Leflunomide derivative FK778 inhibits production of antibodies in an experimental model of alloreactive T-B cell interaction.
The contribution of humoral immune response in allograft and xenograft rejection has been clearly demonstrated in recent years. For this reason, inhibition of alloantibody production has become essential in managing transplanted patients. Here, we assessed the effects of the leflunomide derivative FK778 (FK778) in the control of antibody production resulting from semiallogeneic cognate T-B-cell interactions. BALB/c mice were tolerized at birth with semiallogeneic spleen cells from (BALB/c X C57BL/6) F1 mice, with or without overexpression of human bcl-2 transgene in B cells. These tolerized mice were treated with different dosages of FK778, either from birth, or from the third week of age, when autoantibody production was detected. The production of autoantibodies, used as markers of semiallogeneic cognate T-B - cell interactions, was evaluated at different time points during drug administration or after the interruption of treatment. FK778 treatment started at birth inhibited the production of semiallogeneic-driven antibodies in a dose-dependent manner. In addition, FK778 also reduced the levels of preformed circulating autoantibodies in adult mice, although the dosage required was 4 times higher than that used in neonates. However, the levels of IgG antibodies in these tolerized mice increased after FK778 withdrawal, indicating that FK778 failed to induce tolerance to semiallogeneic host CD4+ Th2 and/or donor B cells. Our results demonstrate the efficacy of FK778 in the control of antibody production resulting from semiallogeneic cognate T-B - cell interactions.


Ferrous Iron Up-regulation in Fibroblasts of Patients with Beta Propeller Protein-Associated Neurodegeneration (BPAN).
Mutations in WDR45 gene, coding for a beta-propeller protein, have been found in patients affected by Neurodegeneration with Brain Iron Accumulation, NBIA5 (also known as BPAN). BPAN is a movement disorder with Non Transferrin Bound Iron (NTBI) accumulation in the basal ganglia as common hallmark between NBIA classes (Hayflick et al., 2013). WDR45 has been predicted to have a role in autophagy, while the impairment of iron metabolism in the different NBIA subclasses has not currently been clarified. We found the up-regulation of the ferrous iron transporter (-)IRE/Divalent Metal Transporter1 and down-regulation of Transferrin receptor in the fibroblasts of two BPAN affected patients with splicing mutations 235+1G>A (BPAN1) and 517_519ΔVal 173 (BPAN2). The BPAN patients showed a concomitant increase of intracellular ferrous iron after starvation. An altered pattern of iron transporters with iron overload is highlighted in BPAN human fibroblasts, supporting for a role of DMT1 in NBIA. We here present a novel element, about iron accumulation, to the existing knowledge in field of NBIA. Attention is focused to a starvation-dependent iron overload, possibly accounting for iron accumulation in the basal ganglia. Further investigation could clarify iron regulation in BPAN.


CD38 and CD157 ectoenzymes mark cell subsets in the human corneal limbus.
Nicotinamide adenine dinucleotide (NAD(+)), a precursor of molecules involved in cell regulatory processes, is released in extra-cellular compartments after stress or inflammation.This study investigates the expression in the human cornea of CD38 and CD157, two NAD(+)-consuming ectoenzymes and surface receptors. The analysis in corneal epithelial and stromal cells was performed by means of multiple approaches, which included immunofluorescence, reverse transcriptase polymerase chain reaction (RT-PCR), Western blot, and confocal microscopy. The presence of enzymatically active NAD(+)-consumers in intact corneal cells was analyzed by high performance liquid chromatography (HPLC)-based assays. The results obtained show that CD38 and CD157 are expressed constitutively by corneal cells: CD38 appears as a 45-kDa monomer, while CD157 is a 42- to 45-kDa doublet. The molecules are enzymatically active, with features reminiscent of those observed in human leukocytes. CD38 is expressed by cells of the suprabasal limbal epithelium, whereas it is not detectable in central corneal epithelium and stroma. CD157 is expressed by basal limbal clusters, a p63(+)/cytokeratin 19(+) cell subset reported to contain corneal stem cells, and by stromal cells. The results of the work indicates that the human cornea is equipped with molecular tools capable of consuming extracellular NAD(+), and that CD157 is a potential marker of corneal limbal cells in the stem cell niche. The presence and characteristics of these ectoenzymes may be exploited to design drugs for wound repair or for applications in tissue transplantation.


Receptor-mediated regulation of neuropeptide gene expression in astrocytes.
One of the functions of glial receptors is to regulate synthesis and release of a variety of neuropeptides and growth factor peptides, which in turn act on neurons or other glia. Because of the potential importance of these interactions in injured brain, we have examined the role of two different receptors in the regulation of astrocyte neuropeptide synthesis. Stimulation of beta-adrenergic receptors on type 1 astrocytes resulted in increased mRNA and protein for the proenkephalin (PE) and somatostatin genes. This receptor also increased expression of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF). The potential role of opiate receptors was examined in several ways. Treatment of newborn rats for 7 days with the opiate antagonist naltrexone, prior to preparation of astrocytes, had no effect on PE mRNA or met-enkephalin content but resulted in a significant increase in NGF content. However, treatment of astrocytes in culture with met-enkephalin, morphine, or naltrexone had no effect on any of these parameters. No opiate binding could be detected, using either etorphine or bremazocine, to membranes of astrocytes prepared from cortex, cerebellum, striatum, or hippocampus of 1-day, 7-day, or 14-day postnatal rats. Thus we conclude that type 1 astrocytes do not express opiate receptors and that the in vivo effects of naltrexone are mediated indirectly via some other cell type/receptor.


Disseminated mucormycosis due to Cunninghamella bertholletiae in a liver transplant recipient.
Disseminated mucormycosis occurred in a 19 year old female following orthotopic liver transplantation for fulminant Wilson's disease. The causative organism Cunninghamella bertholletiae has previously been described in ten clinical cases, but never before in this setting.


TRP Channels as Sensors of Chemically-Induced Changes in Cell Membrane Mechanical Properties.
Transient Receptor Potential ion channels (TRPs) have been described as polymodal sensors, being responsible for transducing a wide variety of stimuli, and being involved in sensory functions such as chemosensation, thermosensation, mechanosensation, and photosensation. Mechanical and chemical stresses exerted on the membrane can be transduced by specialized proteins into meaningful intracellular biochemical signaling, resulting in physiological changes. Of particular interest are compounds that can change the local physical properties of the membrane, thereby affecting nearby proteins, such as TRP channels, which are highly sensitive to the membrane environment. In this review, we provide an overview of the current knowledge of TRP channel activation as a result of changes in the membrane properties induced by amphipathic structural lipidic components such as cholesterol and diacylglycerol, and by exogenous amphipathic bacterial endotoxins.


Syntactic Patterns Improve Information Extraction for Medical Search.
Medical professionals search the published literature by specifying the type of patients, the medical intervention(s) and the outcome measure(s) of interest. In this paper we demonstrate how features encoding syntactic patterns improve the performance of state-of-the-art sequence tagging models (both linear and neural) for information extraction of these medically relevant categories. We present an analysis of the type of patterns exploited, and the semantic space induced for these, i.e., the distributed representations learned for identified multi-token patterns. We show that these learned representations differ substantially from those of the constituent unigrams, suggesting that the patterns capture contextual information that is otherwise lost.


[Beta blockers and corneal sensitivity].
We performed several placebo-controlled, randomized, prospective, parallel, double-masked studies in ophthalmologically healthy people to determine the local anesthetic effect and the topical subjective tolerability of beta blockers with similar potency of action that are available in the Federal Republic of Germany. There are marked differences in the local anesthetic effects of the different beta blockers. Depending on the type of beta blockers the local anesthetic effect ranges from practically none up to anesthesia allowing contact glass examinations and removal of foreign bodies after application of only one drop. The subjective tolerability becomes worse with increasing strength of local anesthetic effect of the beta-blocker. There is a statistically significant correlation between the subjective tolerability and local anesthetic effect and the octanol-water distribution coefficient. Because of the local anesthetic effect, we recommend repeated checks of corneal sensitivity during beta blocker therapy. This should be done more frequently in people with sensitive eyes, "responders," contact lens wearers, and people with reduced corneal sensitivity for other reasons, especially during therapy with beta-blockers with marked local anesthetic effects. In addition to systemic side effects, cardioselectivity, influence on tear film, intrinsic sympathomimetic activity and topical tolerance, etc., the local anesthetic effect of beta blockers is a further important aspect in the "differential indications" of beta-blockers.


Cardiovascular prognosis in patients admitted to an emergency department with hypertensive emergencies and urgencies.
At present, few data are available on the prognosis of hypertensive emergencies and urgencies admitted to emergency departments. The aim of our study was to evaluate the incidence of total and cardiovascular events during follow-up in hypertensive patients admitted to the emergency departments of Brescia Hospital (Northern Italy) with hypertensive emergencies or urgencies from 1 January to 31 December 2015. Medical records of patients aged more than 18 years, admitted to the emergency department with SBP values at least 180 mmHg (SBP) and/or DBP values at least 120 mmHg (DBP) were collected and analysed (18% of patients were classified as 'hypertensive emergency' and 82% as 'hypertensive urgency'). Data in 895 patients (385 men and 510 women, mean age 70. 5 ± 15 years) were analysed; the mean duration of follow-up after admission to the emergency department was 12 ± 5 months. During the follow-up, 96 cardiovascular events (28 fatal) occurred (20 cardiac events, 30 cerebrovascular events, 26 hospital admission for heart failure, 20 cases of new onset kidney disease). In 40 patients (4.5%), a new episode of acute blood pressure rise with referral to the emergency department was recorded. Cardiovascular mortality and morbidity were greater in patients with a previous hypertensive emergency (14.5 vs. 4.5% in patients with hypertensive emergency and urgency, respectively, chi-square, P < 0.0001). Similar results were obtained when the occurrence of cerebrovascular or renal events were considered separately. Admission to the emergency department for hypertensive emergencies and urgencies identifies hypertensive patients at increased risk for fatal and nonfatal cardiovascular events. Our findings add some new finding suggesting that further research in this field should be improved aiming to define, prevent, treat and follow hypertensive urgencies and emergencies.


Talent identification and selection process of outfield players and goalkeepers in a professional soccer club.
Abstract The aim of this study was to analyse the talent identification process of a professional soccer club. A preselection of players (n = 64) aged 9-10 years and a final selection (n = 21) were performed by the technical staff through the observation during training sessions and matches. Also, 34 age-matched players of an open soccer camp (CampP) acted as controls. All participants underwent anthropometric, maturity and performance measurements. Preselected outfield players (OFs) were older and leaner than CampP (P < 0.05). Besides, they performed better in velocity, agility, endurance and jump tests (P < 0.05). A discriminant analysis showed that velocity and agility were the most important parameters. Finally, selected OFs were older and displayed better agility and endurance compared to the nonselected OFs (P < 0.05). Goalkeepers (GKs) were taller and heavier and had more body fat than OFs; also, they performed worse in the physical tests (P < 0.05). Finally, selected GKs were older and taller, had a higher predicted height and advanced maturity and performed better in the handgrip (dynamometry) and jump tests (P < 0.05). Thus, the technical staff selected OFs with a particular anthropometry and best performance, particularly agility and endurance, while GKs had a different profile. Moreover, chronological age had an important role in the whole selection process.


Abscisic acid - an anti-angiogenic phytohormone that modulates the phenotypical plasticity of endothelial cells and macrophages.
Abscisic acid (ABA) has shown anti-inflammatory and immunoregulatory properties in preclinical models of diabetes and inflammation. Herein, we studied the effects of ABA on angiogenesis, a strictly controlled process that, when dysregulated, leads to severe angiogenic disorders including vascular overgrowth, exudation, cellular inflammation and organ dysfunction. By using a 3D sprouting assay, we show that ABA effectively inhibits migration, growth and expansion of endothelial tubes without affecting cell viability. Analyses of the retinal vasculature in developing normoxic and hyperoxic mice challenged by oxygen toxicity reveal that exogenously administered ABA stunts the development and regeneration of blood vessels. In these models, ABA downregulates endothelial cell (EC)-specific growth and migratory genes, interferes with tip and stalk cell specification, and hinders the function of filopodial protrusions required for precise guidance of vascular sprouts. In addition, ABA skews macrophage polarization towards the M1 phenotype characterized by anti-angiogenic marker expression. In accordance with this, ABA treatment accelerates macrophage-induced programmed regression of fetal blood vessels. These findings reveal protective functions of ABA against neovascular growth through modulation of EC and macrophage plasticity, suggesting the potential utility of ABA as a treatment in vasoproliferative diseases.


Burden on family members of the mentally ill: a naturalistic study in Japan.
People with mental disorders often cause distress among their family members. We examined a total of 25 pairs of newly referred psychiatric patients and their family members to investigate the correlations between family burden and patient diagnosis (using the Structured Clinical Interview for DSM-III-R [SCID] axis I disorders), symptomatic severity (Positive and Negative Symptoms Scales [PANSS]), global function (Global Assessment of Functioning [GAF]), and the general level of family function (Family Adaptability and Cohesion Evaluation Scale [FACES]). The subjective and objective burdens on the family were assessed by self-report. The subjective and objective burdens were significantly predicted only by the GAF score.


A novel glucosylation enzyme: molecular cloning, expression, and characterization of Trichoderma viride JCM22452 alpha-amylase and enzymatic synthesis of some flavonoid monoglucosides and oligoglucosides.
It was found that commercial cellulase preparations from Trichoderma viride showed transglucosylation activity toward (+)-catechin and (-)-epigallocatechin gallate (EGCG) using dextrin as a glucosyl donor. To isolate the enzyme exhibiting transglucosylation activity toward (+)-catechin and EGCG, the present study isolated the cDNA encoding the T. viride JCM22452 alpha-amylase homologue (TRa2), which showed high amino acid sequence identity to functionally uncharacterized alpha-amylase homologues from other ascomycetes, which also produced some (+)-catechin and EGCG glucosides. TRa2 was able to glucosylate a wide range of natural flavonoids, particularly (+)-catechin and EGCG, and to hydrolyze maltooligosaccharides (k(cat)/K(m) for maltotriose, maltotetraose, maltopentaose, maltohexaose, and maltoheptaose were 1.98, 45.2, 58.3, 97.4, and 92.6 s(-1) mM(-1), respectively) except maltose but could not transfer the monoglucosyl residue to maltooligosaccharides. By (1)H NMR and (13)C NMR, the structures of several novel glucosides obtained by commercial cellulase preparations from T. viride and TRa2 were determined as (+)-catechin 5-O-alpha-D-glucopyranoside, (+)-catechin 5-O-alpha-D-maltoside, (+)-catechin 4'-O-alpha-D-maltoside, EGCG 5-O-alpha-D-glucopyranoside, and EGCG 7-O-alpha-D-maltoside. One of these glucosides, EGCG 5-O-alpha-D-glucopyranoside, showed higher heat stability and solubility and lower astringency and astringent stimulation than its aglycon, suggesting that EGCG glucosides are functionally superior to EGCG as food additives.


Fast and accurate measurement of the polarization-dependent detection efficiency of superconducting nanowire single photon detectors.
Superconducting nanowire single photon detectors (SNSPDs) have been extensively investigated due to their superior characteristics, including high system detection efficiency, low dark count rate and short recovery time. The polarization sensitivity introduced by the meandering-type superconductor nanowires is an intrinsic property of SNSPD, which is normally measured by sweeping hundreds of points on the Poincaré sphere to overcome the unknown birefringent problem of the SNSPD's delivery fiber. In this paper, we propose an alternative method to characterize the optical absorptance of SNSPDs, without sweeping hundreds of points on the Poincaré sphere. It is shown theoretically that measurements on the system detection efficiencies (SDEs) subject to cases of four specific photon polarization states are sufficient to reveal the two eigen-absorptances of the SNSPD. We validate the proposed method by comparing the measured detection spectra with the spectra attained from sweeping points on the Poincaré sphere and the simulated absorption spectra.


Purification of native Arp2/3 complex from bovine thymus.
The Arp2/3 complex is an actin filament nucleator involved in cell motility and vesicle trafficking. Owing to the role the complex plays in important and fundamental cell biological processes, the purified complex is used in biochemical assays, reconstituted motility assays, and structural biology. As this is a eukaryotic complex assembled from seven polypeptides, the complex is purified from eukaryotic sources. Described here is a detailed method for purification of the complex from a mammalian tissue, bovine thymus.


Variation in Fungicide Sensitivity Among Rhizoctonia Isolates Recovered from Potatoes in South Africa.
Rhizoctonia is a major pathogen of potato causing substantial yield losses worldwide. Control of Rhizoctonia diseases is based predominantly on the application of fungicides. However, little is known about the fungicide response variability of different Rhizoctonia anastomosis groups associated with potato diseases in South Africa. A total of 131 Rhizoctonia isolates were obtained from potato growing regions of South Africa from 2012 to 2014 and evaluated for sensitivity to fungicides in vitro and in vivo. The fungicides comprised six chemical formulations and one bio-fungicide representing seven Fungicide Resistance Action Committee groups. All Rhizoctonia anastomosis groups were sensitive to tolclofos-methyl (EC50: 0.001 to 0.098 μg a.i. ml-1) and fludioxonil (EC50: 0.06 to 0.09 μg a.i. ml-1) and showed variation in sensitivity to pencycuron, iprodione, benomyl, and Bacillus subtilis QST 713. However, for azoxystrobin, Rhizoctonia isolates exhibited variable sensitivity ranging from sensitivity (EC50: <0.09 μg a.i. ml-1) to insensitivity with EC50 values exceeding 5 μg a.i. ml-1. In greenhouse and field trials, tolclofos-methyl and fludioxonil exhibited significantly greater control of stem and black scurf whereas azoxystrobin was the least effective. This work demonstrated variable sensitivity within and among anastomosis groups of R. solani and binucleate Rhizoctonia to different fungicides. Information on fungicide sensitivity of Rhizoctonia isolates is crucial in the development of effective Rhizoctonia control strategies and facilitates monitoring of fungicide insensitive isolates in the pathogen population.


Molecular analysis of xenograft models of human cancer cachexia--possibilities for therapeutic intervention.
Approximately 50% of all cancer patients develop cachexia, a paraneoplastic syndrome that is characterized by wasting of adipose tissue and skeletal muscle mass. Cytokines, including TNF-alpha, interleukins-1, -6, and interferon-A are known mediators of the cachectic process. The latter however represent only one of many imbalanced systems in cancer cachexia. The aim of this study was to further delineate the pathogenesis of cachexia by molecular profiling. Human renal cancer xenografts that do and do not induce cachexia in mice were used as disease models. Cachexia-associated gene expression was studied on Human Genome U95 Affymetrix arrays and revealed several new genes such as TNF-alpha ligand superfamily protein, interferon-A treatment inducible protein, and DKFZ5641I1922. The expression of the IL-8 gene was also elevated in cachexia inducing xenografts (CIX). At the protein level, TNF-alpha was found expressed only in CIX, whereas IL-1 and IL-6 were not cachexia specific. Levels of parathyroid hormone-related protein were elevated in CIX and accompanied by hypercalcemia. COX-2 and prostaglandin E2 were also found to be over expressed. By using the COX-2 inhibitors rofecoxib and nimesulide, we were able to delay tumor-mediated wasting in vivo. Overall, our results suggest that cachexia is a multigenetic disease that will require complex combinations of drugs for an effective therapeutic intervention.


RNA Sequencing Reveals the Regulation Mechanism of Yunnan Baiyao in Treating Skin Infection Caused by Staphylococcus aureus.
Yunnan Baiyao is a well-known traditional Chinese medicine that can be formulated into a powder or capsule form. The mechanism by which it exerts its anti-inflammation effect, which is used in skin care products, needs to be further explored. In this study, we established the Staphylococcus aureus-induced mouse skin inflammatory model to investigate the effects of Yunnan Baiyao by the method of RNA-sequencing technology. The mice were randomly assigned to three groups, and those were control, model, and the Yunnan Baiyao-treated (YNtreated) group. Key genes and pathways were identified using bioinformatics analyses. In the study, we obtained 1,053 differentially expressed genes (DEGs) induced by Yunnan Baiyao. The 233 upregulated genes were enriched in 32 GO terms and 5 KEGG pathways, focused on the items, such as wound healing, cell metabolism, and proliferation, indicating the accelerating effects of Yunnan Baiyao on these aspects. The 820 downregulated genes were enriched mainly in the items, including the regulation of inflammation factor production, immune responses, and regulation of structure dermal components. Besides, Yunnan Baiyao reversed the expressions of 277 (201 decreased and 76 increased DEGs, respectively) induced by S. aureus. Ten key regulatory nodes (MMP2, PLK1, CCNB1, TLR4, CDK1, CCNA2, CDC25C, PDGFRA, MYOC, and KNG1) were identified by the construction of the protein interaction network, half of which were related to cell proliferation. VAV1 was another hub node that was affected by Yunnan Baiyao (Top 20). In the study, VAV1 and TLR4 can be considered key module genes in inflammation regulation. In conclusion, this study found that Yunnan Baiyao can significantly relieve inflammatory symptoms by regulating genes and pathways involved in the regulation of inflammation and immune response and also helped to deepen our understanding of the associated molecular mechanisms.


Neutrophil function in elderly persons assessed by flow cytometry.
It is well known that the immune response declines with senescence and it is suggested that these changes render an individual susceptible to infection, autoimmune phenomena and cancer. Bacterial and viral infections are a major cause of illness and death amongst aged subjects, and once infection is established, the elderly also have a diminished capacity to prevent its spread (1). The cellular and molecular basis for this age-related decline in immunocompetence are still unknown and, possibly, are related to an alteration in cell transduction mechanisms (2).


T-Cell Dynamics in Chronic Lymphocytic Leukemia under Different Treatment Modalities.
Using next-generation sequencing (NGS), we recently documented T-cell oligoclonality in treatment-naïve chronic lymphocytic leukemia (CLL), with evidence indicating T-cell selection by restricted antigens. Here, we sought to comprehensively assess T-cell repertoire changes during treatment in relation to (i) treatment type [fludarabine-cyclophosphamide-rituximab (FCR) versus ibrutinib (IB) versus rituximab-idelalisib (R-ID)], and (ii) clinical response, by combining NGS immunoprofiling, flow cytometry, and functional bioassays. T-cell clonality significantly increased at (i) 3 months in the FCR and R-ID treatment groups, and (ii) over deepening clinical response in the R-ID group, with a similar trend detected in the IB group. Notably, in constrast to FCR that induced T-cell repertoire reconstitution, B-cell receptor signaling inhibitors (BcRi) preserved pretreatment clones. Extensive comparisons both within CLL as well as against T-cell receptor sequence databases showed little similarity with other entities, but instead revealed major clonotypes shared exclusively by patients with CLL, alluding to selection by conserved CLL-associated antigens. We then evaluated the functional effect of treatments on T cells and found that (i) R-ID upregulated the expression of activation markers in effector memory T cells, and (ii) both BcRi improved antitumor T-cell immune synapse formation, in marked contrast to FCR. Taken together, our NGS immunoprofiling data suggest that BcRi retain T-cell clones that may have developed against CLL-associated antigens. Phenotypic and immune synapse bioassays support a concurrent restoration of functionality, mostly evident for R-ID, arguably contributing to clinical response.


Importance of the urinary excretion in the total elimination of SC4453 in the guinea-pig.
Administration of SC4453 (30 micrograms/kg) in the guinea-pig increases the rate of urinary excretion from 35 ml/kg/24 h to 55 ml/kg/24 h. Ligature of ureters modifies parameters of SC4453 ; t 1/2 beta increases from 4.2 h to 5.6 h and the body clearance diminishes from 622 ml/h/kg (normal guinea-pig) to 530 ml/h/kg in the ligatured ureters guinea-pig. Analysis of the radioactive material excreted in the urine after administration of tritiated SC4453 shows that 58.4% is dichloromethane (DCM) soluble and 41.6% is water soluble. Thin layer chromatography (TLC) of the DCM soluble fraction shows that 74.0% is unchanged SC4453 . After enzymatic hydrolysis of the water soluble fraction, it is shown that 28% is again extractable by DCM of which 57% consists of unchanged SC4453 which is therefore the main partner of conjugated products. Following an i.v. injection of tritiated SC4453 , only 17% of the dose was recovered in urine, against 14% after an intragastric administration. Gastro-intestinal absorption of SC4453 was estimated to be 82.2%.


Cholesterol ester transfer mediated by lipid transfer protein as influenced by changes in the charge characteristics of plasma lipoproteins.
The relationship between the cholesterol ester (CE) transfer activity of lipid transfer protein (LTP) and its affinity with lipid and lipoprotein particles was investigated. The study of the effects of chemical modification of low density lipoprotein (LDL) amino groups and carboxyl groups on the CE transfer activity showed that the maximal activity is obtained upon succinylation or acetylation of approximately 7% of LDL amino groups. Further increases in the extent of modification progressively reduced the transfer activity. The treatment of LDL with fatty acids gave results comparable to the chemical modification of LDL amino groups. The addition of low concentrations of fatty acids was stimulatory, while that of high concentrations was inhibitory. Although increases in the positive charges of LDL by the carboxyl group modification did not appreciably influence the CE transfer, the addition of cationic detergents gave a profound effect on the CE transfer. A maximal CE transfer activity was obtained upon addition of very small amounts of the detergents, with the higher concentrations sharply reducing the transfer activity. We also studied the effects of the concentrations of phosphate buffer and various salts on the CE transfer as well as the affinity of LTP for very low density lipoproteins, low density lipoproteins, high density lipoproteins 3, and high density lipoproteins 2. It appeared that the affinity of LTP for various lipoproteins is governed by a delicate balance of electrostatic and hydrophobic interactions. Optimal degrees of the interaction of LTP with both donor and acceptor particles seem to be required for the maximal degree of CE transfer.


Generation and Characterization of Virus-Enhancing Peptide Nanofibrils Functionalized with Fluorescent Labels.
Retroviral gene transfer is the method of choice for the stable introduction of genetic material into the cellular genome. However, efficient gene transfer is often limited by low transduction rates of the viral vectors. We have recently described a 12-mer peptide, termed EF-C, that forms amyloid-like peptide nanofibrils (PNF), strongly increasing viral transduction efficiencies. These nanofibrils are polycationic and bind negatively charged membranes of virions and cells, thereby overcoming charge repulsions and resulting in increased rates of virion attachment and gene transfer. EF-C PNF enhance vector transduction more efficiently than other soluble additives and offer prospects for clinical applications. However, while the transduction-enhancing activity of PNF has been well-characterized, the exact mechanism and the kinetics underlying infection enhancement as well as the cellular fate of the fibrils are hardly explored. This is partially due to the fact that current labeling techniques for PNF rely on amyloid probes that cause high background staining or lose signal intensities after cellular uptake. Here, we sought to generate EF-C PNF covalently coupled with fluorescent labels. To achieve such covalent bioconjugates, the free amino groups of the EF-C peptide were coupled to the ATTO 495 or 647N NHS ester dyes. When small amounts of the labeled peptides were mixed with a 100- to 10 000-fold excess of the native peptide, PNF formed that were morphologically indistinguishable from those derived from the unlabeled peptide. The fluorescence of the fibrils could be readily detected using fluorescence spectroscopy, microscopy, and flow cytometry. In addition, labeled and nonlabeled fibrils captured viral particles and increased retroviral transduction with similar efficacy. These covalently fluorescence-labeled PNF are valuable tools with which to elucidate the mechanism(s) underlying transduction attachment and the fate of the fibrils in cells, tissues, and animal models.


Neural-Network-Based Robust Optimal Tracking Control for MIMO Discrete-Time Systems With Unknown Uncertainty Using Adaptive Critic Design.
This paper is concerned with the robust optimal tracking control strategy for a class of nonlinear multi-input multi-output discrete-time systems with unknown uncertainty via adaptive critic design (ACD) scheme. The main purpose is to establish an adaptive actor-critic control method, so that the cost function in the procedure of dealing with uncertainty is minimum and the closed-loop system is stable. Based on the neural network approximator, an action network is applied to generate the optimal control signal and a critic network is used to approximate the cost function, respectively. In contrast to the previous methods, the main features of this paper are: 1) the ACD scheme is integrated into the controllers to cope with the uncertainty and 2) a novel cost function, which is not in quadric form, is proposed so that the total cost in the design procedure is reduced. It is proved that the optimal control signals and the tracking errors are uniformly ultimately bounded even when the uncertainty exists. Finally, a numerical simulation is developed to show the effectiveness of the present approach.


Food group consumption in an Italian population using the updated food classification system FoodEx2: Results from the Italian Nutrition & HEalth Survey (INHES) study.
Dietary habits evolve over time, being influenced by many factors and complex interactions. This work aimed at evaluating the updated information on food group consumption in Italy. A total of 8944 (4768 women and 4176 men) participants aged >18 years from all over Italy recruited in 2010-13 (Italian Nutrition & HEalth Survey, INHES) was analyzed. The recruitment was performed using computer-assisted-telephone-interviewing and one-day 24-h dietary recall retrieved from all participants. The updated, second version, of FoodEx2 food classification system was applied to extract data on food group consumption. The participation rate was 53%; 6.2% of the participants declared to follow a special diet, the most prevalent being hypo-caloric diets (55.7% of special diets). Men compared to women presented significantly higher intakes of "grains and grain-based products", "meat and meat products", "animal and vegetable fats and oils and primary derivatives" and "alcoholic beverages" (P for all<0.001); moreover, men had lower intakes of "milk and dairy products", "water and water-based beverages" and "products for non-standard diets, food imitates and food supplements" (P for all<0.001). Differences in food group intake among age groups, geographical regions and educational level groups were also identified (P for all<0.05). Data on the consumption of more than 70 food groups and sub-groups were illustrated in different strata. The present analysis could be considered as an updated source of information for future nutrition research in Italy and in the EU.


Significance of TERT and ATRX mutations in glioma.
Mutations of telomerase reverse transcriptase (TERT) and the α thalassemia/mental retardation syndrome X-linked (ATRX) genes have been the subject of numerous studies on the classification and prognosis of glioma. However, the association between TERT and ATRX in World Health Organization (WHO) grade II to IV glioma remains unclear. The present study utilized Sanger sequencing and immunohistochemical methods to detect the expression of the TERT promoter region, ATRX mutations and proliferation marker protein Ki-67 (Ki-67) protein expression in 179 cases of glioma. The current study analyzed these variables and their association with clinicopathological characteristics to further basic research and provide a theoretical basis for the clinical diagnosis and treatment of this type of tumor (1). The results demonstrated that TERT promoter mutations were negatively associated with ATRX. Additionally, Ki-67 protein expression in TERT wild-type samples was higher compared with samples with ATRX deletion. Overall, the results demonstrated, for the first time to the best of the authors' knowledge, that TERT promoter mutations are negatively associated with ATRX expression in WHO grade II to IV gliomas. These findings provide a theoretical basis for further basic research and may improve clinical diagnosis and treatment of glioma in the future.


Modulating the proliferative and cytotoxic properties of patient-derived TIL by a synthetic immune niche of immobilized CCL21 and ICAM1.
A major challenge in developing an effective adoptive cancer immunotherapy is the ex-vivo generation of tumor-reactive cells in sufficient numbers and with enhanced cytotoxic potential. It was recently demonstrated that culturing of activated murine CD8+ T-cells on a "Synthetic Immune Niche" (SIN), consisting of immobilized CCL21 and ICAM-1, enhances T-cell expansion, increases their cytotoxicity against cultured cancer cells and suppresses tumor growth in vivo. In the study reported here, we have tested the effect of the CCL21+ICAM1 SIN, on the expansion and cytotoxic phenotype of Tumor Infiltrating Lymphocytes (TIL) from melanoma patients, following activation with immobilized anti-CD3/CD28 stimulation, or commercial activation beads. The majority of TIL tested, displayed higher expansion when cultured on the coated SIN compared to cells incubated on uncoated substrate and a lower frequency of TIM-3+CD8+ cells after stimulation with anti-CD3/CD28 beads. Comparable enhancement of TIL proliferation was obtained by the CCL21+ICAM1 SIN, in a clinical setting that included a 14-day rapid expansion procedure (REP). Co-incubation of post-REP TIL with matching target cancerous cells demonstrated increased IFNγ secretion beyond baseline in most of the TIL cultures, as well as a significant increase in granzyme B levels following activation on SIN. The SIN did not significantly alter the relative frequency of CD8/CD4 populations, as well as the expression of CD28, CD25, several exhaustion markers and the differentiation status of the expanded cells. These results demonstrate the potential capacity of the CCL21+ICAM1 SIN to reinforce TIL-based immunotherapy for cancer patients.


Pyogenic granulomas after silicone punctal plugs: a clinical and histopathologic study.
To describe clinical findings, histopathologic changes, and risk factors for pyogenic granuloma formation complicating silicone punctal plug therapy. Retrospective observational case series. Between November 2000 and April 2004, 903 silicone punctal plugs of the same brand were inserted in 404 subjects. Cases associated with pyogenic granuloma formation were identified and reviewed. Granulation tissue was obtained from 10 patients for histopathologic examination. Multiple risk regression analyses identified factors related to pyogenic granuloma development and factors associated with histologic patterns. Pyogenic granuloma development led to the extrusion of 4.2% of all plugs placed in a median time period of 141 days. All patients presented with varying degrees of plug extrusion. Similar distributions of partial and complete plug extrusions, as well as bilateral and unilateral plug extrusions, were seen. Findings at presentation ranged from a subclinical pyogenic granuloma causing partial plug extrusion to a pyogenic granuloma in the punctum with a ring of fibrovascular tissue retaining a completely extruded plug. Histopathologic examination revealed two patterns, representing either acute pyogenic granuloma or involuting pyogenic granuloma. Pyogenic granulomas resolved after 3.1 +/- 1.3 weeks in all patients after plug removal. Multiple regression analysis revealed that large plug size was associated with increased pyogenic granuloma formation (P < .0001). Partial or complete plug extrusion was associated with active or involuting pyogenic granuloma, respectively (P = .023). Pyogenic granuloma-related spontaneous plug extrusions may be more common than previously thought and can present with a range of clinical findings. The degree of plug extrusion correlates with the histopathologic pattern. Larger plug size and sharp edges in plug geometry may be responsible for pyogenic granuloma formation.


Characterization of polyclonal autoantibodies specific for beta 2-microglobulin in multiple myeloma sera.
Among 185 sera from 62 patients with multiple myeloma, two serum samples with high beta 2-microglobulin (beta 2m) binding activity (S beta 2m-BA) were investigated. The S beta 2m-BA was shown to be distinct from the monoclonal component and to be represented by autoantibody of the IgG class. These antibodies were specific for beta 2m. They formed macromolecular complexes with beta 2m, indicating that at least two distinct epitopes of beta 2m can be recognized by these antibody molecules. The association-dissociation constants and antigen binding capacities of these autoantibodies were compared with that of monoclonal or polyclonal heterologous antibodies.


Gowning does not affect colonization or infection rates in a neonatal intensive care unit.
To study the effect of gowning in a neonatal intensive care unit on colonization patterns, necrotizing enterocolitis, respiratory syncytial virus and other infections, mortality, and traffic and handwashing patterns. Alternate 2-month gowning and no-gowning cycles were established in a 24-bed level III neonatal intensive care unit for 8 months, with respiratory site, umbilical, and stool surveillance cultures done weekly on all patients. Traffic flow and handwashing compliance were evaluated by direct observation. Demographic data did not differ between periods. There were no significant differences between the gowning and no-gowning periods in the rates of bacterial colonization, any type of infection, or mortality. There was no effect on traffic flow or handwashing compliance. Gowning in the neonatal intensive care unit is an unnecessary custom without benefit in neonatal colonization, infection rates, mortality, traffic patterns, and handwashing behavior.


Etiology and pathogenesis of Parkinson's disease.
Parkinson's disease (PD) is an age-related neurodegenerative disorder that affects approximately 1 million persons in the United States. It is characterized by resting tremor, rigidity, bradykinesia or slowness, gait disturbance, and postural instability. Pathological features include degeneration of dopaminergic neurons in the substantia nigra pars compacta coupled with intracytoplasmic inclusions known as Lewy bodies. Neurodegeneration and Lewy bodies can also be found in the locus ceruleus, nucleus basalis, hypothalamus, cerebral cortex, cranial nerve motor nuclei, and central and peripheral components of the autonomic nervous system. Current treatment consists of a dopamine replacement strategy using primarily the dopamine precursor levodopa. While levodopa provides benefit to virtually all PD patients, after 5-10 years of treatment the majority of patients develop adverse events in the form of dyskinesia (involuntary movements) and fluctuations in motor response. Further, disease progression is associated with the development of dementia, autonomic dysfunction, and postural instability, which do not respond to levodopa therapy. Accordingly, research efforts have been directed toward understanding the etiology and pathogenesis of PD in the hope of developing a more effective therapy that will slow or halt the natural progression of PD. This paper reviews recent advances.


Characterization of Pasteurella multocida isolated from rabbits in Canada.
In a survey for the somatic and capsular serotypes of Pasteurella multocida present in domestic rabbits in Canada, but mainly in Ontario, samples were obtained from research facilities, commercial rabbitries and from abattoir and necropsy specimens. Sources of isolates were upper respiratory tract infections, localized bronchopneumonias , acute fibrinous pneumonias, abscesses and otitis media. Of 59 isolates obtained, 47.0% were type 12:A, 30.5% 3:D and 12.0% were 3:A. Less common types were 12(4):A, 12:D, 4(12):A and 3:untypable. Somatic group 3 was most commonly isolated from acute pneumonic disease, while serogroup 12:A was most commonly found in upper respiratory tract infections and in localized chronic bronchopneumonia. Two serotypes of P. multocida were isolated from four pneumonic lungs collected from abattoir specimens. Most isolates were susceptible to the commonly used antibiotics.


[Scintigraphy with thallium chloride in a case of clear cell sarcoma of the tendons and aponeurosis].
Clear cell sarcoma of the soft parts (CCSSP) is a rare and highly malignant tumor. This is a case report of a 31 years old woman who presented with a tumor in the internal face of left thigh. The tumor biopsy was suggestive of a possible malignant tumor of the peripheral nerve sheath (malignant schwannoma). Biochemical analyses, computed tomography and magnetic resonance were performed and reported a sarcoma of soft parts (CCSSP) without abnormal inguinal lymph nodes. With this diagnostic suspicion, the patient was sent to the Nuclear Medicine Service of our center where a 201Thallium scintigraphy study was performed. In this study, it showed the primary tumor together with a hot spot in the homolateral inguinal region, suggestive of the presence of a metastastic lymph node. This finding would change the surgical attitude in the patient, including inguinal lymphadenectomy. The post-surgical histological study confirmed a clear cell sarcoma of soft parts (CCSSP) with a metastatic inguinal lymph node.


Umbilical arterial-venous blood gas difference: a novel expression of placental respiratory function.
We applied the concept of umbilical arterial and venous (A-V) blood gas difference in evaluating placental respiratory function to establish normative values for these relationships. Umbilical A-V cord blood gas samples were obtained in standardized fashion in uncomplicated singleton pregnancies. A-V pO (2) and A-V pCO (2) differences were related to obstetric factors, placental weight, and gestational age. Mean absolute differences in uncomplicated vaginal deliveries between umbilical artery and umbilical vein were: pO (2) 11.4 +/- 7.0 mm Hg and pCO (2) 11.9 +/- 6.8 mm Hg. Values in nonlabor cesarean section cases did not differ significantly from those of cases delivered vaginally. A-V pO (2) differences did not vary significantly across gestation (Pearson's R = 0.05, P = 0.43) and did not correlate with birth weight ( R = 0.06, P = 0.30). A-V pCO (2) differences showed increases with gestational age ( R = 0.15, P = 0.008) and with birth weight ( R = 0.18, P = 0.002). Results similar to those of women delivered vaginally were seen in nonlabor cesarean deliveries. Duration of labor did not impact the A-V blood gas relationship. Umbilical A-V oxygen differences reflect placental respiratory function independent of gestational age, birth weight, duration of labor, and mode of delivery. They are easy to collect and represent a potentially valuable resource for assessment of placental function.


Young children's interest in live animals.
Animals are important stimuli for humans, and for children in particular. In three experiments, we explored children's affinity for animals. In Experiment 1, 11- to 40-month-old children were presented with a free-play session in which they were encouraged to interact with several interesting toys and two live animals - a fish and a hamster. Experiment 2 used the same methodology with 18- to 36-month-old children and two additional animals - a snake and a spider - to examine whether children's behaviours would differ for benign and potentially threatening animals. Finally, in Experiment 3, a more controlled paired-preference paradigm was employed to assess 18- to 33-month-old children's interactions with three live animals - a fish, hamster, and gecko - versus three physically similar toy animals. Across all three experiments, children interacted with the animals more often than with the toys. Further, they behaved differently towards the animals than the toys, talking about the animals more than the toys and asking more questions about them. The parents of the children also spent more time interacting with the animals, directing their children's attention more towards the animals than the toys. This research supports the idea that humans have an affinity for animals that draws their attention to animals, even when attractive toys are present.


Inhibitors of peptidases: how they influence the biological activities of substance P, neurokinins, bradykinin and angiotensin in guinea pig, hamster and rat urinary bladders.
Neurokinins, bradykinin and angiotensins were tested in isolated urinary bladder of the guinea pig, the hamster and the rat, in the absence and in presence of a variety of peptidase inhibitors in order to establish if peptide degradation interferes with the bladder contractions elicited by the three types of peptides. Indeed, the effects of neurokinins, bradykinin and angiotensin I in the guinea pig bladder were significantly enhanced by captopril (4.6 x 10(-6) mol/l), chymostatin (1 mg/l), phosphoramidon (4.6 x 10(-6) mol/l) and thiorphan (1.0 x 10(-6) mol/l), while only captopril was found to potentiate the effects of the same peptides in the rat bladder. The four peptidase inhibitors, as well as bacitracin were found to modify the responses of the hamster urinary bladder to one or another or to all three groups of peptides and to DiMeC7. The present results suggest that the urinary bladders of various species have different types of active proteolytic enzymes: only the angiotensin-converting enzyme appears to be present in the rat bladder, while the same enzyme and possibly two additional endopeptidases interfere with the myotropic effects of neurokinins, kinins and angiotensins in the guinea pig and the hamster bladder.


Comparative nutrient analysis of four species of cockroaches used as food for insectivores by life stage, species, and sex.
A variety of insects are fed to insectivorous animals; however, nutritional analyses are often limited to adult life stages. Four species of nymph and adult female and male cockroaches (Blaberus giganteus, Blaptica dubia, Blatta lateralis, and Gromphadorhina portentosa) were analyzed for moisture, crude protein (CP), acid detergent fiber (ADF), fat, ash, and mineral content. The small sample size of this study precluded statistical analyses, however comparatively, CP in adult B. lateralis and B. dubia was greater than the CP in nymphs of the same species. Adult and nymph B. dubia had the greatest CP (96.6% and 65.3%, respectively) compared to the other three species. Inversely, fat content in adult B. lateralis and B. dubia was lower than that of nymphs of the same species. All adults contained similar levels of ADF, yet adult B. giganteus had greater ADF than nymphs; conversely, B. lateralis and B. dubia adults had less ADF than nymphs. There were differences noted in mineral parameters among the four species and life stages. Adult B. giganteus had less Ca than G. portentosa, and the lowest Ca content of the four species of nymphs and adults analyzed. This study underscores the differences in nutrient content with respect to life stage and species in previously understudied cockroaches to improve nutrient intake in captive insectivores.


The effect of dietary and topical celecoxib on 4-nitroquinoline-1-oxide-induced lingual epithelium alternations in rat.
To compare the preventive effect of dietary and topical Celecoxib (CCB), a potent inhibitor of COX-2 on 4-NQO-induced tongue SCC in rat. Fifty male Sprague Dawley adult 3- 3.5 months old rats were used as animal model in this study. The tongue SCC was induced by a daily administration of 30 ppm 4-NQO, in drinking water, for 8 months. The rats in case groups received dietary or topical CCB. Tongue Specimens were prepared for histopathological and immunohistochemical (Ki-67) staining and or TUNEL assay were examined. Values are expressed as mean +/- SEM and analyzed with Npar Kruscal Wallis and one-way ANOVA tests. p < 0.05 was considered significant. The incidence of tongue precancer lesions, judged by morphological and morhometrical criteria and apoptosis/proliferation ratio, was significantly (p < 0.01) reduced by CCB. The effect of topical CCB use, at high doses, was comparable to the effect of dietary CCB. Both topical and dietary CCB have inhibitory effect on 4-NQO induced SCC on tongue. The effect of CCB is probably mediated by suppression of cell proliferation and induction of apoptosis.


[Comparison of antibacterial potencies of oral and parenteral antibiotic preparations against Escherichia coli, Klebsiella, Citrobacter, and Proteus isolated from urinary tract infections (3: 1981). 2. Changes in bacterial sensitivities].
Since 1979 the antibacterial activity of antibiotics against E. coli, Klebsiella, Citrobacter and Proteus isolated from patients with urinary tract infections has been investigated. The serious transition of susceptibilities of E. coli and Klebsiella could not be recognized in these antibiotics (MPC, ABPC, NA, PPA, CEX, CEZ, CTM, CMZ and CFX). However, a few resistant organisms against the third generation's antibiotics (CTX, CMX, CZX, LMOX and CPZ) have already been appeared, we have to observe these results, continuously.


The role of the mesangium in glomerular function.
When discussing glomerular function, one cell type is often left out, the mesangial cell (MC), probably since it is not a part of the filtration barrier per se. The MCs are instead found between the glomerular capillaries, embedded in their mesangial matrix. They are in direct contact with the endothelial cells and in close contact with the podocytes and together they form the glomerulus. The MCs can produce and react to a multitude of growth factors, cytokines, and other signaling molecules and are in the perfect position to be a central hub for crosstalk communication between the cells in the glomerulus. In certain glomerular diseases, for example, in diabetic kidney disease or IgA nephropathy, the MCs become activated resulting in mesangial expansion. The expansion is normally due to matrix expansion in combination with either proliferation or hypertrophy. With time, this expansion can lead to fibrosis and decreased glomerular function. In addition, signs of complement activation are often seen in biopsies from patients with glomerular disease affecting the mesangium. This review aims to give a better understanding of the MCs in health and disease and their role in glomerular crosstalk and inflammation.


Changing relations between phonological processing abilities and word-level reading as children develop from beginning to skilled readers: a 5-year longitudinal study.
Relations between phonological processing abilities and word-level reading skills were examined in a longitudinal correlational study of 216 children. Phonological processing abilities, word-level reading skills, and vocabulary were assessed annually from kindergarten through 4th grade, as the children developed from beginning to skilled readers. Individual differences in phonological awareness were related to subsequent individual differences in word-level reading for every time period examined. Individual differences in serial naming and vocabulary were related to subsequent individual differences in word-level reading initially, but these relations faded with development. Individual differences in letter-name knowledge were related to subsequent individual differences in phonological awareness and serial naming, but there were no relations between individual differences in word-level reading and any subsequent phonological processing ability.


Replication pattern of a large homogenously staining chromosome region in antifolate-resistant Chinese hamster cell lines.
We have investigated the replication pattern of a large, homogenously staining chromosome region (HSR) in two antifolate-resistant Chinese hamster cell lines. This region is believed to be the location of an amplified genetic sequence which includes at least the gene coding for dihydrofolate reductase and which may be present in as many as 200 copies. It is shown that the HSR in both cell lines is among the first chromosome regions to begin DNA synthesis after reversal of an early G1 block. In cells synchronized in the S period with hydroxyurea, it is also clear that the HSR in both cell lines begins replication at many sites within its length in early S. The replicons comprising the HSR therefore may respond to a common initiation signal in early S. In on cell line (A3), replication of the HSR requires, at most, 3 hours of a 7-hour S period; in a second line (MQ19), replication proceeds for approximately 5 hours. In neither line does replication of the HSR occur concomitantly with synthesis of characteristic late replicating regions. These results were confirmed in exponential cultures using a retroactive labeling technique. The significance of these findings is discussed with reference to the possible origin and arrangement of the amplified sequence in these two cell lines.


Brain scintigraphy in recurrent medulloblastoma.
Twenty-three patients with previously treated medulloblastoma had brain scintigrams revealed posterior fossa recurrence in 15 patients and supratentorial metastases in 9; only 1 of the latter was clinically suspected. Two subdural hematomas and one calvarial metastasis were also detected. Comparison with other diagnostic evidence shows agreement in 13 out of 14 cases (93%) for posterior fossa recurrence and 8/10 (80%) for supratentiorial masses. Brain scintigraphy appears to be a sensitive and specific test for the detection of recurrence and metastasis in patients with treated medulloblastoma.


Case report: impressive response to pembrolizumab in a patient with mismatch-repair deficient metastasized colorectal cancer and bulky disease.
Here, we report the history of a 42-year-old female patient with sporadic mismatch-repair-deficient metastatic colorectal cancer and abdominal bulky disease, who received pembrolizumab (200 mg every 3 weeks) after the failure of third-line treatment. Restaging 3 months after initiation of treatment revealed a striking response with shrinkage of the bulky peritoneal tumour mass (baseline size 11×11×14 cm) to nearly 25% of the original tumour volume (6.2×7.1×10.4 cm). Restaging 8 months after initiation showed further downsizing of the tumour mass (5.5×7.0×8.0 cm). Tumour markers CEA and CA 19-9 decreased to normal levels, haemoglobin level increased from 8 to 13 mg/dL and her overall clinical performance status increased from ECOG 3 to 1 within 3 months. Therapy with pembrolizumab was continued and is still ongoing. We emphasise the importance of testing for mismatch-repair status in metastatic disease.


Patient Perception of Telemedicine in Maternal-Fetal Medicine.
Background: Maternal-fetal medicine (MFM) is a medical subspecialty that cares for patients with high-risk pregnancies. Methods: An IRB-approved survey was offered to patients in MFM offices of a tertiary health care system from March 2022 to May 2022. Demographics and responses to 15 statements about telemedicine were collected via a 5-point Likert scale. Exploratory factor analysis was performed to identify factors affecting patient perception of the telemedicine experience. We sought to examine perception of telemedicine in MFM, identify factors affecting perception, and identify whether specific demographics were associated with patients who view telemedicine appointments favorably. Results: Surveys were completed by 327 of 347 (94%) patients. A total of 233 (71%) patients felt that the telemedicine experience was equal in quality to in-person appointments, and 257 (79%) were open to telemedicine appointments in the future. Exploratory factor analysis yielded two factors: "physician attentiveness" and "technology comfort." Telemedicine was viewed favorably or neutrally for both factors. Education (lower) and marital status (single) were associated with a favorable perception of physician attentiveness. Ethnicity (Hispanic), employment status (employed), and prior telemedicine experience were associated with a favorable perception of technology comfort. Conclusion: Most patients felt the quality of telemedicine appointments was equal to those completed in person. Physician attentiveness and technology comfort affect telemedicine perception. Specific patient demographic characteristics were associated with differing perceptions of telemedicine in MFM. Our findings suggest that telemedicine is positively viewed for MFM and can be used for improving health care delivery efficiency in MFM.


Increased milk protein synthesis in response to exogenous growth hormone is associated with changes in mechanistic (mammalian) target of rapamycin (mTOR)C1-dependent and independent cell signaling.
The objective of this study was to determine if increased milk protein synthesis observed in lactating dairy cows treated with growth hormone (GH) was associated with mechanistic (or mammalian) target of rapamycin complex 1 (mTORC1) regulation of downstream factors controlling nucleocytoplasmic export and translation of mRNA. To address this objective, biochemical indices of mammary growth and secretory activity and the abundance and phosphorylation status of mTORC1 pathway factors were measured in mammary tissues harvested from nonpregnant lactating dairy cows 6 d after treatment with a slow-release formulation of GH or saline (n=4/group). Treatment with GH increased mammary parenchymal weight and total protein content and tended to increase ribosome number and cell size, whereas protein synthetic efficiency, capacity, and cell number were unchanged. Cellular abundance of the mTORC1 components mTOR and (phosphorylated) mTOR(Ser2448) increased, as did complex eukaryotic initiation factor 4E:eukaryotic initiation factor 4E binding protein 1 (eIF4E:4EBP1), whereas no change was observed for mTORC1-downstream targets 4EBP1, 4EBP1(Ser65), p70/p85(S6K) and p70(S6K)Thre389/p85(S6K)Thre412. Changes in activation were not observed for any of the targets measured. These results indicate that GH treatment influences signaling to mTORC1 but not downstream targets involved in the nucleocytoplasmic export and translation of mRNA. Increased eIF4E:4EBP1 complex formation indicates involvement of the mitogen-activated protein kinase (MAPK) pathway. Abundance of MAPK pathway components eIF4E, eIF4E(Ser209), eIF4E:eIF4G complex, MAP kinase-interacting serine/threonine-protein kinase 1 (MKNK1), MKNK1(Thr197202), and ribosomal protein S6 kinase, 90kDa, polypeptide 1 (RPS6KA1) increased significantly in response to GH, whereas relative activation of the proteins was unchanged. Expression of IGFBP3 and IGFBP5 increased, that of IGF1R decreased, and that of IGF1 remained unchanged in response to GH. PatSearch analysis of the milk caseins αS1-casein, αS2-casein, and β-casein, MAPK signaling target RPS6KA1, and proliferation gene IGFBP3 mRNA indicated that all contained putative eIF4E-sensitivity elements. In response to GH, these genes were all upregulated, suggesting that increased abundance of eIF4E and eIF4E(Ser209) plays a role in mediating their nucleocytoplasmic export. We propose that, in response to GH, the IGF1-IGF1R-MAPK signaling cascade regulates eIF4E-mediated nucleocytoplasmic export and translation of mRNA, whereas mTOR controls cell renewal, cell turnover, and rRNA transcription through an alternative signaling cascade.


Expression of Ras GTPases in normal kidney and in glomerulonephritis.
Small monomeric Ras GTPases play critical and specific roles in the control of cellular proliferation and apoptosis but the expression of the three Ras isoforms (Ha-Ras, Ki-Ras and N-Ras) in human renal tissue is unknown. This work is an immunohistochemical study of Ras expression in normal renal tissue and in membranous glomerulonephritis (MGN), IgA nephropathy (IgAN) and IgA-negative mesangioproliferative glomerulonephritis (MPGN). Formalin-fixed, paraffin-embedded tissue was stained using pan-Ras monoclonal antibody (mAb) and Ras isoform-specific mAb. Detection employed a (DAKO Envision) modified polymer system. The expression of Ras isoforms in normal human kidney was cell-specific. For example, N-Ras was detected in tubule epithelial cells but not in glomerular or interstitial cells. Ki-Ras was expressed in mesangial cells, interstitial cells and in proximal convoluted tubule cells (PCT) (particularly localized at brush borders) and in collecting duct cells (CD) (localized to cell membranes) but not in podocytes. Cytoplasmic Ha-Ras was detected in all the above cell types except podocytes. MGN was associated with podocyte expression of all three Ras isoforms and with reduced mesangial cell expression of Ha-Ras and Ki-Ras. IgAN was characterized by podocyte expression of Ha-Ras (but not Ki-Ras) and reduced mesangial cell expression of Ki-Ras without alterations in mesangial Ha-Ras expression. MPGN was associated with reduced mesangial cell Ha-Ras and Ki-Ras expression without significant podocyte Ras expression. These disease-specific and isoform-specific alterations in Ras expression may be of significance in pathogenesis and warrant further functional investigation.


Clinical and pathological features and imaging manifestations of bronchial anthracofibrosis: the findings in 15 patients.
Bronchial anthracofibrosis (BAF) has been defined as a luminal narrowing associated with anthracotic pigmentation on bronchoscopy without a relevant history of pneumoconiosis or smoking. The aim of the study is to study the clinical features and imaging manifestations of BAF, and to promote the awareness of this disease. Between October 2006 and January 2012, 15 patients were diagnosed at our department as BAF that showed a narrowing or obliteration of lobar or segmental bronchi with anthracotic pigmentation in the overlying mucosa on bronchoscopy. The medical records including clinical features, imaging manifestations, electronic bronchoscopic findings, and pathological features were analysed, and the literature was reviewed. A total of 15 patients were analyzed; 13 were female (86.7%) and two were male (13.3%) and the age range was from 62 to 86 years with a mean age of 74.5 years. Three cases (20.0%) had a history of tuberculosis. The most common clinical symptoms of BAF were cough (100%), expectoration (73.3%), dyspnea (60.0%), and fever (46.7%). Twelve cases displayed mild to moderate obstructive ventilatory dysfunction. In the electronic bronchoscopic evaluation, the most common findings were black bronchial mucosal pigmentation, bronchial stenosis, bronchial occlusion, and bronchial mucosal folds. Pathological evaluation revealed chronic inflammation of the bronchial mucosa, submucosal carbon particle deposition, and mucosal or submucosal fibrosis. Chest CT scans showed that 15 patients had bronchial stenosis or obstruction (direct signs) with the right middle lobe being the most common site (11 cases, 73.3%). The indirect sign was mainly the presence of bronchial obstructive diseases (including secondary infection), represented by 11 cases of pulmonary consolidation (73.3%), seven cases of atelectasis (46.7%), and five cases of nodules (33.3%). The CT mediastinal window showed bronchial lymph node lesions, mediastinal lymph node calcification (12 cases, 80.0%), and enlargement of multiple mediastinal lymph nodes. The diagnosis of BAF was mainly based on bronchoscopic evaluation. Its pathogenesis is currently unclear, although it may be related to tuberculosis or bio-fuel inhalation. The diagnosis of BAF has important clinical significance, and improved awareness of this disease will contribute to prevention of unnecessary thoracotomies.


Management of jugular bulb injury during retrosigmoid transmeatal resection of vestibular schwannoma.
Inadvertent laceration of the jugular bulb is a potentially serious complication of the retrosigmoid transmeatal approach to vestibular schwannomas. Here, the authors present the case of a 51-year-old woman with a right Hannover T4a vestibular schwannoma and bilateral high-riding jugular bulb, which was opened during drilling of the internal auditory canal (IAC). They highlight the immediate management of this complication, technical nuances for closing the defect without occluding the jugular bulb, and modifications of the standard technique needed to continue surgical resection. The video can be found here: https://stream.cadmore.media/r10.3171/2021.7.FOCVID2155.


Ability of Massachusetts-type infectious bronchitis virus to increase colibacillosis susceptibility in commercial broilers: a comparison between vaccine and virulent field virus.
The abilities of Massachusetts-type vaccine virus and virulent infectious bronchitis (IB) field virus to increase colibacillosis susceptibility were compared. In four experiments, 29-day-old female commercial broilers housed in isolators, were infected intratracheally and oculonasally with IB vaccine strains (H120 and H52) or virulent IB field strains (D387 and M41) (4.8 or 6.8 log(10) median embryo infective dose, per broiler). Five days later, Escherichia coli 506 strain was given intratracheally (5.6 to 8.8 log(10) colony forming units/broiler). The incidence of nasal discharge at 3 and 5 days after IB virus infection was used to assess the clinical effect of the IB infection, while mortality, body weight uniformity and E. coli lesions at 7 days following E. coli inoculation were used as parameters for colibacillosis. Nasal discharge was observed in 6/117 (5%), 26/119 (22%), 35/119 (29%) and 115/120 (96%) of broilers infected with H120, H52, D387 and M41 virus, respectively. Apart from H52 and D387, differences between IBV strains were significant. IB vaccine and virulent IB viruses did not generally differ significantly in their ability to induce colibacillosis susceptibility. Mean colibacillosis lesion scores of H52-infected birds even significantly exceeded those of birds infected with the other IB viruses. The ability of H120 virus to induce colibacillosis susceptibility tended to be the weakest. The practical consequences of these findings are discussed.


[Not Available].
Protecting healthcare staff against the risk of infection caused by injuries when using sharp or pointed instruments has been placed - as a special, common and difficult type of "biological risk" - at the centre of attention in the investigations, studies and research conducted by various countries and subjects that have led to a convergence between the results able to influence European and supranational legislation. This convergence of results is headed towards the recognition of the great potential of the role of technology and, more specifically, of the "protection devices" of paramount and essential importance, as "technical measures", over the other, nevertheless important "organisational" and "personal" measures. The national and international results of the studies and research, as well as their corresponding conclusions, deserve special space in this journal, given the supranational legislative recognition they received in Directive 2010/32/EU. Indeed, this latter Directive, set forth and classified, with a system of absolutely mandatory regulations, the "Framework agreement" concluded between the organisations that are most representative of the hospital and healthcare sector for the "prevention of cut and prick injuries". On the basis of the "European Union" treaty, Directive 2010/32/EU must be assimilated in the legal system of all the member States of this "Union" with appropriately sanctioned legal provisions in the event of breach by "employers" and/or "directors". In this sense, with Legislative Decree no. 19 of 2014, Italy introduced a dedicated "criminal offence of danger" for the breach of "prevention" regulations intended to prevent the specific "biological risk" in question, regulations that have been incorporated into Legislative Decree 81/08 - "The Occupational Safety and Health Consolidation Act" with a special "Section" (X bis) dedicated exclusively to them.


L-648,051, sodium 4-[3-(4-acetyl-3-hydroxy-2-propylphenoxy)- propylsulfonyl]-gamma-oxo-benzenebutanoate: a leukotriene D4 receptor antagonist.
L-648,051, sodium 4-[3-(4-acetyl-3-hydroxy-2-propylphenoxy) propylsulfonyl]-gamma-oxo-benzenebutanoate is a selective and competitive inhibitor of [3H]leukotriene D4 (KB value of 4.0 microM) and to a lesser extent [3H]leukotriene C4 (Ki value of 36.7 microM) binding in guinea pig lung homogenates. Functionally, it selectively antagonized contractions of guinea pig trachea induced by leukotrienes C4, D4, E4, and F4 in concentrations that did not antagonize contractions induced by acetylcholine, histamine, serotonin, prostaglandin F2 alpha, or U-44069 (endoperoxide analogue). Schild plot analysis indicated that L-648,051 competitively antagonized contractions of guinea pig ileum induced by leukotriene D4 (pA2 7.7) and contractions of trachea induced by leukotrienes D4, E4, and F4 (pA2 7.3, 7.4, and 7.5, respectively). Contractions of guinea pig trachea induced by leukotriene C4 were inhibited in a noncompetitive fashion (Schild plot slope, 0.45). Developed contractions of trachea induced by the leukotrienes were rapidly reversed by L-648,051 greater than FPL-55712 greater than L-649,923. Intravenous L-648,051 selectively blocked bronchoconstriction induced in anaesthetized guinea pigs by intravenous leukotrienes C4, D4, and E4 but not that induced by arachidonic acid, serotonin, U-44069, or acetylcholine. The compound displayed poor activity following intraduodenal administration. The profile of activity for L-648,051 indicates that it may be a useful topical agent for studying the role of leukotrienes in diseases such as bronchial asthma.


Low temperature bioremediation of oil-contaminated soil using biostimulation and bioaugmentation with a Pseudomonas sp. from maritime Antarctica.
To identify native Antarctic bacteria capable of oil degradation at low temperatures. Oil contaminated and pristine soils from Signy Island (South Orkney Islands, Antarctica) were examined for bacteria capable of oil degradation at low temperatures. Of the 300 isolates cultured, Pseudomonas strain ST41 grew on the widest range of hydrocarbons at 4 degrees C. ST41 was used in microcosm studies of low temperature bioremediation of oil-contaminated soils. Microcosm experiments showed that at 4 degrees C the levels of oil degradation increased, relative to the controls, with (i) the addition of ST41 to the existing soil microbial population (bioaugmentation), (ii) the addition of nutrients (biostimulation) and to the greatest extent with (iii) a combination of both treatments (bioaugmentation and biostimulation). Addition of water to oil contaminated soil (hydration) also enhanced oil degradation, although less than the other treatments. Analysis of the dominant species in the microcosms after 12 weeks, using temporal temperature gradient gel electrophoresis, showed Pseudomonas species to be the dominant soil bacteria in both bioaugmented and biostimulated microcosms. Addition of water and nutrients may enhance oil degradation through the biostimulation of indigenous oil-degrading microbial populations within the soil. However, bioaugmentation with Antarctic bacteria capable of efficient low temperature hydrocarbon degradation may enhance the rate of bioremediation if applied soon after the spill. In the future, native soil bacteria could be of use in bioremediation technologies in Antarctica.


Abnormal Leydig cell development at puberty in the androgen-resistant Tfm mouse.
Leydig cells are the major source of androgens in the male, and it is likely that these cells are also targets for androgen action. The role of androgen action in regulating the development of Leydig cell function has been determined using the testicular feminized (Tfm/Y) mouse, which lacks androgen receptors. In Tfm mice, the testes fail to descend at the normal time (25 days), and testicular descent was, therefore, surgically prevented in control (+/Y) animals. The activity of 3 beta-hydroxysteroid dehydrogenase increased during development from 5-40 days in control and Tfm mice, with no significant difference between the groups. In control animals, 17 alpha-hydroxylase activity was low from 5-25 days, at which time there was a 26-fold increase up to 40 days. In the Tfm group, 17 alpha-hydroxylase activity was 4-fold higher than that in controls on day 5, but showed no change in activity after 25 days and remained at neonatal levels up to 40 days. The activity of 17-ketosteroid reductase in the control animals showed a developmental pattern similar to that of 17 alpha-hydroxylase, with a marked increase in activity after 25 days. In the Tfm group, 17-ketosteroid reductase was normal on day 5, but failed to show any significant change thereafter and remained at neonatal levels on day 40. Serum LH levels in control animals increased from 5 days to a peak at 30 days. In Tfm mice, LH levels were significantly increased on days 20 and 40, but did not differ from controls on days 5, 25, and 30. In control +/Y animals, in which normal testicular descent was allowed to proceed at 25 days, the pattern of development was similar to that in the cryptorchid +/Y animals, although the increase in 17 alpha-hydroxylase activity at 30 and 40 days was significantly greater. The results show 1) that fetal Leydig cell steroidogenesis is enhanced in the absence of androgen receptors, but 2) that adult Leydig cells require receptor-mediated androgen activity around day 25 for normal functional development. In addition, 3) the lack of testicular descent at 25 days reduces the pubertal rise in 17 alpha-hydroxylase activity.


Effects of water concentration on the free volume of amino acid ionic liquids investigated by molecular dynamics simulations.
Amino acid ionic liquids (AAILs) are gaining attention because of their potential in CO2 capture technology. Molecular dynamics simulations of AAILs tetramethylammonium glycinate ([N1111][Gly]), tetrabutylammonium glycinate ([N4444][Gly]), and 1,1,1-trimethylhydrazinium glycinate ([aN111][Gly]) and their corresponding mixtures with water were performed to investigate the effect of water concentration on the cation-anion interactions. The water content significantly influenced the free volume (FV) and fractional free volume (FFV) of the AAILs that varied with the hydrophobic and hydrophilic nature of the ion pairs. Under dry conditions, the FFV increased with increasing cation molecular sizes, indicative of proportional adsorption of any inert gases, such as N2, as consistent with experimental observations. Furthermore, the polarity of the cation played an important role in FFV and hence the diffusion of the AAILs. Density functional theory calculations suggested that hydrophilic [aN111][Gly] featured stronger interactions in the presence of water, whereas the hydrophobic IL showed weaker interactions. The carboxylate group of glycinate displayed stronger interactions with water than the cation. The computational study provided qualitative insight into the role of FV of the AAILs on CO2 and N2 absorption and suggests that [aN111][Gly] has CO2 adsorption capacity in the presence of water superior to that of other studied AAILs.


Neuroendocrinoma of the jejunum: electron microscopic and biochemical analysis.
A 56-year-old woman presented with a sudden, severe hemorrhage per rectum. Angiography localized a jejunal tumor, which was excised. Light microscopy suggested a neuroendocrine tumor, but neither a smooth muscle tumor nor a lymphoma could be excluded. Electron microscopy showed dense cored, single membrane bound secretory granules 150--220 nm in diameter; myofilaments were not observed. Biochemical analysis of tumor tissue yielded considerable amounts of catecholamines. VMA, 5-HIAA, and metanephrines. These combined ultrastructural and biochemical observations establish the diagnosis of neuroendocrine tumor; however, in this case neither type of information is sufficiently specific to define the tumor as either a paraganglioma or a carcinoid. Although paraganglia and mucosal endocrine cells in the GI are currently thought to constitute distinct cell types, they share numerous structural and functional properties, and they are both thought to be part of the APUD cell system. These parallels and similarities are shared by the neoplasms derived from them which often display features of both. In the absence of specific granule types or specific substances isolated from tumor tissue, only the application of specific immunocytochemistry techniques may allow the precise "functional" classification of such tumors.


REST Inhibits Direct Reprogramming of Pancreatic Exocrine to Endocrine Cells by Preventing PDX1-Mediated Activation of Endocrine Genes.
The emerging appreciation of plasticity among pancreatic lineages has created interest in harnessing cellular reprogramming for β cell replacement therapy of diabetes. Current reprogramming methodologies are inefficient, largely because of a limited understanding of the underlying mechanisms. Using an in vitro reprogramming system, we reveal the transcriptional repressor RE-1 silencing transcription factor (REST) as a barrier for β cell gene expression in the reprogramming of pancreatic exocrine cells. We observe that REST-bound loci lie adjacent to the binding sites of multiple key β cell transcription factors, including PDX1. Accordingly, a loss of REST function combined with PDX1 expression results in the synergistic activation of endocrine genes. This is accompanied by increased histone acetylation and PDX1 binding at endocrine gene loci. Collectively, our data identify a mechanism for REST activity involving the prevention of PDX1-mediated activation of endocrine genes and uncover REST downregulation and the resulting chromatin alterations as key events in β cell reprogramming.


Elastic resistance training improved glycemic homeostasis, strength, and functionality in sarcopenic older adults: a pilot study.
The purpose of this study was to verify the effects of 12 weeks of elastic resistance training on the glucose homeostasis, strength and functionally in sarcopenic older adults. Seven sarcopenic subjects (age, 70.71± 8.0 years; body mass index, 22.75±3.1 kg/m2) participated of training protocol with 12 weeks of elastic resistance training. The oral glucose tolerance test, handgrip strength, sit-to-stand test, 4-m walk test, and coordination test were measured at baseline and after training. According to the results, baseline values of area under the curve of glucose and homeostatic model assessment-insulin resistance were significantly lower than after 12 weeks, respectively (808.2±185.0 mmol/L vs. 706.6±114.8 mmol/L, P=0.049; 1.44±0.48 vs. 0.73±0.32, P=0.040). There were a significant improve of HGS (24.3±5.7 kg vs. 27.3±7.3 kg, P=0.01), 4-m walking test (3.64±0.4 sec vs. 3.23±0.3 sec, P=0.04), and STS (10.2±2.3 sec vs. 9.0±1.9 sec, P=0.04) compared with baseline. In conclusion, these findings suggest that elastic resistance training improved glucose homeostasis, strength, and functionality in sarcopenic older adults.


Bilateral otorrhagia: a rare complication of laparoscopic abdominopelvic surgery.
An 80-year-old woman without any previous otological symptoms underwent laparoscopic abdominoperineal resection for T3N0M0 low rectal carcinoma 4-5 cm from the anal verge. The total operative time was 6 h, of which she spent long hours in the Trendelenburg (35°) position due to difficult pelvic dissection. Midway through the procedure, she developed spontaneous non-traumatic bilateral otorrhagia. This case highlights the potential risk of increased intracranial pressure during prolonged periods of being in a steep Trendelenburg position caused either by the position itself or in combination with carbon dioxide pneumoperitoneum. We also consider the effect of a sudden change from this position to supine as a potential risk.


Plant Specialized Metabolites in Hazelnut (Corylus avellana) Kernel and Byproducts: An Update on Chemistry, Biological Activity, and Analytical Aspects.
Corylus avellana (hazelnut) is one of the most popular tree nuts on a worldwide basis. The main products of C. avellana are kernels, a nutritious food, with a high content of healthy lipids, contained in a hard shell. In recent years, along with the ongoing research carried out on hazelnut kernels, a growing interest has been addressed to the hazelnut byproducts including hazelnut skin, hazelnut hard shell, and hazelnut green leafy cover as well as hazelnut tree leaf. These byproducts deriving from the roasting, cracking, shelling/hulling, and harvesting processes have been found as a source of "phytochemicals" with biological activity. The aim of this review is to provide a comprehensive and critical update on the chemistry and biological activity of specialized metabolites occurring in hazelnut kernels and byproducts. Phenolics are the most abundant phytochemicals not only in the kernels, but also in other processing byproducts. Attention has been also devoted to taxane derivatives isolated from C. avellana leaves. An overview on the biological activity, mainly antioxidant, antiproliferative, and antimicrobial along with less common biological effects, has been provided, contributing to highlight C. avellana as a source of bioactive phytochemicals with the potential to exert beneficial effects on human health. Finally, analytical techniques for the quali-quantitative analysis of specialized metabolites occurring in the different parts of C. avellana have been reviewed.


[Tests for anthropozoonoses in sera of farmers in eastern-Austria. I. Communication (author's transl)].
Because of human disease and the evidence of antibodies in domestic- and wild animal-sera, human sera were tested for various anthropozoonoses in three districts of the so called "Seewinkel" (Eastern-Austria). Positive reactions were found for listeriosis, tularemia, leptospirosis, ornithosis, Q-fever, rickettsiae of the RMSF-group "Stamm B", mycoplasmosis, TBE-(FSME-), Sindbis-, West Nil-VIRUS. These results are indicators for the occurrence of the pathogens and are supported by the positive results already found at serological tests of domestic and wild animals.


[Training of inspiratory muscles in chronic obstructive lung disease. Its impact on functional changes and exercise tolerance].
The aim of this study was to evaluate the impact of inspiratory muscle training on lung function and exercise tolerance in patients with chronic obstructive pulmonary disease (COPD). Thirty-five patients with stable COPD were enrolled. We measured lung function variables and peak inspiratory and expiratory pressures (PImax and PEmax). Tests of progressive maximal exercise tolerance and stable submaximal exercise tolerance were administered. Two study groups were formed. Group A patients (n = 20) were enrolled in a respiratory muscle training program lasting four months. Group B (n = 15) was the control group. At the end of the study period the patients underwent testing similar to the first battery of tests. All showed moderate to severe obstruction with no significant differences between groups (FEV1: group A 37.6 +/- 13%, group B 36.6 +/- 12%; FVC: group A 80.4 +/- 15%, group B 80 +/- 12%). Nor were there any significant differences between the two groups in initial results of either maximal respiratory pressures or exercise tolerance. No lung function changes were observed in either group. PImax in group A increased significantly at the end of the study (from 54 +/- 9 to 78 +/- 16 cmH2O; p < 0.001); there were no changes in group B. No changes were seen in VO2max or ventilatory response and/or gasometry during exercise in any of the groups. The trained group, on the other hand, experienced a significant decrease in dyspnea evaluated on the Borg scale exercise in maximal (5.7 +/- 1.1 versus 4.7 +/- 1.2, p < 0.005) and submaximal (5.9 +/- 0.9 versus 4.9 +/- 1.3, p < 0.005) and an increase in time of submaximal exercise tolerance (5.5 +/- 2 versus 7 +/- 3 min, p < 0.05), changes that were not observed in the control group. Based on these results, and although specific training of inspiratory muscles does not appear to improve lung function in patients with COPD, it is accompanied by a decreased sense of dyspnea during exercise and greater tolerance.


[Abnormal hemoglobinopathy (HbE) diagnosed from microcytic hypochromic red blood cells in a 31 year-old Bangladeshian male].
Microcytic hypochromic red blood cells (RBC) were discovered in a 31 year-old Bangladeshi man. Additional laboratory data revealed only slight elevations of LDH and transaminase activities. The patient was clinically asymptomatic and showed no signs of anemia. On this basis, along with biochemical genetic analysis of hemoglobin, a diagnosis of hemoglobinopathy (HbE) was made. HbE is common in southeast Asia with over 30% of the population affected. Most people are asymptomatic. In northern Asia, including Japan, HbE is found in only 0.001% of the population. With ther recent influx of southeast Asians to Japan, care must be taken not to overlook the diagnosis of hemoglobinopathies.


An open clinical trial of cognitive therapy for borderline personality disorder.
Although borderline personality disorder (BPD) is a major public health concern, psychotherapeutic trials have been limited. The present uncontrolled clinical trial examines whether cognitive therapy for BPD is associated with significant improvement on measures of psychopathology. A total of 32 patients with BPD, who also reported suicide ideation or who engaged in self-injury behavior, received weekly cognitive therapy sessions over a 1-year period as described by Layden et al. (1993). The results revealed significant and clinically important decreases on measures of suicide ideation, hopelessness, depression, number of borderline symptoms and dysfunctional beliefs at termination and 18-month assessment interviews. Implications for further research with this difficult-to-treat patient population are discussed.


Drug Use on Mont Blanc: A Study Using Automated Urine Collection.
Mont Blanc, the summit of Western Europe, is a popular but demanding high-altitude ascent. Drug use is thought to be widespread among climbers attempting this summit, not only to prevent altitude illnesses, but also to boost physical and/or psychological capacities. This practice may be unsafe in this remote alpine environment. However, robust data on medication during the ascent of Mont Blanc are lacking. Individual urine samples from male climbers using urinals in mountain refuges on access routes to Mont Blanc (Goûter and Cosmiques mountain huts) were blindly and anonymously collected using a hidden automatic sampler. Urine samples were screened for a wide range of drugs, including diuretics, glucocorticoids, stimulants, hypnotics and phosphodiesterase 5 (PDE-5) inhibitors. Out of 430 samples analyzed from both huts, 35.8% contained at least one drug. Diuretics (22.7%) and hypnotics (12.9%) were the most frequently detected drugs, while glucocorticoids (3.5%) and stimulants (3.1%) were less commonly detected. None of the samples contained PDE-5 inhibitors. Two substances were predominant: the diuretic acetazolamide (20.6%) and the hypnotic zolpidem (8.4%). Thirty three samples were found positive for at least two substances, the most frequent combination being acetazolamide and a hypnotic (2.1%). Based on a novel sampling technique, we demonstrate that about one third of the urine samples collected from a random sample of male climbers contained one or several drugs, suggesting frequent drug use amongst climbers ascending Mont Blanc. Our data suggest that medication primarily aims at mitigating the symptoms of altitude illnesses, rather than enhancing performance. In this hazardous environment, the relatively high prevalence of hypnotics must be highlighted, since these molecules may alter vigilance.


Children's Personal Exposure Measurements to Extremely Low Frequency Magnetic Fields in Italy.
Extremely low frequency magnetic fields (ELF-MFs) exposure is still a topic of concern due to their possible impact on children's health. Although epidemiological studies claimed an evidence of a possible association between ELF-MF above 0.4 μT and childhood leukemia, biological mechanisms able to support a causal relationship between ELF-MF and this disease were not found yet. To provide further knowledge about children's ELF-MF exposure correlated to children's daily activities, a measurement study was conducted in Milan (Italy). Eighty-six children were recruited, 52 of whom were specifically chosen with respect to the distance to power lines and built-in transformers to oversample potentially highly exposed children. Personal and bedroom measurements were performed for each child in two different seasons. The major outcomes of this study are: (1) median values over 24-h personal and bedroom measurements were <3 μT established by the Italian law as the quality target; (2) geometric mean values over 24-h bedroom measurements were mostly <0.4 μT; (3) seasonal variations did not significantly influence personal and bedroom measurements; (4) the highest average MF levels were mostly found at home during the day and outdoors; (5) no significant differences were found in the median and geometric mean values between personal and bedroom measurements, but were found in the arithmetic mean.


Preoperative informational video reduces preoperative anxiety and postoperative negative behavioral changes in children.
Anesthesia and surgery can lead to major distress for children. Sedative premedication and preoperative preparation techniques are available to reduce preoperative anxiety in children. We aimed to assess the effect of informational video based on role-play modelling on preoperative anxiety and postoperative behavior changes in children undergoing surgery. Forty-two children aged 5-12 years, with American Society of Anesthesiologist physical status I-II, scheduled for elective outpatient surgery, were enrolled in this study. Patients were randomly allocated to one group with or without information video presentation. In group V, patients watched the information video (N.=21). Other children were verbally informed in a standard care (group C, N.=21). We recorded patient's demographics (age, birth order, surgery time, surgery type, history of previous surgery, parent's age, parental working status, parental education level), the preoperative anxiety level using Modified Yale Preoperative Anxiety Scale (MYPAS) and at 1 week after discharge, new developing postoperative maladaptive behaviors (POMB) using the Post Hospitalization Behavioral Questionnaire by telephone interview. Patient's demographics were similar in both groups. Total MYPAS scores were found to be lower in group V as compared to group C P=0.0001). Difficulty getting to sleep, nocturnal enuresis, fear of dark, to object to go to bed at night and decreased appetite of new developing POMB were found to be lower in group V than group C. We also found a correlation between anxiety scores and POMB. The presentation of an informational video based on model making reduces preoperative anxiety at the time of placement of the facemask and postoperative negative behavioral changes in children.


Tooth wear: prevalence and associated factors in general practice patients.
To estimate the prevalence of tooth wear and to investigate factors associated with tooth wear in patients from general practices in the Northwest United States. Data on the diagnosis and treatment of oral diseases during the previous year were collected in a survey with a systematic random sample of patients (n= 1530) visiting general dentists from the Northwest Practice-based REsearch Collaborative in Evidence-based DENTistry (PRECEDENT) (n=80). Prevalence ratios (PRs) of moderate to severe occlusal and incisal tooth wear by patient characteristics were estimated using cluster-adjusted multiple binomial regression for adults (18+ years) and children/adolescents (3-17 years). For adults, the mean number of teeth with wear facets was 5.4 [95% confidence interval (CI) =4.6-6.2] and 51% of the adults had four or more teeth with wear. Participants 45-64 and 65+ years old were 1.3 (95% CI=1.1-1.6) and 1.4 (95% CI=1.1-1.8) times as likely to have 4+ teeth with moderate to severe wear facets as participants 18-44 years old. Adult males had a 20% (PR=1.2; 95% CI=1.1-1.4) higher prevalence of wear than adult females. Adults who were using, or had ever used occlusal splints had higher prevalence of tooth wear compared to those who never used such appliances (PR=1.3; 95% CI=1.0-1.5). Adults with any periodontal bone loss also had a 20% higher prevalence of wear than adults without periodontal disease (PR=1.2; 95% CI=1.0-1.4). For children/adolescents, the mean number of teeth with moderate to severe wear facets was 1.6 (95% CI=0.9-2.6) and 31% of the children had one or more teeth with wear facets. The adjusted prevalence ratio of tooth wear (1+ teeth with wear facets) for boys was 1.6 times as high (95% CI=1.1-2.4) as compared with girls. The prevalence of wear for children 12+ years old was 50% (PR=0.5; 95% CI=0.3-0.8) lower than that of children <12 years old. Angle's class II was associated with higher tooth wear prevalence (PR= 1.8; 95% CI=1.3-2.6) than class I. Children with posterior or anterior open bite had lower prevalence of wear than their counterparts (PR=0.6; 95% CI=0.3-1.0). No associations were observed between tooth wear and orthodontic treatment, missing teeth, and race/ethnicity. Tooth wear is a prevalent condition in this population. Among adults, higher prevalences of tooth wear were observed among those who were older, males, had used occlusal splints and had periodontal disease. Among children, higher prevalences were associated with younger age, male gender, class II malocclusion and the absence of open bite. Submitted on behalf of the Northwest PRECEDENT network, with support from NIDCR grants DE016750 and DE016752.