Use of Extracorporeal Membrane Oxygenation Prior to Lung Transplantation Does Not Jeopardize Short-term Survival.

BACKGROUND: The use of pretransplantation extracorporeal membrane oxygenation (ECMO) has been considered to be a relative contraindication and a risk factor associated with poor outcomes in lung transplantation. However, with a donor shortage, use of ECMO before transplantation is often unavoidable. This study aimed to review our experiences of lung transplantation outcome with regards to the use of pretransplantation ECMO. METHODS: We retrospectively reviewed the clinical data of patients who underwent lung transplantation at our institution. Clinical variables as well as ECMO-related data were analyzed with surgical outcomes. RESULTS: From 2006 to 2014, 27 patients underwent lung transplantation: 26 bilateral sequential lung transplants and 1 right-side single lung transplant. Of these, 12 (44.4%) received ECMO treatment during the pretransplantation waiting period. Pretransplantation ECMO patients showed higher body mass index scores (P = .047) and mechanical ventilation support (P < .001) than the non-ECMO group. All ECMO patients were weaned from ECMO after transplantation. The median ECMO runtime was 12 days. The survival-to-discharge rates of the 2 groups did not differ. Survival after lung transplantation at 1, 6, 12, and 24 months was 100%, 73.3%, 61.1%, and 61.1% in the ECMO group and 100%, 86.7%, 86.7%, and 66.0% in the non-ECMO group, respectively (P = .540). CONCLUSIONS: Use of pretransplantation ECMO did not jeopardize survival-to-discharge and short-term survival rates in our experience. Our result suggests pretransplantation ECMO can provide a chance of receiving lung transplantation to those who were classified as "too sick to be transplanted."

Receptor tyrosine kinase Axl is required for resistance of leukemic cells to FLT3-targeted therapy in acute myeloid leukemia.

In acute myeloid leukemia (AML), about 25-30% of patients harbor a constitutively active receptor tyrosine kinase (RTK) FLT3 encoded by a FLT3 allele harboring internal tandem duplication (FLT3-ITD) mutation. The presence of FLT3-ITD correlates with poor prognosis in AML and it makes FLT3 an attractive therapeutic target in AML. Unfortunately, to date small-molecule inhibitors of FLT3 have resulted in only partial and transient clinical responses with residual leukemic blasts resistant to FLT3 inhibitors detected in blood or bone marrow. In this study, we investigated whether the RTK Axl is responsible for resistance of FLT3-ITD(+) AML cells to PKC412 and AC220, FLT3 inhibitors currently under clinical trials for FLT3-ITD(+) AML patients. Upon treatment with PKC412 or AC220, phosphorylation of Axl was significantly enhanced in the FLT3-ITD(+) MV4-11 AML cell line and in primary blasts from a FLT3-ITD(+) AML patient. Consistently, a PKC412-resistant AML cell line and PKC412-resistant primary blasts from FLT3-ITD(+) AML patients had significantly higher levels of constitutively phosphorylated Axl and total Axl when compared with a PKC412-sensitive AML cell line and PKC412-sensitive primary blasts from FLT3-ITD(+) AML patients. We also found that resistance of AML cells against the FLT3 inhibitor PKC412 and AC220 was substantially diminished by the inhibition of Axl via a small-molecule inhibitor TP-0903, a soluble receptor Axl fusion protein Axl-Fc or knockdown of Axl gene expression by shRNA. Collectively, our study suggests that Axl is required for resistance of FLT3-ITD(+) AML cells against the FLT3 inhibitor PKC412 and AC220, and that inhibition of Axl activation may overcome resistance to FLT3-targeted therapy in FLT3-ITD(+) AML.

A randomised controlled trial comparing incentive spirometry with the Acapella® device for physiotherapy after thoracoscopic lung resection surgery.

Lung resection surgery has been associated with numerous postoperative complications. Seventy-eight patients scheduled for elective video-assisted thoracoscopic lung resection were randomly assigned to receive standard postoperative care with incentive spirometry or standard care plus positive vibratory expiratory pressure treatment using the Acapella(®) device. There was no significant difference between incentive spirometry and the Acapella device in the primary outcome, forced expiratory volume in 1 s, on the third postoperative day, mean (SD) 53% (16%) vs 59% (18%) respectively, p = 0.113. Patients treated with both devices simultaneously found incentive spirometry to be less comfortable compared with the Acapella device, using a numeric rating scale from 1 to 5 with lower scores indicating higher comfort, median (IQR [range]) 3 (2-3 [2-4]) vs 1 (1-2 [1-3]) respectively, p < 0.001. In addition, 37/39 patients (95%) stated a clear preference for the Acapella device. Postoperative treatment with the Acapella device did not improve pulmonary function after thoracoscopic lung resection surgery compared with incentive spirometry, but it may be more comfortable to use.

MR traceable delivery of p53 tumor suppressor gene by PEI-functionalized superparamagnetic iron oxide nanoparticles.

Cancer gene therapy involves the replacement of missing or altered genes with healthy ones. In this paper, we have proposed tumor suppressor gene-carrying superparamagnetic iron oxide nanoparticles (SPIONs) for anti-cancer gene therapy. Thermally crosslinked SPIONs (TCL-SPIONs) were conjugated with branched polyethylenimine (PEI 1800 Da) by EDC-NHS chemistry for p53 plasmid DNA delivery. The morphology of the bPEI conjugated TCL-SPIONs (bPEI-TCL-SPION) and pDNA-loaded bPEI-TCL-SPION nanoparticles was measured using transmission electron microscopy (TEM). The particle sizes of the pDNA-loaded bPEI-TCL-SPION nanoparticles were also confirmed by dynamic light scattering, and ranged from 100 to 130 nm, depending on the molar charge ratio. The fluorescently labeled pDNA was complexed with bPEI-TCL-SPION and its intracellular internalization was investigated using confocal microscopy. The p53 plasmid-loaded bPEI-TCL-SPION nanoparticles achieved significantly higher p53 tumor suppressor gene expression and cellular viability compared to positive controls. The expressed wild-type p53 protein suppressed tumor cell proliferation as compared to the mutant control. When transgene expression of the p53 tumor suppressor gene was evaluated at the mRNA level and quantified using real-time PCR, the results were highly dependent on the molar charge ratio (N/P) as well as the cancer cell type. SPIONs internalized within cancer cells were tracked by magnetic resonance (MR) imaging. It was concluded that bPEI-TCL-SPION could be used as efficient gene delivery carriers that can be tracked by MR imaging.

Oesophagography and oesophagoscopy are not necessary in patients with spontaneous pneumomediastinum.

BACKGROUND: Because the condition is rare, the proper assessment of spontaneous pneumomediastinum (SPM) remains controversial. The purpose of this study was to determine whether additional oesophageal investigations beyond chest x ray and chest computed tomography (CT) scan are necessary for the diagnosis of SPM. METHODS: The medical records of 25 patients diagnosed and treated for SPM from March 1986 to December 2007 were retrospectively reviewed. RESULTS: There were 22 men and 3 women, with a median age of 19 years (range 15-57 years). All patients received chest x rays, which revealed air shadows within the mediastinum or subcutaneous emphysema in 24 patients. Twenty-two patients underwent chest CT scans, which showed pneumomediastinum in all cases. Oesophagography was performed in 14 patients and oesophagoscopy in three. All oesophagographies and oesophagoscopies were clear. Despite conservative treatment, no patients developed mediastinitis or complications associated with oesophageal injury. CONCLUSIONS: Chest x ray and CT scan are sufficient to diagnose SPM. Additional diagnostic assessments such as oesophagography and oesophagoscopy are not necessary in patients without evidence of mediastinitis or a history of oesophageal injury.

Peptide-modified vectors for nucleic acid delivery to neurons.

Neuron-targeted nucleic acid delivery systems are important technologies for realizing the potential of gene therapy for nervous system disorders. However, neurons are difficult cells to transfect using non-viral vectors due in part to the specific and unique delivery challenges present in these cells. We have investigated several bioactive peptides for their ability to assist in overcoming delivery barriers in mammalian cells. We summarize here our recent progress in developing and applying peptide-modified polycations for nucleic acid delivery. In addition, we present data demonstrating the potential of using multicomponent, peptide-modified polycations for nucleic acid delivery to neurons.

Galactose-carrying polymers as extracellular matrices for liver tissue engineering.

Extracellular matrix (ECM) plays important roles in tissue engineering because cellular growth and differentiation, in the two-dimensional cell culture as well as in the three-dimensional space of the developing organism, require ECM with which the cells can interact. Especially, the bioartificial liver-assist device or regeneration of the liver-tissue substitutes for liver tissue engineering requires a suitable ECM for hepatocyte culture because hepatocytes are anchorage-dependent cells and are highly sensitive to the ECM milieu for the maintenance of their viability and differentiated functions. Galactose-carrying synthetic ECMs derived from synthetic polymers and natural polymers bind hepatocytes through a receptor-mediated mechanism, resulting in enhanced hepatocyte functions. Attachment and functions of hepatocytes were affected by physico-chemical properties including ECM geometry as well as the type, density and orientation of galactose. Also, cellular environment, medium composition and dynamic culture system influenced liver-specific functions of hepatocytes beside ECM.

Phase II study of concurrent chemoradiotherapy with capecitabine and cisplatin in patients with locally advanced squamous cell carcinoma of the head and neck.

We aimed to evaluate the efficacy and safety of concurrent chemoradiotherapy with capecitabine and cisplatin in patients with locally advanced squamous cell carcinoma of the head and neck (SCCHN). In total, 37 patients with stage III or IV SCCHN were enrolled on the study. The chemotherapy consisted of two cycles of intravenous cisplatin of 80 mg m(-2) on day 1 and oral capecitabine 825 mg m(-2) twice daily from day 1 to day 14 at 3-week intervals. The radiotherapy (1.8-2.0 Gy 1 fraction day(-1) to a total dose of 70-70.2 Gy) was delivered to the primary tumour site and neck. The primary tumour sites were as follows: oral cavity (n=6), oropharynx (n=11), hypopharynx (n=8), larynx (n=3), nasopharynx (n=6), and paranasal sinus (n=3). After the chemoradiotherapy, 29 complete responses (78.4%) and 6 partial responses (16.2%) were confirmed. Grade 3 or 4 neutropenia occurred only in two patients, plus grade 3 febrile neutropenia was observed only in one patient. At a median follow-up duration of 19.8 months, the estimated overall survival and progression-free survival rate at 2-year was 76.8 and 57.9%, respectively. Concurrent chemoradiotherapy with capecitabine and cisplatin was found to be well tolerated and effective in patients with locally advanced SCCHN.

Influence of air exposure and storage condition on serum ionized magnesium level.

The aim of this study was to evaluate whether reporting serum level of ionized magnesium (iMg) is appropriate when affected by various conditions such as exposure to air and delayed measurement. Serum levels of pH, iMg and normalized magnesium (nMg, normalized or adjusted concentration of iMg to pH 7.40) from 28 inpatients were measured at intervals of 3 min after exposing the samples to air at room temperature. Serum from 30 inpatients was stored in closed tubes at 4 degrees C and -20 degrees C and iMg and nMg levels were measured after 2 days. It was found that serum iMg and nMg concentrations exposed to air were decreased by 0.0023 mmol/l and 0.0001 mmol/l per minute, respectively. nMg did not display any significant changes compared with iMg at 0 min, whereas iMg showed significant changes, which exceeded between-day precision. For the stored serum, only iMg of serum at -20 degrees C showed no statistically significant changes (p = 0.169). It is concluded that to report the result as iMg, the sample should be kept anaerobically, and if exposed to air, the result should be reported as nMg. For storage, iMg of serum kept anaerobically at -20 degrees C is reliable.

A case report: isolated a heavy chain monoclonal gammopathy in a patient with polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy and skin change syndrome.

A 45-year-old South-Korean man presented with abdominal distension, progressive paresthesia and motor weakness of both lower extremities. Our case was identified as polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy and skin change (POEMS) syndrome based on diagnostic criteria. Circulating M components of POEMS syndrome consist mainly of IgG or IgA-lambda and rarely IgM-lambda, IgG-kappa or isolated light chains. In this case, the M-band on serum protein electrophoresis and isolated IgA heavy chain on serum immunofixation electrophoresis were demonstrated, but there was no abnormal light chain. We suggest that this case may be associated with a pattern of abnormal secretion of monoclonal protein or a coincidence of a heavy chain disease in POEMS syndrome, even though the latter possibility may be very rare.

Visualization of transfection of hepatocytes by galactosylated chitosan-graft-poly(ethylene glycol)/DNA complexes by confocal laser scanning microscopy.

Dual-labeled galactosylated chitosan-graft-poly(ethylene glycol) (PEG) (GCP)/DNA complexes were prepared and their hepatocyte-specific delivery and cellular distribution were investigated by confocal laser scanning microscopy (CLSM). The complexes were transfected into hepatocyte through specific interaction of galactose moiety of the GCP and asialoglycoprotein receptors (ASGPR) of the hepatocytes. The GCP/DNA complexes taken up by the hepatocytes were rapidly released into the cytoplasm, but nuclear trafficking of the released complexes was slow and rate-limiting process. The more efficient transfection of the complex occurred in the human-derived HepG2 cells than in primary hepatocytes.

Galactosylated chitosan (GC)-graft-poly(vinyl pyrrolidone) (PVP) as hepatocyte-targeting DNA carrier. Preparation and physicochemical characterization of GC-graft-PVP/DNA complex (1).

Galactosylated chitosan was conjugated with poly(vinyl pyrrolidone) (PVP) as a hydrophilic group. The complex formation of GC-graft-PVP (GCPVP)/DNA complexes was confirmed by agarose gel electrophoresis. The morphology of the complex observed by atomic force microscopy had a compact and spherical shape, around 40 nm particle sizes at a charge ratio of 3. The binding strength of GCPVP 10K/DNA complex measured by ethidium bromide binding assay was superior to that of the GCPVP 50K/DNA one, probably attributable to its higher flexibility due to the smaller size, whereas the DNase I protection of GCPVP 10K/DNA complex was inferior to that of the GCPVP 50K/DNA one. This indicated that effective complex formation required both higher binding strength and minimal molecular weight of polycation enough to induce the condensation of DNA. The DNA-binding property of GCPVP mainly depended on the molecular weight of chitosan and composition of PVP.

Receptor-mediated delivery of all trans-retinoic acid to hepatocyte using poly(L-lactic acid) nanoparticles coated with galactose-carrying polystyrene.

All trans-retinoic acid (RA)-loaded poly(L-lactic acid) (PLA) nanoparticles coated with galactose-carrying polymer, as hepatocyte-specific targeting material using galactose ligands as recognition signals to asialoglycoprotein receptors were prepared by the diafiltration method. Effects of released RA from its loaded nanoparticles on morphology and DNA synthesis of hepatocytes were studied. Receptor-mediated endocytosis of the nanoparticles was checked by fluorescence and confocal laser microscopy. It was found that the shapes of most hepatocytes attached onto polystyrene dish precoated with collagen solution were flat and spreading at low concentration of RA for the RA-loaded nanoparticles, whereas their shapes were round at even low concentration of RA when RA was mixed with the nanoparticles. From the fluorescence and confocal laser microscopic studies, it was suggested that the nanoparticles coated with galactose-carrying polymers were internalized by the hepatocytes through the receptor-mediated mechanism. The RA-loaded nanoparticles were more potent stimulators of hepatocyte DNA synthesis than the free RA system in the presence of epidermal growth factor (EGF) owing to the controlled release of RA from the RA-loaded nanoparticles.

Galactosylated chitosan-graft-poly(ethylene glycol) as hepatocyte-targeting DNA carrier.

Lactobionic acid bearing galactose group was coupled with chitosan for liver specificity, and poly(ethylene glycol) (PEG) was grafted to galactosylated chitosan (GC) for stability in water and enhanced cell permeability. Complex formation of galactosylated chitosan-graft-PEG (GCP)/DNA complexes was confirmed by agarose gel electrophoresis. Compared to GC/DNA complex, the stability of GCP/DNA complex could be enhanced. Particle sizes of GCP/DNA complexes decreased as the charge ratio of GCP to DNA increased and had a minimum value around 27 nm at the charge ratio of 5. Conformational change of DNA did not occur after complex formation with GCP compared to conformation of DNA itself. GCP/DNA complexes were only transfected into Hep G2 having asialoglycoprotein receptors (ASGR), indicative of specific interaction of ASGR on cells and galactose ligands on GCP.

Effects of neurotoxin 6-aminonicotinamide on levels of enzyme activities and metabolites in quail plasma.

Effects of 6-aminonicotinamide (6-AN) on the levels of proteins, metabolites and enzyme activities in the plasma of Japanese quail were investigated. The concentrations of soluble proteins in the pectoral and hindlimb muscle of the 6-AN treated and the pair-fed groups were significantly reduced compared to the control group. In the plasma, the levels of total proteins and albumin were not affected, but the levels of globulin were significantly lower than those of the control and pair-fed groups. In contrast, the levels of glucose and creatine were significantly elevated. Cellulose acetate gel electrophoresis showed that 6-AN induced a new synthesis of prealbumin and also increased the levels of beta-globulin relative to the control and pair-fed groups. In contrast, the levels of gamma-globulin were markedly lower than those of the control group, whereas the levels of alpha-globulin were not affected. The specific activity of alkaline phosphatase of the 6-AN group was significantly lower than that of the control and pair-fed groups and that of aspartate aminotransferase only lower than that of the control group but not the pair-fed group. The specific activities of creatine phosphokinase and lactate dehydrogenase of the 6-AN group were the greatest among the three groups, whereas those of the pair-fed group were greater than those of the control group. The results suggest that 6-AN may interfere with the proper maintenance of energy charges and the immune system function.

The clinical and ocular manifestations of Takayasu arteritis.

PURPOSE: To evaluate the clinical and ocular manifestations of Takayasu arteritis and the fundus fluorescein angiography (FFA) characteristics of Takayasu retinopathy (TR). PATIENTS AND METHODS: Medical records and fundus fluorescein angiograms of 156 eyes of 78 patients with Takayasu arteritis were reviewed. Fundus FA using a wide-field fundus camera (60 degrees) was performed in 19 patients, and conventional angiography or spiral computed tomographic angiography was performed in all 78 patients. RESULTS: The series included 67 female and 11 male patients; mean age at time of diagnosis was 26.7 years (range, 4-61 years). Hypertension was found in 44 (56.4%) patients, ischemic cerebrovascular symptoms in 18 (23.1%) patients, and amaurosis fugax in 20 (25.6%) patients. On fundus examination, no retinopathy was found in 87 (55.8%) eyes; hypertensive retinopathy was found in 48 (30.8%) eyes; and TR was found in 21 (13.5%) eyes. Patients with TR had carotid artery or aortic arch involvement, and patients with hypertensive retinopathy had involvement of the descending aorta or renal artery and sparing of the carotids. Best-corrected visual acuity in TR Stage 1 to 3 ranged from 20/15 to 20/30, but in Stage 4, it ranged from 20/200 to hand motions because of secondary ocular complications. On FFA, the arm-to-retina circulation time was prolonged in all 21 eyes with TR (mean, 22.7+/-8.9 seconds), but only 14 eyes showed delayed arteriovenous filling time, which was mainly found in chronic, moderate to severe TR, Stage 3 or 4. Arteriovenous anastomosis was found in all 12 eyes with Stage 3 and 4 TR. CONCLUSIONS: Delayed arm-to-retina circulation time is shown in all cases of TR, but delayed arteriovenous filling time is mostly found in moderate and severe TR. During ophthalmic examination, the delay of arteriovenous filling time and formation of arteriovenous anastomosis must be examined carefully to prevent visual deterioration.

NAD+ inhibits the self-splicing of the group I intron.

We investigated the effects of the coenzyme NAD+ (nicotinamide adenine dinucleotide) and its analogs on the self-splicing of primary transcripts of the phage T4 thymidylate synthase gene (td). Of all the nicotinamide coenzymes and analogs tested, NADP+ was the strongest inhibitor, with a potency approximately threefold that of NAD+. Kinetic analysis demonstrated that NAD+ acts as a mixed type noncompetitive inhibitor for the td intron RNA with a K(i) of 4.1 mM. The splicing specificity inhibition by NAD+ is predominantly due to changes in Km and kcat, and was Mg2+ concentration dependent. The results suggest that both the ADP and nicotinamide moieties are the key structural features in NAD+ responsible for the inhibition of splicing.