Circulating Donor Lung-specific Exosome Profiles Enable Noninvasive Monitoring of Acute Rejection in a Rodent Orthotopic Lung Transplantation Model.
There is a critical need for development of biomarkers to noninvasively monitor for lung transplant rejection. We investigated the potential of circulating donor lung-specific exosome profiles for time-sensitive diagnosis of acute rejection in a rat orthotopic lung transplant model. Left lungs from Wistar transgenic rats expressing human CD63-GFP, an exosome marker, were transplanted into fully MHC-mismatched Lewis recipients or syngeneic controls. Recipient blood was collected between 4 h and 10 d after transplantation, and plasma was processed for exosome isolation by size exclusion column chromatography and ultracentrifugation. Circulating donor exosomes were profiled using antihuman CD63 antibody quantum dot on the nanoparticle detector and via GFP trigger on the nanoparticle flow cytometer. In syngeneic controls, steady-state levels of circulating donor exosomes were detected at all posttransplant time points. Allogeneic grafts lost perfusion by day 8, consistent with acute rejection. Levels of circulating donor exosomes peaked on day 1, decreased significantly by day 2, and then reached baseline levels by day 3. Notably, decrease in peripheral donor exosome levels occurred before grafts had histological evidence of acute rejection. Circulating donor lung-specific exosome profiles enable an early detection of acute rejection before histologic manifestation of injury to the pulmonary allograft. As acute rejection episodes are a major risk factor for the development of chronic lung allograft dysfunction, this biomarker may provide a novel noninvasive diagnostic platform that can translate into earlier therapeutic intervention for lung transplant patients.


The branchpoint residue is recognized during commitment complex formation before being bulged out of the U2 snRNA-pre-mRNA duplex.
We have analyzed the mechanism of branchpoint nucleotide selection during the first step of pre-mRNA splicing. It has previously been proposed that the branchpoint is selected as an adenosine residue bulged out of an RNA helix formed by the U2 snRNA-pre-mRNA base pairing. Although compatible with this bulge hypothesis, available data from both yeast and mammalian systems did not rule out alternative structures for the branch nucleotide. Mutating the residue preceding the branchpoint nucleotide in our reporter construct conferred a splicing defect that was suppressed in vivo by the complementary U2 snRNA mutants. In contrast, substitutions on the 3' side of the branchpoint could be suppressed by complementary U2 snRNA mutants only in a weakened intron context. To test why the identity of the branch nucleotide was important for its selection, we analyzed the effect of substitutions at this position on spliceosome assembly. We observed that these mutations block the formation of one of the two commitment complexes. Our results demonstrate that yeast branchpoint selection occurs in multiple steps. The nature of the branch residue is recognized, in the absence of U2 snRNA, during commitment complex formation. Then, base pairing with U2 snRNA constrains this residue into a bulge conformation.


Enhanced controlled release of loratadine from the ethylene-vinyl acetate matrix containing plasticizer.
An ethylene-vinyl acetate (EVA) matrix containing plasticizer was prepared as a potential controlled release system for loratadine. The EVA matrix containing loratadine was prepared as the transdermal device using casting methods. The solubility of loratadine according to the volume fraction of PEG 400 was determined. The effects of the drug concentration, temperature, and plasticizers on the release of the drug were determined at 37 degrees C using 40% PEG 400 solution as the receptor medium using the modified Keshary-Chien cell. Some types of plasticizers. such as citrates and phthalates, were used to prepare the pores and increase the flexibility of the EVA matrix. The solubility test according to the PEG 400 volume fraction revealed the highest solubility in the 40% PEG 400 solution. The rate of drug released from the EVA matrix increased with increasing temperature and drug loading. There was a linear relationship between the release rate and the square root of the loading dose. The activation energy for drug release from the EVA matrix with a loading dose of 1%, 2%, 3%, 4%, and 5% was estimated to be 6.83, 6.80, 6.77, 6.71, and 6.65 kcal/mol, respectively Among the plasticizers used, diethyl phthalate showed the highest level of loratadine release. In conclusion, an EVA matrix containing plasticizer could be used to enhance the controlled release of loratadine.


[Role of immunological factors in the diagnosis and prediction of intrauterine fetal infections].
Comparative analysis of the data of immunological examination of pregnant women with markers of genitourinary infections (GUI), by taking into account the etiological factor, has established unidirectional changes in the levels cytokines and neopterin (NP). At the same time, the level of NP in all the groups of examinees with GUI was higher than that of cytokines. The highest content of NP in blood and cervical canal mucus (CCM) was noted in pregnant women with GUI of viral etiology. The changes in NP levels in infections caused by various viruses were similar. The study of the levels of cytokines and NP in the peripheral blood and CCM samples yielded information on the functional activity of different types of immunocompetent cells, the magnitude of an inflammatory process, its transition to the systemic level, and on the prediction of infection development.


Multifunctional nanorods serving as nanobridges to modulate T cell-mediated immunity.
Electrodeposited nanorods serving as multivalent bridges were fabricated and surface-decorated with ligands for immune cells. Gold and nickel solutions were sequentially electrodeposited on nanoporous anodized disc templates and the template was dissolved to retrieve bisegmented nanorods with different lengths. Gold and nickel segmented nanorods were surface-immobilized with mannose and RGD peptides to prepare immune-cell recruiting nanorods. Surface-functionalization of nanorods were confirmed by fluorescence-labeling of each ligands and confocal microscopy. Dendritic cells and T cells were co-incubated with the surface-functionalized nanorods, and the proximity between the nanorods and the immune cells was visualized by variable pressure scanning electron microscopy and confocal microscopy. The long nanorods were associated with the immune cells, whereas the shorter nanorods were rather endocytosed by cells, suggesting a feasibility of the longer nanorods as bridging for the cells. Cytokine releases from the immune cells were monitored by cultivating lipopolysaccharide-activated dendritic cells with T cells. Interleukine-2 and interferon-γ release profiles showed a strong correlation with the length of the nanorod, where the 4 μm nanorods induced the highest levels of cytokine release compared to 1 or 2 μm nanorods. Thus, we concluded that the proximity of the immune cells increased by bridging the immune cells with the nanobridging system, which subsequently increased cytokine release by facilitating the antigen presentation process.


Sling compared with plate osteosynthesis for treatment of displaced midshaft clavicular fractures: a randomized clinical trial.
Few randomized controlled trials have compared operative with nonoperative treatment of clavicular fractures. Patients with displaced midshaft clavicular fractures were randomized either to nonoperative treatment with a sling or to operative treatment with a stainless steel 3.5-mm reconstruction plate. Outcome measures were the Constant shoulder score, DASH (Disabilities of the Arm, Shoulder and Hand) score, pain, fracture-healing, and complications. The null hypothesis was that the Constant and DASH scores would not differ between the groups at the one-year follow-up evaluation. In accordance with the power analysis, we included sixty patients in the study; thirty-two were randomized to the nonoperative group and twenty-eight to the operative group. We found no difference in the Constant score (p = 0.75), the DASH score (p = 0.89), or pain (p = 0.98) between the groups at the one-year follow-up evaluation. All fractures in the operative group healed, but six nonunions (24%) occurred in the nonoperative group. One year after a displaced midshaft clavicular fracture, nonoperative treatment resulted in a higher nonunion rate but similar function and disability compared with operative treatment.


Genipin Reverses HFD-Induced Liver Damage and Inhibits UCP2-Mediated Pyroptosis in Mice.
Liver damage is a typical manifestation of nonalcoholic fatty liver disease (NAFLD). It originates from excessive fat accumulation, leading to hepatocyte death, inflammation, and fibrosis. Nonalcoholic steatohepatitis (NASH) is a type of NAFLD with a prevalence of 49% in morbidly obese patients. Pyroptosis plays an important role in the development of NASH; thus, it is important to elucidate the effect of lipid accumulation on pyroptosis. Genipin (GNP), a natural water-soluble cross-linking agent, has hepatoprotective effects and decreases lipid accumulation in the liver; however, the mechanisms underlying these effects are unknown. In this study, qPCR and Western blot were used to examine pyroptotic gene expression in high-fat diet (HFD) induced obese mice and free fatty acids (FFAs) treated hepatocytes. At the same time, relative lactate dehydrogenase (LDH) release and Hoechst & propidium iodide (PI) staining were done to verify cell death. To explore the molecular mechanism, cell transfection were constructed with siRNA or plasmid to obtain knockdown or overexpression hepatocytes. We found that HFD-fed mice and FFAs-treated hepatocytes had obvious pyroptosis, and addition of GNP reversed liver damage and inhibited pyroptosis both in vitro and in vivo. Besides, UCP2 knockdown cells showed suppressed FFAs-mediated pyroptosis, as determined by decreased pyroptotic gene expression, reduced lactate dehydrogenase (LDH) release, and reduced cell death. Consistent with this, cells transfected with UCP2 had upregulated pyroptotic gene expression, increased LDH release, and increased cell death. GNP reverses HFD-induced liver damage and inhibits UCP2-mediated pyroptosis. Thus, GNP may serve as a potential therapeutic candidate for NAFLD.


Cardiovascular Outcomes among Combustible-Tobacco and Electronic Nicotine Delivery System (ENDS) Users in Waves 1 through 5 of the Population Assessment of Tobacco and Health (PATH) Study, 2013-2019.
Prior studies have not clearly established risk of cardiovascular disease (CVD) among smokers who switch to exclusive use of electronic nicotine delivery systems (ENDS). We compared cardiovascular disease incidence in combustible-tobacco users, those who transitioned to ENDS use, and those who quit tobacco with never tobacco users. This prospective cohort study analyzes five waves of Population Assessment of Tobacco and Health (PATH) Study data, Wave 1 (2013-2014) through Wave 5 (2018-2019). Cardiovascular disease (CVD) incidence was captured over three intervals (Waves 1 to 3, Waves 2 to 4, and Waves 3 to 5). Participants were adults (40+ years old) without a history of CVD for the first two waves of any interval. Change in tobacco use status, from exclusive past 30 day use of any combustible-tobacco product to either exclusive past 30 day ENDS use, dual past 30 day use of ENDS and combustible-tobacco, or no past 30 day use of any tobacco, between the first two waves of an interval was used to predict onset of CVD between the second and third waves in the interval. CVD incidence was defined as a new self-report of being told by a health professional that they had congestive heart failure, stroke, or a myocardial infarction. Generalized estimating equation (GEE) analyses combined 10,548 observations across intervals from 7820 eligible respondents. Overall, there were 191 observations of CVD among 10,548 total observations (1.7%, standard error (SE) = 0.2), with 40 among 3014 never users of tobacco (1.5%, SE = 0.3). In multivariable models, CVD incidence was not significantly different for any tobacco user groups compared to never users. There were 126 observations of CVD among 6263 continuing exclusive combustible-tobacco users (adjusted odds ratio [AOR] = 1.44; 95% confidence interval (CI) 0.87-2.39), 15 observations of CVD among 565 who transitioned to dual use (AOR = 1.85; 0.78-4.37), and 10 observations of CVD among 654 who quit using tobacco (AOR = 1.18; 0.33-4.26). There were no observations of CVD among 53 who transitioned to exclusive ENDS use. This study found no difference in CVD incidence by tobacco status over three 3 year intervals, even for tobacco quitters. It is possible that additional waves of PATH Study data, combined with information from other large longitudinal cohorts with careful tracking of ENDS use patterns may help to further clarify this relationship.


Congenital Brucellosis: A Case Report.
Background: Brucellosis is the most common zoonotic infection worldwide, and is caused by bacterial genus Brucella. The disease is rarely transmitted through human-to-human transmission. Few cases have been reported about vertical transmission of human brucellosis. Herein, we reported a case of congenital brucellosis, with clear evidence of pathogen detected in mother's placental specimen. Case Presentation: A 34-day-old girl was admitted to the department of pulmonology with fever for 8 days. Three blood samples and one sample of cerebrospinal fluid were positive for Brucella melitensis. The diagnosis of brucellosis and B. melitensis meningitis were established, along with hyperbilirubinemia and liver dysfunction. Treatment of rifampicin (for 6 weeks) and meropenem (for 2 weeks) was administered. However, the disease relapsed within 18 days. Thereafter, a combination therapy of rifampicin and sulfamethoxazole/trimethiprim (SMZ/TMP) was administered for 8 weeks. The disease relapsed again in 42 days. For chronic brucellosis, three courses of combination therapy of rifampicin and SMZ/TMP was administered. The mother had fatigue and arthralgia for 2 weeks, fever and membrane rupture 1 day before the baby was born. B. melitensis DNA was detected in the mother's placental specimen by next-generation sequencing and bacterial identification under microscope proved chorioamnionitis. Conclusions: We reported a confirmed case of congenital brucellosis. This disease should be closely monitored even in nonepidemic areas. The treatment of brucellosis in infancy faces challenges of drug choice and disease relapse.


How prenatal cortisol levels may differentially affect the neurodevelopment of boys and girls.
Prenatal stress could have serious consequences on maternal and fetal health. In this sense, some studies have stated that maternal HCC during pregnancy could contribute to sex-specific effects on infant neurodevelopment, following the Developmental Origins of Health and Disease Hypothesis. This study aimed to determine whether maternal hair cortisol concentration (HCC) during each trimester of pregnancy and postpartum could predict the neurodevelopmental outcomes of their 12-month-old offspring, with sex-specific differences considered. longitudinal. The study involved 93 pregnant women and their babies. Hair samples collected during each trimester and postpartum and The Bayley Scales for Infant Development III was used to assess the infants' abilities. The results showed that maternal HCC during the first and second trimesters could predict language and motor abilities. However, when discriminated by sex, only females' cognitive, expressive language, and fine and gross motor skills were predicted by cortisol, not males. These findings support the idea that non-toxic levels of cortisol can positively influence infants' neurodevelopment.


Parallux beta-lactam: a capillary-based fluorescent immunoassay for the determination of penicillin-G, ampicillin, amoxicillin, cloxacillin, cephapirin, and ceftiofur in bovine milk.
An analytical system was developed for detection of antibiotic residues in bovine milk. The method is based on competitive fluorescent immunoassays in glass capillary tubes (U.S. Patent No. 5,624,850). The system consists of an assay cartridge containing 4 glass capillaries, a reagent tray with 4 wells of dried reagents, and a Parallux processor, which processes the assay, reads fluorescent output, and reports test results. Minimum sensitivity for detection of 6 beta-lactam antibiotics in bovine milk was determined to be penicillin-G, 3.2 ppb; ampicillin, 2.9 ppb; amoxicillin, 3.6 ppb; cloxacillin, 7.4 ppb; cephapirin, 16.3 ppb; and ceftiofur, 33.7 ppb. The assay system was also specific and sensitive for detection of incurred residues at U.S. Food and Drug Administration tolerance levels: penicillin-G, 5 ppb; ampicillin, 10 ppb; amoxicillin, 10 ppb; cloxacillin, 10 ppb; cephapirin, 20 ppb; and ceftiofur, 50 ppb. There was no interference in detection of minimum sensitivity levels of antibiotic by the presence of somatic cells at approximately 1 x 10(6) cells/mL. Milk containing 3 x 10(6) cells/mL bacteria commonly found in mastitic milk also showed no interference when tolerance levels of antibiotic were present. There was no detectable interference on results by a wide variety of non-beta-lactam drugs.


An insight into the mechanism of inhibition and reactivation of the F(1)-ATPases by tentoxin.
The mechanism of inhibition and reactivation of chloroplast ATP-synthase by the fungal cyclotetrapeptide tentoxin was investigated by photolabeling experiments, binding studies, and kinetic analysis using synthetic analogues of tentoxin. The alpha-subunit of chloroplast F(1)-ATPase (CF(1)) was specifically labeled by a photoactivatable tentoxin derivative, providing the first direct evidence of tentoxin binding to the alpha-subunit, and 3D homology modeling was used to locate tentoxin in its putative binding site at the alpha/beta interface. The non-photosynthetic F(1)-ATPase from thermophilic bacterium (TF(1)) proved to be also tentoxin-sensitive, and enzyme turnover dramatically increased the rate of tentoxin binding to its inhibitory site, contrary to what was previously observed with epsilon-depleted CF(1) [Santolini, J., Haraux, F., Sigalat, C., Moal, G., and André, F. (1999) J. Biol. Chem. 274, 849-858]. We propose that tentoxin preferentially binds to an ADP-loaded alpha beta pair, and mechanically blocks the catalytic cycle, perhaps by the impossibility of converting this alpha beta pair into an ATP-loaded alpha beta pair. Using (14)C-tentoxin and selected synthetic analogues, we found that toxin binding to the tight inhibitory site of CF(1) exerts some cooperative effect on the loose reactivatory site, but that no reciprocal effect exists. When the two tentoxin-binding sites are filled in reactivated F(1)-ATPase, they do not exchange their role during catalytic turnover, indicating an impairment between nucleotide occupancy and the shape of tentoxin-binding pocket. This analysis provides a mechanical interpretation of the inhibition of F(1)-ATPase by tentoxin and a clue for understanding the reactivation process.


A novel microsurgical nerve implantation technique preserving outer nerve layers.
A novel epineural tube implantation paradigm in the adult rat was designed for the analysis of regulatory cell interactions in peripheral nerves and for the development of therapeutic implants. The aim was to allow the integration of synthetic regenerative structures and cells into the nerve interior while preserving an outer nerve tissue layer with a supportive vasculature. The microsurgical technique allowed us to remove the interfascicular epineurium, leaving behind an epineural tube with an intact tissue wall of about 0.1-0.2mm. The resulting tube was filled with hundreds of bioengineered bands of Büngner which were composed of resorbable polymer filaments seeded with Schwann cells. Alternatively, purified cells to be analyzed or different types of growth matrices were injected into the epineural tube. Such manipulations will allow to generate and investigate concentration gradients of biological factors or to analyse cell-matrix interactions under defined conditions in a supportive in vivo environment. Our current aim is to evaluate bioengineered neural implants. In summary, a microsurgical in vivo paradigm has been developed to address multiple aspects of peripheral nerve regeneration.


Antisecretory factor as a potential health-promoting molecule in man and animals.
Antisecretory factor (AF) is a protein secreted in plasma and other tissue fluids in mammals with proven antisecretory and anti-inflammatory activity; its immunohistological distribution suggests a role in the immune system. The expression level and the distribution of AF protein are altered during an immunological response. Exposure to bacterial toxins induces secretion of AF in plasma, probably reflecting a natural defence mechanism to agents causing diarrhoea, thereby contributing to a favourable clinical outcome and disease termination. An increase of AF levels in plasma by dietary means, such as specially processed cereals (SPC), has been demonstrated in human subjects and animals. Administration of SPC to patients affected by inflammatory bowel disease, gastroenteritis and Ménière's disease relieved symptoms and improved quality of life. A recent study showed the positive effect of SPC diet supplementation on prevention of the effects of exposure to low levels of blast overpressure in rats, reducing the extent of intracranial pressure increase and cognitive function impairment. AF-rich egg yolk powder improved health status in children suffering acute and chronic diarrhoea, reducing the frequency and increasing the consistency of stools. This kind of functional food could be used for prophylaxis in populations exposed to a high risk of morbidity and mortality caused by diarrhoea and as a complementary therapy in patients affected by chronic intestinal inflammatory disease to improve well-being. In pig husbandry AF-inducing diets, owing to their antisecretory activity and anti-inflammatory action, are a suitable option as an alternative to antibiotic growth promoters to counteract post-weaning diarrhoea.


[The short-term and long-lasting changes in DNA and RNA synthesis in the cerebral cortex cells of animals subjected to hypoxia and nerve tissue transplantation].
The DNA and RNA synthesis in the cells of the brain cortex of intact rats and animals subjected to hypoxia, hypoxia with subsequent transplantation or by the local brain injury has been investigated. The DNA synthesis changes insignificantly in the case of hypoxia, it enhances slightly in the area of the injury and increases much more after transplantation. The RNA synthesis decreases considerably immediately after hypoxia and decreases much more 120 days later. Using the ultracentrifuge method it has been found that under the effect of hypoxia the number of nervous cells decreases, the number of glial cells does not change. The local injury in the nervous tissue enhances abruptly the synthesis in neurons and glial cells in the hypoxia-exposed animals, the embryonic nervous tissue transplantation normalizes the number of neurons in the specimens under study and the RNA synthesis in the neurons and glial cells.


Y1 signalling has a critical role in allergic airway inflammation.
Asthma affects 300 million people worldwide, yet the mechanism behind this pathology has only been partially elucidated. The documented connection between psychological stress and airway inflammation strongly suggests the involvement of the nervous system and its secreted mediators, including neuropeptides, on allergic respiratory disease. In this study, we show that neuropeptide Y (NPY), a prominent neurotransmitter, which release is strongly upregulated during stress, exacerbates allergic airway inflammation (AAI) in mice, via its Y1 receptor. Our data indicate that the development of AAI was associated with elevated NPY expression in the lung and that lack of NPY-mediated signalling in NPYKO mice or its Y1 receptor in Y1KO mice significantly improved AAI. In vivo, eosinophilia in the bronchoalveolar fluid as well as circulating immunoglobulin E in response to AAI, were significantly reduced in NPY- and Y1-deficient compared with wild-type mice. These changes correlated with a blunting of the Th2 immune profile that is characteristic for AAI, as shown by the decreased release of interleukin-5 during ex vivo re-stimulation of T cells isolated from the thoracic draining lymph nodes of NPY- or Y1-deficient mice subjected to AAI. Taken together this study demonstrates that signalling through Y1-receptors emerges as a critical pathway for the development of airway inflammation and as such potentially opens novel avenues for therapeutic intervention in asthma.


[Plasma procalcitonin in rheumatic diseases].
Data on the origin and biological function of procalcitonin, the pro-hormone of calcitonin, are scarce. Since this peptide can be induced in bacterial invasive infections, serum procalcitonin levels may be useful in differential diagnosis of systemic inflammatory response syndrome. This review will focus on the clinical significance of changes in serum procalcitonin levels in patients with connective tissue diseases and other rheumatic disorders.


[Giant colonic diverticulum. Report of a case and review of the literature].
We report about a 67 year-old male patient with a giant colonic diverticulum and multiple comorbid conditions. The unusual clinical presentation and high surgical risk in this case required challenging therapeutic decisions that eventually lead to a left colon resection and complementary surgical procedures, despite the patient had been considered non-surgical candidate by two surgical teams before. A literature review from the last ten years was performed using Medline and Pubmed databases, looking for "giant colonic diverticulum". We found 43 case reports, one case series and one review article including 135 patients. Discussion is based on the information obtained from the literature review and the particular aspects of this interesting case.


Lumbar microdiscectomy and post-operative activity restrictions: a protocol for a single blinded randomised controlled trial.
Lumbar microdiscectomy is the most commonly performed spinal surgery procedure, with over 300,000 cases performed annually in the United States alone. Traditionally, patients were advised to restrict post-operative activity as this was believed to reduce the risk of disc reherniation and progressive instability. However, this practice would often delay patients return to work. In contemporary practice many surgeons do not restrict patient post-operative activity due to the perception this practice is unnecessary. We describe a randomised controlled trial to assess the impact of activity restrictions on clinical outcome following lumbar discectomy. The lumbar microdiscectomy and post-operative activity restriction trial is a multi-centre, randomised, controlled single blinded trial. Two hundred ten patients due to undergo single level lumbar microdiscectomy without a history of previous spine surgery, infection or fracture are randomised to be advised either restricted or unrestricted activity for a period of 30 days following lumbar microdiscectomy. Actual adherence with trial allocation will be monitored bioelectronically via a wearable device. Outcome assessment at follow up will occur at 1, 3, 6 and 12 months. The primary outcome will be a composite endpoint comprising changes in Visual Analogue Scale (Leg and Back), Oswestry Disability Index and the absence of intervertebral disc reherniation or secondary intervention. This randomised controlled trial will directly compare post-operative protocols of activity restrictions and no restrictions following lumbar discectomy with adherence monitored bioelectronically. Australian New Zealand Clinical Trials Registry: ACTRN12616001360404 (retrospectively registered 30/09/2016).


Radiation-Induced Transformations of C6H6 Molecules in Solid Noble-Gas Matrices: Is Benzene Intrinsically Resistant in Condensed Media?
The radiation resistance of aromatic compounds is one of the key concepts of basic and applied radiation chemistry in condensed phases. Usually, it is attributed to the intrinsic radiation stability of the benzene ring. In this work, we have demonstrated for the first time that the isolated benzene molecules undergo rather efficient radiation-induced degradation in rigid inert media at cryogenic temperatures (comparable to that of aliphatic hydrocarbons), and their stability is essentially determined by the intermolecular relaxation correlating with matrix polarizability. The principal primary products of benzene radiolysis in matrices are phenyl radicals and fulvene. The matrix environment strongly affects the proportion of these species because of external heavy atom effect on the intersystem crossing, which may trigger further reaction pathways. The obtained results may have important implications for the prediction of radiation stability of complex organic systems and polymers. Furthermore, they may contribute to a better understanding of the radiation-induced evolution of aromatic species in cold interstellar media.


A rapid fingerprinting approach to distinguish between closely related strains of Shewanella.
One of the big operational problems facing laboratories today is the ability to rapidly distinguish between strains of bacteria that, while physiologically distinct, are nearly impossible to separate based on 16S rRNA gene sequence differences. Here we demonstrate that ITS-DGGE provides a convenient approach to distinguishing between closely related strains of Shewanella, some of which were impossible to separate and identify by 16 rRNA gene sequence alone. Examined Shewanella genomes contain 8-11 copies of rrn (ribosomal RNA gene) operons, and variable size and sequence of 16S-23S ITS (intergenic transcribed spacer) regions which result in distinct ITS-DGGE profiles. Phylogenetic constructions based on ITS are congruent with the genomic trees generated from concatenated core genes as well as with those based on conserved indels, suggesting that ITS patterns appear to be linked with evolutionary lineages and physiology. In addition, three new Shewanella strains (MFC 2, MFC 6, and MFC 14) were isolated from microbial fuel cells enriched from wastewater sludge and identified by ITS-DGGE. Subsequent physiological and electrochemical studies of the three isolates confirmed that each strain is phenotypically/genotypically distinct. Thus, this study validates ITS-DGGE as a quick fingerprint approach to identifying and distinguishing between closely related but novel Shewanella ecotypes.


The differential transcription network between embryo and endosperm in the early developing maize seed.
Transcriptome analysis of early-developing maize (Zea mays) seed was conducted using Illumina sequencing. We mapped 11,074,508 and 11,495,788 paired-end reads from endosperm and embryo, respectively, at 9 d after pollination to define gene structure and alternative splicing events as well as transcriptional regulators of gene expression to quantify transcript abundance in both embryo and endosperm. We identified a large number of novel transcribed regions that did not fall within maize annotated regions, and many of the novel transcribed regions were tissue-specifically expressed. We found that 50.7% (8,556 of 16,878) of multiexonic genes were alternatively spliced, and some transcript isoforms were specifically expressed either in endosperm or in embryo. In addition, a total of 46 trans-splicing events, with nine intrachromosomal events and 37 interchromosomal events, were found in our data set. Many metabolic activities were specifically assigned to endosperm and embryo, such as starch biosynthesis in endosperm and lipid biosynthesis in embryo. Finally, a number of transcription factors and imprinting genes were found to be specifically expressed in embryo or endosperm. This data set will aid in understanding how embryo/endosperm development in maize is differentially regulated.


Chemical weathering and progressing alteration as possible controlling factors for creeping landslides.
Landslides can behave as dynamic processes, which emerge from the complex interplay of tectonics, erosion, weathering and gravitational influences, triggered by various hydrological, mineralogical, biological and geotechnical factors. Integral studies to assess the mechanisms underlying landslide initiation and progression are mainly focussed on specific cases with high geohazard potential. The landslide near Stadtschlaining (Austria) represents a key study site to elucidate the impacts of pelitic sediment composition, weathering regime, alteration patterns and hydrochemistry on recurrent damage progression in the local infrastructure. Based on field work, soil-mechanical logging (Atterberg limits, undrained strength, friction angles), water chemistry (ICP-OES, IC, hydrochemical modeling), solid-phase characterization (XRD, XRF, SEM) and sorption experiments we establish a conceptual model for initiating and progressing of landslides: Infiltration of low mineralized meteoric water (EC: <200 μS/cm) in permeable limonitic gravels triggers chemical weathering of greenschist-derived detritus and promotes its transformation into kaolinite and smectite. The clayey strata (>50 wt% of clay minerals) create zones of mechanical and chemical weakness in the underground (~4-6 m below ground level), which are characterized by particle disintegration/delamination, slip bedding and deformations, and development of porous layers depicting water flow paths. Subsequent Na+ exchange for bivalent ions in the smectite interlayer delivered by percolating, highly mineralized water (EC: 1600-5100 μS/cm) is caused by de-icing salt and fertilizer applications during winter and late summer, and yield in i) decohesion and physical breakdown of the particle aggregates and ii) swelling of the clay matrix in early spring and autumn. These processes reduce the shear strength of the pelitic sediments, resulting in failure and initiation of landslides (deformation: ~500 mm within a month) and subsequent steady creeping motion (deformation: ~100 mm in 6 months). Customized engineered solutions to prevent landslides in this area are presented, which can be conveyed to analogous landslide-affected areas worldwide.


Acalculous cystinuria.
We compared quantitative urinary cystine values in stone-forming family members and unaffected relatives based on an index stone-forming patient. A family pedigree of 39 members was identified. Quantitative urinalyses and serum biochemical profiles were collected with the subjects on similar diets over 72 hours. Detailed medical histories were obtained. Three family members are stone-forming cystinuric patients with a mean urinary cystine excretion of 496 mg/g of creatinine per day. Six persons without a history of stones (mean age 50) had a mean urinary cystine excretion of 364 mg/g of creatinine per day. Thirty-four family members had elevated urinary cystine concentrations (mean 151 mg/g of creatinine per day), and only two family members had normal urinary cystine excretion. Fifteen subjects with elevated urinary cystine values (> 150 mg/g of creatinine per day) had no history of urinary stones despite a mean age of 44 years. Urinary electrolytes and volumes were similar in the stone-forming and non-stone-forming groups. Thirty-nine per cent of the persons from this family pedigree had elevated quantitative urinary cystine concentrations and no history of urinary stones despite a mean age of 44 years. Acalculous cystinuria is an elevated urinary cystine concentration without evidence of urinary stone disease. Despite abnormal urinary cystine excretion, acalculous cystinuria does not necessarily culminate in clinical urinary stone disease.


Filterability of blood cells: methods and clinical applications.
It could be claimed that expansion in clinical aspects of haemorheology has largely been fueled by the development of a simple test of blood flow properties, i.e., analysis of filterability. With time, the level of sophistication in equipment, theory and sample preparation has increased. Theories for the development of flow in the filter have been described and these enable cellular parameters, such as transit times, to be calculated from experimental data. These theories can be quite general, and applied to filtration of red or white cells. Ideally, experimental design requires an understanding of cell behaviour at the filter and of the effects of factors such as the sample concentration and volume, and the presence of different types of cells or subpopulations. Otherwise, results are susceptible to misinterpretation, particularly if impurities or mixed populations of cells are present. It is thus very important to know accurately the constituents of the test suspension. In clinical applications the trend has been to move away from whole blood filtration, toward use of relatively pure suspension of separated red cells and white cells. In the area of red cell filtration this has led to reappraisal of some previously reported abnormalities. The relatively new study of white cell filtration should benefit from previous experience, but there is the added problem of the reactivity of the cells under test, and their mixed nature. In any case, critical evaluation of the meaning and clinical relevance of results is necessary.


Rituximab and minimal change nephrotic syndrome: a therapeutic option.
Minimal change nephrotic syndrome (MCNS) usually responds to steroids but frequently relapses, requiring additional treatment with immunosuppressive agents. Rituximab is a chimeric murine/human monoclonal immunoglobulin G1 antibody that targets CD20, a B-cell differentiation marker. B-cell recovery begins at approximately 6 months following the completion of treatment. Rituximab has a beneficial effect, with the sustained remission or reduction of proteinuria in patients with steroid-dependent MCNS. Relapses are thought to be associated with an increase in CD19 cells. The mean serum half-life of rituximab was reported to be 10-15 days in patients with steroid-dependent nephrotic syndrome. Only infusion reactions, such as rash and chills, occurred after single-dose rituximab infusion and can be managed by pre-medication or infusion rate adjustments. Even though severe adverse effects of rituximab are not expected, physicians must be aware of potentially life-threatening adverse effects. Controlled randomized trials that include adult patients with steroid-dependent or steroid-resistant MCNS are required to prove the efficacy and safety of rituximab and to evaluate the cost-effectiveness of rituximab treatment.


Behavioral therapy for childhood constipation: a randomized, controlled trial.
It has been suggested that the addition of behavioral interventions to laxative therapy improves continence in children with functional fecal incontinence associated with constipation. Our aim was to evaluate the clinical effectiveness of behavioral therapy with laxatives compared with conventional treatment in treating functional constipation in childhood. In this randomized, controlled trial conducted in a tertiary hospital in The Netherlands, 134 children aged 4 to 18 years with functional constipation were randomly assigned to 22 weeks (12 visits) of either behavioral therapy or conventional treatment. Primary outcomes were defecation frequency, fecal incontinence frequency, and success rate. Success was defined as defecation frequency of > or = 3 times per week and fecal incontinence frequency of < or = 1 times per 2 weeks irrespective of laxative use. Secondary outcomes were stool-withholding behavior and behavior problems. Outcomes were evaluated at the end of treatment and at 6-months follow-up. All of the analyses were done by intention to treat. Defecation frequency was significantly higher for conventional treatment. Fecal incontinence frequency showed no difference between treatments. After 22 weeks, success rates did not differ between conventional treatment and behavioral therapy (respectively, 62.3% and 51.5%), nor did it differ at 6 months of follow-up (respectively, 57.3% and 42.3%). The proportion of children withholding stools was not different between interventions. At follow-up, the proportion of children with behavior problems was significantly smaller for behavioral therapy (11.7% vs 29.2%). Behavioral therapy with laxatives has no advantage over conventional treatment in treating childhood constipation. However, when behavior problems are present, behavioral therapy or referral to mental health services should be considered.


Identification of Novel Protein Targets of Dimethyl Fumarate Modification in Neurons and Astrocytes Reveals Actions Independent of Nrf2 Stabilization.
The fumarate ester dimethyl fumarate (DMF) has been introduced recently as a treatment for relapsing remitting multiple sclerosis (RRMS), a chronic inflammatory condition that results in neuronal demyelination and axonal loss. DMF is known to act by depleting intracellular glutathione and modifying thiols on Keap1 protein, resulting in the stabilization of the transcription factor Nrf2, which in turn induces the expression of antioxidant response element genes. We have previously shown that DMF reacts with a wide range of protein thiols, suggesting that the complete mechanisms of action of DMF are unknown. Here, we investigated other intracellular thiol residues that may also be irreversibly modified by DMF in neurons and astrocytes. Using mass spectrometry, we identified 24 novel proteins that were modified by DMF in neurons and astrocytes, including cofilin-1, tubulin and collapsin response mediator protein 2 (CRMP2). Using an in vitro functional assay, we demonstrated that DMF-modified cofilin-1 loses its activity and generates less monomeric actin, potentially inhibiting its cytoskeletal remodeling activity, which could be beneficial in the modulation of myelination during RRMS. DMF modification of tubulin did not significantly impact axonal lysosomal trafficking. We found that the oxygen consumption rate of N1E-115 neurons and the levels of proteins related to mitochondrial energy production were only slightly affected by the highest doses of DMF, confirming that DMF treatment does not impair cellular respiratory function. In summary, our work provides new insights into the mechanisms supporting the neuroprotective and remyelination benefits associated with DMF treatment in addition to the antioxidant response by Nrf2.


Clinical total hip replacement with hydroxyapatite from 1984 to 1991.
Hydroxyapatite (HA) has been used for revision and primary total hip replacement surgery. HA has been used on both textured and porous coated implants for both primary and revision surgery. The results with textured implants have been compared with those of textured press-fit implants; with both revision and primary surgery the HA-coated prostheses were superior both clinically and roentgenographically. The results with porous coated implants have shown better bone remodeling and less radiolucent lines around the implants. The author's conclusions are that HA is effective in providing earlier stability of the implant and demonstrates better remodeling of the bone around the implant than do implants without HA. The author recommends the use of HA on porous coated implants so that if resorption of the HA occurs, the fixation of the porous coating will be present to provide durability.


Longterm follow-up of electroencephalographic and clinical findings of a case with Gaucher's disease type 3a.
Among three recognised clinical phenotypes, type 3a Gaucher's disease is characterised by mild to severe systemic disease, neurological manifestations and myoclonic seizures. We report the long term clinical and electrophysiological follow-up of a 27-year old man with a diagnosis of type 3a Gaucher's disease, which was confirmed by bone marrow biopsy examination and leukocyte glucocerebrosidase level measurement. His neurological examination was normal throughout the follow-up period. EEG examination, recorded five days after the first seizure, revealed generalised nonrhythmic paroxysmal rapid spikes with occipital predominance increased by photic stimulation and normal background activity. The frequency of seizures increased from 3-4/year to 1-2/month within a follow-up period of 12 years and a repeat EEG examination on the eight year of diagnosis revealed additional background slowing. A giant potential was obtained in somatosensory evoked potential (SEP) examination. EEG findings of this case demonstrate a specific pattern with rapid spike activity, photosensitivity, eye closure sensitivity and gradual background slowing.


Lymphoproliferative syndrome with autoimmunity: A possible genetic basis for dominant expression of the clinical manifestations.
Fas (CD95/Apo-1) mutations were previously reported as the genetic defect responsible for human lymphoproliferative syndrome associated with autoimmune manifestations (also known as autoimmune lymphoproliferative syndrome or Canale-Smith syndrome). We have identified 14 new heterozygous Fas mutations. Analysis of patients and families allow us to further dissect this syndrome with regards to the relationship between Fas mutations, inheritance pattern, and phenotype as observed on long-term follow-up. In vitro studies show that lymphocytes from all Fas mutant carriers exhibit a Fas-antibody-induced apoptosis defect. However, among the 8 inherited mutations, 4 of 4 Fas missense mutations were associated with high clinical penetrance, whereas 3 of 4 mutations leading to a truncated Fas product were associated with variable clinical penetrance. This suggests that a second defect, in another yet undefined factor involved in apoptosis and/or lymphoproliferation control, is necessary to induce full clinical expression of the disease. These results also indicate that the currently available antibody-mediated in vitro apoptosis assay does not necessarily reflect the in vivo ability of abnormal Fas molecules to trigger lymphocyte death. In addition, we found that lymphoproliferative manifestations resolved with age, whereas immunological disorders [ie, hypergammaglobulinemia and detection of TcR alphabeta(+) CD4(-) CD8(-) lymphocytes] persisted. This observation suggests that Fas-mediated apoptosis plays a more important role in lymphocyte homeostasis in early childhood than later on in life.


Diagnosis and Antibiotic Treatment of Urinary Tract Infections in Danish General Practice: A Quality Assessment.
Rational antibiotic treatment of urinary tract infections (UTI) is important. To improve the quality of antibiotic treatment of UTI, it is essential to obtain insight into diagnostic approaches and prescribing patterns in general practice. The aim of this study was to investigate the quality of diagnostics and treatment of UTI in general practice by means of quality indicators (QIs). QIs provide a quantitative measure of quality and are defined by a numerator (the number of patients receiving a specific investigation or treatment) and a denominator (the number of patients included in the quality assessment). For adult patients with suspected UTI, practices registered the following: age, sex, risk factors, symptoms and signs, examinations, diagnosis and treatment. The levels of the QIs were compared with their corresponding standards. Half of the patients diagnosed with lower UTI or pyelonephritis fulfilled the diagnostic criteria for UTI: characteristic UTI symptoms and clear signs of bacteriuria, respectively. Urinalysis was performed for nearly all patients, including patients without characteristic symptoms of UTI. One-fourth of the patients with suspected lower UTI were treated with antibiotics despite no urinalysis and nearly half received antibiotics despite an inconclusive dipstick test. Pivmecillam was the preferred antibiotic. The findings of this study indicate that there is room for improvement in the management of UTI in Danish general practice.


Phylogenetic analysis of Helicobacter species based on partial gyrB gene sequences.
Analysis of 16S rRNA gene sequences is one of the most common methods for investigating the phylogeny and taxonomy of bacteria. However, several studies have indicated that the 16S rRNA gene does not distinguish between certain Helicobacter species. We therefore selected for phylogenetic analysis an alternative marker, gyrB, encoding gyrase subunit B. The aim of this investigation was to examine the applicability of gyrB gene fragments (approximately 1100 bp) for the phylogenetic study of 16 Helicobacter species and a total of 33 Helicobacter strains included in this study. Based on the sequenced fragments, a phylogenetic tree was obtained that contained two distinct clusters, with gastric species forming one cluster and enterohepatic species the other. The only exception was the gastric species Helicobacter mustelae, which clustered with the enterohepatic species. The calculated similarity matrix revealed the highest interspecies similarity between Helicobacter salomonis and Helicobacter felis (89 %) and the lowest similarity between Helicobacter pullorum and H. felis (60 %). The DNA G+C content of the sequenced fragments was < or =40 mol% in enterohepatic species and >46 mol% in gastric species, excluding Helicobacter pylori and H. mustelae, with G+C contents of 34 and 42 mol%, respectively. In summary, the gyrB gene fragments provided superior resolution and reliability to the 16S rRNA gene for differentiating between closely related Helicobacter species. A further outcome of this study was achieved by designing gyrB gene-based species-specific PCR primers for the identification of Helicobacter bizzozeronii.


Evaluation of media for selective isolation of yeasts from oral, rectal, and burn wound specimens.
Six media were evaluated to determine their ability to isolate yeasts and inhibit bacteria. The media included the following: Snyder, Snyder tellurite, Sabouraud tellurite, Littman-gentamicin, molybdate, and Mycosel (BBL). Doses of mixed intestinal gram-negative bacilli and enterococci were most effectively inhibited by Snyder tellurite agar. Klebsiella pneumoniae was the most common bacterial contaminant of the other media. All six media were comparable in isolating yeasts while preventing the growth of the oral bacterial flora. The selection of a basal fungal growth medium for tellurite incorporation to inhibit bacteria but permit growth of yeasts was affected by pH. The bacteriostatic effect of tellurite was decreased with increasing pH of media while fungistatic action was increased. The arbitrary selection of Snyder and Littman agars to isolate yeast from burn wound cultures demonstrated the need to include a selective medium for these specimens. Blood, phenylethyl alcohol blood agar, and Columbia CN blood agar were all inadequate for isolating yeasts from burns. Growth of a variety of filamentous saprophytic and pathogenic dimorphic fungi grew adequately on four of five selective media tested.


Association between tension and orientation of periodontal ligament fibroblasts and exogenous collagen fibres in collagen gels in vitro.
The relationship between the development of tension in sheets of fibroblasts and the orientation of these cells and collagen fibres in collagen gels was examined. Cell-containing, three-dimensional collagen gels were established in agarose-coated Epon dies measuring 10 mm X 4 mm X 4 mm, to which pieces of demineralized tooth and bone had been attached at opposite ends. Contraction of the gel into an opaque structure suspended between the two particles occurred over 24 h and resulted in concave upper and lateral surfaces and a flat to slightly concave lower surface. Initial orientation of the fibres along the tooth-bone axis was followed by similar orientation of the cells. Gels cast without cells exhibited no change in dimensions. Release of the tooth particle after 12 or 24 h of incubation led to shortening of the contracted gels 5 min following release. This shortening was significantly greater (P less than 0.001) than that of uncontracted or slightly contracted gels (1 h and 3 h incubation). Gels attached at one end only compacted around the site of attachment but did not show orientation of cells or fibres. Gels containing colcemid or cytochalasin D were only slightly compacted and did not develop tension. Collagen fibres, but not cell in colcemid-containing gels, showed some alignment; neither were aligned in the presence of cytochalasin D. These data suggest that both microtubules and microfilaments are necessary for alignment of cells and the establishment of tension between two points of attachment in collagen gels. Furthermore, they lend support to our previously advanced hypothesis that the development of tension between two points can result in the orientation of the cells along an axis connecting the points of attachment. This could provide a mechanism for the development of oriented fibre systems in vivo.


Monoclonal gammopathy of undetermined significance: a consensus statement.
On February 25, 2009, a panel of international experts on plasma cell dyscrasia and skeletal disease met to discuss monoclonal gammopathy of undetermined significance (MGUS). This non-malignant B-cell disorder is the most common plasma cell dyscrasia and is associated with an increased risk of developing serious B-cell disorders. Individuals with MGUS also have an increased risk of osteoporosis and osteopenia associated with an increased likelihood of developing fractures especially in the vertebral column, peripheral neuropathy and thromboembolic events. The goal of the meeting was to develop a consensus statement regarding the appropriate tests to screen, evaluate and follow-up patients with MGUS. The panel also addressed the identification and treatment of MGUS-related skeletal problems, thromboembolic events and neurological complications. The following consensus statement outlines the conclusions and marks the first time that a consensus statement for the screening and treatment of MGUS has been clearly stated.


One-Year Committed Exercise Training Reverses Abnormal Left Ventricular Myocardial Stiffness in Patients With Stage B Heart Failure With Preserved Ejection Fraction.
Individuals with left ventricular (LV) hypertrophy and elevated cardiac biomarkers in middle age are at increased risk for the development of heart failure with preserved ejection fraction. Prolonged exercise training reverses the LV stiffening associated with healthy but sedentary aging; however, whether it can also normalize LV myocardial stiffness in patients at high risk for heart failure with preserved ejection fraction is unknown. In a prospective, randomized controlled trial, we hypothesized that 1-year prolonged exercise training would reduce LV myocardial stiffness in patients with LV hypertrophy. Forty-six patients with LV hypertrophy (LV septum >11 mm) and elevated cardiac biomarkers (N-terminal pro-B-type natriuretic peptide [>40 pg/mL] or high-sensitivity troponin T [>0.6 pg/mL]) were randomly assigned to either 1 year of high-intensity exercise training (n=30) or attention control (n=16). Right-heart catheterization and 3-dimensional echocardiography were performed while preload was manipulated using both lower body negative pressure and rapid saline infusion to define the LV end-diastolic pressure-volume relationship. A constant representing LV myocardial stiffness was calculated from the following: P=S×[Exp {a (V-V0)}-1], where "P" is transmural pressure (pulmonary capillary wedge pressure - right atrial pressure), "S" is the pressure asymptote of the curve, "V" is the LV end-diastolic volume index, "V0" is equilibrium volume, and "a" is the constant that characterizes LV myocardial stiffness. Thirty-one participants (exercise group [n=20]: 54±6 years, 65% male; and controls (n=11): 51±6 years, 55% male) completed the study. One year of exercise training increased max by 21% (baseline 26.0±5.3 to 1 year later 31.3±5.8 mL·min-1·kg-1, P<0.0001, interaction P=0.0004), whereas there was no significant change in max in controls (baseline 24.6±3.4 to 1 year later 24.2±4.1 mL·min-1·kg-1, P=0.986). LV myocardial stiffness was reduced (right and downward shift in the end-diastolic pressure-volume relationship; LV myocardial stiffness: baseline 0.062±0.020 to 1 year later 0.031±0.009), whereas there was no significant change in controls (baseline 0.061±0.033 to 1 year later 0.066±0.031, interaction P=0.001). In patients with LV hypertrophy and elevated cardiac biomarkers (stage B heart failure with preserved ejection fraction), 1 year of exercise training reduced LV myocardial stiffness. Thus, exercise training may provide protection against the future risk of heart failure with preserved ejection fraction in such patients. Registration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT03476785.


Comparison of several point-of-care testing (POCT) glucometers with an established laboratory procedure for the diagnosis of type 2 diabetes using the discordance rate. A new statistical approach.
The applicability of point-of-care testing (POCT) glucometers for monitoring blood glucose concentrations has been demonstrated. However, their use in diagnosing type 2 diabetes is still debated. Therefore, a new statistical procedure for estimating discordance rates (DRs) was applied in comparing a well-established laboratory method (Ebio) with another laboratory method (Cobas Integra 700) and with several POCT glucometers (Accu-Chek, Accutrend, Elite, HemoCue, Omni) in detecting glucose intolerance states. All procedures led to parallel glucose concentration patterns in capillary blood, venous plasma, and venous blood during oral glucose tolerance tests. However, the mean concentrations differed more or less. The Ebio and Integra results agreed within a maximal deviation of 3%. In blood samples, the HemoCue and Accutrend results were closest to the laboratory procedures (Ebio and Integra) and the highest differences were obtained with the Elite. Comparing whole blood values with those obtained in the aqueous blood compartment (Omni), even greater differences were observed. When all procedures were referred to the same glucose standard, the Ebio, Integra, Accutrend, and Omni results remained almost unchanged, whereas the Elite "moved" toward the Ebio results, and the Accu-Chek results toward the Omni results. Thus, traceability to an aqueous standard was observed with the Ebio, Integra, Accutrend, and Elite in all three sample systems. The Accu-Chek was only traceable in the presence of albumin, and HemoCue was not traceable at all. The clinical relevance of the differences observed between Ebio and POCT glucometers was tested by comparing the relative number of discordant classifications. The highest DRs were observed in the fasting state. They were higher in capillary blood than in the other sample systems. The DRs were found higher with POCT glucometers than with the other established laboratory procedure (Integra). Thus, at least in the fasting state, all POCT glucometers were less reliable than the established laboratory procedures and above the chosen criteria of clinical acceptability (DR < or = 5%). After transforming all glucometer results with a regression function (bias correction), the DRs were less than 5% if compared with the Ebio procedure in all sample systems. In conclusion, the WHO recommendation not to use POCT glucometers for diagnosing type 2 diabetes must be supported. However, after proper recalibration, the tested systems were acceptable. Therefore, manufacturers should reconsider their calibration procedure. Those POCT procedures should be preferred that can be referred to aqueous glucose solutions.


A review of problems, obstacles and sequelae encountered during femoral lengthening : uniplanar versus circular external fixator.
There is currently a consensus regarding the superiority of circular type external fixators over uniplanar fixators for lengthening of the tibia, but femoral lengthening is still subject to the surgeon's preference. This study compares the occurrence rates of significant problems, obstacles and sequelae between these two techniques. Fifty patients (29 male, 21 female), with a mean age of 20 years were assigned to a circular type fixator group (54 lengthening segments), whereas 60 patients (29 male, 31 female), with a mean age of 20 years were assigned to a uniplanar fixator group (67 lengthening segments). The incidence of knee stiffness was significantly higher in the circular external fixator group (031 per segment) compared to the uniplanar external fixator group (0.13 per segment) (p < 0.05). The incidence of pain during lengthening was higher in the circular external fixator group, and patient satisfaction was higher in the uniplanar external fixator group. We recommend the uniplanar external fixator as a preferable device for femoral lengthening.


[Differential experience of anxiety conditions: a contribution to the diagnosis and pathogenesis of anxiety attacks. A case report].
By means of a case report about a depressive female patient suffering from clusters of attacks of various psychosomatic disorders associated with anxiety states of different degrees problems of diagnostic classification are discussed with regard to the DSM-III. The subjective experiences of anxiety during the various episodes of the patient's disease are described. A phenomenological difference between fear of death with preserved relation to reality on the one hand and "calamity anxiety" with a short-lasting breakdown of reality testing on the other hand is indicated. The neurobiological model of noradrenergic dysregulation is taken on trial to understand the various phenomenons of the disease.


Enhanced osteogenic and selective antibacterial activities on micro-/nano-structured carbon fiber reinforced polyetheretherketone.
Carbon fiber reinforced polyetheretherketone (CFRPEEK) possesses a similar elastic modulus to that of human cortical bone and is considered as a promising candidate to replace metallic surgical implants. However, the bioinertness and deficiency of antibacterial activities impedes the application of CFRPEEK as orthopedic and especially oral implants. In this work, dual zinc and oxygen plasma immersion ion implantation (Zn/O-PIII) is applied to modify CFRPEEK, achieving both zinc incorporation and introduction of unique micro-/nano-structures on the surface. Scanning electron microscopy (SEM) observation reveals the formation of micro-pits with the size of around 800 nm on the CFRPEEK surface. X-ray photoelectron spectroscopy (XPS) test results confirm the existence of ZnO on the near surface. In vitro cell adhesion, viability assay, and real-time PCR analyses disclose enhanced adhesion, proliferation, and osteo-differentiation of mouse osteoblasts (MC3T3-E1) and rat bone mesenchymal stem cells (bMSCs) on the structured surface. Furthermore, the multilevel structures on CFRPEEK exhibit great antibacterial activity to biofilm-positive Staphylococcus aureus (ATCC 25923), MRSA (ATCC 43300) and Staphylococcus epidermidis (ATCC 35984) while show no obvious inhibition to Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853) and biofilm-negative Staphylococcus epidermidis (ATCC 12228). The obtained results indicate that the CFRPEEK surface with specific biological properties can be enhanced by zinc incorporation and multilevel structure introduction, through which the application of CFRPEEK to orthopedic and dental implants can be further broadened.


Investigating the use and awareness of artificial sweeteners among diabetic patients in Bangladesh.
As with many countries around the world, the incidence of diabetes in Bangladesh is increasing significantly. Whilst there is controversy in the field regarding the health impact of artificial sweeteners in Western communities, the link between sweetener consumption and awareness in Bangladesh has not been established. In the present study, 260 diabetic patients completed a questionnaire survey to investigate the use and awareness of sweeteners and how this links to demographics and potential co-morbidities. Findings show that daily artificial sweetener consumption is significantly associated with hypertension but not other co-morbidities such as kidney disease or obesity. We further demonstrate that there is limited checking of artificial sweeteners in food or drink products by participants. the rurality of diabetic participants was found to significantly correlates with lower awareness of any health impact of artificial sweeteners. The findings from this study demonstrate that there is a need to increase the awareness of artificial sweetener use in diabetic patients in Bangladesh. Combined with a more robust understanding of the health impact of artificial sweeteners, these findings suggest that there is potential to improve outcomes for diabetic patients by improving this awareness.


Influence of temperature gradients on charge transport in asymmetric nanochannels.
Charge selective asymmetric nanochannels are used for a variety of applications, such as nanofluidic sensing devices and energy conversion applications. In this paper, we numerically investigate the influence of an applied temperature difference over tapered nanochannels on the resulting charge transport and flow behavior. Using a temperature-dependent formulation of the coupled Poisson-Nernst-Planck and Navier-Stokes equations, various nanochannel geometries are investigated. Temperature has a large influence on the total ion transport, as the diffusivity of ions and viscosity of the solution are strongly affected by temperature. We find that the selectivity of the nanochannels is enhanced with increasing asymmetry ratios, while the total current is reduced at higher asymmetry cases. Most interestingly, we find that applying a temperature gradient along the electric field and along the asymmetry direction of the nanochannel enhances the selectivity of the tapered channels even further, while a temperature gradient countering the electric field reduces the selectivity of the nanochannel. Current rectification is enhanced in asymmetric nanochannels if a temperature gradient is applied, independent of the direction of the temperature difference. However, the degree of rectification is dependent on the direction of the temperature gradient with respect to the channel geometry and the electric field direction. The enhanced selectivity of nanochannels due to applied temperature gradients could result in more efficient operation in energy harvesting or desalination applications, motivating experimental investigations.


[Anesthetic management of Y-shaped tracheobronchial stent insertion via tracheostomy site].
We report two cases of Y-shaped tracheobronchial stent insertion via tracheostomy site. A 62-year-old woman diagnosed with tracheo-gastric roll fistula after subtotal esophagectomy and a 47-year-old woman diagnosed tracheobronchial stenosis due to lung cancer underwent Y-shaped tracheobronchial stent insertion. In both cases, anesthesia was induced and maintained by propofol, remifentanil, and rocuronium. After general anesthesia was induced, oral tracheal tube was inserted and Y-shaped tracheobronchial stent was inserted via tracheostomy site. Patients were apneic during surgical procedure and ventilated via oral tracheal tube in case of Sp(O2) decrease. During ventilation, the operator closed tracheostomy by hand. We used oral tracheal tube with short cuff which enabled surgical procedure without extubation. Compared with metal stent, silicone Y-shaped tracheobronchial stent needs much more time for insertion. Therefore, oral insertion needs frequent intubation and extubation which causes airway mucosal edema or damage. Our method, stent insertion via tracheostomy site and ventilation via oral tracheal tube, was compatible with safer airway management and surgical procedure.


Spinal Cord Stimulation in Chronic Pain: Mode of Action.
Literature review. A review of the literature that presents a perspective on mechanisms of actions behind spinal cord stimulation (SCS) therapy for chronic pain. SCS is an effective therapeutic alternative for the treatment of intractable chronic pain. Its application has been mostly based on the gate control theory of pain. Computational models have been fundamental on the understanding of clinical observations and the design of therapies that provide optimal neuromodulation. Research has provided insight into the involvement of specific neurotransmitters that support segmental and supraspinal mechanisms of action. A literature review was performed with emphasis on mechanisms of action for SCS including the effects of electrical fields on spinal cord structures based on computational models and preclinical and clinical explorations. This review provides background on the development of SCS, which has been driven around a paresthesia-based paradigm as a result of the gate control theory. A review of computational models emphasizes their importance on our current understanding of the mechanism of action and clinical optimization of therapy. Electrophysiology and molecular biology have provided a closer, yet narrow, view of the effect of SCS on neurotransmitters and their receptors, which have led to the formulation of segmental and supraspinal mechanisms. Literature supporting the involvement of glial cells in chronic pain and their characteristic response to electrical fields should motivate further investigation of mechanisms involving neuroglia. Finally, a review of recent results paresthesia-free strategies should encourage research on mechanisms of action. The mechanisms of SCS have been extensively studied and several consistent phenomena have emerged. The activation of A-beta fibers to induce paresthesia also involve neurotransmitter release via segmental and supraspinal pathways. Despite advancements, much remains to be understood, particularly as new stimulation strategies are developed. N /A.


Bisphenol A: an endocrine and metabolic disruptor.
Bisphenol A (BPA), initially designed, like diethylstilbestrol, as a synthetic estrogen, has been rapidly and widely used for its cross-linking properties in the manufacture of polycarbonate plastics and epoxy resins. Because of incomplete polymerization and degradation of the polymers by exposure to higher than usual temperatures, BPA leaches out from food and beverage containers, as well as from dental sealants. In humans, free active unconjugated BPA is metabolized by rapid glucurono- or sulfo-conjugation and eliminated via renal clearance. However, exposure to environmental nanomolar concentrations of BPA is ubiquitous and continuous via different routes: oral, air, skin. In rodents, fetal and perinatal exposure to such environmentally relevant doses of BPA has been shown to affect the brain, liver, gut, adipose tissue, endocrine pancreas, mammary gland and reproductive tract and function. Similar concentrations are also able in vitro to impact human malignant breast, prostate, male germ or adipocyte cell lines (with a promoting effect and by interfering with chemotherapy drugs), or to stimulate pancreatic β cell insulin secretion. High levels of BPA have recently been correlated with obesity, diabetes, cardiovascular diseases, polycystic ovarian disease or low sperm count. However, before the real impact of BPA on human health can be clearly assessed, prospective longitudinal epidemiological studies are needed as well as characterization of selective biomarkers to verify long-term exposure and selective imprinting.


[Determination of HIV-RNA using in situ hybridization on myocardial tissue. Role of HIV in cardiomyopathy in AIDS].
Aim of this study was to evaluate the presence of HIV-1 and CMV genomic sequences in myocardial tissue from autopsy of AIDS patients by in situ hybridization technique and to make a correlation with clinical and/or pathological signs of myocarditis. Seven patients out of 12 (58.3%) were positive for HIV-1 (6/7 patients with pathological signs of myocarditis and 1/5 without pathological abnormalities; p = 0.029). In situ hybridization for CMV was positive in 5/12 patients (41.7%) (3 with myocarditis and 2 without myocarditis); three clinically symptomatic cases were contemporary positive for HIV-1 and CMV. Our results seem to confirm emerging data about the presence of HIV-1 in myocardial tissue and about the possible role of HIV-1 in determining cardiac pathologies. Furthermore we suggest the hypothesis of an interaction of HIV-1 and CMV in latest of HIV infection.


Integrin beta1 mediates hepatocellular carcinoma cells chemotaxis to laminin.
Chemotaxis is an important step during the invasion of carcinoma cells. And integrins are most important receptors mediating interaction between cells and extracellular matrix (ECM). This study was designed to study integrin beta1 mediating chemotaxis of hepatocellular carcinoma (HCC) cells to laminin(LN). A micropipette technique was adopted to investigate the effect of blockade of integrin beta1 on pseudopod protrusion of HCC cells in response to LN stimulation. Chemotactic pseudopod protrusion of a HCC cell was evaluated using a dual-pipette set-up, in which two pipettes filled with LN solution were positioned in close contact with the same cell, and pseudopod protrusion into each pipette was viewed dynamically and recorded with a tape recorder. The lengths of pseudopods were measured and plotted against time to obtain a pseudopod growth curve. The integrin beta1 subunit on the surfaces of HCC cells were analyzed by flow cytometry. In dual pipette chemotaxis experiment, when the two pipettes were filled with LN(50 microg/ml, 200 microg/ml), pseudopods extended from the HCC cell into each of the pipettes nearly symmetrically, ie, with nearly identical maximum pseudopod length and similar pseudopod growth curves. Upon addition of anti-CD29 (20 microg/ml) to one of the pipettes, pseudopod protrusion was blocked nearly completely while protrusion into the opposite pipette became more evidently, with a larger maximum length. Expression of integrin beta1 was up to 95.78% to cells chosen in the experiment. Integrin beta1 subunit was an important constituent receptor subunit for mediating chemotactic pseudopod protrusion of HCC cell to LN.


Gal-NCAM is a differentially expressed marker for mature sensory neurons in the rat olfactory system.
A new monoclonal antibody, 2E11, was produced by immunizing mice with the microsomal fraction of rat accessory olfactory bulb cells. This IgM recognizes a previously described complex alpha-galactosyl containing glycolipid, as well as N-linked glycoproteins at 170 and 210 kD. These proteins correspond to a new nerve cell adhesion molecule (NCAM) glycoform, Gal-NCAM, which contains a blood group B-like oligosaccharide. During embryonic development, the 2E11 epitope is expressed by a subset of mature olfactory sensory neurons randomly dispersed throughout the olfactory epithelium, whereas in the olfactory bulb, immunostaining is restricted to medial areas of the nerve layer. When compared to PSA-NCAM, another NCAM glycoform, Gal-NCAM has a mutually exclusive distribution pattern both in the olfactory epithelium and in the olfactory bulb. We propose a model for the hierarchy of neuronal maturation in the olfactory epithelium, including a switch from PSA-NCAM expression by immature neurons to the expression of Gal-NCAM by mature neurons.


Public funding for research on antibacterial resistance in the JPIAMR countries, the European Commission, and related European Union agencies: a systematic observational analysis.
Antibacterial resistant infections are rising continuously, resulting in increased morbidity and mortality worldwide. With no new antibiotic classes entering the market and the possibility of returning to the pre-antibiotic era, the Joint Programming Initiative on Antimicrobial Resistance (JPIAMR) was established to address this problem. We aimed to quantify the scale and scope of publicly funded antibacterial resistance research across JPIAMR countries and at the European Union (EU) level to identify gaps and future opportunities. We did a systematic observational analysis examining antibacterial resistance research funding. Databases of funding organisations across 19 countries and at EU level were systematically searched for publicly funded antibacterial resistance research from Jan 1, 2007, to Dec 31, 2013. We categorised studies on the basis of the JPIAMR strategic research agenda's six priority topics (therapeutics, diagnostics, surveillance, transmission, environment, and interventions) and did an observational analysis. Only research funded by public funding bodies was collected and no private organisations were contacted for their investments. Projects in basic, applied, and clinical research, including epidemiological, public health, and veterinary research and trials were identified using keyword searches by organisations, and inclusion criteria were based on the JPIAMR strategic research agenda's six priority topics, using project titles and abstracts as filters. We identified 1243 antibacterial resistance research projects, with a total public investment of €1·3 billion across 19 countries and at EU level, including public investment in the Innovative Medicines Initiative. Of the total amount invested in antibacterial resistance research across the time period, €646·6 million (49·5%) was invested at the national level and €659·2 million (50·5%) at the EU level. When projects were classified under the six priority topics we found that 763 (63%) of 1208 projects funded at national level were within the area of therapeutics, versus 185 (15%) in transmission, 131 (11%) in diagnostics, 53 (4%) in interventions, and only 37 (3%) in environment and 39 (3%) in surveillance. This was the first systematic analysis of research funding of antibacterial resistance of this scale and scope, which relied on the availability and accuracy of data from organisations included. Large variation was seen between countries both in terms of number of projects and associated investment and across the six priority topics. To determine the future direction of JPIAMR countries a clear picture of the funding landscape across Europe and Canada is needed. Countries should work together to increase the effect of research funding by strengthening national and international coordination and collaborations, harmonising research activities, and collectively pooling resources to fund multidisciplinary projects. The JPIAMR have developed a publicly available database to document the antibacterial resistance research collected and can be used as a baseline to analyse funding from 2014 onwards. JPIAMR and the European Commission.


Continuous extracorporeal clearance in metformin-associated lactic acidosis and metformin-induced lactic acidosis: a systematic review.
Metformin poisoning with lactic acidosis is an uncommon yet clinically serious condition related to the inhibition of normal aerobic metabolism. Toxicity may occur after an acute overdose although it is much more common after a systemic insult, such as acute kidney injury, in the setting of chronic use. Hemodialysis is currently the preferred extracorporeal treatment modality (Grade 1D evidence) although some patients may be too hemodynamically unstable to tolerate it. Continuous renal replacement therapy is considered an alternative if hemodialysis is unavailable but an evaluation of survival amongst this specific treatment class is lacking. To assess overall survival and provide an updated review of the toxicokinetic elimination parameters of patients receiving continuous renal replacement therapy for metformin poisoning. A comprehensive search was performed using the EMBASE and MEDLINE libraries from inception until November 30, 2021. Data was extracted and findings were summarized. Toxicokinetic parameters were analyzed and confirmed for accuracy when data permitted. Eighty-three reports met inclusion criteria. These consisted of only low-quality evidence including 75 case reports, four case series, and four descriptive retrospective reviews. Overall survival among patients suffering from metformin toxicity who received continuous extracorporeal treatment was 85.8%. When stratified between metformin-induced lactic acidosis and metformin-associated lactic acidosis, survival was 75.0% and 87.4%, respectively. Available continuous renal replacement therapy toxicokinetic parameters were quite heterogeneous. Errors in previously published toxicokinetic calculations were noted in only two instances. The overall average and median peak metformin concentrations were 70.5 mg/L and 41.9 mg/L, respectively. The average and median extracorporeal clearance rates were 39.0 mL/min and 42.1 mL/min (range 9.0-58.7 mL/min). The average and median elimination half-life parameters were 27.5 h and median 23.0 h. Elimination half-life ranged from seven to 74 h. There was no meaningful relationship between peak metformin concentration and continuous extracorporeal treatment half-life at lower concentrations, though at very high concentrations (over 200 mg/L), there was a trend towards a half-life below 20 h. There is insufficient data to robustly evaluate overall survival in relation to the extracorporeal clearance rate. Finally, there was no relevant relationship between maximal lactate concentration and survival, nor nadir pH and survival, for patients with either type of metformin toxicity. This retrospective systematic analysis of published cases treating metformin related lactic acidosis with continuous renal replacement therapy notes an overall slightly greater survival percentage compared to previous publications of individuals requiring any modality of renal replacement therapy. Because of publication bias, these results should be interpreted with caution and serve as hypothesis generating for future research. Prospective study focusing on the most clinically meaningful endpoint - survival - will help elucidate if continuous modalities are non-inferior to intermittent hemodialysis in metformin toxicity.


Hydrogen isotope variability in prairie wetland systems: implications for studies of migratory connectivity.
Hydrogen isotopes (delta2H) are often used to infer the origins of migratory animals based on the strong correlation between deuterium content of tissues and long-term patterns of precipitation. However, the extreme flood and drought dynamics of surface waters in prairie wetland systems could mask these expected correlations. We investigated H isotopic variability in an aquatic food web associated with Tree Swallows (Tachycineta bicolor) that rely heavily on wetland-derived aerial insects for food. We evaluated isotopic turnover and incorporation of environmental water into tissue, processes that could affect H isotopic composition. Wetland water and aquatic invertebrates showed intra- and interannual H isotopic variation mainly related to evaporation and the amount and timing of precipitation. Snails showed rapid turnover of tissue deuterium and a large contribution of environmental water to their tissues. Swallow feather deuterium (delta2Hf) was variable but did not clearly follow changes in any of the food web compartments measured. Instead, isotopic variability may have been driven by shifts in the type or relative amounts of grey consumed and types of wetlands used. Nevertheless, despite relatively high variance in delta2Hf, the majority of birds fell within the predicted range of delta2Hf for the study area, revealing that significant trophic averaging occurred. However, both (presumed) diet shifts and variable hydrological conditions have the potential to greatly increase variance that must be considered when assigning origins of migratory animals based on delta2H.


Characterization of energy-efficient and colorless ONUs for future TWDM-PONs.
The Full Services Access Network group has recently selected the time and wavelength division multiplexed passive optical network (TWDM-PON) as the base technology solution for next-generation PON stage-2 (NG-PON2). Meeting the core requirements of NG-PON2 necessitates the following additional features in the transceivers of the optical network unit (ONU) that is located at subscriber premises: (a) legacy system compliant; (b) wavelength tunable; (c) cost-efficient; and (d) energy-efficient. To address these features, we investigate the properties of short-cavity vertical-cavity surface-emitting lasers (SC-VCSELs) for implementation as colorless ONU transmitters in future TWDM-PONs. Specifically, we investigate the tunability and transmission performance of the SC-VCSEL across the C-minus wavelength band for legacy system compliance. We report on error-free transmission across a 800 GHz tuning range with a potential aggregate upstream capacity of 80 Gbps over a system reach of 40 km and with a split ratio of 1:128 per wavelength channel. Results were achieved without dispersion compensation and electronic equalization. We also evaluate the energy efficiency of the SC-VCSEL in active, doze, and sleep mode. When in active mode, the SC-VCSEL transmitter block consumes 91.7% less power than a distributed feedback (DFB) laser transmitter block. When transitioning between doze and active modes, the transmitter block has a short settling time of only 205 ns, thus increasing the power-saving duration and consequently reducing the overall power consumption of the ONU. Through numerical analysis, evaluation of the energy-savings of the SC-VCSEL ONU over the DFB ONU under various modes of operation, demonstrates up to 84% of energy-savings. The capacity, tuning range, split ratio, system reach, and energy-savings arising from SC-VCSEL ONU implementation as reported in this work, exceed the minimum requirements of NG-PON2 for future TWDM-PON deployments.


Prevalence, incidence, mortality and co-morbidities amongst human immunodeficiency virus (HIV) patients in Stockholm County, Sweden - the Greater Stockholm HIV Cohort Study.
Our aim was to study the prevalence and incidence of patients with human immunodeficiency virus (HIV) in the general population in Stockholm, Sweden. We also aimed to study mortality among individuals with HIV and to explore co-morbidities. The study population included all living persons who resided in Stockholm County, Sweden, as of 31 December 2012 (N = 2,212,435). Information on all consultations between 2007 and 2012 was obtained from primary health care, specialist outpatient care and inpatient care. Analyses were done by age and gender. All patients with a recorded diagnosis of HIV were included. The prevalence of HIV was calculated using 2012 data. The prevalence of HIV in Stockholm area as per end of December 2012 was as low as 0.1% in females and 0.2% in males, and the annual incidence of HIV continued to decline over the years. In recent years, cancers, diabetes and hypertension were about as common in individuals with HIV as in the general population. Males with HIV had 3- to 4-fold higher age-adjusted odds of being diagnosed with depression and 3-fold higher odds of anxiety disorders and women had 1.6 to 2-fold higher age-adjusted odds of depression and anxiety disorders, than males and females in the general population, respectively. The relatively good somatic health observed in this study could be attributed to nearly optimal HIV therapy in Sweden. The mental health of HIV patients was significantly worse than that in the general population and needs further attention.


Clinical Assessment of Judgment in Adults and the Elderly: Development and Validation of the Three Domains of Judgment Test-Clinical Version (3DJT-CV).
(1) Background: This article discusses the first two phases of development and validation of the Three Domains of Judgment Test (3DJT). This computer-based tool, co-constructed with users and capable of being administered remotely, aims to assess the three main domains of judgment (practical, moral, and social) and learn from the psychometric weaknesses of tests currently used in clinical practice. (2) Method: First, we presented the 3DJT to experts in cognition, who evaluated the tool as a whole as well as the content validity, relevance, and acceptability of 72 scenarios. Second, an improved version was administered to 70 subjects without cognitive impairment to select scenarios with the best psychometric properties in order to build a future clinically short version of the test. (3) Results: Fifty-six scenarios were retained following expert evaluation. Results support the idea that the improved version has good internal consistency, and the concurrent validity primer shows that 3DJT is a good measure of judgment. Furthermore, the improved version was found to have a significant number of scenarios with good psychometric properties to prepare a clinical version of the test. (4) Conclusion: The 3DJT is an interesting alternative tool for assessing judgment. However, more studies are needed for its implementation in a clinical context.


Fat-associated lymphoid clusters control local IgM secretion during pleural infection and lung inflammation.
Fat-associated lymphoid clusters (FALC) are inducible structures that support rapid innate-like B-cell immune responses in the serous cavities. Little is known about the physiological cues that activate FALCs in the pleural cavity and more generally the mechanisms controlling B-cell activation in FALCs. Here we show, using separate models of pleural nematode infection with Litomosoides sigmodontis and Altenaria alternata induced acute lung inflammation, that inflammation of the pleural cavity rapidly activates mediastinal and pericardial FALCs. IL-33 produced by FALC stroma is crucial for pleural B1-cell activation and local IgM secretion. However, B1 cells are not the direct target of IL-33, which instead requires IL-5 for activation. Moreover, lung inflammation leads to increased IL-5 production by type 2 cytokine-producing innate lymphoid cells (ILC2) in the FALC. These findings reveal a link between inflammation, IL-33 release by FALC stromal cells, ILC2 activation and pleural B-cell activation in FALCs, resulting in local and antigen-specific IgM production.


Skin as an immune organ and clinical applications of skin-based immunotherapy.
The prevalence of food allergy is increasing, and allergen avoidance continues to be the main standard of care. There is a critical need for safe and effective forms of immunotherapy for patients with food allergy as well as other allergic diseases. The skin is a multifunctional organ with unique immunologic properties, making it a favorable administration route for allergen-specific immunotherapy. Epicutaneous immunotherapy (EPIT) takes advantage of the skin's immune properties to modulate allergic responses and is thus one of the allergen-specific immunotherapy approaches currently being investigated for food allergy. Advances made in the understanding of how epicutaneously applied proteins interact with the immune system and in the technology for facilitating such interactions offer many opportunities for clinical application. Research has shown that allergen delivered to intact skin via EPIT is taken up in the superficial layers of the skin by Langerhans cells, avoiding passive movement of allergen through the dermis and limiting systemic circulation. EPIT brings about allergen desensitization by activating a population of regulatory T cells (Tregs) with unique properties and the potential for inducing a sustained effect as well as the possibility (seen in animal models) for protection against further sensitizations. Several clinical trials investigating the therapeutic efficacy of EPIT for treatment of peanut allergy have been completed, as well as a Phase 2 trial for treatment of milk allergy. Taken together, the reviewed literature supports the concept that EPIT activates the natural desensitization pathway of the skin, offering a progressive, possibly sustained response. EPIT offers a potential alternative for allergen immunotherapy that is less invasive and carries a lower risk for systemic reactions than oral immunotherapy.


Y65C missense mutation in the WW domain of the Golabi-Ito-Hall syndrome protein PQBP1 affects its binding activity and deregulates pre-mRNA splicing.
The PQBP1 (polyglutamine tract-binding protein 1) gene encodes a nuclear protein that regulates pre-mRNA splicing and transcription. Mutations in the PQBP1 gene were reported in several X chromosome-linked mental retardation disorders including Golabi-Ito-Hall syndrome. The missense mutation that causes this syndrome is unique among other PQBP1 mutations reported to date because it maps within a functional domain of PQBP1, known as the WW domain. The mutation substitutes tyrosine 65 with cysteine and is located within the conserved core of aromatic amino acids of the domain. We show here that the binding property of the Y65C-mutated WW domain and the full-length mutant protein toward its cognate proline-rich ligands was diminished. Furthermore, in Golabi-Ito-Hall-derived lymphoblasts we showed that the complex between PQBP1-Y65C and WBP11 (WW domain-binding protein 11) splicing factor was compromised. In these cells a substantial decrease in pre-mRNA splicing efficiency was detected. Our study points to the critical role of the WW domain in the function of the PQBP1 protein and provides an insight into the molecular mechanism that underlies the X chromosome-linked mental retardation entities classified globally as Renpenning syndrome.


Outcomes of complete vs targeted approaches to endoscopic sinus surgery.
Functional endoscopic sinus surgery (FESS) was historically predicated on targeted widening of narrow anatomic structures that caused postobstructive persistent sinus inflammation. It is now clear that chronic rhinosinusitis (CRS) is a multifactorial disease with subsets of patients which may require a more extensive surgical approach. This study compares quality-of-life (QOL) and disease severity outcomes after FESS based on the extent of surgical intervention. Participants with CRS were prospectively enrolled into an ongoing, multi-institutional, observational, cohort study. Surgical extent was determined by physician discretion. Participants undergoing bilateral frontal sinusotomy, ethmoidectomy, maxillary antrostomy, and sphenoidotomy were considered to have undergone "complete" surgery, whereas all other participants were categorized as receiving "targeted" surgery. Improvement was evaluated between surgical subgroups with at least 6-month follow-up using the 22-item Sino-Nasal Outcome Test (SNOT-22) and the Brief Smell Inventory Test (B-SIT). A total of 311 participants met inclusion criteria with 147 subjects undergoing complete surgery and 164 targeted surgery. A higher prevalence of asthma, acetylsalicylic acid (ASA) sensitivity, nasal polyposis, and a history of prior sinus surgery (p ≤ 0.002) was present in participants undergoing complete surgery. Mean improvement in SNOT-22 (28.1 ± 21.9 vs 21.9 ± 20.6; p = 0.011) and B-SIT (0.8 ± 3.1 vs 0.2 ± 2.4; p = 0.005) was greater in subjects undergoing complete surgery. Regression models demonstrated a 5.9 ± 2.5 greater relative mean improvement on SNOT-22 total scores with complete surgery over targeted approaches (p = 0.016). Complete surgery was an independent predictor of greater postoperative SNOT-22 score improvement, yet did not achieve clinical significance. Further study is needed to determine the optimal surgical extent.


Phenotype and function of T cells infiltrating visceral metastases from gastrointestinal cancers and melanoma: implications for adoptive cell transfer therapy.
Adoptive cell transfer of tumor-infiltrating lymphocytes (TILs) can mediate cancer regression in patients with metastatic melanoma, but whether this approach can be applied to common epithelial malignancies remains unclear. In this study, we compared the phenotype and function of TILs derived from liver and lung metastases from patients with gastrointestinal (GI) cancers (n = 14) or melanoma (n = 42). Fewer CD3(+) T cells were found to infiltrate GI compared with melanoma metastases, but the proportions of CD8(+) cells, T cell differentiation stage, and expression of costimulatory molecules were similar for both tumor types. Clinical-scale expansion up to ~50 × 10(9) T cells on average was obtained for all patients with GI cancer and melanoma. From GI tumors, however, TIL outgrowth in high-dose IL-2 yielded 22 ± 1.4% CD3(+)CD8(+) cells compared with 63 ± 2.4% from melanoma (p < 0.001). IFN-γ ELISA demonstrated MHC class I-mediated reactivity of TIL against autologous tumor in 5 of 7 GI cancer patients tested (9% of 188 distinct TIL cultures) and in 9 of 10 melanoma patients (43% of 246 distinct TIL cultures). In these assays, MHC class I-mediated up-regulation of CD137 (4-1BB) expression on CD8(+) cells suggested that 0-3% of TILs expanded from GI cancer metastases were tumor-reactive. This study implies that the main challenge to the development of TIL adoptive cell transfer for metastatic GI cancers may not be the in vitro expansion of bulk TILs, but the ability to select and enrich for tumor-reactive T cells.