HIF2α-dependent Dock4/Rac1-signaling regulates formation of adherens junctions and cell polarity in normoxia.

Hypoxia-inducible factors (HIF) 1 and 2 regulate similar but distinct sets of target genes. Although HIFs are best known for their roles in mediating the hypoxia response accumulating evidence suggests that under certain conditions HIFs, particularly HIF2, may function also under normoxic conditions. Here we report that HIF2α functions under normoxic conditions in kidney epithelial cells to regulate formation of adherens junctions. HIF2α expression was required to induce Dock4/Rac1/Pak1-signaling mediating stability and compaction of E-cadherin at nascent adherens junctions. Impaired adherens junction formation in HIF2α- or Dock4-deficient cells led to aberrant cyst morphogenesis in 3D kidney epithelial cell cultures. Taken together, we show that HIF2α functions in normoxia to regulate epithelial morphogenesis.

Long-term skeletal, cephalometric, and volumetric changes in two Helsinki bimaxillary face transplant patients.

Analysis of skeletal, cephalometric, and volumetric changes and occlusion during long-term follow-up was performed for two patients who underwent bimaxillary facial transplantation (FT). The study material consisted of the follow-up data of two bimaxillary composite FT performed in Helsinki University Hospital, one in 2016 and the other in 2018. Serial three-dimensional computed tomography scans obtained during follow-up (6 years for patient 1, 4 years for patient 2) were analyzed. The position of the maxilla remained stable in both patients. At 4 and 6 years, the changes in the anterior maxilla were ≤1 mm, while the anterior mandible had moved 2.6-4 mm anteriorly and the mandibular midline 0.4-3.7 mm to the left side. Patient 1 underwent re-osteosynthesis 4 months after transplantation due to mandibular non-union. Patient 2 had a sagittal mandibular osteotomy at 15 months after FT due to lateral crossbite and tension created by temporomandibular joint rotation. Thereafter both patients had a stable occlusion. A continuous bone volume reduction in the mandible was noticed in both patients (6% and 9% reduction of the transplanted volume). The volume of the transplanted maxilla decreased during the early postoperative period but increased back to the original transplanted volume during the follow-up.

Variability in PET image quality and quantification measured with a permanently filled 68Ge-phantom: a multi-center study.

BACKGROUND: This study evaluated, as a snapshot, the variability in quantification and image quality (IQ) of the clinically utilized PET [18F]FDG whole-body protocols in Finland using a NEMA/IEC IQ phantom permanently filled with 68Ge. METHODS: The phantom was imaged on 14 PET-CT scanners, including a variety of models from two major vendors. The variability of the recovery coefficients (RCmax, RCmean and RCpeak) of the hot spheres as well as percent background variability (PBV), coefficient of variation of the background (COVBG) and accuracy of corrections (AOC) were studied using images from clinical and standardized protocols with 20 repeated measurements. The ranges of the RCs were also compared to the limits of the EARL 18F standards 2 accreditation (EARL2). The impact of image noise on these parameters was studied using averaged images (AVIs). RESULTS: The largest variability in RC values of the routine protocols was found for the RCmax with a range of 68% and with 10% intra-scanner variability, decreasing to 36% when excluding protocols with suspected cross-calibration failure or without point-spread-function (PSF) correction. The RC ranges of individual hot spheres in routine or standardized protocols or AVIs fulfilled the EARL2 ranges with two minor exceptions, but fulfilling the exact EARL2 limits for all hot spheres was variable. RCpeak was less dependent on averaging and reconstruction parameters than RCmax and RCmean. The PBV, COVBG and AOC varied between 2.3-11.8%, 9.6-17.8% and 4.8-32.0%, respectively, for the routine protocols. The RC ranges, PBV and COVBG were decreased when using AVIs. With AOC, when excluding routine protocols without PSF correction, the maximum value dropped to 15.5%. CONCLUSION: The maximum variability of the RC values for the [18F]FDG whole-body protocols was about 60%. The RC ranges of properly cross-calibrated scanners with PSF correction fitted to the EARL2 RC ranges for individual sphere sizes, but fulfilling the exact RC limits would have needed further optimization. RCpeak was the most robust RC measure. Besides COVBG, also RCs and PVB were sensitive to image noise.

ESTIMATION OF HP(3) AMONG STAFF MEMBERS IN TWO NUCLEAR MEDICINE UNITS IN FINLAND.

The eye lens exposure among 16 technicians in two nuclear medicine departments at university hospitals in Finland was investigated by measuring the operational quantity Hp(3) using EYE-D dosemeters. For all workers, the annual mean Hp(3) was estimated to be 1.1 mSv (max. 3.9 mSv). The relation between Hp(3) to routinely monitored personal dose equivalent Hp(10) was clearly correlated. Considering individual dose measurement periods (2-4 weeks), the Hp(3)/Hp(10) ratio was 0.7 (Pearson's coefficient r = 0.90, p < 0.001, variation of ratio 0.1-2.3). The variation decreased considerably with increasing Hp(10) (σ2 = 0.04 vs. 0.43 for Hp(10) > 0.1 mSv vs. < 0.1 mSv, respectively), i.e. higher Hp(10) predicts Hp(3) more reliably. Moreover, annual Hp(10) data from national dose register during 2009-2018 were used to derive the annual Hp(3) applying the Hp(3)/Hp(10) ratio. The data from Finnish nuclear medicine departments imply that routine measurements of Hp(3) among nuclear medicine technicians are not justified.

Identification of the Nef-associated kinase as p21-activated kinase 2.

The Nef protein of primate immunodeficiency viruses plays an important role in the pathogenesis of acquired immunodeficiency syndrome (AIDS) [1] [2]. The interaction of Nef with the Nef-associated kinase (NAK) is one of the most conserved properties of different human and simian immunodeficiency virus (HIV and SIV) Nef alleles. The role of NAK association is currently not known but it has been implicated in enhanced viral infectivity in cell culture and in disease progression in SIV-infected macaques [3]. Previous studies have indicated that NAK shares many features with the p21-activated kinases (PAKs) [3], but the molecular identity of NAK has remained unknown. We have generated specific antisera against PAKs 1-3, and expressed these kinases individually as epitope-tagged proteins. By using these reagents in experiments involving partial proteolytic mapping, and exploiting the unique ability of PAK2 to serve as a caspase substrate, we have positively identified NAK as PAK2. Interestingly, although ectopic PAK2 overexpression efficiently replaced endogenous PAK2 from the complex with Nef, the total Nef-associated PAK2 activity was not increased, indicating the abundance of another cellular factor(s) as the limiting factor in Nef-PAK2 complex formation. Identification of NAK as PAK2 should now facilitate elucidation of its role as a mediator of the pathogenic effects of Nef.

Oncogenic K-Ras upregulates ITGA6 expression via FOSL1 to induce anoikis resistance and synergizes with αV-Class integrins to promote EMT.

In many cancer types, integrin-mediated signaling regulates proliferation, survival and invasion of tumorigenic cells. However, it is still unclear how integrins crosstalk with oncogenes to regulate tumorigenesis and metastasis. Here we show that oncogenic K-RasV12 upregulates α6-integrin expression in Madin-Darby canine kidney (MDCK) cells via activation of the mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK)/Fos-related antigen 1-signaling cascade. Activated α6-integrins promoted metastatic capacity and anoikis resistance, and led to perturbed growth of MDCK cysts. Transcriptomic analysis of K-RasV12-transformed MDCK cells also revealed robust downregulation of αV-class integrins. Re-expression of αV-integrin in K-RasV12-transformed MDCK cells synergistically upregulated the expression of Zinc finger E-box-binding homeobox 1 and Twist-related protein 1 and triggered epithelial-mesenchymal transition leading to induced cell motility and invasion. These results delineate the signaling cascades connecting oncogenic K-RasV12 with α6- and αV-integrin functions to modulate cancer cell survival and tumorigenesis, and reveal new possible strategies to target highly oncogenic K-RasV12 mutants.

Hyperinsulinaemia, hyperaminoacidaemia and post-exercise muscle anabolism: the search for the optimal recovery drink.

Dietary supplements and other ergogenic aids are popular among athletes. Recent studies have shown that nutritional mixtures containing protein hydrolysates, added leucine, and high-glycaemic carbohydrates greatly augment insulin secretion compared with high-glycaemic carbohydrates only. When post-exercise hyperinsulinaemia is supported by hyperaminoacidaemia induced by protein hydrolysate and leucine ingestion, net protein deposition in muscle should occur. Thus, consumption of post-exercise recovery drinks containing these nutrients in conjunction with appropriate resistance training may lead to increased skeletal muscle hypertrophy and strength. However, the long-term effects on body composition and exercise performance remain to be determined.

Reducing current noise in cryogenic experiments by vacuum-insulated cables.

We measure the current noise of several cryogenic cables in a pulse tube based dilution refrigerator at frequencies between about 1 mHz and 50 kHz. We show that vibration-induced noise can be efficiently suppressed by using vacuum-insulated cables between room temperature and the 2nd pulse tube stage. A noise peak below 4 fA at the 1.4 Hz operation frequency of the pulse tube and a white noise density of 0.44 fA/Hz in the millihertz range are obtained.

Validation of a MOSFET dosemeter system for determining the absorbed and effective radiation doses in diagnostic radiology.

This study aimed to validate a MOSFET dosemeter system for determining absorbed and effective doses (EDs) in the dose and energy range used in diagnostic radiology. Energy dependence, dose linearity and repeatability of the dosemeter were examined. The absorbed doses (ADs) were compared at anterior-posterior projection and the EDs were determined at posterior-anterior, anterior-posterior and lateral projections of thoracic imaging using an anthropomorphic phantom. The radiation exposures were made using digital radiography systems. This study revealed that the MOSFET system with high sensitivity bias supply set-up is sufficiently accurate for AD and ED determination. The dosemeter is recommended to be calibrated for energies <60 and >80 kVp. The entrance skin dose level should be at least 5 mGy to minimise the deviation of the individual dosemeter dose. For ED determination, dosemeters should be implanted perpendicular to the surface of the phantom to prevent the angular dependence error.

Inhalation of swine-house dust increases the concentrations of interleukin-1 beta (IL-1 beta) and interleukin-1 receptor antagonist (IL-1ra) in peripheral blood.

Inhalation of dust in swine confinement buildings causes airway inflammation and systemic symptoms. The proinflammatory cytokines interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha) increase in bronchoalveolar and nasal lavage fluid, and in serum. The aim of this investigation was to study changes in the IL-1 family of cytokines in peripheral blood in 36 healthy volunteers exposed to swine house dust for 3 h. Interleukin (IL-1 beta) was measured in platelet poor plasma and in a mononuclear cell fraction (PBMC) and interleukin-1 receptor antagonist (IL-Ira), IL-6 and TNF-alpha were measured in serum 4 and 7 h after the start of 3 h exposure. Lung function and a methacholine bronchial provocation test were performed before and 7 h after the start of exposure. The leukocyte count in whole blood and the mononuclear cell count in PBMC were examined before, and 4 and 7 h after the start of exposure. The concentration of airborne inhalable dust and endotoxin were measured using personal samples. The concentration of inhalable dust was 23 (20-30) mg m-3 (median 25th-75th percentile) endotoxin was 1.1 (0.8-1.4) micrograms m-3 and respirable dust (n = 8) was 1.0 (0.7-1.2) mg m-3. IL-1 beta increased from < 0.125 to 0.9 (0.5-1.3) ng l-1 in plasma and from 1.6 to 2.7 (1.1-4.4) ng l-1 in PBMC (P < 0.01). IL-1 ra, IL-6 and TNF-alpha increased 2-, 12- and 2-fold in serum after exposure, respectively. Changes in IL-1 ra correlated with changes in FEV1, bronchial responsiveness, oral temperature (P < 0.01) and blood white cell count (P < 0.05). IL-1 beta correlated significantly with temperature (P < 0.05). These results indicate that IL-1 beta and IL-1 ra increase in peripheral blood following inhalation of swine house dust and may participate in and modulate the inflammatory response together with IL-6 and TNF-alpha.

Maternal and neonatal essential fatty acid status in phospholipids: an international comparative study.

OBJECTIVE: To investigate whether the steady decline in the maternal essential fatty acids (EFA) status during pregnancy observed in Dutch pregnant women is a local or general phenomenon. DESIGN: The EFA status was measured during uncomplicated, singleton pregnancy of healthy women from the Netherlands, Hungary, Finland, England and Ecuador. In addition, the EFA status of their neonates were measured at birth. Fatty acid profiles were analyzed in phospholipids isolated from maternal plasma and from umbilical plasma and cord vessel walls. RESULTS: Considerable differences between these centers were observed in the maternal EFA levels and EFA status indexes. However, the change in the absolute as well as relative amounts of the EFAs followed a similar course in the five populations during pregnancy. The neonatal EFA profiles reflected the differences found in maternal plasma during pregnancy and shortly after delivery. Comparable correlations were found, particularly, between the neonatal and the maternal n-3 fatty acids in the participating groups. CONCLUSIONS: It seems that the reduction in maternal EFA status during pregnancy is a general phenomenon, and is largely independent of differences in dietary habits and ethnic origin. Since the lowest values for certain maternal EFAs in a given country were significantly higher than the highest value of these EFAs throughout pregnancy in other countries, the functional implications of the pregnancy-associated reduction in the maternal EFA status for the fetal and neonatal development is not obvious and needs to be further elucidated.

Ammonia, hydrogen sulphide and methyl mercaptides in Finnish municipal sewage plants and pumping stations.

Ammonia, methane, hydrogen sulphide and methyl mercaptides were analyzed in the atmospheres of 16 Finnish municipal waste water treatment plants and in 18 pumping stations. The sulphides were analyzed on site as it was found that, with the standard 0.5-l laminated plastic bags, significant amounts of the vapours were lost from the gas phase. The relative humidity of the sampled air was the most likely cause. Under normal operating conditions, sulphides varied from less than 0.07 to 53 micrograms l-1, with highest concentrations found at the sludge presses. Pumping stations had sulphide vapours from 0.07 to 0.5 microgram l-1. The presence of ammonia (0.007-3.5 micrograms l-1) and methane (0.7-18 micrograms l-1) confirmed the hypothesis of the anaerobic origin of the offensive gases. Although more effective ventilation without scrubbing the emission gases would reduce the health hazard in the plants it would burden the environment.

Nifedipine concentrations in maternal and umbilical serum, amniotic fluid, breast milk and urine of mothers and offspring.

Eleven hypertensive pregnant patients were treated with nifedipine orally 10 mg thrice daily. At steady state, nifedipine concentration, measured by the high-performance liquid chromatographic method, was 4.3 +/- 1.0 ng/ml (mean +/- s.e.m.) in maternal serum in the third trimester of pregnancy. During delivery, the parent drug levels in maternal and umbilical serum and in amniotic fluid were 12.4 +/- 4.0, 8.6 +/- 2.3 and 2.5 +/- 1.2 ng/ml, respectively. Small amounts of nifedipine were found also in urine of the neonates. In breast milk, the nifedipine concentration was 4.1 +/- 0.8 ng/ml on the third day after delivery. The antihypertensive effect of nifedipine did not correlate with the serum drug concentration. The outcome of pregnancy was favourable in all cases.

Blood pressure, plasma renin activity and calcium metabolism in hypertensive pregnancy: the effect of nifedipine.

The effects of nifedipine on blood pressure, plasma renin activity and calcium metabolism were studied in nine hypertensive pregnant patients. Nifedipine (10 mg thrice daily per os) reduced blood pressure from 158/103 +/- 4/1 to 150/96 +/- 4/2 mmHg (mean +/- s.e.m.) during 4 to 5 days' treatment (p less than 0.05). The percentage change in diastolic blood pressure correlated negatively with the initial ambulatory (p less than 0.001) and rest (p less than 0.01) plasma renin activity and the initial daily urine calcium excretion (p less than 0.01). Calcium excretion in urine correlated positively with the initial ambulatory and rest plasma renin activity (p less than 0.01). The blood pressure reduction did not correlate with serum ionized, or total calcium, or the initial blood pressure. In six non-pregnant women, the rest plasma renin activity increased (p less than 0.05) after four days' administration of nifedipine. In the patients with hypertensive pregnancy, no changes in plasma renin activity were found during the treatment. The results indicated that the initial plasma renin activity, but neither the serum ionized calcium, nor the initial blood pressure, predicted the blood pressure-lowering effect of nifedipine in hypertensive pregnancy.

Secondary metabolite concentrations and terpene emissions of Scots pine xylem after long-term forest fertilization.

Secondary compounds are known to be associated with the resistance of conifer xylem against insects and fungi. The effects of long-term forest fertilization with nitrogen (N) or with N, calcium (Ca), and phosphorus (P) on secondary compounds in the xylem of 50-yr-old Scots pine (Pinus sylvestris L.) trees were examined. Xylem samples were collected from trees growing in three locations in southern Finland: Vilppula, Padasjoki, and Punkaharju. Forests were fertilized every fifth (Vilppula and Padasjoki) or tenth (Punkaharju) year since the 1950s. We compared concentrations of individual and total monoterpenes and resin acids in the heartwood and sapwood of Scots pine. Terpene emissions were analyzed from the sapwood and total phenolics from the heartwood. Fertilization did not have any significant effect on the concentrations and emissions of xylem monoterpenes. Concentrations of several individual terpenes in sapwood were positively correlated with the corresponding terpene emission. The concentrations of individual resin acids (i.e., abietic and dehydroabietic) decreased significantly in Punkaharju, but increased in the sapwood of N-fertilized trees compared with control ones at Padasjoki and Vilppula. The concentrations of resin acids in the heartwood were not significantly affected by fertilization. Both fertilization treatments decreased the total phenolic concentrations in the heartwood of trees growing in Padasjoki. There was a significant positive correlation between the total phenolics and total resin acid concentration. Overall, resin acids and phenolics seemed be more responsive than monoterpenes to N treatment. These results suggest that forest fertilization might cause slight changes in secondary compound concentrations of xylem, and thus might have significance in the decay resistance of wood.

The applicability of radiophotoluminescence dosemeter (RPLD) for measuring medical radiation (MR) doses.

The applicability of radiophotoluminescence dosimetry was determined by assessing various radiophotoluminescence dosemeter (RPLD) properties for measuring medical radiation doses from radiation sources of a continuous spectrum. The RPLD was found to be accurate for measuring doses in diagnostics (50-125 keV) and radiation therapy (6, 10 and 18 MV photons, 6 and 15 MeV electrons). The RPLD shows excellent dose linearity (R(2) > 0.99), reproducibility and batch uniformity, and minimal fading and accurate accumulated dose measurement. The dosemeter material is independent of photon energy in the diagnostic range; however, the dosemeter requires additional calibration in the mammography energy range and also for accurate dose measurement with photon or electron energies in radiation therapy. RPLD measurements with a tin filter show considerable angular dependence at angles exceeding 50° between the photon beam and the normal to the long axis of the dosemeter. The RPLD measurement accuracy at high doses can be improved with optimised pre-heating schemes.

A comparison of radiation exposure between diagnostic CTA and DSA examinations of cerebral and cervicocerebral vessels.

BACKGROUND AND PURPOSE: While the number of CTA examinations is continually increasing compared with DSA examinations, there is little comparative dose information about the different imaging techniques. We compared patient radiation exposure resulting from diagnostic CTA and DSA examinations for both cerebral and cervicocerebral vessels. MATERIALS AND METHODS: An anthropomorphic phantom was irradiated by using typical diagnostic CTA and DSA setups and imaging parameters. For both imaging techniques, the imaging area of cerebral vessels included intracranial vessels only, while the imaging area of cervicocerebral vessels included both cervical and intracranial vessels from the aortic arch to the vertex. The effective dose was determined by using RPLDs. The DSA examination was simulated by using a biplane angiography system, and the CTA examination, by using a 64-row multidetector CT scanner. RESULTS: For the imaging of cerebral vessels, the effective dose according to ICRP 103 was 0.67 mSv for CTA and 2.71 mSv for DSA. For the imaging of cervicocerebral vessels, the effective dose was 4.85 mSv for CTA and 3.60 mSv for DSA. The maximum absorbed dose (milligray) for skin, brain, salivary glands, and eyes was 166.2, 73.5, 35.6, and 21.8 mGy for DSA and 19.0, 16.9, 20.4, and 14.8 mGy for CTA, respectively. The conversion factors from DAP and DLP to effective dose were calculated. CONCLUSIONS: The effective dose for CTA assessment of cerebral vessels was approximately one-fifth the dose compared with DSA. In the imaging of cervicocerebral vessels, the effective dose for CTA was approximately one-third higher compared with DSA.

Instrumentation for measuring fluorescence cross sections from airborne microsized particles.

An experimental instrument for measuring a laser-induced fluorescence spectrum from a single aerosol particle is described. As a demonstration of instrument capabilities, the results of monodisperse 4.7 microm sodium chloride particles doped with fluorescent riboflavin, produced with an inkjet aerosol generator, are presented. The fluorescence of the aerosol particles is excited in the wide range from 210 to 419 nm using a pulsed, tunable optical parametric oscillator laser. The maximum of the fluorescence emission of separately measured particles is detected at 560 nm. The dependence of the fluorescence on the excitation wavelength is studied and fluorescence cross sections are estimated. Agreement between the measured fluorescence data and the literature data for riboflavin is observed.

Nifedipine reduces thromboxane A2 production by platelets without changing aggregation in hypertensive pregnancy.

Platelet aggregation and thromboxane A2 generation were studied in hypertensive pregnant women using normotensive non-pregnant and pregnant controls. In hypertensive pregnancy, adrenaline- and adenosine diphosphate-induced platelet aggregation was at the non-pregnant level and lower than in normotensive pregnancy. Collagen-induced aggregation was at a lower level in hypertensive pregnancy than in both control groups. Thromboxane generation during spontaneous clotting and in platelet-rich plasma did not differ between the three groups. However, thromboxane generation was low during aggregation induced by small collagen concentrations in hypertensive pregnancy, but at higher collagen concentrations the difference between the groups disappeared. During nifedipine treatment (10 mg t.i.d.), aggregation and thromboxane production in platelet-rich plasma induced by the three stimuli remained unaltered in hypertensive pregnancy, while thromboxane synthesis during spontaneous clotting was reduced. In nifedipine-treated non-pregnant controls, only EC80 for adrenaline-induced aggregation decreased. In vitro, pharmacological concentrations of nifedipine inhibited platelet aggregation and thromboxane production. In conclusion, nifedipine reduces thromboxane generation in spontaneous clotting, without inhibiting platelet aggregation and thromboxane production in platelet-rich plasma in hypertensive pregnancy. Reduced aggregability of platelet ex vivo may reflect their continuous activation and desensitization in vivo in hypertensive pregnancy.