Undifferentiated murine embryonic stem cells cannot induce portal tolerance but may possess immune privilege secondary to reduced major histocompatibility complex antigen expression.
Induction of donor-specific tolerance using embryonic stem (ES) cells followed by transplantation of ES cell-derived tissues from the same allogeneic strain could theoretically engender successful transplantation without immunosuppression. We sought to induce tolerance using bona fide murine ES cells in immunocompetent mice. ES cells were evaluated for the expression of markers restricted to undifferentiated cells [stage-specific embryonic antigen-1 (SSEA-1) and OCT-4] and the ability to form teratomas in immunodeficient mice. BALB/cByJ mice underwent intraportal inoculation with YC5-EYFP ES cells (129 strain; R1-derived) or saline followed by transplantation with 129X1/SvJ, CBA/J, or BALB/cByJ nonvascularized, neonatal cardiac grafts. Mice were sacrificed at graft failure and underwent histologic evaluation of transplanted grafts and lymphoid organs. ES cells and early differentiated progeny underwent real time (RT)-PCR and fluorescence-activated cell sorting (FACS) analysis to detect major histocompatibility complex (MHC) gene transcription and antigen expression. ES cells expressed markers restricted to undifferentiated cells while maintaining the ability to form teratomas in immunodeficient mice. No prolongation of allograft survival or evidence of lymphoid chimerism was observed in immunocompetent recipient mice despite hepatic teratoma formation. MHC class I, class II, and nonclassical antigens were undetectable on ES cells and early differentiated progeny despite the presence of mRNA transcripts. Class I expression was strongly upregulated upon exposure to gamma-interferon. Intraportal inoculation with murine ES cells does not produce lymphoid chimerism or induce donor-specific unresponsiveness to neonatal cardiac grafts in unmanipulated immunocompetent hosts. However, specific differentiated cell types such as ES cellderived dendritic cells, or alternate routes of ES cell administration, may be effective. ES cells appear to have immune privilege, allowing them to form teratomas in immunocompetent mice.


Optical generation of a precise microwave frequency comb by harmonic frequency locking.
A semiconductor laser under negative optoelectronic feedback is applied to the generation of a microwave frequency comb through the nonlinear dynamics. The laser system is operated in a harmonic frequency-locked pulsing state, where its power spectrum is a microwave frequency comb that consists of multiples of a locking frequency. Every frequency component of the comb can be simultaneously stabilized by simply injecting an external microwave modulation at any component of the comb. This phenomenon can be viewed as a kind of microwave injection locking of the laser dynamics.


Pentoxifylline attenuated the renal disease progression in rats with remnant kidney.
Previous studies have reported that pentoxifylline, a phosphodiesterase inhibitor, attenuates experimental mesangial proliferative glomerulonephritis. This study hypothesized that pentoxifylline could also attenuate the renal disease progression in rats with remnant kidney. After 5/6 subtotal nephrectomy, rats developed progressively elevated proteinuria and plasma creatinine, glomerulosclerosis, interstitial inflammation, and fibrosis, all of which were attenuated by 40 to 60% by pentoxifylline. However, the elevated BP was not changed by pentoxifylline. Pentoxifylline reduced the upregulation of monocyte chemoattractant protein-1 gene by 60% in the cortex of remnant kidney, as well as in a dose-dependent manner in the albumin- or angiotensin II-stimulated proximal tubular cells. It also reduced the upregulation of mitogenic and profibrogenic genes by 50%, including platelet-derived growth factor, fibroblast growth factor-2, transforming growth factor-beta(1), connective tissue growth factor, and types I and III collagen in the cortex of remnant kidney. Furthermore, pentoxifylline was found to decrease the numbers of interstitial myofibroblasts by 60% in the cortex of remnant kidney and suppress the proliferation of cultured interstitial fibroblasts. It also reduced the angiotensin II-induced or transforming growth factor-beta(1)-induced expression of connective tissue growth factor gene in cultured fibroblasts and mesangial cells. Combining pentoxifylline with an angiotensin-converting enzyme inhibitor, cilazapril, almost completely attenuated the renal disease progression in rats with remnant kidney. In conclusion, pentoxifylline alone can attenuate the chronic renal disease progression. Its combination with cilazapril has the potential to prevent the renal disease progression almost completely.


Social support for elderly patients with chronic wounds.
Few attempts have been made to measure the social support received by elderly patients with chronic wounds. To focus research on these issues, an established model integrating the various roles played by social support in the adaptation of patients to stressful situations was applied. Two questionnaires were used to measure perceived social support and coping in a sample of patients with leg ulcers (N = 15, mean age 70.4 years) or diabetic foot ulcers (N = 15, mean age 63.6 years) at two time-points over a four-month period. The results indicate that there were no statistical differences between the groups. The overall levels of social support were low, with emotional support recorded most frequently. The standardised scores for types of coping indicate no unusual patterns, although the scores for logical analysis were low. However, there was considerable variation in the types of coping strategies used by individuals.


Silicon oxides: a promising family of anode materials for lithium-ion batteries.
Silicon oxides have been recognized as a promising family of anode materials for high-energy lithium-ion batteries (LIBs) owing to their abundant reserve, low cost, environmental friendliness, easy synthesis, and high theoretical capacity. However, the extended application of silicon oxides is severely hampered by the intrinsically low conductivity, large volume change, and low initial coulombic efficiency. Significant efforts have been dedicated to tackling these challenges towards practical applications. This Review focuses on the recent advances in the synthesis and lithium storage properties of silicon oxide-based anode materials. To present the progress in a systematic manner, this review is categorized as follows: (i) SiO-based anode materials, (ii) SiO2-based anode materials, (iii) non-stoichiometric SiOx-based anode materials, and (iv) Si-O-C-based anode materials. Finally, future outlook and our personal perspectives on silicon oxide-based anode materials are presented.


The WHO construct of health-related functioning (HrF) and its implications for health policy.
The International Classification of Functioning (ICF) has acquired a central role in the WHO Family of International Classifications and it has been extensively adopted as the reference framework for health-related functioning (HrF). This review aims to provide a description of the ICF/HrF to contextualise ICF/HrF in relation to other approaches to health functioning and to describe its application in policy and legislation with a special focus on Spain. Narrative review based on the scientific literature and prior expert knowledge. ICF is both a coding system and a conceptual framework of HrF, which is framed as a unidimensional, bipolar and asymmetric construct with a negative pole (disability) and a positive pole (good functioning) with higher complexity. Other models of HrF include health promotion, quality of life and activities of daily living (ADL). The curtailed taxonomy of ICF and its unclear distinction from other approaches have had significant implications for research, policy and legislation, as illustrated by the case of the legislation and services for functional dependency in Spain and other examples. The ICF model of functioning is more comprehensive and usable than previous alternatives, but a full taxonomy of the HrF construct is needed to avoid further confusions in this field. This should also comprise harmonization with other classifications of the WHO Family of International Classifications and other models of health functioning.


Utilization of Mind-Body Intervention for Integrative Health Care of COVID-19 Patients and Survivors.
Recent findings suggest a correlation between COVID-19 and diabetes, although the underlying causes are still little understood. COVID-19 infection tends to induce severe symptoms in patients with underlying diabetes, increasing their mortality rate. Moreover, COVID-19 itself appears to be a diabetogenic factor. In addition, mental health conditions, such as depression due to lockdown and anxiety about infection, were found to affect glycemic control and immunity, highlighting the importance of mental health care during the pandemic. Mind-Body Intervention (MBI), which includes meditation, yoga, and qigong, has emerged as a tool for mental health management due to its effects on stress reduction and the promotion of mental and physical well-being. Here, we review the latest randomized controlled trials to determine the effects of MBI on glycemic control and the immune system and discuss the underlying mechanisms by which MBI facilitates the virtuous cycle of stress management, glycemic control, and immune modulation. Furthermore, we examine the actual utilization of MBI during the COVID-19 pandemic era through recent studies. With proper online education, non-pharmacological MBI may be more widely used as an important tool for self-health care that complements the usual treatment of COVID-19 patients and survivors.


Effects of apomorphine on sexual behavior in male quail.
In the rat, dopamine (DA) facilitates male copulatory behavior. Indirect evidence based largely on neuroanatomical data suggest that in quail DA is also implicated in the control of male reproductive behavior but there is no pharmacological evidence to support this conclusion. To test this idea, castrated testosterone (T)-treated male quail were injected with various doses of the dopaminergic agonist apomorphine (APO) in the range 1-10,000 micrograms/kg. The sexual behavior of birds was recorded starting 15 min after APO injection for a duration of 30 min. A dose-dependent inhibition of male reproductive behavior that lasted for the entire duration of the test was observed. In a second experiment, gonadectomized T-treated male Japanese quail were injected daily with APO (0, 10, or 1,000 micrograms/kg) during 8 days. Their sexual interactions with a partner were quantified either 24 h or 15 min after the last injection. No influence of the treatment on copulatory behavior was observed 24 h after the last injection, but a strong inhibition was present when the test was performed 15 min after. To research whether the inhibitory effects of APO were due to a preferential action on D2 presynaptic autoreceptors, male quail were pretreated with two different D2 antagonists (spiperone or pimozide; 0.5 or 2 mg/kg) before being injected with APO (100 micrograms or 1 mg/kg). Spiperone facilitated male sexual behavior but did not suppress the inhibitory effect of APO. No significant effect of pimozide was observed. These results support the notion that DA modulates male sexual activity in the Japanese quail. The specific role of the different dopaminergic receptor subtypes remains, however, to be elucidated.


Interleukin 3-dependent mediator release in basophils triggered by C5a.
The anaphylatoxin C5a is a potent trigger for basophil degranulations, but in contrast to IgE-dependent basophil activation, it does not result in the synthesis of sulfidoleukotrienes (leukotriene C4/D4/E4). Thus, degranulation and the generation of lipid mediators are separately regulated cellular responses. Exposure of human blood basophils to the cytokine IL-3 alone does not induce the release of histamine in cells from most donors and never leads to the generation of LTC4, indicating that IL-3 is not a direct agonist for basophil mediator release. However, preincubation of basophils with IL-3 enhances the degranulation response to C5a. Most importantly, IL-3 "primes" basophils to release large amounts of leukotriene C4 after challenge with C5a (mean of 50 gp LTC4 per nanograms cellular histamine), while neither peptide alone is capable of inducing the formation of bioactive lipids. This effect is dose dependent, occurring at IL-3 concentrations considerably lower than are required to stimulate the growth of bone marrow progenitor cells. IL-3 affects the extent but not the time course of basophil degranulation, and leukotriene release of cells sequentially exposed to IL-3 and C5a occurs very rapidly concomitant with degranulation. A preincubation of the basophils with IL-3 is strictly required for C5a-induced LTC4 synthesis, but not for an enhancement of degranulation. Priming for C5a-induced lipid mediator generation occurs rapidly after exposure of the cells to IL-3, starting at 1 min and reaching maximal effects at 5 min, but this altered state of responsiveness is relatively long lasting. Cell fractionation studies indicate that the basophil is the source of lipid mediators and that IL-3 affects the basophil response directly. This study demonstrates that IL-3 is a potent modifier of effector functions of mature basophils; this is possibly of greater in vivo significance than its growth factor properties. The large amounts of LTC4 formed after triggering of IL-3-primed basophils may not only enhance but also qualitatively change the pathophysiological consequences of complement activation, and this might be important in the pathogenesis of immediate type hypersensitivity reactions, shock syndromes, and inflammation.


Evolution of antimicrobial resistance among Enterobacteriaceae (focus on extended spectrum β-lactamases and carbapenemases).
Bacteria within the family Enterobacteriaceae are important pathogens in nosocomial and community settings. Over the past two decades, antimicrobial resistance among Enterobacteriaceae dramatically escalated worldwide. The authors review the mechanisms of antimicrobial resistance among Enterobacteriaceae, epidemiology and global spread of resistance elements and discuss therapeutic options. An exhaustive search for literature relating to Enterobacteriaceae was performed using PubMed, using the following key words: Enterobacteriaceae; Klebsiella pneumoniae; Escherichia coli; antimicrobial resistance; plasmids; global epidemiology; carbapenemases (CPEs); extended spectrum β-lactamases (ESBLs) and multidrug resistance (MDR). Enterobacteriaceae are inhabitants of intestinal flora and spread easily among humans (via hand carriage, contaminated food or water or environmental sources). Antimicrobial resistance may develop via plasmids, transposons or other mobile resistance elements. Mutations conferring resistance typically increase over time; the rate of increase is amplified by selection pressure from antibiotic use. Factors that enhance spread of antimicrobial resistance include: crowding; lack of hygiene; overuse and over-the-counter use of antibiotics; tourism; refugees and international travel. Clonal spread of resistant organisms among hospitals, geographic regions and continents has globally fueled the explosive rise in resistance. The emergence and widespread dissemination of MDR clones containing novel resistance elements (particularly ESBLs and CPEs) has greatly limited therapeutic options. In some cases, infections due to MDR Enterobacteriaceae are untreatable with existing antimicrobial agents. The authors discuss current and future therapeutic options for difficult-to-treat infections due to these organisms.


Evidence for a structural role for acid-fast lipids in oocyst walls of Cryptosporidium, Toxoplasma, and Eimeria.
Coccidia are protozoan parasites that cause significant human disease and are of major agricultural importance. Cryptosporidium spp. cause diarrhea in humans and animals, while Toxoplasma causes disseminated infections in fetuses and untreated AIDS patients. Eimeria is a major pathogen of commercial chickens. Oocysts, which are the infectious form of Cryptosporidium and Eimeria and one of two infectious forms of Toxoplasma (the other is tissue cysts in undercooked meat), have a multilayered wall. Recently we showed that the inner layer of the oocyst walls of Toxoplasma and Eimeria is a porous scaffold of fibers of β-1,3-glucan, which are also present in fungal walls but are absent from Cryptosporidium oocyst walls. Here we present evidence for a structural role for lipids in the oocyst walls of Cryptosporidium, Toxoplasma, and Eimeria. Briefly, oocyst walls of each organism label with acid-fast stains that bind to lipids in the walls of mycobacteria. Polyketide synthases similar to those that make mycobacterial wall lipids are abundant in oocysts of Toxoplasma and Eimeria and are predicted in Cryptosporidium. The outer layer of oocyst wall of Eimeria and the entire oocyst wall of Cryptosporidium are dissolved by organic solvents. Oocyst wall lipids are complex mixtures of triglycerides, some of which contain polyhydroxy fatty acyl chains like those present in plant cutin or elongated fatty acyl chains like mycolic acids. We propose a two-layered model of the oocyst wall (glucan and acid-fast lipids) that resembles the two-layered walls of mycobacteria (peptidoglycan and acid-fast lipids) and plants (cellulose and cutin). Oocysts, which are essential for the fecal-oral spread of coccidia, have a wall that is thought responsible for their survival in the environment and for their transit through the stomach and small intestine. While oocyst walls of Toxoplasma and Eimeria are strengthened by a porous scaffold of fibrils of β-1,3-glucan and by proteins cross-linked by dityrosines, both are absent from walls of Cryptosporidium. We show here that all oocyst walls are acid fast, have a rigid bilayer, dissolve in organic solvents, and contain a complex set of triglycerides rich in polyhydroxy and long fatty acyl chains that might be synthesized by an abundant polyketide synthase. These results suggest the possibility that coccidia build a waxy coat of acid-fast lipids in the oocyst wall that makes them resistant to environmental stress.


Leptin inhibits prehibernation hyperphagia and reduces body weight in arctic ground squirrels.
The ob gene product leptin is thought to play a physiological role in the fine tuning of a homeostatic mechanism regulating satiety and adiposity. Mouse recombinant leptin was administered to seasonally hyperphagic arctic ground squirrels as a first step in demonstrating the evolutionary conservation of leptin function and the potential involvement of leptin in the seasonal regulation of adiposity in hibernators. Continuous infusion of leptin for 3 wk via miniosmotic pumps resulted in a reduction in food intake and body weight in a manner consistent with its proposed role as a satiety hormone. During the recovery period after leptin administration, squirrels that had received leptin became hyperphagic relative to controls. Percent body fat was estimated at weekly intervals by measuring total body electrical conductivity and decreased after 3 wk of leptin administration. Our observations support the role of leptin as a regulatory hormone involved in the control of satiety, adiposity, and possibly energy expenditure in hibernating mammals.


Ethyl pyruvate ameliorates endotoxin-induced corneal inflammation.
The purpose of this study was to evaluate the anti-inflammatory effect of ethyl pyruvate (EP) in a mouse model of lipopolysaccharide (LPS)-induced corneal inflammation. LPS was injected intrastromally into the corneas of C57BL/6 mice followed by treatment with a solution of 2.5% EP in 0.2% hydroxypropyl methylcellulose (HPMC) every 90 minutes during the course of 12 hours. Prednisolone acetate 1% solution (PRED FORTE) was used as a positive control. Mice were sacrificed after 3 days, and corneas were examined by in vivo confocal microscopy and analyzed for infiltrated cells by flow cytometry. Gr-1, TNF-α, and pNF-κB-p65 were detected immunohistochemically, and TNF-α, IL-6, and IL-1β levels were quantified by ELISA. LPS-induced haze in mice corneas was decreased by 2-fold upon EP treatment; however, it was not changed upon PRED FORTE treatment. Flow cytometry and immunohistochemistry showed infiltration of leukocytes in the LPS-treated corneas; among the infiltrated cells, neutrophils (Gr-1+ and CD11b+) and macrophages (F4/80+ and CD11b+) were 3403.4- and 4.5-fold higher in number, respectively, than in vehicle-treated control corneas. EP or PRED FORTE treatment of LPS-injected corneas decreased the number of neutrophils 7.5- and 7.2-fold and macrophages by 5.6- and 3.5-fold, respectively. Both EP and PRED FORTE decreased TNF-α and IL-6 expression considerably, and to a lesser extent IL-1β expression, in the LPS-treated corneas. The present study demonstrated that EP reduces LPS-induced inflammation in the cornea and thus may have a potential therapeutic application in the inhibition of corneal inflammation.


Lost in the present but confident of the past: experiences of being in a psycho-geriatric unit as narrated by persons with dementia.
To illuminate meanings of being in a psycho-geriatric unit. Background. There are known risks associated with moving persons with dementia from home to an institution, but little is known about how they experience being in psycho-geriatric units. Using open-ended research interviews, six persons with mild to severe dementia were asked to narrate about their experiences in the hospital. The interviews were interpreted using a phenomenological hermeneutical method of analysis. The comprehensive understanding of being in a psycho-geriatric unit points towards an understanding of being lost in the present but confident of the past. The analysis showed that the participants appeared lost as they could not narrate where they were and why, but that they became confident when narrating about their previous life. The analysis also showed that being in the hospital meant sharing living space with strangers, invasions of private space and establishing new acquaintances. Being in the unit could also mean boredom and devaluation for participants. The interviews were interpreted in the light of narrative theory of identity: persons with dementia narrating about previous life experiences as to make claims of how to be interpreted by others; as persons instead of merely as 'demented' patients. Experiences of care narrated by persons with dementia present meaningful and useful information that can provide feedback to inform care practice. Experiences of care from persons with dementia provide meaningful information about care and the doing and being of staff. Creating time for conversations with these persons may facilitate well-being.


Genomic copy number alterations in 33 malignant peritoneal mesothelioma analyzed by comparative genomic hybridization array.
Malignant peritoneal mesotheliomas (MPM) are rare, accounting for approximately 8% of cases of mesothelioma in France. We performed comparative genomic hybridization (CGH) on frozen MPM samples using the Agilent Human Genome CGH 180 K array. Samples were taken from a total of 33 French patients, comprising 20 men and 13 women with a mean (range) age of 58.4 (17-76) years. Asbestos exposure was reported in 8 patients (24.2%). Median (range) overall survival (OS) was 39 (0-119) months. CGH analysis demonstrated the presence of chromosomal instability in patients with MPM, with a genomic pattern that was similar to that described for pleural mesothelioma, including the loss of chromosomal regions 3p21, 9p21, and 22q12. In addition, novel genomic copy number alterations were identified, including the 15q26.2 region and the 8p11.22 region. Median OS was associated with a low peritoneal cancer index (P=.011), epithelioid subtype (P=.038), and a low number of genomic aberrations (P=.015), all of which constitute good prognostic factors for MPM. Our results provide new insights into the genetic and genomic background of MPM. Although pleural and peritoneal mesotheliomas have different risk factors, different therapeutics, and different prognosis; these data provide support to combine pleural and peritoneal mesothelioma in same clinical assays.


Multivesicular droplets: a cell model system to study compartmentalised biochemical reactions.
Multivesicular vesicles (MVVs) are artificial liposomal structures widely used as a platform to study the compartmentalisation of cells and as a scaffold for artificial cell/protocell models. Current preparation techniques for MVVs, however, offer poor control on the size, lamellarity, and loading of inner lipid vesicles. Here, we introduce a microfluidic device for the production of multivesicular droplets (MVDs): a novel model system combining the ease of use and control of droplet microfluidics with the biological relevance of MVVs. We use a perfluorinated carrier phase with a biocompatible surfactant to generate monodisperse droplets of an aqueous giant unilamellar lipid vesicle suspension. The successful on-chip formation and stability of MVDs is verified through high-speed microscopy. For bright field or fluorescence microscopy inspection, the MVDs are trapped in an array where the integrity of both lipid vesicles and droplets is preserved for up to 15 minutes. Finally, we show a two-step enzymatic reaction that takes place across the lipid vesicle membranes; the second reaction step occurs in the vesicle's interior, where the enzyme is encapsulated, while both the substrate and fluorescent product permeate across the membrane. Our approach opens the possibility to mimic artificial organelles with optimised reaction parameters (pH, ions, etc.) in each compartment.


Unearthing a sesterterpene biosynthetic repertoire in the Brassicaceae through genome mining reveals convergent evolution.
Sesterterpenoids are a rare terpene class harboring untapped chemodiversity and bioactivities. Their structural diversity originates primarily from the scaffold-generating sesterterpene synthases (STSs). In fungi, all six known STSs are bifunctional, containing C-terminal trans-prenyltransferase (PT) and N-terminal terpene synthase (TPS) domains. In plants, two colocalized PT and TPS gene pairs from Arabidopsis thaliana were recently reported to synthesize sesterterpenes. However, the landscape of PT and TPS genes in plant genomes is unclear. Here, using a customized algorithm for systematically searching plant genomes, we reveal a suite of physically colocalized pairs of PT and TPS genes for the biosynthesis of a large sesterterpene repertoire in the wider Brassicaceae. Transient expression of seven TPSs from A. thaliana, Capsella rubella, and Brassica oleracea in Nicotiana benthamiana yielded fungal-type sesterterpenes with tri-, tetra-, and pentacyclic scaffolds, and notably (-)-ent-quiannulatene, an enantiomer of the fungal metabolite (+)-quiannulatene. Protein and structural modeling analysis identified an amino acid site implicated in structural diversification. Mutation of this site in one STS (AtTPS19) resulted in premature termination of carbocation intermediates and accumulation of bi-, tri-, and tetracyclic sesterterpenes, revealing the cyclization path for the pentacyclic sesterterpene (-)-retigeranin B. These structural and mechanistic insights, together with phylogenetic analysis, suggest convergent evolution of plant and fungal STSs, and also indicate that the colocalized PT-TPS gene pairs in the Brassicaceae may have originated from a common ancestral gene pair present before speciation. Our findings further provide opportunities for rapid discovery and production of sesterterpenes through metabolic and protein engineering.


Differential effects of stromal derived factor-1 alpha (SDF-1 alpha) on early and late stages of human megakaryocytic development.
Stromal derived factor-1 alpha (SDF-1 alpha), the high-affinity ligand of CXC-chemokine receptor 4 (CXCR4), was added to human CD34(+) hematopoietic progenitor cells that can be induced to differentiate along the monocytic or megakaryocytic lineages. In control liquid cell cultures supplemented with two different cytokine cocktails: stem cell factor (SCF), interleukin-3 (IL-3), macrophage-colony stimulating factor (M-CSF), and 10% fetal calf serum (FCS), or, SCF and thrombopoietin (TPO), the expression of surface CXCR4 progressively increased in both the CD14(+) monocytic and CD41(+) megakaryocytic lineages. While SDF-1 alpha caused only modest effects on cells of the monocytic lineage, it induced profound down-regulation of CXCR4 in megakaryocytic cells at all stages of differentiation. Moreover, while SDF-1 alpha initially up-regulated the early megakaryocytic antigen CD41, at later time points (days 12-16) it induced down-regulation of the late megakaryocytic antigen CD42b. Consistently, at day 16, the number of mature megakaryocytes was significantly decreased in cultures supplemented with SDF-1 alpha. These findings indicate that, besides its primary role in regulating the retention of precursor cells in hematopoietic tissues, the SDF-1 alpha/CXCR4 system participates in the regulation of megakaryocytic development by stimulating the formation of immature megakaryoblasts and inhibiting the formation of mature megakaryocytes.


Single vs. Multi-Joint Resistance Exercises: Effects on Muscle Strength and Hypertrophy.
Some authors suggest that single joint (SJ) exercises promote greater muscle hypertrophy because they are easier to be learned and therefore have less reliance on neural factors. On the other hand, some authors recommend an emphasis on multi-joint (MJ) exercises for maximizing muscle strength, assuming that MJ exercises are more effective than SJ exercises because they enable a greater magnitude of weight to be lifted. The present study aimed to compare the effects of MJ vs. SJ exercises on muscle size and strength gains in untrained young men. Twenty-nine young men, without prior resistance training experience, were randomly divided into two groups. One group performed (n = 14) only MJ exercises involving the elbow flexors (lat. pull downs), while the other (n = 15) trained the elbow flexors muscles using only SJ exercises (biceps curls). Both groups trained twice a week for a period of ten weeks. The volunteers were evaluated for peak torque of elbow flexors (PT) in an isokinetic dynamometer and for muscle thickness (MT) by ultrasonography. There were significant increases in MT of 6.10% and 5.83% for MJ and SJ, respectively; and there were also significant increases in PT for MJ (10.40%) and SJ (11.87%). However, the results showed no difference between groups pre or post training for MT or PT. In conclusion, the results of the present study suggest that MJ and SJ exercises are equally effective for promoting increases in upper body muscle strength and size in untrained men. Therefore, the selection between SJ and MJ exercises should be based on individual and practical aspects, such as, equipment availability, movement specificity, individual preferences and time commitment.


Ideating Mobile Health Behavioral Support for Compliance to Therapy for Patients with Chronic Disease: A Case Study of Atrial Fibrillation Management.
Poor patient compliance to therapy results in a worsening condition that often increases healthcare costs. In the MobiGuide project, we developed an evidence-based clinical decision-support system that delivered personalized reminders and recommendations to patients, helping to achieve higher therapy compliance. Yet compliance could still be improved and therefore building on the MobiGuide project experience, we designed a new component called the Motivational Patient Assistant (MPA) that is integrated within the MobiGuide architecture to further improve compliance. This component draws from psychological theories to provide behavioral support to improve patient engagement and thereby increasing patients' compliance. Behavior modification interventions are delivered via mobile technology at patients' home environments. Our approach was inspired by the IDEAS (Integrate, Design, Assess, and Share) framework for developing effective digital interventions to change health behavior; it goes beyond this approach by extending the Ideation phase' concepts into concrete backend architectural components and graphical user-interface designs that implement behavioral interventions. We describe in detail our ideation approach and how it was applied to design the user interface of MPA for anticoagulation therapy for the atrial fibrillation patients. We report results of a preliminary evaluation involving patients and care providers that shows the potential usefulness of the MPA for improving compliance to anticoagulation therapy.


Investigation of several virulence properties among Vibrio alginolyticus strains isolated from diseased cultured fish in Tunisia.
We analysed 34 Vibrio alginolyticus strains isolated from gilthead sea bream Sparus aurata L. and sea bass Dicentrarchus labrax L. cultured in fish farms on the Tunisian Mediterranean coast for the presence of several virulence properties such as extracellular products (ECP) production, growth in iron-limiting conditions and survival in fish serum. The results obtained with different substrates showed that ECP of V. alginolyticus were hydrolytic. The virulence was correlated with the ability of strains to grow in the presence of non-immune fish serum or under conditions of iron limitation. We further examined the presence of virulence genes homologous to those in V. cholerae (toxR, toxS, VPI and ace); toxR was found in 16 V. alginolyticus strains and toxS in 17 strains out of 34 analysed. A positive amplification for the virulence pathogenicity island (VPI) was produced by 12 V alginolyticus strains. Finally, the ace expected amplification fragment was found in 7 V. alginolyticus isolates. Thus, the pathogenicity of V. alginolyticus may be the result of a combination of all these factors.


"Bind, cleave and leave": multiple turnover catalysis of RNA cleavage by bulge-loop inducing supramolecular conjugates.
Antisense sequence-specific knockdown of pathogenic RNA offers opportunities to find new solutions for therapeutic treatments. However, to gain a desired therapeutic effect, the multiple turnover catalysis is critical to inactivate many copies of emerging RNA sequences, which is difficult to achieve without sacrificing the sequence-specificity of cleavage. Here, engineering two or three catalytic peptides into the bulge-loop inducing molecular framework of antisense oligonucleotides achieved catalytic turnover of targeted RNA. Different supramolecular configurations revealed that cleavage of the RNA backbone upon sequence-specific hybridization with the catalyst accelerated with increase in the number of catalytic guanidinium groups, with almost complete demolition of target RNA in 24 h. Multiple sequence-specific cuts at different locations within and around the bulge-loop facilitated release of the catalyst for subsequent attacks of at least 10 further RNA substrate copies, such that delivery of only a few catalytic molecules could be sufficient to maintain knockdown of typical RNA copy numbers. We have developed fluorescent assay and kinetic simulation tools to characterise how the limited availability of different targets and catalysts had restrained catalytic reaction progress considerably, and to inform how to accelerate the catalytic destruction of shorter linear and larger RNAs even further.


Changes-of-mind in the absence of new post-decision evidence.
Decisions are occasionally accompanied by changes-of-mind. While considered a hallmark of cognitive flexibility, the mechanisms underlying changes-of-mind remain elusive. Previous studies on perceptual decision making have focused on changes-of-mind that are primarily driven by the accumulation of additional noisy sensory evidence after the initial decision. In a motion discrimination task, we demonstrate that changes-of-mind can occur even in the absence of additional evidence after the initial decision. Unlike previous studies of changes-of-mind, the majority of changes-of-mind in our experiment occurred in trials with prolonged initial response times. This suggests a distinct mechanism underlying such changes. Using a neural circuit model of decision uncertainty and change-of-mind behaviour, we demonstrate that this phenomenon is associated with top-down signals mediated by an uncertainty-monitoring neural population. Such a mechanism is consistent with recent neurophysiological evidence showing a link between changes-of-mind and elevated top-down neural activity. Our model explains the long response times associated with changes-of-mind through high decision uncertainty levels in such trials, and accounts for the observed motor response trajectories. Overall, our work provides a computational framework that explains changes-of-mind in the absence of new post-decision evidence.


SOD1, ANG, VAPB, TARDBP, and FUS mutations in familial amyotrophic lateral sclerosis: genotype-phenotype correlations.
Mutations in SOD1, ANG, VAPB, TARDBP and FUS genes have been identified in amyotrophic lateral sclerosis (ALS). The relative contributions of the different mutations to ALS were estimated by systematically screening a cohort of 162 families enrolled in France and 500 controls (1000 chromosomes) using molecular analysis techniques and performing phenotype-genotype correlations. 31 pathogenic missense mutations were found in 36 patients (20 SOD1, 1 ANG, 1 VAPB, 7 TARDBP and 7 FUS). Surprisingly two FUS mutation carriers also harboured ANG variants. One family of Japanese origin with the P56S VAPB mutation was identified. Seven novel mutations (three in SOD1, two in TARDBP, two in FUS) were found. None of them was detected in controls. Segregation of detected mutations with the disease was confirmed in 11 families including five pedigrees carrying the novel mutations. Clinical comparison of SOD1, TARDBP, FUS and other familial ALS patients (with no mutation in the screened genes) revealed differences in site of onset (predominantly lower limbs for SOD1 and upper limbs for TARDBP mutations), age of onset (younger with FUS mutations), and in lifespan (shorter for FUS carriers). One third of SOD1 patients survived more than 7 years: these patients had earlier disease onset than those presenting with a more typical course. Differences were also observed among FUS mutations, with the R521H FUS mutation being associated with longer disease duration. This study identifies new genetic associations with ALS and provides phenotype-genotype correlations with both previously reported and novel mutations.


Investigation of Occupational Stress and its Relationship with the Demographic Characteristics of Workers in Ilam, Iran.
Workers are more susceptible to stress disorder for several reasons. The aim of this study was to investigate occupational stress among workers and other personnel at the ZardjinBaft Workshop in Ilam, Iran, and to examine the relationship between occupational stress and several parameters, such as age, gender, marital status, educational level, work experience, tobacco use, body mass index (BMI), and the type of work. This was a descriptive, cross-sectional study that was performed in June 2010. The sample population that was studied consisted of all of the workers and other personnel (130 people) at ZardjinBaft Workshop in Ilam, Iran. They were studied using Cooper's standard questionnaire for stress in the work environment, and we examined the relationship between stress, as the dependent variable, and the eight independent variables mentioned in the Abstract. In the sample population that was studied, severe stress was not observed in most of the sections of the workshop. In addition, using the analysis of variance test (ANOVA), no significant relationship was observed between the average degree of stress and the average demographic characteristics. However, the average stress level had significant relationships with the level of education, work experience, and the type of work, suggesting that stress increases as the level of education increases. In addition, in the beginning and early years of employment, medium levels of stress were observed, and the prevalence of stress decreased after the first 10 years of employment. In addition, workers in the ventilation section of the factory had the highest stress levels. The fear of losing their jobs was the main factor that caused stress among the workers at ZardjinBaft Workshop in Ilam. We suggest that managers consider the high importance of stress in the work environment. They should be able to recognize when workers are stressed and offer them assistance and training. They also can eliminate the factors that cause stress in the work environment by taking preventive action. In parallel, workers should try to interact well with their managers and talk to her or him about the conditions of their jobs.


Development of an optimised 1:1 physiotherapy intervention post first-time lumbar discectomy: a mixed-methods study.
To develop an optimised 1:1 physiotherapy intervention that reflects best practice, with flexibility to tailor management to individual patients, thereby ensuring patient-centred practice. Mixed-methods combining evidence synthesis, expert review and focus groups. Secondary care involving 5 UK specialist spinal centres. A purposive panel of clinical experts from the 5 spinal centres, comprising spinal surgeons, inpatient and outpatient physiotherapists, provided expert review of the draft intervention. Purposive samples of patients (n=10) and physiotherapists (n=10) (inpatient/outpatient physiotherapists managing patients with lumbar discectomy) were invited to participate in the focus groups at 1 spinal centre. A draft intervention developed from 2 systematic reviews; a survey of current practice and research related to stratified care was circulated to the panel of clinical experts. Lead physiotherapists collaborated with physiotherapy and surgeon colleagues to provide feedback that informed the intervention presented at 2 focus groups investigating acceptability to patients and physiotherapists. The focus groups were facilitated by an experienced facilitator, recorded in written and tape-recorded forms by an observer. Tape recordings were transcribed verbatim. Data analysis, conducted by 2 independent researchers, employed an iterative and constant comparative process of (1) initial descriptive coding to identify categories and subsequent themes, and (2) deeper, interpretive coding and thematic analysis enabling concepts to emerge and overarching pattern codes to be identified. The intervention reflected best available evidence and provided flexibility to ensure patient-centred care. The intervention comprised up to 8 sessions of 1:1 physiotherapy over 8 weeks, starting 4 weeks postsurgery. The intervention was acceptable to patients and physiotherapists. A rigorous process informed an optimised 1:1 physiotherapy intervention post-lumbar discectomy that reflects best practice. The developed intervention was agreed on by the 5 spinal centres for implementation in a randomised controlled trial to evaluate its effectiveness.


Differential diagnosis of bipolar disorder and major depressive disorder.
Patients with bipolar disorder spend approximately half of their lives symptomatic and the majority of that time suffering from symptoms of depression, which complicates the accurate diagnosis of bipolar disorder. Challenges in the differential diagnosis of bipolar disorder and major depressive disorder are reviewed, and the clinical utility of several screening instruments is evaluated. The estimated lifetime prevalence of major depressive disorder (i.e., unipolar depression) is over 3 and one-half times that of bipolar spectrum disorders. The clinical presentation of a major depressive episode in a bipolar disorder patient does not differ substantially from that of a patient with major depressive disorder (unipolar depression). Therefore, it is not surprising that without proper screening and comprehensive evaluation many patients with bipolar disorder may be misdiagnosed with major depressive disorder (unipolar depression). In general, antidepressants have demonstrated little or no efficacy for depressive episodes associated with bipolar disorder, and treatment guidelines recommend using antidepressants only as an adjunct to mood stabilizers for patients with bipolar disorder. Thus, correct identification of bipolar disorder among patients who present with depression is critical for providing appropriate treatment and improving patient outcomes. Clinical characteristics indicative of bipolar disorder versus major depressive disorder identified in this review are based on group differences and may not apply to each individual patient. The overview of demographic and clinical characteristics provided by this review may help medical professionals distinguish between major depressive disorder and bipolar disorder. Several validated, easily administered screening instruments are available and can greatly improve the recognition of bipolar disorder in patients with depression.


Characterization of Thermo-Physical Properties of EVA/ATH: Application to Gasification Experiments and Pyrolysis Modeling.
The pyrolysis of solid polymeric materials is a complex process that involves both chemical and physical phenomena such as phase transitions, chemical reactions, heat transfer, and mass transport of gaseous components. For modeling purposes, it is important to characterize and to quantify the properties driving those phenomena, especially in the case of flame-retarded materials. In this study, protocols have been developed to characterize the thermal conductivity and the heat capacity of an ethylene-vinyl acetate copolymer (EVA) flame retarded with aluminum tri-hydroxide (ATH). These properties were measured for the various species identified across the decomposition of the material. Namely, the thermal conductivity was found to decrease as a function of temperature before decomposition whereas the ceramic residue obtained after the decomposition at the steady state exhibits a thermal conductivity as low as 0.2 W/m/K. The heat capacity of the material was also investigated using both isothermal modulated Differential Scanning Calorimetry (DSC) and the standard method (ASTM E1269). It was shown that the final residue exhibits a similar behavior to alumina, which is consistent with the decomposition pathway of EVA/ATH. Besides, the two experimental approaches give similar results over the whole range of temperatures. Moreover, the optical properties before decomposition and the heat capacity of the decomposition gases were also analyzed. Those properties were then used as input data for a pyrolysis model in order to predict gasification experiments. Mass losses of gasification experiments were well predicted, thus validating the characterization of the thermo-physical properties of the material.


A protocol for simultaneous high-sensitivity genotyping and chromatin accessibility profiling in single cells.
Single-cell assay for transposase-accessible chromatin with sequencing (scATAC-seq) resolves the heterogeneity of epigenetic states across cells but does not typically capture exonic mutations, which limits our knowledge of how somatic mutations alter chromatin landscapes. Here, we present a plate-based approach coupling high-sensitivity genotyping of genomic loci with high-content scATAC-seq libraries from the same single cells. We first describe steps for optimization of genotyping primers, followed by detailed guidance on the preparation of both scATAC-seq and single-cell genotyping libraries, fully automated on high-throughput liquid handling platforms. For complete details on the use and execution of this protocol, please refer to Turkalj, Jakobsen et al.1.


Non-motor predictors of 36-month quality of life after subthalamic stimulation in Parkinson disease.
To identify predictors of 36-month follow-up quality of life (QoL) outcome after bilateral subthalamic nucleus deep brain stimulation (STN-DBS) in Parkinson's disease (PD). In this ongoing, prospective, multicenter international study (Cologne, Manchester, London) including 73 patients undergoing STN-DBS, we assessed the following scales preoperatively and at 6-month and 36-month follow-up: PD Questionnaire-8 (PDQ-8), NMSScale (NMSS), Scales for Outcomes in PD (SCOPA)-motor examination, -activities of daily living, and -complications, and levodopa equivalent daily dose (LEDD). We analyzed factors associated with QoL improvement at 36-month follow-up based on (1) correlations between baseline test scores and QoL improvement, (2) step-wise linear regressions with baseline test scores as independent and QoL improvement as dependent variables, (3) logistic regressions and receiver operating characteristic curves using a dichotomized variable "QoL responders"/"non-responders". At both follow-ups, NMSS total score, SCOPA-motor examination, and -complications improved and LEDD was reduced significantly. PDQ-8 improved at 6-month follow-up with subsequent decrements in gains at 36-month follow-up when 61.6% of patients were categorized as "QoL non-responders". Correlations, linear, and logistic regression analyses found greater PDQ-8 improvements in patients with younger age, worse PDQ-8, and worse specific NMS at baseline, such as 'difficulties experiencing pleasure' and 'problems sustaining concentration'. Baseline SCOPA scores were not associated with PDQ-8 changes. Our results provide evidence that 36-month QoL changes depend on baseline neuropsychological and neuropsychiatric non-motor symptoms burden. These findings highlight the need for an assessment of a wide range of non-motor and motor symptoms when advising and selecting individuals for DBS therapy.


Trends in net survival from liver cancer in six European Latin countries: results from the SUDCAN population-based study.
Liver cancer represents a major clinical challenge. The aim of the SUDCAN collaborative study was to compare the net survival from liver cancer between six European Latin countries (Belgium, France, Italy, Portugal, Spain and Switzerland) and provide trends in net survival and dynamics of excess mortality rates (EMRs) up to 5 years after diagnosis. The data were extracted from the EUROCARE-5 database. First, net survival was studied over the period 2000-2004 using the Pohar-Perme estimator. For trend analyses, the study period was specific to each country. Results are reported from 1992 to 2004 in France, Italy, Spain and Switzerland and from 2000 to 2004 in Belgium and Portugal. These trend analyses were carried out using a flexible excess-rate modeling strategy. There were little differences between the six countries in the 5-year age-standardized net survival (2000-2004): it ranged from 13% (France and Portugal) to 16% (Belgium). An increase in the net age-standardized survival was observed in all countries between 1992 and 2004, both at 1 year and at 5 years (the highest in Spain, the lowest in France). Generally, patients aged 60 years showed the highest increase. There was a progressive decrease in EMR over the 5-year- period following diagnosis. The study confirmed the poor prognosis of liver cancer. Innovative treatments might improve the prognosis as well as preventive screening of cirrhotic patients with good liver function. Efforts are also needed to improve registration practices.


Measurement and modeling of human T cell kinetics.
The ability to measure, describe and interpret T cell kinetics is pivotal in understanding normal lymphocyte homeostasis and diseases that affect T cell numbers. Following in vivo labeling of dividing cells with 6,6-D(2)-glucose in eight healthy volunteers, peripheral blood T cells were sorted by CD4, CD8 and CD45 phenotype. Enrichment of deuterium in DNA was measured by gas chromatography-mass spectrometry. A novel model of T cell kinetics, allowing for heterogeneity within T cell pools, was used to analyze data on acquisition and loss of label and calculate proliferation and disappearance rates for each subpopulation. Proliferation rates for CD45RO(+)CD8(+) cells and CD45RO(+)CD4(+) cells were 5.1% and 2.7% /day, respectively (equivalent doubling times: 14 and 26 days). CD45RA(+)CD8(+) lymphocytes and CD45RA(+)CD4(+) lymphocytes had slower proliferation rates, 0.5% and 0.6% / day, respectively (doubling time about 4 months). Disappearance rates of labeled cells were similar for all cell types (7%-12% / day) and exceeded corresponding proliferation rates. This disparity may be understood conceptually in terms of either phenotypic heterogeneity (rapid versus slow turnover pools), or history (recently divided cells are more likely to die). The new kinetic model fits the data closely and avoids the need to postulate a large external source of lymphocytes to maintain equilibrium.


Managing dive tourism for the sustainable use of coral reefs: validating diver perceptions of attractive site features.
It has been argued that strategies to manage natural areas important for tourism and recreation should integrate an understanding of tourist preferences for specific natural features. However, the accuracy of tourist recalled perceptions of environmental attributes, which are usually derived from post hoc surveys and used to establish management priorities, is currently unmeasured. We tested the validity of the relationship between tourist-stated preferences and actual condition of coral reefs around the Caribbean island of Bonaire. Using standardized questionnaires, we asked 200 divers to select their most and least favorite dive sites and the attributes that contributed to that selection. We also carried out ecological surveys at 76 of the 81 dives sites around the island to assess the actual conditions of the attributes indicated as important for site selection. Fish- and coral-related attributes were key features affecting dive enjoyment. In general, divers appeared to be able to perceive differences between sites in the true condition of biological attributes such as fish species richness, total number of fish schools, live coral cover, coral species richness, and reef structural complexity, although men and women divers differed in their ability to perceive/recall some of the attributes. Perceived differences in environmental attributes, such as surface conditions, underwater current, and the likelihood of encountering rare fish and sea turtles, were not empirically validated. The fact that divers perceive correctly differences in the condition of some of the key biological attributes that affect dive enjoyment reinforces the need to maintain overall reef condition at satisfactory levels. However, variation in accuracy of perceptions owing to demographic factors and attribute type suggests the need for caution when using public perceptions to develop environmental management strategies, particularly for coral reefs.


Patient satisfaction and use of Veterans Affairs versus non-Veterans Affairs healthcare services by veterans.
Chronically ill patients who are not satisfied with their care may change healthcare providers or systems, which could disrupt continuity of care and impede management of their conditions. We examined whether patient satisfaction affected subsequent use of non-Veterans Affairs (VA) services among chronically ill veterans discharged from VA hospitals. The data used in this study came from a multicenter trial of increased access to primary care. We enrolled patients with diabetes, heart failure, and/or chronic obstructive pulmonary disease who were discharged from 1 of 9 VA medical centers. At baseline, we assessed satisfaction using the Patient Satisfaction Questionnaire. VA and non-VA utilization over the subsequent 6 months were assessed using VA and Medicare administrative data, non-VA billing data, and patient interviews. Using multivariable logistic regression analyses, we examined whether baseline patient satisfaction was associated with non-VA inpatient or outpatient utilization during the next 6 months. We conducted the same analysis for Medicare-eligible veterans, a group with better access to non-VA care. Of 1375 study patients, 174 (13%) used non-VA healthcare. Patients with non-VA utilization were older and lived farther from a VA. The odds of non-VA use decreased by 11% as satisfaction increased (odds ratio 0.89; 95% confidence interval 0.83-0.97; P = 0.005). This relationship was strongest among Medicare-eligible veterans (odds ratio 0.85; 95% confidence interval 0.77-0.93; P = 0.001). Dissatisfied veterans discharged from the hospital were more likely to go outside VA for care. Thus, improvements in patient satisfaction may lead to improvements in continuity of care.


Goal-directed resuscitation for patients with early septic shock.
Early goal-directed therapy (EGDT) has been endorsed in the guidelines of the Surviving Sepsis Campaign as a key strategy to decrease mortality among patients presenting to the emergency department with septic shock. However, its effectiveness is uncertain. In this trial conducted at 51 centers (mostly in Australia or New Zealand), we randomly assigned patients presenting to the emergency department with early septic shock to receive either EGDT or usual care. The primary outcome was all-cause mortality within 90 days after randomization. Of the 1600 enrolled patients, 796 were assigned to the EGDT group and 804 to the usual-care group. Primary outcome data were available for more than 99% of the patients. Patients in the EGDT group received a larger mean (±SD) volume of intravenous fluids in the first 6 hours after randomization than did those in the usual-care group (1964±1415 ml vs. 1713±1401 ml) and were more likely to receive vasopressor infusions (66.6% vs. 57.8%), red-cell transfusions (13.6% vs. 7.0%), and dobutamine (15.4% vs. 2.6%) (P<0.001 for all comparisons). At 90 days after randomization, 147 deaths had occurred in the EGDT group and 150 had occurred in the usual-care group, for rates of death of 18.6% and 18.8%, respectively (absolute risk difference with EGDT vs. usual care, -0.3 percentage points; 95% confidence interval, -4.1 to 3.6; P=0.90). There was no significant difference in survival time, in-hospital mortality, duration of organ support, or length of hospital stay. In critically ill patients presenting to the emergency department with early septic shock, EGDT did not reduce all-cause mortality at 90 days. (Funded by the National Health and Medical Research Council of Australia and the Alfred Foundation; ARISE ClinicalTrials.gov number, NCT00975793.).


ASSESSING THE BENEFITS OF HAVING A SPECIALIST PAEDIATRIC PHARMACIST AND PHYSIOTHERAPIST IN THE COMMUNITY TO IMPROVE CHILDHOOD ASTHMA OUTCOMES.
The local Clinical Commissioning Group has funded an innovative one-year pilot project to assess the value of providing specialist paediatric pharmacist and physiotherapist support direct to families and health care professionals (GP's, community pharmacists, practice nurses etc.) regarding asthma in the primary care setting. Currently no such support is provided within community setting by physiotherapy or pharmacy. Joint holistic reviews by the clinical specialist physiotherapist and specialist paediatric pharmacist were performed in the patient's home environment or school. The review involved a thorough respiratory review and in-depth medication optimisation review ensuring patients were on appropriate regimes and using devices appropriately. Specifically, if an inhaler was indicated a device that the patient was comfortable using was chosen. Furthermore, parents, patients, teachers and school support workers were counselled on how to self manage asthma exacerbations. In order to review benefits patients answered the five question asthma control test (score out of 25), a standardised quality of life questionnaire (score out of 92) and hospital admissions were monitored. At the six-month stage of the project a total of 42 patients had been reviewed and followed up by the project. During the review period there was a total of 1 hospital admission and 1 attendance to the accident and emergency, this is in comparison to the 8 hospital admissions and 47 accident and emergency attendances with this group of patients in the same period the previous year. All patients had an improvement in outcome measures. The average improvement in asthma control test after intervention was 7 points (30%) and a 30% increase in QoL score. We found that symptomatic children had poor FEF25-75 values (<80%) indicating poor lower airways function possibly due to poor drug deposition. After interventions these scores returned to normal limits (>80%). Compliance to medications regimes was noted to be improved after optimisation. It can be clearly seen that joint multidisciplinary reviews by physiotherapy and pharmacy can help improve the outcomes of asthma patients. The joint review of inhaler technique in particular was key. Pharmacy services will tend to concentrate on the use of the device itself whereas physiotherapy monitor the strength and depth of breathes taken. It is well known that good drug deposition is key to the success of inhaled medications. By combining pharmacy and physiotherapy both the use of the device and breathing patterns were optimised both contributing to better drug deposition and improved FEF 25-75% reading. Optimisation of medication was also vital. By ensuring the patient was happy with the device they were using and that it had little negative impact on their daily routine compliance with medications was increased. At the six month stage of the pilot this aspect has proven vital to outcomes in asthmatic patients.


Video Analysis of Pectoralis Major Injuries in Professional Australian Football Players.
There is little evidence regarding the mechanisms of pectoralis major (PM) injury and player outcomes in Australian Football League (AFL) players. The study aims were to investigate (1) the mechanisms of PM muscle injury in elite AFL players via video analysis and (2) the player profile, method of management, and clinical outcomes of the PM injuries sustained. We hypothesized that the majority of PM tears would occur in outer-range PM positions (hyperextension of the glenohumeral joint). Case series; Level of evidence, 4. We analyzed video of the precipitating event for traumatic PM injuries during AFL competition or training over a 20-year period (2002-2021). The footage was analyzed by 4 experienced assessors, and the following were evaluated: mechanism of injury, injury variables (arm position, initial contact point, visual awareness, and use of taping), player characteristics (age at the time of injury, hand dominance, and history of injury), injury profile (location and size of tear), method of management (operative vs nonoperative), patient outcomes (time to return to full senior training/match play), and complication rates. The mean ± standard deviation age of the players was 26.5 ± 3.1 years (range, 21-32 years). Overall, 22 PM injuries were identified in the AFL injury database for a rate of 1.1 per year; 16 of these injuries had accompanying video footage. We identified 3 mechanisms for PM injury: horizontal hyperextension (62.5%), hyperflexion-abduction (25.0%), and horizontal adduction (sustained tackling; 12.5%). The most common site of the tear was the insertion point of the sternocostal head (91.0%). Twenty players (91.0%) required surgical repair, with 75% undergoing surgery within 1 week (range, 0-26 weeks). The mean return to competition for the surgical repair group was 11.1 weeks (range, 8-15 weeks). The rerupture rate was 5.0% (1 repair; <4 weeks postoperatively in 2004). PM tears in elite male AFL players were due to 1 of 3 distinct mechanisms: horizontal hyperextension, hyperflexion-abduction, and horizontal adduction (sustained tackling). Players returned to play on average 11 weeks after injury. Knowledge regarding mechanisms of injury, player profile, and return-to-sport timelines is important for appropriate medical management and provides potential areas to target for prevention of PM injuries.


The effect of referral to an open-group behavioural weight-management programme on the relative risk of normoglycaemia, non-diabetic hyperglycaemia and type 2 diabetes: Secondary analysis of the WRAP trial.
To examine the impact of open-group behavioural weight-management programmes on the risk of diabetes among those with a body mass index (BMI) of ≥28 kg/m2 and those with non-diabetic hyperglycaemia (NDH). This was a secondary analysis of data from the WRAP trial, in which participants (N = 1267; aged ≥18 years, BMI ≥ 28 kg/m2 ) were randomized to brief intervention (BI; self-help booklet), a weight-management programme (WW; formerly Weight Watchers) for 12 weeks, or WW for 52 weeks. We used multinomial logistic regression to examine the effect of intervention group on the risk of hyperglycaemia and diabetes at 12 months in all participants with glycaemic status at both time points (N = 480; 38%) and those with NDH at baseline (N = 387; 31%). We used mixed effects models and linear fixed effects models to examine the effect of intervention group on body weight and HbA1c at 12 months in people with NDH. There was a 61% relative reduction in the risk of NDH at the 12-month follow-up (12 weeks vs. BI: relative risk ratio [RRR] = 0.39 [95% CI 0.18, 0.87], P = .021; 52 weeks vs. BI: RRR = 0.38 [95% CI 0.17, 0.86], P = .020). For intervention effects on the risk of diabetes, confidence intervals were wide and overlapped 1 [12 weeks vs. BI: RRR = 0.49 [95% CI 0.12, 1.96], P = .312; 52 weeks vs. BI: RRR = 0.40 [95% CI 0.10, 1.63], P = .199). Participants with hyperglycaemia at baseline in the weight-management programme were more probable to have normoglycaemia at the 12-month follow-up [12-week programme vs. BI: RRR = 3.57 [95% CI 1.24, 10.29], P = .019; 52-week programme vs. BI: RRR = 4.14 [95% CI 1.42, 12.12], P = .009). Open-group behavioural weight-management programmes can help to prevent the development of NDH in people with overweight and obesity and to normalize glycaemia in people with NDH.


Discovery of Novel Oral Protein Synthesis Inhibitors of Mycobacterium tuberculosis That Target Leucyl-tRNA Synthetase.
The recent development and spread of extensively drug-resistant and totally drug-resistant resistant (TDR) strains of Mycobacterium tuberculosis highlight the need for new antitubercular drugs. Protein synthesis inhibitors have played an important role in the treatment of tuberculosis (TB) starting with the inclusion of streptomycin in the first combination therapies. Although parenteral aminoglycosides are a key component of therapy for multidrug-resistant TB, the oxazolidinone linezolid is the only orally available protein synthesis inhibitor that is effective against TB. Here, we show that small-molecule inhibitors of aminoacyl-tRNA synthetases (AARSs), which are known to be excellent antibacterial protein synthesis targets, are orally bioavailable and effective against M. tuberculosis in TB mouse infection models. We applied the oxaborole tRNA-trapping (OBORT) mechanism, which was first developed to target fungal cytoplasmic leucyl-tRNA synthetase (LeuRS), to M. tuberculosis LeuRS. X-ray crystallography was used to guide the design of LeuRS inhibitors that have good biochemical potency and excellent whole-cell activity against M. tuberculosis Importantly, their good oral bioavailability translates into in vivo efficacy in both the acute and chronic mouse models of TB with potency comparable to that of the frontline drug isoniazid.


Autoimmune disease aggregation in families with primary Sjögren's syndrome.
Diverse autoimmune diseases may coexist in the same individual and in families, implying a common etiology. We examined the aggregation of autoimmune diseases among first-degree relatives (FDR) of patients with primary Sjögren's syndrome (pSS). This was a population-based case-control family study in which 101 families of women classified as having pSS according to the revised American-European criteria and 124 families of matched controls without autoimmune disease were enrolled to investigate the presence of autoimmune diseases. We performed a genetic analysis that included familial correlation and recurrent risk ratios. In family cases, 38% had at least one FDR with an autoimmune disease, versus 22% in control families [odds ratio (OR) 2.2, 95% confidence interval (CI) 1.2-3.9, p = 0.01]. An autoimmune disease was registered for 7.3% of 876 patients' FDR as compared with 3.85% of 857 controls' FDR (OR 1.97, 95% CI 1.28-3.03, p = 0.002). The most frequent autoimmune diseases registered among the pSS patients' FDR were autoimmune thyroid disease (AITD), systemic lupus erythematosus, and rheumatoid arthritis, which disclosed aggregation. The proband phenotype (i.e., pSS) was correlated with AITD, systemic sclerosis, and all autoimmune diseases when considered together as a trait. Maternal transmission of the autoimmunity trait was observed in cases but not in controls. Our results indicate that autoimmune diseases cluster within families of patients with pSS. This familial aggregation of autoimmune diseases adds further evidence that clinically different autoimmune phenotypes might share common susceptibility gene variants, which acting in epistatic pleitropy may represent risk factors for autoimmunity.


Cross-sectional survey of users of Internet depression communities.
Internet-based depression communities provide a forum for individuals to communicate and share information and ideas. There has been little research into the health status and other characteristics of users of these communities. Online cross-sectional survey of Internet depression communities to identify depressive morbidity among users of Internet depression communities in six European countries; to investigate whether users were in contact with health services and receiving treatment; and to identify user perceived effects of the communities. Major depression was highly prevalent among respondents (varying by country from 40% to 64%). Forty-nine percent of users meeting criteria for major depression were not receiving treatment, and 35% had no consultation with health services in the previous year. Thirty-six percent of repeat community users who had consulted a health professional in the previous year felt that the Internet community had been an important factor in deciding to seek professional help. There are high levels of untreated and undiagnosed depression in users of Internet depression communities. This group represents a target for intervention. Internet communities can provide information and support for stigmatizing conditions that inhibit more traditional modes of information seeking.


Effect of obesity on left ventricular function studied by radionuclide angiocardiography.
Several studies have shown a significant association of obesity with cardiovascular morbidity and mortality. The present study was carried out to investigate central and systemic haemodynamics in overweight and moderate obese, but otherwise healthy subjects, and in a lean control group to determine whether obesity can influence left ventricular performance per se. In this study an attempt has been made to eliminate misleading factors, such as diabetes, lipid abnormalities and hypertension. A total of 67 subjects, 44 with overweight or moderate obesity and 23 lean healthy subjects, were included. Patients were divided into three groups according to BMI levels and Garrow's criteria as follows: lean control group (BMI less than 25 kg/m2); overweight (BMI from 25 to 30 kg/m2); moderate obese (BMI greater than 30 less than 40 kg/m2). Overweight and moderate obese subjects were further subgrouped according to duration of obesity (DO) in subgroup A (DO less than 98 months) and in subgroup B (DO greater than 98 months). Haemodynamic assessment was performed using first pass radionuclide angiocardiography. When compared with lean subjects, overweight and moderate obese subjects were characterized by a significant increase in cardiac output (CO), stroke volume (SV), end diastolic volume (EDV), end systolic volume (ESV), total blood volume (TBV) and total plasma volume (TPV) and by a significant decrease in left ventricular ejection fraction (EF); some of these changes appeared to be related to the degree of obesity. In overweight and moderate obese subjects, total peripheral resistance (TPR) was lower than in lean controls, but this difference was not significant.(ABSTRACT TRUNCATED AT 250 WORDS)


Sorption of arsenic(V) and arsenic(III) to schwertmannite.
This study describes the sorption of As(V) and As(III) to schwertmannite as a function of pH and arsenic loading. In general, sorption of As(V) was greatest at low pH, whereas high pH favored the sorption of As(III). The actual pH of equivalent As(V) and As(III) sorption was strongly loading dependent, decreasing from pH approximately 8.0 at loadings <120 mmol(As) mol(Fe)(-1) to pH approximately 4.6 at a loading of 380 mmol(As) mol(Fe)(-1). Sorption isotherms for As(V) were characterized by strong partitioning to the schwertmannite solid-phase at low loadings and sorption capacities of 225-330 mmol(As(V)) mol(Fe)(-1) at high loadings. In contrast, the As(III) isotherms revealed a weak affinity for sorption of As(III) versus As(V) at low loadings yet a greater affinity for As(III) sorption compared with As(V) at high loadings (when pH > 4.6). Sorption of As(V) and As(III) caused significant release of SO(4)(2-) from within the schwertmannite solid-phase, without major degradation of the schwertmannite structure (as evident by X-ray diffraction and Raman spectroscopy). This can be interpreted as arsenic sorption via incorporation into the schwertmannite structure, rather than merely surface complexation at the mineral-water interface. The results of this study have important implications for arsenic mobility in the presence of schwertmannite, such as in areas affected by acid-mine drainage and acid-sulfate soils. In particular, arsenic speciation, arsenic loading, and pH should be considered when predicting and managing arsenic mobility in schwertmannite-rich systems.


Increasing river discharge to the Arctic Ocean.
Synthesis of river-monitoring data reveals that the average annual discharge of fresh water from the six largest Eurasian rivers to the Arctic Ocean increased by 7% from 1936 to 1999. The average annual rate of increase was 2.0 +/- 0.7 cubic kilometers per year. Consequently, average annual discharge from the six rivers is now about 128 cubic kilometers per year greater than it was when routine measurements of discharge began. Discharge was correlated with changes in both the North Atlantic Oscillation and global mean surface air temperature. The observed large-scale change in freshwater flux has potentially important implications for ocean circulation and climate.


Viable cytophaga-like bacterium in the 0.2 microm-filtrate seawater.
A strain of the Cytophaga-like bacterium (CLB), Nano-1, was isolated from the 0.2 microm-filtrate of natural seawater. Both cellular fatty acid and 16S rDNA sequence analyses indicated that Nano-1 is closely affiliated to the marine gliding CLB genus, Microscilla. Nano-1 was observed to undergo cyclic morphological change typical of the genus Microscilla, and sub-0.2-microm cells were formed in the late stationary phase. The sub-0.2-microm cells were repeatedly revived and subcultured. Formation of the sub-0.2-microm cells seems to be adaptive for oligotrophic growth and starvation survival.


The potential impact of coinfection on antimicrobial chemotherapy and drug resistance.
Across a range of pathogens, resistance to chemotherapy is a growing problem in both public health and animal health. Despite the ubiquity of coinfection, and its potential effects on within-host biology, the role played by coinfecting pathogens on the evolution of resistance and efficacy of antimicrobial chemotherapy is rarely considered. In this review, we provide an overview of the mechanisms of interaction of coinfecting pathogens, ranging from immune modulation and resource modulation, to drug interactions. We discuss their potential implications for the evolution of resistance, providing evidence in the rare cases where it is available. Overall, our review indicates that the impact of coinfection has the potential to be considerable, suggesting that this should be taken into account when designing antimicrobial drug treatments.


Renal glomerular atrial natriuretic factor receptors in one-kidney, one clip rats.
One-kidney, one clip (1K1C) hypertension is often associated with an expanded plasma volume and (once the arterial clip is removed) with natriuresis. Blood pressure (BP), atrial and plasma concentrations of atrial natriuretic factor (ANF), hematocrit, and renal glomerular ANF receptors were therefore studied in 1K1C rats and in their normotensive uninephrectomized controls before and after unclipping. Six hours after removal of the clip, BP was normal in the 1K1C group and plasma ANF presented a sharp decline but was still significantly higher than in the normotensive controls, with a slight difference being evident 24 hours after unclipping. Hematocrit was lower in the 1K1C rats than in their control counterparts, but this difference tended to disappear once the clip was removed, indicating a contraction of plasma volume in these unclipped 1K1C animals. The renal glomerular ANF receptor population was markedly smaller in 1K1C rats than in the uninephrectomized controls but showed a twofold increase in number and affinity 24 hours after unclipping. It is concluded that the up-regulation and enhanced affinity of glomerular ANF receptors (probably secondary to the decrease in plasma levels of ANF) may contribute to the natriuresis reported in hypertensive 1K1C animals on removal of the arterial clip.


Haemodynamic and neurohumoral changes in spontaneously hypertensive rats with aortocaval fistulae.
1. Our aim was to evaluate the effects of an aortocaval fistula (1 mm) on cardiorenal haemodynamics, cardiac hypertrophy and neurohumoral factors in spontaneously hypertensive rats and to compare the results with those observed in Wistar rats at 2 weeks after fistulae placement. Sham-operated spontaneously hypertensive rats and Wistar rats served as controls. 2. Heart weight was significantly increased in spontaneously hypertensive rats (34%) and in Wistar rats (43%) at 2 weeks after fistula creation. Left ventricular systolic pressure and dp/dtmax. were significantly decreased (both P < 0.01) in spontaneously hypertensive rats with fistulae which had higher left ventricular end-diastolic pressure than Wistar rats with fistulae (P < 0.01). Signs of circulatory congestion (ascites, tachypnoea, prostration) were observed only in the overloaded spontaneously hypertensive rats (45%). Cardiac index was comparably increased in both fistulae groups due to an increase in stroke index, since heart rate was not increased. 3. Fistulae placement decreased renal blood flow and kidney weight, and increased blood urea nitrogen to a greater degree in spontaneously hypertensive rats (all P < 0.05); serum creatinine levels were unaltered. Plasma noradrenaline concentration was increased in spontaneously hypertensive rats with fistulae (P < 0.05), whereas plasma renin activity was not changed. 4. Thus, spontaneously hypertensive rats with fistulae developed overt haemodynamic signs of high-output heart failure with frequent ascites and dyspnoea, whereas most of these findings were milder or absent in Wistar rats. This model provides an opportunity to evaluate the pathophysiological and pharmacological responses in high-output heart failure.


Quality assurance for the diagnostics of viral diseases to enhance the emergency preparedness in Europe.
The threat posed by emerging and re-emerging communicable diseases and, more recently, by the intentional release of infectious agents in a susceptible population, has been receiving considerable attention at the national and international levels. Public health efforts to strengthen disease detection, surveillance and control have been intensified. However, clinicians and clinical microbiology laboratories play an important role in the early detection of disease, the identification of the putative agent, and notification of the appropriate authorities. To be effective in this role, laboratories must be specially prepared to handle viral agents safely, and need, among other things, the appropriate rapid and sensitive diagnostic tests. In 1998 the European Network for Diagnostics of 'Imported' Viral Diseases (ENIVD) was established. ENIVD presently comprises, as permanent members, 44 expert laboratories in 21 European Union (EU) member states and 4 non-EU countries and is one of the networks on infectious diseases funded by the European Commission. ENIVD fulfils many of the important tasks required for the surveillance and control of imported, rare and emerging viral infections such as the exchange of expertise and the organisation of external quality assurance (EQA) programmes, both of which are needed to improve diagnostics. Here, we summarise the data generated by recent EQA activities focussed on the diagnostics of infections with hantavirus, dengue virus, filovirus, Lassa virus, orthopox virus and the SARS-coronavirus (SARS-CoV). These were carried out between 1999 and 2004 and involved 93 laboratories from 41 countries, including laboratories from additional countries outside of Europe. Particularly the EU-candidate countries and Eastern neighbouring countries will be invited to join the network in the near future. A public website is available at http://www.enivd.de.


Efficacy of pulsed dye laser plus topical tazarotene versus topical tazarotene alone in psoriatic nail disease: a single-blind, intrapatient left-to-right controlled study.
Nail psoriasis is difficult to treat. The efficacy of pulsed dye laser (PDL) therapy for nail psoriasis has been reported in non-placebo controlled studies. To evaluate the efficacy and safety of PDL with topical retinoid in the treatment of nail psoriasis. The study was an intrapatient, left-to-right controlled trial of PDL in 25 patients with recalcitrant, bilateral fingernail psoriasis recruited between February 2011 and August 2011. We included two groups of patients: (1) patients with severe psoriasis who were receiving stable systemic therapy (phototherapy and systemic medication); (2) patients with mild psoriasis who were not receiving systemic therapy. One hand received the experimental treatment (PDL and tazarotene 0.1% cream) and the other, the control treatment (tazarotene 0.1% cream). All five fingernails of the experimental hand were treated with 595-nm PDL once a month for 6 months. Assessments included the modified Nail Psoriasis Severity Index (NAPSI) at baseline, 3, and 6 months, the Physician's Global Assessment at 3 and 6 months, the patient's global assessment, and adverse events. Nineteen patients completed the 6-month protocol. The mean decrease in modified NAPSI score from baseline to 6 months was significantly more after the experimental treatment than the control treatment. Physician's global assessment showed significantly higher percentage of patients had ≥75% improvement at 6 months in the experimental group than the control group (31.6% vs. 5.3%, P = 0.045). Scores on the patient's global assessment were significantly higher in the experimental group than the control group (P < 0.001). PDL plus topical tazarotene 0.1% cream is an effective and safe therapy in the treatment of nail psoriasis.


Impact of climate change on health: what is required of climate modellers?
The potential impacts of climate change on human health are significant, ranging from direct effects such as heat stress and flooding, to indirect influences including changes in disease transmission and malnutrition in response to increased competition for crop and water resources. Development agencies and policy makers tasked with implementing adaptive strategies recognize the need to plan for these impacts. However at present there is little guidance on how to prioritize their funding to best improve the resilience of vulnerable communities. Here we address this issue by arguing that closer collaboration between the climate modelling and health communities is required to provide the focused information necessary to best inform policy makers. The immediate requirement is to create multidisciplinary research teams bringing together skills in both climate and health modelling. This will enable considerable information exchange, and closer collaboration will highlight current uncertainties and hopefully routes to their reduction. We recognize that climate is only one aspect influencing the highly complex behaviour of health and disease issues. However we are optimistic that climate-health model simulations, including uncertainty bounds, will provide much needed estimates of the likely impacts of climate change on human health.


Morphological and cellular changes in secondary epidermal laminae of horses with insulin-induced laminitis.
To determine cellular changes associated with secondary epidermal laminae (SEL) in forefeet and hind feet of ponies with insulin-induced laminitis. 8 ponies. Laminitis was induced in 4 ponies by IV administration of insulin and glucose; 4 control ponies received saline (0.9% NaCl) solution IV. Laminar tissue samples obtained from the dorsal aspects of the hooves were histologically evaluated. Primary epidermal lamina (PEL) length and width and SEL length, width, and angle were determined. Numbers of epidermal cell nuclei per micrometer and per total length of SEL and numbers of apoptotic and proliferative cells in axial, middle, and abaxial laminar regions were determined. SEL in treatment group ponies were significantly longer, were significantly narrower, and had a smaller angle relative to PEL in all laminar regions versus control ponies. In treatment group ponies, the number of epidermal cell nuclei per SEL was typically higher and the number of cells per micrometer of SEL was lower in laminar regions, apoptotic cell numbers were higher in abaxial and middle regions in forefeet and hind feet, and proliferating cell numbers were higher in axial laminar regions in forefeet and all laminar regions in hind feet, versus control ponies. Results indicated SEL elongation, narrowing, and alteration in orientation developed in all feet of ponies with insulin-induced laminitis. This was primarily attributable to cell stretching that developed at the same time as an accelerated cell death-proliferation cycle; differences in cell cycle responses among laminar regions between forefeet and hind feet may have been attributable to differences in load bearing.


Control of tissue size and development by a regulatory element in the yorkie 3'UTR.
Regulation of the Hippo pathway via phosphorylation of Yorkie (Yki), the Drosophila homolog of human Yes-associated protein 1, is conserved from Drosophila to humans. Overexpression of a non-phosphorylatable form of Yki induces severe overgrowth in adult fly eyes. Here, we show that yki mRNA associates with microsomal fractions and forms foci that partially colocalize to processing bodies in the vicinity of endoplasmic reticulum. This localization is dependent on a stem-loop (SL) structure in the 3' untranslated region of yki. Surprisingly, expression of SL deleted yki in eye imaginal discs also results in severe overgrowth phenotypes. When the structure of the SL is disrupted, Yki protein levels increase without a significant effect on RNA levels. When the SL is completely removed, protein levels drastically increase, but in this case, due to increased RNA stability. In the latter case, we show that the increased RNA accumulation is due to removal of a putative miR-8 seed sequence in the SL. These data demonstrate the function of two novel regulatory mechanisms, both controlled by the yki SL element, that are essential for proper Hippo pathway mediated growth regulation.


Biochemical and biophysical properties of recombinant human interphotoreceptor retinoid binding protein.
Interphotoreceptor retinoid-binding protein (IRBP) binds and transports retinoids and fatty acids in the interphotoreceptor space (IPS). To understand the relationship between the protein structure and its functions requires bulk quantities of human IRBP. The authors sought to produce recombinant human IRBP (rhIRBP), a perfect duplicate in amino acid sequence of the authentic human protein. This material could serve as a supply of the protein and later could be used to make mutants of the protein. The goals of the present study were to produce human IRBP in an expression system and to examine some of its biochemical properties. A cDNA encoding human IRBP was cloned into the transplacement vector, pVL1392, and the plasmid was recombined with linearized baculovirus on cotransfection into Sf9 cells. Viruses containing the human IRBP cDNA were identified by polymerase chain reaction analysis. IRBP was secreted from virus-infected insect cells. rhIRBP was purified from cell medium and was examined by chromatography, N-terminal protein sequencing, immunologic techniques, and fluorometry. Eyecup and retina washes of human donor eyes provided a source of authentic human IRBP (IPS-IRBP). rhIRBP and IPS-IRBP exhibit similar elution profiles on concanavalin A, ion-exchange, and size exclusion chromatography. rhIRBP contains a five-amino-acid propeptide at the N-terminus as deduced from the cDNA sequence. Retinol binding of rhIRBP has been characterized by fluorometric titration. The dissociation constant is approximately 1.04 microM, close to that reported for bovine IRBP. By scanning fluorometry, the emission and excitation maxima are 479 nm and 339 nm, respectively. The baculovirus system provides an excellent method to produce and secrete human IRBP. The recombinant protein can be readily purified from cell culture medium. Its behavior in chromatography and in binding studies suggests that the recombinant protein is virtually identical to the authentic protein. This validates its use in place of IRBP from human donor eyes. Small, but significant, differences in biochemical properties in comparing human and bovine material highlight the significance of studying the human protein.


Aerosol antibiotic therapy in children with chronic upper airway infections: a potential alternative to surgery.
Tonsillectomy and adenoidectomy remain the first choice treatment of chronic or recurrent acute infections of the upper respiratory tract in children. The aim of this study is to investigate the efficacy of the combination of thiamphenicol glycinate acetylcysteinate plus beclomethasone, administered as aerosol, in children awaiting tonsillectomy and/or adenoidectomy. The study comprised 204 children, aged 1 to 12 years, with chronic adenotonsillitis who had been listed for surgery due to obstructive symptoms and recurrent acute infections. Patients were randomized to treatment with thiamphenicol glycinate acetylcysteinate, dosage 250 mg/day in 2 administrations plus beclomethasone with a dosage of 400 microg/day in 2 administrations, or no treatment, control group, unless required. The drugs were administered by aerosol for 10 days/month over a period of 6 months. Clinical visits were at 4, 7 and 12 months after the start of treatment. The primary efficacy outcome was the reduction in the number of patients requiring surgery. Secondary efficacy measures were the reduction of nasal obstruction, the decrease in the number of infectious episodes and the tolerability of the treatment. Aerosol treatment with thiamphenicol glycinate acetylcysteinate plus beclomethasone resulted in a significantly lower proportion of patients requiring surgery (29 of 101; 29 percent) compared to patients in the control group (100 of 103; 97 percent) (p < 0.0001). Treatment was also associated with a reduction of nasal obstruction and a decrease in the number of infectious episodes. No treatment-related adverse events were reported and the aerosol therapy proved easy to administer to children. The aerosol therapy with the combination of thiamphenicol glycinate acetylcysteinate plus beclomethasone was able to prevent or postpone surgery in a substantial percentage of patients, without adverse events. These preliminary results suggest that this novel approach could play a role in the antibiotic prophylaxis of chronic infectious diseases of the upper airways.


Structural basis of tRNA agmatinylation essential for AUA codon decoding.
The cytidine at the first position of the anticodon (C34) in the AUA codon-specific archaeal tRNA(Ile2) is modified to 2-agmatinylcytidine (agm(2)C or agmatidine), an agmatine-conjugated cytidine derivative, which is crucial for the precise decoding of the genetic code. Agm(2)C is synthesized by tRNA(Ile)-agm(2)C synthetase (TiaS) in an ATP-dependent manner. Here we present the crystal structures of the Archaeoglobus fulgidus TiaS-tRNA(Ile2) complexed with ATP, or with AMPCPP and agmatine, revealing a previously unknown kinase module required for activating C34 by phosphorylation, and showing the molecular mechanism by which TiaS discriminates between tRNA(Ile2) and tRNA(Met). In the TiaS-tRNA(Ile2)-ATP complex, C34 is trapped within a pocket far away from the ATP-binding site. In the agmatine-containing crystals, C34 is located near the AMPCPP γ-phosphate in the kinase module, demonstrating that agmatine is essential for placing C34 in the active site. These observations also provide the structural dynamics for agm(2)C formation.


Immune responses following the first dose of the Sputnik V (Gam-COVID-Vac).
As the first dose of Gam-COVID-Vac, is currently used as a single dose vaccine in some countries, we investigated the immunogenicity of this at 4 weeks (327 naïve individuals). 88.7% seroconverted, with significantly lower seroconversion rates in those over 60 years (p = 0.004) and significantly lower than previously seen with AZD1222 (p = 0.018). 82.6% developed ACE2 receptor blocking antibodies, although levels were significantly lower than following natural infection (p = 0.0009) and a single dose of AZD1222 (p < 0.0001). Similar titres of antibodies were observed to the receptor binding domain of WT, B.1.1.7 and B.1.617.2 compared to AZD1222, while the levels for B.1.351 were significantly higher (p = 0.006) for Gam-COVID-Vac. 30% developed ex vivo IFNγ ELISpot responses (significantly lower than AZD1222), and high frequency of CD107a expressing T cells along with memory B cell responses. Although single dose of Gam-COVID-Vac was highly immunogenic, administration of a second dose is likely to be beneficial.


Alfapump® system vs. large volume paracentesis for refractory ascites: A multicenter randomized controlled study.
Patients with refractory ascites (RA) require repeated large volume paracenteses (LVP), which involves frequent hospital visits and is associated with a poor quality-of-life. This study assessed safety and efficacy of an automated, low-flow pump (alfapump® [AP]) compared with LVP standard of care [SoC]. A randomized controlled trial, in seven centers, with six month patient observation was conducted. Primary outcome was time to first LVP. Secondary outcomes included paracentesis requirement, safety, health-related quality-of-life (HRQoL), and survival. Nutrition, hemodynamics, and renal injury biomarkers were assessed in a sub-study at three months. Sixty patients were randomized and 58 were analyzed (27 AP, 31 SoC, mean age 61.9years, mean MELD 11.7). Eighteen patients were included in the sub-study. Compared with SoC, median time to first LVP was not reached after six months in the AP group, meaning a significant reduction in LVP requirement for the AP patients (AP, median not reached; SoC, 15.0days (HR 0.13; 95%CI 13.0-22.0; p<0.001), and AP patients also showed significantly improved Chronic Liver Disease Questionnaire (CLDQ) scores compared with SoC patients (p<0.05 between treatment arms). Improvements in nutritional parameters were observed for hand-grip strength (p=0.044) and body mass index (p<0.001) in the sub-study. Compared with SoC, more AP patients reported adverse events (AEs; 96.3% vs. 77.4%, p=0.057) and serious AEs (85.2 vs. 45.2%, p=0.002). AEs consisted predominantly of acute kidney injury in the immediate post-operative period, and re-intervention for pump related issues, and were treatable in most cases. Survival was similar in AP and SoC. The AP system is effective for reducing the need for paracentesis and improving quality of life in cirrhotic patients with RA. Although the frequency of SAEs (and by inference hospitalizations) was significantly higher in the AP group, they were generally limited and did not impact survival. Lay summary: The alfapump® moves abdominal fluid into the bladder from where it is then removed by urination. Compared with standard treatment, the alfapump reduces the need for large volume paracentesis (manual fluid removal by needle) in patients with medically untreatable ascites. This can improve life quality for these patients. www.clinicaltrials.gov#NCT01528410.


Immunohistochemical analysis of clear cell carcinoma of the gynecologic tract.
Clear cell carcinoma of the gynecologic tract has been defined in terms of its clinical and histologic features; however, its immunophenotypic profile has not been fully characterized. Seventeen cases of primary clear cell carcinoma from various sites within the female genital tract (11 ovary, 5 uterus, 1 vagina) were analyzed by immunohistochemistry. These tumors were assessed for the expression of cytokeratin 7 (CK7), cytokeratin 20 (CK20), low and high molecular weight cytokeratin, (CAM5.2 and 34 beta E12, respectively), carcinoembryonic antigen (CEA), Leu-M1, vimentin, estrogen receptor (ER), progesterone receptor (PR), bcl-2, p53, HER-2/neu, and CA-125. The characteristic immunoprofile for all sites was positivity for CK7, CAM5.2, 34 beta E12, CEA, Leu-M1, vimentin, bcl-2, p53, and CA-125; variably positivity for ER and HER-2/neu; and negativity for CK20 and PR. For comparison, two cases of urologic clear cell carcinoma (1 bladder, 1 urethra) were also studied, and their profile was found to be similar to the gynecologic cases. Aside from minor differences, clear cell carcinoma appears to have the same immunophenotype regardless of whether it originates in the endometrium, ovary, or genitourinary tract. Much of its profile is similar to other gynecologic adenocarcinomas, but some of the markers studied may be useful in the differential diagnosis of this tumor.