Body composition assessed by dual-energy X-ray absorptiometry predicts early infectious complications after liver transplantation.

BACKGROUND: It is challenging to identify malnutrition, which is a risk factor for poor outcome in patients with liver cirrhosis. In the present study, we aimed to investigate the prevalence of malnutrition among patients listed for liver transplantation, as assessed by different methods, and also to relate dual-energy X-ray absorptiometry (DXA) to short-term post-transplant outcomes. METHODS: In this retrospective cohort study, we reviewed the medical records of 106 patients who underwent liver transplantation in 2009-2012. Body composition was assessed by the fat-free mass index (FFMI) and fat mass index (FMI) obtained using DXA. Severe infections within 1 month, length of stay in intensive care unit and length of hospital stay were endpoints of primary interest. RESULTS: The prevalence of malnutrition was 2-20% depending on sex and the assessment method. Thirty-nine (37%) patients developed severe infections within 1 month after liver transplantation. In multivariate analysis with logistic regression, body composition was significantly associated with post-operative infection when measured with FFMI (P = 0.043) but not with FMI (P = 0.087). Post-operative dialysis (P = 0.004) and post-operative infections (P < 0.001) were significantly associated with length of stay in hospital. Post-operative bleeding (P = 0.015), duration of mechanical ventilation (P < 0.001) and the need for dialysis (P < 0.001), but not body composition, were significant predictors of the length of stay in the intensive care unit. CONCLUSIONS: The prevalence of malnutrition depends on assessment method. FFMI is an independent predictor for early post-transplant infections. Body composition measured by DXA during the pretransplant evaluation provides valuable information about nutritional status in patients with liver cirrhosis.

The role of unfinished root canal treatment in odontogenic maxillofacial infections requiring hospital care.

OBJECTIVES: The aim of this study was to evaluate clinical and radiological findings and the role of periapical infection and antecedent dental treatment of infected focus teeth in odontogenic maxillofacial abscesses requiring hospital care. MATERIALS AND METHODS: In this retrospective cohort study, we evaluated medical records and panoramic radiographs during the hospital stay of patients (n = 60) admitted due to odontogenic maxillofacial infection originating from periapical periodontitis. RESULTS: Twenty-three (38 %) patients had received endodontic treatment and ten (17 %) other acute dental treatment. Twenty-seven (45 %) had not visited the dentist in the near past. Median age of the patients was 45 (range 20-88) years and 60 % were males. Unfinished root canal treatment (RCT) was the major risk factor for hospitalisation in 16 (27 %) of the 60 cases (p = .0065). Completed RCT was the source only in 7 (12 %) of the 60 cases. Two of these RCTs were adequate and five inadequate. CONCLUSIONS: The initiation of inadequate or incomplete primary RCT of acute periapical periodontitis appears to open a risk window for locally invasive spread of infection with local abscess formation and systemic symptoms. Thereafter, the quality of the completed RCT appears to have minor impact. However, a considerable proportion of the patients had not received any dental treatment confirming the importance of good dental health. Thus, thorough canal debridement during the first session is essential for minimising the risk for spread of infection in addition to incision and drainage of the abscess. If this cannot be achieved, tooth extraction should be considered. CLINICAL RELEVANCE: Incomplete or inadequate canal debridement and drainage of the abscess may increase the risk for spread of endodontic infection.

Male broad-nosed pipefish Syngnathus typhle do not locate females by smell.

Broad-nosed pipefish Syngnathus typhle were used to investigate whether males used scent in their search for mates. When the males in an experiment had access to olfactory cues only, they did not locate females better than they located males. Thus, S. typhle, was less successful in mate search when visual cues were absent.

Correlations between microbiota and metabolites after faecal microbiota transfer in irritable bowel syndrome.

Faecal microbiota transfer (FMT) consists of the infusion of donor faecal material into the intestine of a patient with the aim to restore a disturbed gut microbiota. In this study, it was investigated whether FMT has an effect on faecal microbial composition, its functional capacity, faecal metabolite profiles and their interactions in 16 irritable bowel syndrome (IBS) patients. Faecal samples from eight different time points before and until six months after allogenic FMT (faecal material from a healthy donor) as well as autologous FMT (own faecal material) were analysed by 16S RNA gene amplicon sequencing and gas chromatography coupled to mass spectrometry (GS-MS). The results showed that the allogenic FMT resulted in alterations in the microbial composition that were detectable up to six months, whereas after autologous FMT this was not the case. Similar results were found for the functional profiles, which were predicted from the phylogenetic sequencing data. While both allogenic FMT as well as autologous FMT did not have an effect on the faecal metabolites measured in this study, correlations between the microbial composition and the metabolites showed that the microbe-metabolite interactions seemed to be disrupted after allogenic FMT compared to autologous FMT. This shows that FMT can lead to altered interactions between the gut microbiota and its metabolites in IBS patients. Further research should investigate if and how this affects efficacy of FMT treatments.

A comprehensive classification of mandibular fractures: a preliminary agreement validation study.

This study evaluates a comprehensive classification system for mandibular fractures based on imaging analysis. The AO/ASIF scheme, defining three fracture types (A, B, C), three groups within each type (e.g. A1, A2, A3) and three subgroups within each group (e.g. A1.1, A1.2, A1.3) with increasing severity from A1.1 (lowest) to C3.3 (highest) was used. The mandible is divided into two vertical units (I and V), two lateral horizontal units (II and IV) and one central unit (III) comprising the symphyseal and parasymphyseal region. Type A fractures are non-displaced, type B are displaced and type C are multifragmentary/defect injuries. Groups and subgroups are further defined in the classification system. Two classification sessions using semi-automatic software with 7 and 9 surgeons were performed to evaluate 100 fracture cases in the first session and 50 in the second. Inter-observer reliability and individual rater's accuracy were evaluated by kappa coefficient and latent class analysis, respectively. The analysis of inter-observer agreement for the detailed coding showed kappa coefficients around 0.50 with higher agreement among raters in the vertical units. This system allows standardization of documentation of mandibular fractures, although improvement in the definition of categories and their application is required.

Differential effects of dietary fibres on colonic barrier function in elderly individuals with gastrointestinal symptoms.

Gastrointestinal problems are common in elderly and often associated with psychological distress and increased levels of corticotrophin-releasing hormone, a hormone known to cause mast cell (MC) degranulation and perturbed intestinal barrier function. We investigated if dietary fibres (non-digestible polysaccharides [NPS]) could attenuate MC-induced colonic hyperpermeability in elderly with gastrointestinal (GI) symptoms. Colonic biopsies from elderly with diarrhoea and/or constipation (n = 18) and healthy controls (n = 19) were mounted in Ussing chambers and pre-stimulated with a yeast-derived beta (ß)-glucan (0.5 mg/ml) or wheat-derived arabinoxylan (0.1 mg/ml) before the addition of the MC-degranulator Compound (C) 48/80 (10 ng/ml). Permeability markers were compared pre and post exposure to C48/80 in both groups and revealed higher baseline permeability in elderly with GI symptoms. ß-glucan significantly attenuated C48/80-induced hyperpermeability in elderly with GI symptoms but not in healthy controls. Arabinoxylan reduced MC-induced paracellular and transcellular hyperpermeability across the colonic mucosa of healthy controls, but did only attenuate transcellular permeability in elderly with GI symptoms. Our novel findings indicate that NPS affect the intestinal barrier differently depending on the presence of GI symptoms and could be important in the treatment of moderate constipation and/or diarrhoea in elderly.

Soft tissue reactions to bioactive glass 13-93 combined with chitosan.

The aim of this study was to evaluate rabbit soft tissue reactions to bioactive glass 13-93 mesh by using a histological and immunohistochemical analysis. Bioactive glass (13-93) mesh fixed with 3 wt % chitosan was implanted into the dorsal subcutaneous space of New Zealand White rabbits (n=18) for six, 12, and 24 weeks, respectively. After 6 weeks the bioactive glass remnants were surrounded by foreign-body granuloma with eosinophilic granulocytes. After 12 and 24 weeks the implanted material was mainly absorbed, but, if any particles still remained the foreign-body reaction was notably milder. Yet, a mild chronic inflammatory infiltrate was present. Matrix metalloproteinase (MMP) -2, -3, -13 and tissue inhibitory protein (TIMP-1 and -2) expressions were studied by immunohistochemistry. MMP-3, -13, TIMP-1, and -2 positivity were detected throughout the follow-up period. MMP-2 positivity was only occasionally seen in the 24 week samples, which is constitutively expressed but is not related to inductive MMP-3 and -13 cascade. The presence of eosinophilic granulocytes in some of the samples raises the possibility of an allergic reaction to the materials. MMP-3 and -13 are suggested to participate in the host reaction to either bioactive glass or chitosan.

A histological and immunohistochemical study of tissue reactions to solid poly(ortho ester) in rabbits.

In many cases only the temporary presence of a biomaterial is needed in tissue support, augmentation or replacement. In such cases biodegradable materials are better alternatives than biostable ones. At present, biodegradable polymers are widely used in the field of maxillofacial surgery as sutures, fracture fixation devices and as absorbable membranes. The most often used polymers are aliphatic polyesters, such as polyglycolic acid (PGA) and polylactic acid (PLA). Poly(ortho ester) is a surface eroding polymer, which has been under development since 1970, but is used mostly in drug delivery systems in semisolid form. The aim of this study was to evaluate the tissue reactions of solid poly(ortho ester) (POE), histologically and immunohistochemically. Resorption times and the effect of 2 different sterilization methods (gamma radiation and ethylene oxide) upon resorption were also evaluated. Material was implanted into the tibia and subcutaneously into the mandibular ramus area of 24 rabbits. Follow-up times were 1-10, 14 and 24 weeks. Histological studies showed that POE induces a moderate inflammation in soft tissue and in bone. At 24 week follow-up, inflammation was mild in soft tissue and moderate in bone. In immunohistochemical studies, no highly fluorescent layer of tenascin or fibronectin was found adjacent to the implant. Resorption of gamma-sterilized rods was faster than ethylene oxide-sterilized rods. The total resorption time was more than 24 weeks in both groups. Clinically the healing was uneventful and the implants the well tolerated by the living tissue. This encourages these authors to continue studies with this interesting new material to search for the ideal material for bone filling and fracture fixation.

Clinical, radiographic and MRI findings of the temporomandibular joint in patients with different rheumatic diseases.

The aim of this study was to investigate the condition of the temporomandibular joint (TMJ) in patients with different rheumatic diseases, and report correlations between the clinical, radiographic and magnetic resonance imaging (MRI) findings. The 67 patients were divided into four groups: 16 with rheumatoid arthritis (RA), 15 with mixed connective tissue disease (MCTD), 18 with ankylosing spondylitis (AS) and 18 with spondyloarthropathy (SPA). They were clinically examined, and panoramic tomography, lateral panoramic radiography and MRI of the TMJ were performed. MRI showed reduced articular cartilage in 25% (4/16) of RA, 0% (0/15) of MCTD, 17% (3/18) of AS and 17% (3/18) of SPA patients. Condylar changes included erosion, osteophytes and abnormal shape. Disc alterations included perforation, abnormal anterior position and decreased movement. These abnormalities were most frequent in RA patients, and least frequent in MCTD and SPA patients. Crepitation and reduced maximum opening of the mouth correlated with abnormalities of the disc and articular cartilage as shown by MRI. Severe condylar erosion in panoramic tomograms significantly correlated with MRI findings of condylar erosion (P<0.01), diminished thickness of condylar cartilage, abnormal condylar shape, and abnormal shape of the temporal surface of the TMJ (P< or =0.001). The presence of crepitation, limited mandibular movement and/or pain on movement of the jaw often indicated structural damage to the TMJ. Panoramic radiographs provide an alternative method to MRI but, to obtain a more detailed anatomic picture, MRI is recommended for patients with acute unexplained pain or as part of preoperative work up. A panoramic recording is not indicated when MRI is planned.

Deep targeted sequencing in pediatric acute lymphoblastic leukemia unveils distinct mutational patterns between genetic subtypes and novel relapse-associated genes.

To characterize the mutational patterns of acute lymphoblastic leukemia (ALL) we performed deep next generation sequencing of 872 cancer genes in 172 diagnostic and 24 relapse samples from 172 pediatric ALL patients. We found an overall greater mutational burden and more driver mutations in T-cell ALL (T-ALL) patients compared to B-cell precursor ALL (BCP-ALL) patients. In addition, the majority of the mutations in T-ALL had occurred in the original leukemic clone, while most of the mutations in BCP-ALL were subclonal. BCP-ALL patients carrying any of the recurrent translocations ETV6-RUNX1, BCR-ABL or TCF3-PBX1 harbored few mutations in driver genes compared to other BCP-ALL patients. Specifically in BCP-ALL, we identified ATRX as a novel putative driver gene and uncovered an association between somatic mutations in the Notch signaling pathway at ALL diagnosis and increased risk of relapse. Furthermore, we identified EP300, ARID1A and SH2B3 as relapse-associated genes. The genes highlighted in our study were frequently involved in epigenetic regulation, associated with germline susceptibility to ALL, and present in minor subclones at diagnosis that became dominant at relapse. We observed a high degree of clonal heterogeneity and evolution between diagnosis and relapse in both BCP-ALL and T-ALL, which could have implications for the treatment efficiency.

Functional analysis of the human alpha 2C-C4 adrenergic receptor in insect cells expressed by a luciferase-based baculovirus vector.

A click beetle luciferase-based baculovirus expression vector is described for functional analysis and high level expression of a human alpha 2-adrenergic receptor (alpha 2AR) in Sf9 insect cells. The resultant recombinant baculovirus construct, AcLucGR-alpha 2(C4), was isolated by utilizing the light emitting properties of luciferase and used for abundant expression of the alpha 2C-C4 receptor protein in this lepidopteran insect cell line. A maximal expression of alpha 2-receptors at a level of 1.370 pmol/mg protein was obtained at 48 h after infection as determined by ligand-binding experiments using the alpha 2-receptor antagonist, [3H]rauwolscine. The receptor agonists, noradrenaline and clonidine, displaced the [3H]rauwolscine binding with Ki values 12.3 +/- 1.54 microM and 1.23 +/- 0.11 microM, respectively. The recombinant receptors were functionally intact since the agonists inhibited forskolin-stimulated cAMP production. Here, however, the maximal inhibition was obtained at 36 h after the infection. The results presented here, suggest that the baculovirus expression vector system (BEVS) provides a simple method for abundant expression of functional alpha 2-receptor subtypes. In addition, co-expression of luciferase proved to be useful for screening and isolation of the recombinant baculovirus.

Biodegradable polydioxanone and poly(l/d)lactide implants: an experimental study on peri-implant tissue response.

Several implants for orbital wall fracture treatment are available at the present, but they have drawbacks: resorption, risk for migration and foreign body reaction. Alloplastic resorbable implants would be advantageous: no removal operation and no donor side morbidity. The purpose of this study was to evaluate the foreign body reaction, capsule formation and mechanical properties of two bioresorbable implants. PDS and SR-P(L/DL)LA mesh sheet (70/30) with solid frame (96/4) implants (SR-P(L/DL)LA 70,96) were placed into subcutaneous tissue of 24 rats. Immunohistochemistry was used to evaluate reactivity for Tn-C, alpha-actin, type I and III collagens and two mononuclear cells: T-cells and monocyte/ macrophage. GPC, DSC and SEM were performed. Student's t-test or nonparametric Kruskall-Wallis test were used for statistical analysis. Histology of peri-implant capsule exhibited an inner cell-rich zone and an outer connective tissue zone around both materials. Tn-C reactivity was high in the inner and alpha-actin in the outer zone. At the end of the study, the difference of type I collagen versus type III collagen reactivity in inner zone was statistically significant (P<0.0001) as was the difference of type I collagen versus type III collagen reactivity in outer zone (P<0.0001). Immunohistochemistry did not reveal any statistical differences of T-cell and monocyte/macrophage reactivity around PDS versus SR-P(L/DL)LA 70,96 implants, nor any differences as a function of time. PDS were deformed totally after 2 months. SR-P(L/DL)LA 70,96 implants were only slightly deformed during the follow up of 7 months. PDS degraded rapidly in SEM observation. Particles were detaching from surface. SEM observation revealed that polylactide implant was degrading from the surface and the inner porous core became visible. The degradation came visible at 7 months. There were cracks in perpendicular direction towards to the long axis of the filaments. M(w) of PDS decreased fast compared to the polylactide implant. Foreign body reaction was minimal to both materials but continued throughout the whole observation period. Mechanically PDS was poor, it looses its shape totally within 2 months. It cannot be recommended for orbital wall reconstruction. New mesh sheet-frame structure (SR-P(L/DL)LA 70,96) approved to be mechanically adequate for orbital wall reconstruction. It seems not to possess intrinsic memory and retains its shape. The resorption time is significantly longer compared to PDS and is comparable to other studied P(L/DL)LA copolymers. Thus, the new polylactide copolymer implant may support the orbital contents long enough to give way to bone growth over the wall defect.

Inhibition of CD3-induced Ca2+ signals in Jurkat T-cells by myristic acid.

Stimulation of T lymphocytes with antibodies against the T cell receptor/CD3 complex induces within seconds a rise in the concentration of intracellular free Ca2+. Here we show that treatment with 20 microM free myristic acid completely inhibits this Ca2+ signal and the cellular proliferation in Jurkat T cells. Also lauric acid inhibited cell growth while its blocking effect on the Ca2+ signal was weaker than that of myristic acid. Other saturated free fatty acids were inactive. The inhibitory effect of myristic acid could be reversed by the addition of fatty acid free albumin, which will bind the fatty acid. Myristic acid, but not its methyl ester, inhibited both the anti-CD3-induced Ca2+ influx across the cell membrane and Ca2+ release from intracellular stores, but not the formation of inositol phosphates. In contrast, thapsigargin-induced release of Ca2+ from the same intracellular stores was unaffected by myristic acid. Thus, myristic acid specifically blocks T cell antigen receptor-CD3 induced Ca2+ mobilization in T cells.

Effect of adenosine on human cerebral blood flow as determined by positron emission tomography.

The effect of intravenous infusion of adenosine on CBF was studied in seven patients with cerebral arteriovenous malformation. The patients were examined with positron emission tomography with controlled ventilation using [15O] water and [11C] fluoromethane as tracers. Total and regional CBF were determined before and during infusion of adenosine at rates producing a reduction of the MABP by approximately 10-40%. Six patients were normoventilated, and one was hyperventilated. Mean CBF in areas with normal brain tissue was 54 ml/100 g/min before adenosine infusion under normoventilation. Adenosine infusion increased mean CBF with 23-85%. Mean CVR was decreased with 43-65% and exceeded the percentage reduction of MABP in all normoventilated subjects. In the hyperventilated patient, the reduction of CVR was similar to the reduction of MABP, and CBF was unaffected, except for the 30% increase in the thalamus. It is concluded that intravenous administration of adenosine produces marked cerebral vasodilation in normoventilated subjects and that this response can be counteracted by hyperventilation.

Improved method for multisample evaluation of ligand binding to cells. Application to IL-2 receptor measurement.

A newly developed assay using microtitre plates with removable wells permits rapid measurements of IL-2 receptors. Triplicate or quadruplicate samples of several ligand dilutions are easily handled, thus giving more reliable results. In addition, measurement of binding parameters may be accomplished simultaneously on several cell lines. Results are presented which show that the binding assay may also be used for measuring binding constants of antibodies.

Time-resolved fluorometric assay for natural killer activity using target cells labelled with a fluorescence enhancing ligand.

A time-resolved fluorometric assay for the measurement of natural killer cell activity against target cells labelled with the acetoxymethyl ester of the fluorescence enhancing ligand 2,2':6',2"-terpyridine-6,6"-dicarboxylic acid (TDA) is described. The hydrophobic esterified form of TDA (bis(acetoxymethyl) 2,2':6',2"-terpyridine-6,6"-dicarboxylate, BATDA) diffuses readily through the cell membrane of viable cells. BATDA is hydrolysed by intracellular esterases resulting in accumulation of membrane impermeable TDA inside the target cells. After incubation of labelled K-562 cells with effector cells the TDA released from lysed cells into the supernatant is chelated with Eu3+. The natural killer cell activity is then quantified by measuring the intense fluorescence of the EuTDA chelates formed. Target cells are rapidly labelled when incubated with BATDA, TDA is released from target cells faster than 51Cr, the spontaneous release permits a short-term release assay to be set up and the detection of EuTDA is fast (5 min/96 well plate). Furthermore, this non-radioactive method permits the use of complex culture media since, in contrast to methods based on prompt fluorometry, the problem with autofluorescence can be avoided by the use of time-resolved fluorometry.

A baculovirus-expressed fusion protein containing the antibody-binding domain of protein A and insect luciferase.

A fusion construct encoding two antibody-binding sites of protein A from Staphylococcus aureus and click beetle, Pyrophorus plagiophthalamus, luciferase (LucGR) was designed and expressed using the baculovirus system. The construct was inserted under the transcriptional regulation of the polyhedrin gene promoter of the Autographa californica nuclear polyhedrosis virus (AcNPV) and expressed in the insect Spodoperta frugiperda cell line during viral infection. The properties of the resultant chimeric protein product, protA-LucGR, were studied both in vivo and in vitro by using i) luminometry, ii) immunoblot analysis, iii) immunoprecipitation, iv) metabolic labeling procedures and v) luminescent immunoassays. Together, the results clearly demonstrate that the light-emitting properties of the fused luciferase construct remain intact. Further, the antibody-binding domain of protein A retains its activity as it binds to both rabbit and goat as well as human immunoglobulins. Due to the dual biological function of this fusion protein, it should provide a potential reagent within the field of molecular biology and diagnostics.

Identification of biotinylated molecules using a baculovirus-expressed luciferase-streptavidin fusion protein.

A genetic fusion between streptavidin of Streptomyces avidinii and luciferase of Pyrophorus plagiophthalamus was constructed. The fusion protein was produced in the Sf9 insect cell line using the baculovirus expression vector system (BEVS). Sodium dodecyl sulfate polyacrylamide gel electrophoresis of the proteins from cells infected with the recombinant virus, VL1393-LucGR-StreptAv, revealed that the fusion protein migrated with an apparent molecular weight of 75 kDa. Light emission measurements showed that the infected cells produced about 255 mg of the chimeric protein per liter of cell culture (127.5 micrograms/1 x 10(6) cells). Precipitation of the LucGR-StreptAv fusion protein with biotinylated acrylic beads as well as immunoblot analyses using biotinylated immunoglobulins indicated that both fusion moieties of the chimeric protein product were functional with respect to their physical and enzymatic activities.

Synthesis and processing of the rubella virus p110 polyprotein precursor in baculovirus-infected Spodoptera frugiperda cells.

In order to study the processing of rubella virus (RV) structural proteins (capsid protein, of 33 kDa; E2 of 42-47 kDa; and E1 of 58 kDa) in Spodoptera frugiperda (fall armyworm) cells, a 24S cDNA encoding the polyprotein precursor, p110, was inserted under the transcriptional regulation of the polyhedrin gene promoter of the Autographa californica nuclear polyhedrosis virus (AcNPV) and expressed during viral infection. By immunoblot analysis using antibodies directed against whole RV and the individual structural proteins, evidence is presented that polypeptides similar to those synthesized in RV-infected B-Vero cells are expressed in this lepidopteran insect cell line infected with the recombinant baculovirus, VL1392-RV24S. The identity of the recombinant proteins was further confirmed using human convalescent sera. By expressing the recombinant proteins in the presence and absence of tunicamycin, we have further demonstrated that the 24S transcription-translation unit of RV, is expressed and proteolytically cleaved similarly, if not identically, in Sf9 cells as compared to B-Vero cells.

Solid phase IL-2-IL-2R alpha assay with time resolved fluorometry.

A time-resolved fluorometric, solid phase, receptor ligand interaction assay is described. The assay consists of wells coated with anti-human IL-2 receptor alpha (hIL-2R alpha) monoclonal antibodies (mAb), europium labelled hIL2 (Eu-IL-2) and human recombinant IL-2 receptor alpha subunits expressed in the baculovirus expression vector system (BEVS). In the assay hIL-2R alpha-Eu-IL-2 complexes bind to the solid phase mAb. Receptor bound Eu is dissociated into an enhancer solution where it forms highly fluorescent complexes. The fluorescence is measured in a time-resolved fluorometer. The Kd value calculated from the saturation curve is in good agreement with previously reported values for the low affinity type of IL-2R, making the described assay a simple and nonradioactive alternative for measurement of soluble hIL-2R alpha in biological systems. Furthermore this assay format provides convenient separation of bound ligand from unbound and is therefore suitable for high throughput screenings.