopanoid precursor with two branches in the side chain. Therefore, this compound seems to have less geochemical significance than the new hexacyclic benzohopanes.

Two previously unidentified dehydroabietane isomers were isolated from Miocene Serbian lignite and Rhaetian (Late Triassic) coaly mudstones from South Sweden and characterized using NMR-spectroscopy as cis- and trans-dehydroicetexane. Both have a 9(10→20)-abeo-abietane or icetexane skeleton consisting of a 6-7-6 tricyclic framework with seven carbons in ring B instead of the usual six in common diterpanes of the abietane-type. Dehydroicetexanes can be detected using GC-MS-MS in m/z 270 → 146 chromatograms without interference from dehydroabietane or other isomers. Dehydroicetexanes are often abundant in high latitude coals and mudstones ranging fromTriassic to Miocene, and in high latitude oils (Canada and Greenland) sourced from terrigenous organic matter. The trans/(cis+ trans) dehydroicetexane ratio is low in immature sediments, but usually around 0.83 in oils and mature sediments with vitrinite reflectance (%Rr) above 0.5 suggesting an equilibrium from the start of the oil window. Dehydroicetexanes are more stable than dehydroabietane and some oils, rich in dehydroicetexanes, contain no dehydroabietane. Precursors could be plant diterpenoids having the icetexane structure, which have been known for more than 40 years and isolated from a variety of higher plant sources, including some angiosperms. Many of the relatively simple icetexanes were isolated for the first time from Chamaecyparis pisifera and related species where they seem to be particularly abundant, suggesting that dehydroicetaxanes may be used as markers for the genus Chamaecyparis or for Cupressoideae in general. This is the peer-reviewed version of the following article: Nytoft, H. P.; Kildahl-Andersen, G.; Lindström, S.; Rise, F.; Bechtel, A.; Mitrović, D.; Đoković, N.; Životić, D.; Stojanović, K. A. Dehydroicetexanes in Sediments and Crude Oils: Possible Markers for Cupressoideae. Organic Geochemistry 2019, 129, 14–23. [https://doi.org/10.1016/j.orggeochem.2019.01.001]

A series of novel C33-C35 hexacyclic benzohopanes have been identified in coals and crude oils of different ages from all over the world [1]. They differ from regular benzohopanes by the presence of methyl group in position C-31. These compounds are more abundant in coals and terrestrial oils. The investigation also showed that mature samples have distributions of benzohopanes distinct from immature ones, which are characterised by a greater number of benzohopane isomers. Although exact structures of the additional isomers were not determined, it seems they were formed by an isomerisation of alkyl groups attached to the aromatic ring. Two novel monoaromatic hydrocarbons (MW = 270; basic fragment ions m/z 255 and 146) were identified in coals, mudstones and crude oils. Their structures were determined as cis- and transicetexa-8,11,13-trienes (or dehydroicetexanes) using NMR spectroscopy. Dehydroicetexanes are potential biomarkers of Cupressaceae (cypress conifers). Benzo[b]naphtho[d]furans (BNFs) have been identified in oils, condensates, source-rocks, coals and coaly shales, being more abundant in the latter. We detected BNFs (m/z 218) in relatively high amounts in the aromatic fractions of pyrolysates of brown coals which attained the maturity corresponding to vitrinite reflectance of 1.80 %Rr [2]. This result suggests that BNFs can be attractive for investigations of mature fluids that originate from gas/condensate prone sources, rich in type III kerogen, which are usually depleted in biomarkers. Recently, it was observed that [2,1]/[1,2]BNF ratio can be used to describe lithofacies [3]. This ratio is much lower in sediments from fluvial-deltaic systems than in clay-depleted sediments from marine environments. 1-Chloro-n-alkanes (m/z 91) have been identified in saltmarsh vegetation, recent sediments from estuarine setting and freshwater lake sediments [4]. Despite their absence in the investigated crude oils, we identified a series of 1-chloro-n-alkanes in pyrolysates of the corresponding asphaltenes, obtained at 250 oC. This result indicates very good preservation of biomarkers occluded inside asphaltenes and announces a possible application of 1-chloro-n-alkanes in correlation studies. Invited Lecture

BACKGROUND Fever of unknown origin and high sedimentation rate are common clinical problems. MATERIAL AND METHODS A middle-aged man with fever of unknown origin, night sweats and high sedimentation rate was referred to our hospital for investigation. RESULTS AND INTERPRETATION The patient was suspected to have mononucleosis or reactivation of infectious mononucleosis because of mild anaemia and thrombocytopenia, a weakly positive IgM antibody test for Epstein-Barr virus and monocytosis (in peripheral blood). Because monocytosis, elevated sedimentation rate and fever persisted, bone marrow smears were prepared and biopsies taken.The third biopsy showed that morphology was consistent with chronic myelomonocytic leukemia (CMML), which was confirmed by two later biopsies. However, a malignant cell population (consisting of blasts in peripheral blood) was only found in one of several flow cytometry assessments of peripheral blood and bone marrow aspirate and cytogenetic analyses of bone marrow cells were normal. The patient's clinical situation has been stable for some years and treatment has not been necessary.

Tricyclic terpanes of the cheilanthane series in crude oils and sediments are routinely used for geochemical correlation. The cheilanthanes are particularly useful to identify crude oils derived from marine-clastic or carbonate source rock as well as input to the source rocks from marine algal or terrigenous organic matter. However, they sometimes have limitations for the correlation of oils expelled from different source rocks that received similar organic matter, (e.g. oils from source rocks dominated by terrigenous organic matter). Novel non-cheilanthane tricyclic terpanes, consisting of C21 and C25 compounds and a C27 tetracyclic terpane were identified in oils from the Assam (India), Beaufort-Mackenzie (Canada), and the Niger (Nigeria), deltas. NMR data reveal that all three compounds have an oleanane-type structure with rings C, D and E intact, while rings A and B have been degraded, leaving a side chain attached to ring C in the case of the tricyclics and only a degraded A ring in the tetracyclic. On the basis of occurrence, NMR structure and co-variance with oleanane abundance, these new tricyclic and tetracyclic compounds are believed to form from A-ring functionalised oleanoid precursors in angiosperm (flowering) plants. The distribution of these compounds in representative oils from the Niger Delta accumulations permits separation of the oils into at least three distinct source rock organofacies.

The three most abundant rearranged oleanoid triterpanes eluting early in gas chromatography (GC) have been isolated from a Niger Delta oil and characterized using nuclear magnetic resonance (NMR) spectroscopy. The NMR data confirmed the identification of two of them, observed previously in the literature, as 1(10 → 5)abeo-3β-methyl-24β-nor-18α(H)-oleanane and 5(4 → 3)abeo-3α(H), 5β(Η), 18α(Η)-oleanane on the basis of X-ray crystallography and comparison with a synthetic standard. The third compound was a novel rearranged oleanane, 3α,5β-dimethyl-23α,25-dinor-10β(H),18α(H)-oleanane, with one of the methyl groups at C-4 in the oleanane structure shifted to C-3 and another shifted from C-10 to C-5. The distribution of these three rearranged oleananes and other, yet unidentified, rearranged oleanoid C 30 triterpanes in 25 Late Cretaceous- or Tertiary-sourced oils from Angola, Canada, Greece, India, Indonesia, Iran, New Zealand, Nigeria and Vietnam was examined by way of reverse phase high pressure liquid chromatography (HPLC), GC–mass spectrometry (GC–MS) and GC–MS–MS. Mass spectra of rearranged oleananes and compounds tentatively assigned as rearranged taraxastanes are presented. Rearranged oleananes and other rearranged triterpanes are probably formed by dehydration and rearrangement of higher plant triterpenoids functionalized at C-3. The distribution of rearranged oleananes and their relative concentration seems to be dependent on the extent of clay catalysis during diagenesis. A diagenetic pathway for formation of the rearranged oleananes is proposed. All 25 oils have almost identical rearranged oleanane/oleanane and diasterane/regular sterane ratio values. Two unidentified early eluting (GC) rearranged oleananes are only abundant in oils with a high proportion of diasteranes. Bicadinanes were found in 20 of the 25 oils.

Microbial culture collections are important resources for isolation of natural compounds with novel properties. In this study, a culture collection of around 1,500 pigmented heterotrophic bacteria was established. The bacteria were isolated from the sea surface microlayer at different sampling sites along the mid-part of the Norwegian coast. The bacterial isolates produced pigments of various coloration (e.g. golden, yellow, red, pink and orange). Methanol extracts of sixteen isolates were characterized with LC-Diodearray-TOF mass spectrometry analysis. The number of pigments per isolate varied considerably, and a tentative identification of the pigments was performed based on UV-absorbance profile and molecular formula assignation based on the accurate mass determination. The LC-MS analyses revealed that most of the pigments probably were carotenoids. Furthermore, we developed a high throughput LC-MS method for characterization and screening of a larger sub-fraction (300 isolates) of the culture collection. The aim was to screen and identify bacterial isolates producing carotenoids that absorb light in the UVA-Blue light. Six of the bacterial strains were selected for detailed investigation, including 16s rRNA sequencing, preparative HPLC for purification of major carotenoids and subsequent structural elucidation with NMR. Among the identified carotenoids were zeaxanthin, nostoxanthin and sarcinaxanthin, some with novel glycosylation patterns.

A 70-year-old man with a seronegative rheumatoid arthritis showed some peculiar lamellar crystalline deposits mainly at the Descement/endothelial level in both eyes. Serum protein analysis revealed a monoclonal gammopathy with increased amounts of IgG and K-light chains along with traces of the latter in the urine. Corneal biopsy was not available, but electron microscopy of the conjunctival specimen demonstrated crystalline deposits within the endoplasmic reticulum of plasma cells lying within the connective tissue. This material is supposed to be of protein nature. It seems reasonable that the conjunctival deposits are identical to those seen in the cornea by slit-lamp examination, and it is also suggested that they represents paraprotein aggregates. After treatment with melphalan (Alkeran) and prednisone systemically, the corneal deposits decreased over a period of 9 months with corresponding improvement of the visual acuity.

Two new C 15 sesquiterpanes have been isolated from a Niger Delta oil. Based on nuclear magnetic resonance (NMR) data, their structures are similar to the ring D and E part of oleanane. The mass spectra show an m/z 193 base peak typical of rearranged drimanes. They have been found only in oleanane-containing oils with a Cretaceous or Tertiary source. Data are presented for 34 such oils from Angola, Canada, Greece, India, Indonesia, Iran, New Zealand, Nigeria, Thailand and Vietnam. The ratio between the new sesquiterpanes and rearranged drimanes roughly follows the oleanane index. They are absent from marine oils whose source rock age (Jurassic or older) predates the evolution and proliferation of angiosperms. They are probably formed via degradation of functionalized oleanoids or by cleavage of seco -oleananes. These sesquiterpanes may be useful as markers of angiosperm input in light oils (jet fuel, diesel or condensates) usually devoid of higher molecular weight markers for higher plant input.

In a randomized study of 92 previously untreated patients with multiple myeloma, the intention was to document the possible beneficial effect of combination chemotherapy including vincristine, carmustine, alkylating agents and prednisone, as compared to conventional therapy with melphalan and prednisone. Major prognostic factors did not differ significantly between the treatment groups. With the 2-drug therapy and 5-drug combination therapy, 48 and 54% of the patients achieved remission, respectively. Median survival for patients treated with the 2-drug regimen and 5-drug regimen was 29 and 33.5 months, respectively. No significant difference was found between the survival curves for stage III patients treated with the two regimens. After 12 months, patients who had achieved remission were randomized to have treatment discontinued or to have maintenance treatment. The numbers of relapses, remission duration and survival of the two groups were similar.

67 previously untreated patients with multiple myeloma were entered on a randomized clinical trial to determine whether the use of combination chemotherapy including vincristine, carmustine, alkylating agents, and prednisone was more effective than conventional therapy with melphalan and prednisone. The treatment groups did not show significant differences with respect to major prognostic factors. With the 2-drug combination therapy and 5-drug combination therapy, 67 and 74% of the patients achieved remission, respectively. Moreover, no significant difference was found between the two treatment schedules in terms of median survival (30 + months). The survival curves for stage III patients treated with the two regimens did not differ significantly. After 12 months, patients who had achieved remission were randomized to have treatment discontinued or to have maintenance treatment. 7 of 15 patients on maintenance therapy relapsed, whereas 9 of 14 patients who had their therapy discontinued relapsed, and the survival of the two groups was similar.