Lipid-modified G4-decoy oligonucleotide anchored to nanoparticles: delivery and bioactivity in pancreatic cancer cells.

KRAS is mutated in >90% of pancreatic ductal adenocarcinomas. As its inactivation leads to tumour regression, mutant KRAS is considered an attractive target for anticancer drugs. In this study we report a new delivery strategy for a G4-decoy oligonucleotide that sequesters MAZ, a transcription factor essential for KRAS transcription. It is based on the use of palmitoyl-oleyl-phosphatidylcholine (POPC) liposomes functionalized with lipid-modified G4-decoy oligonucleotides and a lipid-modified cell penetrating TAT peptide. The potency of the strategy in pancreatic cancer cells is demonstrated by cell cytometry, confocal microscopy, clonogenic and qRT-PCR assays.

Haemolysis following rapid experimental red blood cell transfusion--an evaluation of two infusion pumps.

BACKGROUND: The vast majority of infusion pumps used for rapid transfusion of large amounts of blood have never been properly examined regarding their influence on the quality of the red blood cells (RBCs) infused. In this study, we evaluated the effect of two different infusion pumps on the degree of RBC destruction following rapid experimental blood transfusion. METHODS: Divided into 2 groups according to age, 30 u of SAGM RBCs were infused through an experimental transfusion model by either a manual roller pump (MRP) or a pressure infusor pump (PIP). Fresh (i.e stored for 8-11 d) RBCs, 20 u, and 10 u of older (i.e. stored for 25-33 d) RBCs were randomly allocated to infusion with either of the two pumps. The rate of infusion was as fast as possible with the MRP, and with the PIP adjusted with an external applied pressure of 300 mm Hg. RBC samples collected before and after infusion were analyzed for total haemoglobin, free haemoglobin, haematocrit, total free potassium, lactate dehydrogenase (LDH) and the percentage of haemolysis. The time spent for each transfusion was measured by a stop watch. RESULTS: Following infusion, a marginal increase (i.e. considerably below 0.8%) in the percentage of haemolysis and LDH content was seen with both pumps. This increase was only statistically significant when RBCs stored for 8-11 d were used (P = 0.002 for both parameters). Irrespective of the age of the RBCs, no differences between the two pumps could be detected. Compared to the PIP, infusion with the MRP could be accomplished significantly faster, i.e. median 5.9 ml/s (5.2-6.4 ml/s) versus 2.9 ml/s (2.5-3.2 ml/s), (P < 0.0001). CONCLUSIONS: Both the pumps used in this study are safe alternatives for rapid transfusion of RBCs; however, with MRP this can be accomplished approximately twice as fast as with the PIP.

The weight of the normal spleen.

The aim of the present study was to determine the weight of the normal spleen based upon a retrospective material of 539 medico-legal autopsies. Only cases above 20 years of age without indication of disease or conditions known to influence the spleen weight were included. It is found that the spleen weight is positively correlated to height, body weight and degree of acute congestion but not to sex or age. A table in which the spleen weight is correlated to height and weight is presented. The table can be used for comparison with the actual findings at an autopsy.

Preparation of white cell-reduced red cells by filtration: comparison of a bedside filter and two blood bank filter systems.

BACKGROUND: Concern has been raised about the quality of white cell (WBC)-reduced red cells (RBCs) obtained by bedside filtration. The bedside performance and workload of a routine bedside filter have been compared to the laboratory performance and workload of two blood bank filter systems. STUDY DESIGN AND METHODS: Buffy coat-depleted saline-adenine-glucose-mannitol (SAGM) RBCs (90 units) were prepared. Thirty units were filtered with each of the two blood bank filter systems, and 30 units were filtered (but not transfused) with the bedside filter in a clinical department after 8 to 24 days of storage. The RBCs lost and the postfiltration WBC content (Nageotte chamber) were determined for all filtered units, and the workload associated with filtration by each of the filter systems/filter was assessed. Units with a postfiltration content of > or = 2 x 10(6) WBCs were regarded as filtration failures. RESULTS: Four (13%) of the 30 units filtered at the bedside were filtration failures, compared to no failures with either of the blood bank filter systems. In addition, the median WBC content (0.14 x 10(6)) of the units filtered at the bedside (2 units/filter) was significantly higher than that of the units filtered in the blood bank (0.05 x 10(6)). The RBC loss was significantly higher with the filter systems than with the bedside filter, provided 2 units per filter were processed with the latter. The timed workload of the filter systems was 45 to 75 minutes per 12 units, which was similar to the time required for bedside filtration. CONCLUSION: Bedside filtration of 2 units of stored buffy coat-depleted SAGM RBCs per filter resulted in a higher incidence of filtration failure and higher postfiltration WBC content than did laboratory filtration of 1 unit of fresh buffy coat-depleted SAGM RBCs per filter with either of two blood bank filter systems.

Three-colour flow cytometric immunophenotyping in HIV-patients; comparison to dual-colour protocols.

Flow cytometric measurement of circulating CD4+ lymphocytes is important in the evaluation of disease progression in HIV-infected patients. Development of dyes that can be exited at 488 nm and have emission maximum in the far red area has made three-colour protocols, together with fluorescein isothiocyanate (FITC) and R-phycoerythrin (PE), possible in most clinical flow cytometers. We report here the comparison of a two-tube, three-colour protocol (including CD45/CD4/ CD3 and CD8/CD4/CD3) with our conventional dual-colour protocol. No significant differences were found between percentage of CD3+ lymphocytic cells determined with three different antibody combinations. When the CD8/CD4/CD3 combination was used a systematic overestimation of CD3+ CD4+% cells was found. This turned out to be caused by the formation of 'CD8-escapees'. These are clumps of CD8+ cells that fall outside the lymphocyte gating region, principally because of high side scatter. The problem can be overcome by rigorous vortexing to loosen aggregates. The lymphocyte gating principle used in this protocol (gating on a side scatter/CD45 dot plot) is readily applicable to other antibody combinations. This was demonstrated by measuring CD5+ B lymphocytes, a subset receiving increasing attention in the study of HIV-induced immune deviations. We conclude that our three-colour protocol for CD4+ T-lymphocyte determinations offers significant advantages to the conventional dual-colour method, and we suggest that when possible anti-CD45 be added to dual-colour combinations in order to improve lymphocyte gating.

Is standard 555 MBq 131I-therapy of hyperthyroidism ablative?

The effect of a standard 555 MBq 131I dose in ablating the thyroid gland was investigated in 116 consecutive hyperthyroid patients. Fifty-one had Graves' disease, 50 a multinodular toxic goitre and 15 had a solitary toxic nodule. 555 MBq 131I was given regardless of size or type of the gland and severity of the disease. Within one year after this dose hypothyroidism was induced in 41% of patients with Graves' disease, but in only 13% with a solitary toxic adenoma, and 6% with a multinodular gland. Forty-eight percent of the patients with a multinodular gland, 33% with Graves' disease and 13% with a solitary toxic nodule were still hyperthyroid. Since this so called ablative treatment only accomplishes hypothyroidism in 26/116 (23%) of our patients and results seem unpredictable 131I treatment adjusted according to gland size and type aiming at achieving euthyroidism could be contemplated.

[Bronchial asthma treated with long-acting beta 2 agonist. Comparison between formoterol (12 mu/g) inhaled twice daily and salbutamol (200 mu/g) inhaled 4 times daily]. / Asthma bronchiale behandlet med langtidsvirkende beta 2-agonist. En sammenligning mellem formoterol (12 micrograms) inhaleret to gange daglig og salbutamol (200 micrograms) inhaleret fire gange daglig.

Forty patients with stable asthma and daily need for inhaled beta 2-agonist, were included in a randomized double-blind study. They were treated for six weeks with inhaled beta 2-agonist, either salbutamol, 4 x 200 micrograms daily, of formoterol, 2 x 12 micrograms and 2 x placebo daily. This was preceded by a run-in period, where all patients received terbutalin-inhalation, 4 x 500 micrograms daily. Twenty patients were given formoterol and 18 salbutamol. One patient in the salbutamol-treated group discontinued treatment after three weeks, because of deterioration of asthma. On a diary card, patients recorded peak expiratory flow rate (PEFR) morning and evening before medication, score of asthma symptoms (scale 0-3; 0 = no symptoms, and 3 = severe symptoms) and use of additional doses of beta 2-agonist. Forced expiratory volume in 1 sec. (FEV1), forced vital capacity (FVC) and PEFR were obtained after 0, three and six weeks of treatment. Blinded global assessment of the treatment was performed by both patient and physician at the end of the study. During run-in the two groups of patients were different. The group subsequently treated with salbutamol had a statistical significant (ss) higher morning-PEFR, ss fewer asthma-symptom scores than one during night and ss less need for additional puffs of beta 2-agonist. During treatment, the formoterol-treated group showed an ss increase in morning-PEFR, as compared to run-in. Furthermore this group had ss fewer nocturnal symptom scores than one and ss less need for extra beta 2-agonist during night, than the salbutamol-treated group.(ABSTRACT TRUNCATED AT 250 WORDS)

[Benign pancreatic glucagonoma and recurrent pulmonary embolism with fatal course in a 36-year old man]. / Benignt pancreasglukagonom og recidiverende lungeembolier med letalt forløb hos 36-årig mand.

A case of glucagonoma of the pancreas, without necrolytic migratory erythema, but accompanied by lethal pulmonary embolism in a 36 year old man is presented. The pathology, epidemiology and treatment of the glucagonoma syndrome is briefly described.