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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Comparing the variation of needle and wood terpenoids in Scots pine provenances.

We determined variation in both the concentration and composition of terpenoids in needles and wood within nine Scots pine (Pinus sylvestris) provenances. Seedlings of different provenances representing a 1200-km N-S transect from Estonia to northern Finland were cultivated in Suonenjoki nursery field, central Finland, for seven years. Growth of seedlings and the number of vertical resin ducts in wood were also determined. alpha-Pinene and 3-carene were the major monoterpenes both in the needles and wood. The total monoterpene concentration was about five times higher in the needles than in the wood. A strong positive correlation was found between proportional quantities of several terpenes of the needles and wood, particularly for 3-carene, sabinene, and terpinolene. The needles contained both labdane-type and tricyclic resin acids, whereas the wood contained only tricyclic ones. The wood had a four times higher tricyclic resin acid concentration than the needles. The highest total monoterpene concentration in the needles and in the wood occurred in the most northern Muonio provenance and in the most southern Saaremaa provenance plants, respectively. The amount of high 3-carene genotype trees decreased among the northern provenances. The wood of the most northern Muonio provenance showed the lowest total resin acid concentration, but provenance did not affect total tricyclic resin acids in the needles. Korpilahti provenance trees from central Finland had the best growth in height. In addition, Korpilahti and Ruokolahti provenance trees showed largest radial growth of stem and smallest number of vertical resin ducts. The results suggest that especially the proportional quantity of 3-carene in the needles could be used in estimating the amount of this compound in the wood and vice versa.

Inhibition of cellular functions of HIV-1 Nef by artificial SH3 domains.

SH3 domains regulate many normal and pathological cellular processes by guiding specific protein interactions. Studies on binding of HIV-1 Nef to the SH3 domain of the Hck tyrosine kinase have indicated an important role for the SH3 RT-loop region in ligand binding. Here we have tested the potential of artificial Hck-derived SH3 domains carrying tailored RT-loops providing high affinity for Nef as intracellular inhibitors of Nef. These artificial SH3 domains efficiently associated with Nef in cells and thereby potently inhibited SH3-dependent Nef functions, such as association with p21-activated kinase-2 and induction of the transcription factor NFAT. On the other hand, biochemical and functional data indicated that the Nef-targeted SH3 domains were not prone to compete with normal SH3-mediated processes. Thus, RT-loop-modified SH3 domains represent a novel approach for selectively interfering with cellular signaling events, which could be exploited in research as well as in therapeutic applications.

Human immunodeficiency virus type 1 Nef selectively associates with a catalytically active subpopulation of p21-activated kinase 2 (PAK2) independently of PAK2 binding to Nck or beta-PIX.

We have recently identified the Nef-associated serine-threonine kinase (NAK) as the p21-activated kinase 2 (PAK2). Here we have taken advantage of the possibility to manipulate the functional properties of NAK by transfecting PAK2 cDNA or its mutant derivatives in order to further characterize the Nef-NAK complex. To exclude the possibility that some Nef variants might interact with PAK1 instead of PAK2, we also examined the identity of NAK complexed with divergent human immunodeficiency virus type 1 HIV-1 Nef proteins. All tested Nef proteins, including SF2, NL4-3, BH10, and HAN-2, associated with PAK2 but not with PAK1. By exchanging different regions between these two PAK proteins, the selective ability of PAK2 to associate with Nef could be mapped to the carboxy-terminal part of its regulatory domain. Binding of PAK2 with the adapter protein Nck or beta-PIX was found to be dispensable for the assembly of the Nef-PAK2 complex, whereas an intact Cdc42-Rac1 interactive binding motif was required. Most importantly, we found that NAK represented a distinct subpopulation of the total cellular PAK2 characterized by a high specific kinase activity. Thus, although only a small fraction of cellular PAK2 could be found in complex with Nef, NAK represented a major part of cellular PAK2 activity.

Activation of NFAT-dependent gene expression by Nef: conservation among divergent Nef alleles, dependence on SH3 binding and membrane association, and cooperation with protein kinase C-theta.

Here we show that the potential to regulate NFAT is a conserved property of different Nef alleles and that Nef residues involved in membrane targeting and SH3 binding are critical for this function. Cotransfection of an activated protein kinase C-theta (PKC-theta) with Nef implicated PKC-theta as a possible physiological cofactor of Nef in promoting NFAT-dependent gene expression and T-cell activation.

Synergistic activation of NFAT by HIV-1 nef and the Ras/MAPK pathway.

Nef is a lentiviral protein involved in pathogenesis of AIDS, but its molecular mechanisms of action remain incompletely understood. Here we report a novel effect of Nef on lymphocyte signaling, which is mediated via a T cell receptor (TCR)-independent contribution of Nef to induction of nuclear factor of activated T cells (NFAT), a transcription factor that plays a central role in coordinating T cell activation. Expression of Nef did not significantly alter the basal level of NFAT activity in Jurkat cells nor the increased activity following T cell receptor stimulation by anti-CD3 or anti-CD3 + anti-CD28. We also mimicked NFAT induction by TCR triggering by simultaneous activation of the Ras and calcium signaling pathways with phorbol 12-myristate 13-acetate and ionomycin, respectively. Strikingly, whereas activation of either of these pathways individually did not induce NFAT activity in control cells, in Nef-expressing cells phorbol 12-myristate 13-acetate treatment alone resulted in a 100-fold increase in NFAT-directed gene expression. Experiments with different dominant negative mutant signaling proteins, inhibitory chemicals, and Lck-deficient Jurkat cells revealed that this effect was mediated via activation of calcineurin by Nef-induced changes in calcium metabolism, but was independent of TCR-associated signaling events. This ability of Nef to substitute for triggering of the calcium pathway in induction of NFAT could promote activation of human immunodeficiency virus (HIV)-infected T cells in response to stimuli mediated via TCR or other cell surface receptors under conditions when activation of Ras rather than calcium signaling would otherwise predominate.

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.

SH3-Domain binding function of HIV-1 Nef is required for association with a PAK-related kinase.

HIV-1 Nef has previously been shown to bind to Src homology-3 (SH3) domains of a subset of Src family tyrosine kinases. In addition, Nef has been reported to coprecipitate with a serine/threonine kinase activity termed NAK (for Nef-associated kinase). The identity of NAK remains uncertain, but it has been suggested to represent a novel member of the p21-activated kinase (PAK) family. We report here that NAK autophosphorylation is increased not only by an activated form of the p21-family GTPase cdc42 but also by a plasma membrane-targeted fragment of the adapter protein Nck, thus providing further evidence that NAK is related to PAKs. A detailed structure-based mutational analysis of Nef revealed that all amino acid changes that inhibited the Nef/Hck-SH3 interaction, as measured by surface-plasmon resonance, also abolished coprecipitation of NAK. As PAK family proteins do not contain SH3 domains, these observations are best explained by a protein complex in which Nef, NAK, and an SH3-protein all contact each other. In addition, a number of conserved amino acids in Nef that are not involved in SH3 binding were also found to be crucial for association with NAK. Molecular modeling suggests that these residues are involved in formation of an adjacent binding surface for NAK or another critical component of the NAK/Nef complex.

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.