Transcriptomic response and immunological responses to chimpanzee adenovirus- and MVA viral-vectored vaccines for RSV in healthy adults.

Cohorts of healthy younger adults (18-50yrs) and healthy older adults (60-75yrs) were immunized intramuscularly or intranasally with an adenovirus-vectored RSV vaccine (PanAd3-RSV) as a prime dose and boosted with PanAd3-RSV or a poxvirus-vectored vaccine (MVA-RSV) encoding the same insert. Whole blood gene expression was measured at baseline, 3- and 7-days post vaccination. Intramuscular prime vaccination with PanAd3-RSV induced differential expression of 643 genes (DEGs, FDR < 0.05). Intranasal prime vaccination with PanAd3-RSV did not induce any differentially expressed genes (DEGs) in blood samples at 3 days post vaccination. Intranasally primed participants showed greater numbers of DEGS on boosting than intramuscularly primed participants. The most highly enriched biological processes related to DEGs after both prime and boost vaccination were type-1 interferon related pathways, lymphocytic and humoral immune responses.

IFITM3 restricts virus-induced inflammatory cytokine production by limiting Nogo-B mediated TLR responses.

Interferon-induced transmembrane protein 3 (IFITM3) is a restriction factor that limits viral pathogenesis and exerts poorly understood immunoregulatory functions. Here, using human and mouse models, we demonstrate that IFITM3 promotes MyD88-dependent, TLR-mediated IL-6 production following exposure to cytomegalovirus (CMV). IFITM3 also restricts IL-6 production in response to influenza and SARS-CoV-2. In dendritic cells, IFITM3 binds to the reticulon 4 isoform Nogo-B and promotes its proteasomal degradation. We reveal that Nogo-B mediates TLR-dependent pro-inflammatory cytokine production and promotes viral pathogenesis in vivo, and in the case of TLR2 responses, this process involves alteration of TLR2 cellular localization. Nogo-B deletion abrogates inflammatory cytokine responses and associated disease in virus-infected IFITM3-deficient mice. Thus, we uncover Nogo-B as a driver of viral pathogenesis and highlight an immunoregulatory pathway in which IFITM3 fine-tunes the responsiveness of myeloid cells to viral stimulation.

Parental TB associated with offspring asthma and rhinitis.

BACKGROUND: Infections in early life are associated with asthma and allergies in one-generation settings; however, the link between parental infection and offspring phenotype is rarely investigated. We aim to study the association of parental TB before conception of the offspring with offspring asthma and rhinitis.METHODS: We included 2,965 offspring born in 1985-2004 and registered in the Norwegian prescription database to 1,790 parents born after 1960 with a history of TB, and included in the Norwegian TB registry. Offspring asthma (n = 582) and rhinitis (n = 929) were defined based on diagnosis, type of medication and prescribed medication ≥1 year. Associations of parental TB <8 years, ≥8 years but before offspring´s birth year and after birth (reference category) with offspring asthma and rhinitis were analysed using logistic regression.RESULTS: Asthma risk was higher in persons with parental TB in childhood (OR 1.73, 95% CI 1.20-2.50) or later preconception (OR 1.38, 95% CI 1.00-1.91) than in persons with parental TB after offspring´s birth; this was significant only in the maternal line (childhood: OR 1.95, 95% CI 1.13-3.37; later preconception: OR 1.74, 95% CI 1.08-2.80). Associations with rhinitis were not identified.CONCLUSIONS: Parental childhood TB was associated with higher asthma risk in future offspring. We speculate that TB impacts maternal immunity and dysregulates the offspring´s type 2 immunity, and that TB-induced epigenetic reprograming of immune defences are transferred to the offspring.

Alteration of gene expression profile in mouse embryonic stem cells and neural differentiation deficits by ethephon.

Ethephon, a member of the organophosphorus compounds, is one of the most widely used plant growth regulators for artificial ripening. Although million pounds of this chemical is being used annually, the knowledge regarding its molecular toxicity is yet not sufficient. The purpose of this study was to evaluate the potential developmental toxicity of ethephon using embryonic stem cell model. The mouse embryonic stem cells (mESCs) were exposed to various concentrations of ethephon and the viability, cell cycle alteration and changes in the gene expression profile were evaluated using high-throughput RNA sequencing. Further, the effect of ethephon on neural differentiation potential was examined. The results showed that ethephon at noncytotoxic doses induced cell cycle arrest in mESCs. Gene ontology enrichment analysis showed that terms related to cell fate and organismal development, including neuron fate commitment, embryo development and cardiac cell differentiation, were markedly enriched in ethephon-treated cells. Neural induction of mESCs in the presence of ethephon was inhibited and the expression of neural genes was decreased in differentiated cells. Results obtained from this work clearly demonstrate that ethephon affects the gene expression profile of undifferentiated mESCs and prevents neural differentiation. Therefore, more caution against the frequent application of ethephon is advised.

TNF receptor type 1 and TNF receptor type 2 mRNA expression was not associated with coronary artery disease in a group of Iranian Turks.

OBJECTIVES: Present investigation was carried out to evaluate the mRNA level of TNF receptor type 1 (TNFR1) and TNF receptor type 2 (TNFR2) in peripheral-blood cells in patients with premature CAD over healthy controls. BACKGROUND: TNFα as a pleiotropic cytokine could be concerned in cardiovascular pathophysiology regarding its special effects on endothelial cells. TNFα exerts its activities through its receptors, TNFR1 and TNFR2. METHODS: Totally, 40 patients with premature CAD and 40 healthy controls were studied. The qRT-PCR technique was used to determine the mRNA level of TNFR1 and TNFR2 in tested groups. RESULTS: The results of this study show that the relative expression was 1.32 ± 0.34 in cases and 1.11 ± 0.16 in controls for TNFR1. The relative expression was 0.96 ± 0.13 in cases and 1.49 ± 0.41 in controls for TNFR2. There is no significant difference in the level of gene expression in the studied groups regarding TNFR1 and TNFR2 genes (p > 0.05). CONCLUSION: It can be concluded that the mRNA levels of TNFR1 and TNFR2 were not associated with CAD risk. Studies with more details, larger sample size, and new risk loci are necessary to reveal disease-causing mechanisms in the pathogenesis of CAD (Fig. 3, Ref. 21).

Spatio-temporal dynamics of calcium electrotransfer during cell membrane permeabilization.

Pulsed electric fields (PEFs) are applied as physical stimuli for DNA/drug delivery, cancer therapy, gene transformation, and microorganism eradication. Meanwhile, calcium electrotransfer offers an interesting approach to treat cancer, as it induces cell death easier in malignant cells than in normal cells. Here, we study the spatial and temporal cellular responses to 10 µs duration PEFs; by observing real-time, the uptake of extracellular calcium through the cell membrane. The experimental setup consisted of an inverted fluorescence microscope equipped with a color high-speed framing camera and a specifically designed miniaturized pulsed power system. The setup allowed us to accurately observe the permeabilization of HeLa S3 cells during application of various levels of PEFs ranging from 0.27 to 1.80 kV/cm. The low electric field experiments confirmed the threshold value of transmembrane potential (TMP). The high electric field observations enabled us to retrieve the entire spatial variation of the permeabilization angle (θ). The temporal observations proved that after a minimal permeabilization of the cell membrane, the ionic diffusion was the prevailing mechanism of the delivery to the cell cytoplasm. The observations suggest 0.45 kV/cm and 100 pulses at 1 kHz as an optimal condition to achieve full calcium concentration in the cell cytoplasm. The results offer precise levels of electric fields to control release of the extracellular calcium to the cell cytoplasm for inducing minimally invasive cancer calcium electroporation, an interesting affordable method to treat cancer patients with minimum side effects.

Implementing the WHO Safe Childbirth Checklist: lessons from a global collaboration.

The WHO Safe Childbirth Checklist (SCC) was developed to ensure the delivery of essential maternal and perinatal care practices around the time of childbirth. A research collaboration was subsequently established to explore factors that influence use of the Checklist in a range of settings around the world. This analysis article presents an overview of the WHO SCC Collaboration and the lessons garnered from implementing the Checklist across a diverse range of settings. Project leads from each collaboration site were asked to distribute two surveys. The first was given to end users, and the second to implementation teams to describe their respective experiences using the Checklist. A total of 134 end users and 38 implementation teams responded to the surveys, from 19 countries across all levels of income. End users were willing to adopt the SCC and found it easy to use. Training and the provision of supervision while using the Checklist, alongside leadership engagement and local ownership, were important factors which helped facilitate initial implementation and successful uptake of the Checklist. Teams identified several challenges, but more importantly successfully implemented the WHO SCC. A critical step in all settings was the adaptation of the Checklist to reflect local context and national protocols and standards. These findings were invaluable in developing the final version of the WHO SCC and its associated implementation guide. Our experience will provide useful insights for any institution wishing to implement the Checklist.

Impulse-powered needle-free syringe for vaccine/drug injection.

A needle-free vaccine/drug injector that works by virtue of the impulse of a moving shock wave is presented in this communication. The device can deliver controlled micro-volumes of liquid vaccines into skin and soft tissue targets in human with minimal invasion. The operation of the injector was investigated by delivering a dyed liquid into human skin samples and soft tissue models. The depth of penetration of the liquid was examined by histology of the targeted human skin samples. The delivery mechanics and the depth of penetration were analyzed theoretically with an elastic model for the skin and a viscoelastic model for the soft tissue targets, and a good agreement with experiments was observed. The current liquid vaccine/drug delivery technique can reduce pain, trauma and contamination, and can offer a cost-effective, needle-free, health-care solution.

Acute effects of sono-activated photocatalytic titanium dioxide nanoparticles on oral squamous cell carcinoma.

Sonodynamic therapy (SDT) is a new treatment modality using ultrasound to activate certain chemical sensitizers for cancer therapy. In this study, effects of high intensity focused ultrasound (HIFU) combined with photocatalytic titanium dioxide (TiO2) nanoparticles on human oral squamous cell line HSC-2 were investigated. Viability of HSC-2 cells after 0, 0.1, 1, or 3s of HIFU irradiation with 20, 32, 55 and 73Wcm(-2) intensities in the presence or absence of TiO2 was measured immediately after the exposures in vitro. Immediate effects of HIFU (3s, 73Wcm(-2)) combined with TiO2 on solid tumors were also examined by histological study. Cytotoxic effect of HIFU+TiO2in vitro was significantly higher than that of TiO2 or HIFU alone with the tendency to increase for higher HIFU intensity, duration, and TiO2 concentration in the suspension. In vivo results showed significant necrosis and tissue damage in HIFU and HIFU+TiO2 treated samples. However, penetration of TiO2 nanoparticles into the cell cytoplasm was only observed in HIFU+TiO2 treated tissues. In this study, our findings provide a rational basis for the development of an effective HIFU based sonodynamic activation method. This approach offers an attractive non-invasive therapy technique for oral cancer in future.

Human colon cancer HT-29 cell death responses to doxorubicin and Morus Alba leaves flavonoid extract.

The mechanistic basis for the biological properties of Morus alba flavonoid extract (MFE) and chemotherapy drug of doxorubicin on human colon cancer HT-29 cell line death are unknown. The effect of doxorubicin and flavonoid extract on colon cancer HT-29 cell line death and identification of APC gene expression and PARP concentration of HT-29 cell line were investigated. The results showed that flavonoid extract and doxorubicin induce a dose dependent cell death in HT-29 cell line. MFE and doxorubicin exert a cytotoxic effect on human colon cancer HT-29 cell line by probably promoting or induction of apoptosis.

Reparable Cell Sonoporation in Suspension: Theranostic Potential of Microbubble.

The conjunction of low intensity ultrasound and encapsulated microbubbles can alter the permeability of cell membrane, offering a promising theranostic technique for non-invasive gene/drug delivery. Despite its great potential, the biophysical mechanisms of the delivery at the cellular level remains poorly understood. Here, the first direct high-speed micro-photographic images of human lymphoma cell and microbubble interaction dynamics are provided in a completely free suspension environment without any boundary parameter defect. Our real-time images and theoretical analyses prove that the negative divergence side of the microbubble's dipole microstreaming locally pulls the cell membrane, causing transient local protrusion of 2.5 µm in the cell membrane. The linear oscillation of microbubble caused microstreaming well below the inertial cavitation threshold, and imposed 35.3 Pa shear stress on the membrane, promoting an area strain of 0.12%, less than the membrane critical areal strain to cause cell rupture. Positive transfected cells with pEGFP-N1 confirm that the interaction causes membrane poration without cell disruption. The results show that the overstretched cell membrane causes reparable submicron pore formation, providing primary evidence of low amplitude (0.12 MPa at 0.834 MHz) ultrasound sonoporation mechanism.

Studying the calcium serum level in patients suffering from psoriasis.

The recent success of vitamin D and its analogues in the treatment of psoriasis has generated extensive research into the role of vitamin D and calcium in this hyperproliferative skin disease. This study aimed at studying the calcium serum level in patients suffering from psoriasis. In this case control studies, 98 hospitalized cases with psoriasis were compared with 100 patients who were hospitalized due to other diseases. Two groups were matched for age and gender. The type of psoriasis, age and sex of patients and serum calcium and albumin levels in two groups were documented in a especial questionnaire. Of all 98 patients with psoriasis, 37.2% were hypocalcemic and 63.7% had normal serum calcium. There was no hypercalcemia. In other group 9% were hypocalcemic, 89 and 2% were normocalcemic and hypercalcemic respectively. In 64.9% of hypocalcemic psoriatic patients, low serum albumin was noted. But all of control group had normal levels. Hypocalcemia is a risk factor of psoriasis. It is better to include dairy as calcium resource in daily diet of patients suffering from psoriasis.

The role of a dedicated staff psychiatrist in modern burn centers.

Psychiatric disorders may be more common in burn-injured subjects than in the general population, and oftentimes contribute to the injury itself. Even in the absence of underlying psychiatric illnesses, burn patients may still benefit from a psychiatric evaluation during and after their hospitalization. In this regard, we included a dedicated psychiatry service in our multidisciplinary burn team. We review herein the course of burn patients that were offered psychiatric evaluation and highlight the benefits of such a program. We conducted an IRB-approved retrospective chart review of burn subjects admitted to our institution between June 15, 2009 and April 30, 2010 and identified 83 patients that were examined by our psychiatrist. Indications for consultation, history of psychiatric illness and substance abuse, as well as administered drugs, were recorded. Among the 83 evaluated patients, 48 (57.8%) had a preexisting psychiatric disorder and 36 (43.4%) suffered from substance abuse. The most common indications for consultation were pain (28.1%), alcohol dependence (25.8%), anxiety (24.7%), illicit drug abuse (16.8%), depression (15.7%), post-traumatic stress disorder (8.9%), and sleep disturbances (8.9%). Pharmacotherapy was initiated in 75 patients (90.3%). 31 (37.3%) had neither a psychiatric disorder nor a history of substance abuse, although 26 of them (83.9%) still received drugs for psychiatric conditions. The inclusion of a dedicated psychiatrist as part of our burn team has improved our comprehensive burn care. In the overwhelming majority of cases, even in the absence of preexisting psychiatric illnesses, consultation resulted in pharmacologic intervention and enhanced patient care.

Les troubles psychiatriques peuvent être plus fréquents chez les patients brûlés que dans la population générale, et contribuent souvent à la blessure elle-même. Même en l'absence de maladies psychiatriques sous-jacents, les patients brûlés peuvent encore bénéficier d'une évaluation psychiatrique pendant et après leur hospitalisation. À cet égard, nous avons inclus un service de psychiatrie dédié à notre équipe multidisciplinaire pour la gestion des brûlures. Nous examinons ici les cours de patients brûlés à qui une évaluation psychiatrique a été proposée et nous mettons en évidence les avantages d'un tel programme. Nous avons effectué un examen rétrospectif - approuvé par les CPP - des patients brûlés admis dans notre institution entre le 15 Juin 2009 et le 30 Avril, 2010, à partir de lequel nous avons identifié 83 patients qui ont été examinés par notre psychiatre. Nous avons enregistré les indications pour la consultation, les antécédents de maladie psychiatrique et la toxicomanie, ainsi que les médicaments administrés. Parmi les 83 patients évalués, 48 (57,8 %) avaient un trouble psychiatrique préexistante et 36 (43,4%) a souffert de l'abus de substances. Les indications les plus fréquentes de consultation étaient la douleur (28,1%), la dépendance à l'alcool (25,8%), l'anxiété (24,7%), l'abus de drogues illicites (16,8%), la dépression (15,7%), les troubles de stress post-traumatique (8,9%), et troubles du sommeil (8,9%). La pharmacothérapie a été instaurée dans 75 patients (90,3%). 31 (37,3%) ne présentaient pas de troubles psychiatriques ni une histoire d'abus de substance mais quand même 26 d'entre eux (83,9 %) ont reçu des médicaments pour des troubles psychiatriques. L'inclusion d'un psychiatre spécialisé dans le cadre de notre équipe a amélioré notre système complet de soins aux brulés. Dans l'écrasante majorité des cas, même en l'absence des maladies psychiatriques préexistantes, la consultation a donné lieu à une intervention pharmacologique et a amélioré les soins aux patients.

Effect of chemical chaperones on glucose-induced lysozyme modifications.

Nonenzymatic glycation of biomacromolecules occurs due to the diabetes mellitus and ageing. A number of small molecules, known as chemical chaperones, stabilize protein conformation against thermal and chemically induced denaturation. These compounds are including: polyamines (e.g. spermine and spermidine), amino acids (e.g. lysine) and polyols (e.g. glycerol). In this study the effect of spermidine (Spd), spermine (Spm), and glycerol on glycation, structure and function of lysozyme (LZ), as an extra-cellular protein, by different techniques is investigated. LZ is incubated with or without glucose (50 or 100 mM) in the absence or presence of Spd/Spm/glycerol at 37 °C up to 16 weeks. All the observed changes of glycated-LZ in comparison with the native protein, including: increased fluorescence emission, alteration in the secondary and tertiary structure, and reduced electrophoretic mobility- indicate its structural changes that are accompanied with its reduced activity. Glucose in the presence or absence of Spd induces the protein dimerization, but glucose plus Spm induces its trimmerization. In contrast, glycerol inhibits the LZ glycation and prevents the large changes on its structure and function. Glucose binds lysine residues, decreases the protein positive charges and induces some alterations in its structure and activity. Polyamines also directly bind to LZ, increase its positive charges and hence induce more glycation; more conformational changes, oligomerization and its inactivation in the presence of glucose, but glycerol affect the protein environment and preserve protein from these harmful effects.

Promoting hand hygiene in healthcare through national/subnational campaigns.

The World Health Organization (WHO) First Global Patient Safety Challenge conducted a baseline survey of coordinated large-scale activities in improving hand hygiene in healthcare in 2007. The survey was repeated in early 2009 to assess current status and generate information on factors contributing to success. Coordinated activities were identified through WHO regional offices and experts in the field. An online survey using a structured questionnaire was conducted during March-April 2009. Personnel involved in all 38 campaigns/programmes in 2009 completed the survey. Of these, 29 were active national/subnational-level initiatives and 22 (75.8%) were initiated after the Challenge launch in October 2005. Main targets were general, district, and university hospitals with increasing coverage of long-term care facilities and primary care. The scope varied from awareness-raising to formal scaled-up activities with ongoing evaluation. Most initiatives (20/29) obtained funding from multiple sources with governments among the main funders; governments also initiated 25/29 (86.2%) programmes. The facilitator role played by the Challenge in initiating and supporting activities with tools and recommendations was clearly identified. The perceived significance of specific barriers varied considerably across initiatives. Those related to commitment (priority and support) and resource availability were important across all regions. Hand hygiene is being promoted in healthcare in many nations/subnations with clear objectives, strategies, and governmental support through policies and resource allocation. While this is important for sustainability, further action is required to initiate coordinated activities across the world, including countries with limited resources.

Osteoarthritis-like damage of cartilage in the temporomandibular joints in mice with autoimmune inflammatory arthritis.

OBJECTIVE: To study temporomandibular joint (TMJ) involvement in an autoimmune murine model of rheumatoid arthritis (RA), a disease characterized by inflammatory destruction of the synovial joints. Although TMJ dysfunction is frequently found in RA, TMJ involvement in RA remains unclear, and TMJ pathology has not been studied in systemic autoimmune animal models of RA. METHODS: Proteoglycan (PG) aggrecan-induced arthritis (PGIA) was generated in genetically susceptible BALB/c mice. TMJs and joint tissues/cartilage were harvested for histological and immunohistochemical analyses and RNA isolation for quantitative polymerase chain-reaction. Serum cytokine levels were measured in mice with acute or chronic arthritis, and in non-arthritic control animals. RESULTS: Despite the development of destructive synovitis in the limbs, little or no synovial inflammation was found in the TMJs of mice with PGIA. However, the TMJs of arthritic mice showed evidence of aggrecanase- and matrix metalloproteinase-mediated loss of glycosaminoglycan-containing aggrecan, and in the most severe cases, structural damage of cartilage. Serum levels of pro-inflammatory cytokines, including interleukin (IL)-1ß, were elevated in arthritic animals. Expression of the IL-1ß gene was also high in the inflamed limbs, but essentially normal in the TMJs. Local expression of genes encoding matrix-degrading enzymes (aggrecanases and stromelysin) was upregulated to a similar degree in both the limbs and the TMJs. CONCLUSION: We propose that constantly elevated levels of catabolic cytokines, such as IL-1ß, in the circulation (released from inflamed joints) create a pro-inflammatory milieu within the TMJ, causing local upregulation of proteolytic enzymes and subsequent loss of aggrecan from cartilage.

Morphological variation in the horse: defining complex traits of body size and shape.

Horses, like many domesticated species, have been selected for broad variation in skeletal size. This variation is not only an interesting model of rapid evolutionary change during domestication, but is also directly applicable to the horse industry. Breeders select for complex traits like body size and skeletal conformation to improve marketability, function, soundness and performance in the show ring. Using a well-defined set of 35 measurements, we have identified and quantified skeletal variation in the horse species. We collected measurements from 1215 horses representing 65 breeds of diverse conformation such as the American Miniature, Shetland Pony, Arabian Horse, Thoroughbred, Shire and Clydesdale. Principal components analysis has identified two key dimensions of skeletal variation in the horse. Principal component 1 is positively correlated with every measurement and quantifies overall body size. Principal component 2 captures a pattern of bone widths vs. lengths and thus quantifies variation in overall bone thickness. By defining these complex skeletal traits, we have created a framework for whole genome association studies to identify quantitative trait loci that contribute to this variation.

Comparison of national and subnational guidelines for hand hygiene.

Hand hygiene promotion is considered as the cornerstone for healthcare-associated infection prevention. Over the past years, hand hygiene guidelines have been developed by different agencies at international, national and subnational levels. A comparison of these documents could help in understanding recommendations in different parts of the world and the methods used for their development. Guidelines were identified through search engines, electronic libraries, and personal contacts, and their content was analysed using an adapted version of a tool from the European DG XII-funded HARMONY project. Twenty-two guidelines were retrieved and 21 were evaluated. Documents varied in scope, approach, content and terminology. Some were primarily advisory directives, whereas others focused on the technical issues of why, when, and how to perform hand hygiene. The extent to which evidence was collected and assessed varied considerably and details were provided only in very few. Grading systems and definitions to indicate the strength of evidence and recommendations also differed. The intended outcome was to improve hand hygiene practices in healthcare, thus leading to a reduction of healthcare-associated infections and/or antimicrobial resistance. Although overall agreement on indications and procedures was noted, the range and depth of recommendations on best practices and implementation varied. Essential aspects such as compliance measurement and audits to assess guideline effectiveness were neglected in most documents. In conclusion, there is a need for a more consistent approach leading to recommendations based on a thorough evaluation of evidence and applicable worldwide. Aspects related to implementation and impact monitoring deserve greater attention.

African partnerships for patient safety: a vehicle for enhancing patient safety across two continents. [corrected].

African Partnerships for Patient Safety (APPS) aims to develop sustainable partnerships between hospitals in Africa and Europe to create a network of beacon hospitals for patient safety. The three core APPS objectives are focused on building strong patient safety partnerships between hospitals in Africa and Europe, implementing patient safety improvements in each partnership hospital on 12 patient safety action areas, and facilitating spread of patient safety improvements. APPS is working with six first wave hospital partnerships and will capture and report learning from implementation. A range of APPS resources will shortly be available to hospitals working on patient safety systems.