Implant survival and success rates in patients with risk factors: results from a long-term retrospective study with a 10 to 18 years follow-up.
Risk factors for implant therapy are represented by all general and local conditions that through various mechanisms can increase either short-term and long-term failure risk. The aim of this study is to assess the implant survival and implant success rates with single and multiple risk factors. To address the research purpose, a retrospective cohort study was designed and implemented, including a sample of 225 patients with a total of 871 implants placed. The following risk factors were considered: smoking, bruxism, bone augmentation procedures and the presence of load risk (implants with crown/implant relation > 0.8; angulation > 25°; presence of cantilever). Follow-up ranged from 10 years to 18 years (average follow-up 13.6 years). Failures were subdivided into short-term failures, before the prosthetic phase, and long-term failures, after definitive prosthesis. The success criteria published by Albrektsson and Zarb were adopted. A Cox proportional hazard regression model was used to calculate hazard ratio, with a statistically significant p-value <0.05. Out of the 871 implants placed, 138 did not meet the success criteria, (success rate 84.16%), sixty (43.47%) were classified as "early failure" and seventy-eight as "late failure" (56.53%). A total of 70 dental implants were removed, with a survival rate of 91.96%. The presence of a single risk factor does not imply a marked increase of failure risk. Among the analyzed factors, the one that proved to be the most dangerous was bruxism, even when presented as the only risk factor. Bruxism with load risk proved to be the most dangerous association (success rate 69.23%) and could be included among the absolute contraindications for implant treatment.


RISK OF RETINAL VEIN OCCLUSION IN PATIENTS WITH PSORIASIS: A Population-Based Cohort Study.
To investigate the risk of developing retinal vein occlusion (RVO) in patients with psoriasis. In this retrospective population-based cohort study, 30,198 patients with psoriasis (Psoriasis((+)) group) and 30,198 controls without psoriasis (Psoriasis((−)) group) between 2001 and 2006 from the Taiwan National Health Insurance Research Database were selected. The incidence of RVO was 1.46 times higher in the Psoriasis((+)) group than in the Psoriasis((−)) group (3.61 vs. 2.47/10,000 person-years) (adjusted hazard ratio = 1.50; 95% confidence interval = 1.07-2.10) calculated using Cox proportional hazard regression. Age was an independent risk factor for RVO (adjusted hazard ratio: 11.9 for patients 65 years or older vs. 1.00 for those 0-49 years old). In the 65 years or older Psoriasis((+)) group, the incidence of developing RVO was 1.97 times higher (95% confidence interval = 1.19-3.26) than in the 65 years or older Psoriasis((−)) group. In Psoriasis((+)) women, the incidence of developing RVO was 1.82 times higher (95% = 1.05-3.14) than in Psoriasis((−)) women. For the subgroup with comorbid hypertension, the incidence of developing RVO was 2.07 times higher (95% confidence interval = 1.22-3.50) in the Psoriasis((+)) group than in the Psoriasis((−)) group. Psoriasis was significantly associated with a higher risk of developing RVO.


Prescription of analgesia in emergency medicine (POEM) secondary analysis: an observational multicentre comparison of pain relief provided to adults and children with an isolated limb fracture and/or dislocation.
Acute pain is a common reason for emergency department (ED) attendance. Royal College of Emergency Medicine (RCEM) pain management audits have shown national variation and room for improvement. Previous evidence suggests that children receive less satisfactory pain management than adults. Prescription of analgesia in emergency medicine is a cross-sectional observational study of consecutive patients presenting to 12 National Health Service EDs with an isolated long bone fracture and/or dislocation, and was carried out between 2015 and 2017. Using the recommendations in the RCEM Best Practice Guidelines, pain management in ED was assessed for differences of age (adults vs children) and hospital type (children's vs all patients). From the total 8346 patients, 38% were children (median age 8 years). There was better adherence to the RCEM guidance for children than adults (24% (766/3196) vs 11% (579/5123)) for the combined outcome of timely assessment, pain score and appropriate analgesia. In addition, children were significantly more likely than adults to receive analgesia appropriate to the pain score (of those with a recorded pain score 67% (1168/1744) vs 52% (1238/2361)). Children's hospitals performed much better across all reported outcomes compared with general hospitals. In contrast to previous studies, children with a limb fracture/dislocation are more likely than adults to have a pain score documented and to receive appropriate analgesia. Unexpectedly, children's EDs performed better than general EDs in relation to timely and appropriate analgesia but the reasons for this are not apparent from the present study.


The composition of the microbiota modulates allograft rejection.
Transplantation is the only cure for end-stage organ failure, but without immunosuppression, T cells rapidly reject allografts. While genetic disparities between donor and recipient are major determinants of the kinetics of transplant rejection, little is known about the contribution of environmental factors. Because colonized organs have worse transplant outcome than sterile organs, we tested the influence of host and donor microbiota on skin transplant rejection. Compared with untreated conventional mice, pretreatment of donors and recipients with broad-spectrum antibiotics (Abx) or use of germ-free (GF) donors and recipients resulted in prolonged survival of minor antigen-mismatched skin grafts. Increased graft survival correlated with reduced type I IFN signaling in antigen-presenting cells (APCs) and decreased priming of alloreactive T cells. Colonization of GF mice with fecal material from untreated conventional mice, but not from Abx-pretreated mice, enhanced the ability of APCs to prime alloreactive T cells and accelerated graft rejection, suggesting that alloimmunity is modulated by the composition of microbiota rather than the quantity of bacteria. Abx pretreatment of conventional mice also delayed rejection of major antigen-mismatched skin and MHC class II-mismatched cardiac allografts. This study demonstrates that Abx pretreatment prolongs graft survival, suggesting that targeting microbial constituents is a potential therapeutic strategy for enhancing graft acceptance.


Assessing the roles of temperature, precipitation, and ENSO in dengue re-emergence on the Texas-Mexico border region.
The goal of this study was to assess linkages between microclimate and longer-term ENSO-related weather forcing on the week-to-week changes in dengue prevalence in Matamoros, Tamaulipas, Mexico, over a recent decade of dengue observations. An auto-regressive model to evaluate the role of climatic factors (sea-surface temperature) and weather (maximum temperature, minimum temperature, precipitation) on dengue incidence over the period 1995-2005, was developed by conducting time-series analysis. Dengue incidence increased by 2.6% (95% CI: 0.2-5.1) one week after every 1 degree Celsius increase in weekly maximum temperature and increased 1.9% (95% CI: -0.1-3.9) two weeks after every 1 cm increase in weekly precipitation. Every 1 masculineC increase in sea surface temperatures (El Niño region 3.4 ) was followed by a 19.4% (95% CI: -4.7-43.5) increase in dengue incidence (18 weeks later). Climate and weather factors play a small but significant role in dengue transmission in Matamoros, Mexico. This study may provide baseline information for identifying potential longer-term effects of global climate change on dengue expected in the coming decades. To our knowledge, this is the first study to investigate the potential associations between climate and weather events and dengue incidence in this geographical area.


On the programming and reprogramming of actions.
Actions are often selected in the context of ongoing movement plans. Most studies of action selection have overlooked this fact, implicitly assuming that the motor system is passive prior to presentation of instructions triggering movement selection. Other studies addressed action planning in the context of an already present motor plan, but focused mostly on inhibition of a prepotent response under fierce time pressure. Under these circumstances, inhibition of previous motor plans and selection of a new response become temporally intermingled. Here, we explore how the presence of earlier motor plans influences cerebral effects associated with action selection, separating in time movement programming, reprogramming, and execution. We show that portions of parietofrontal circuits, including intraparietal sulcus and left dorsal premotor cortex, are systematically involved in programming motor responses, their activity being indifferent to the presence of earlier motor plans. We identify additional regions recruited when a motor response is programmed in the context of an existing motor program. We found that several right-hemisphere regions, previously associated with response inhibition, might be better characterized as involved in response selection. Finally, we detail the specific role of a right precentral region in movement reprogramming that is involved in inhibiting not only actual responses but also motor representations.


Concerted loss of TGFβ-mediated proliferation control and E-cadherin disrupts epithelial homeostasis and causes oral squamous cell carcinoma.
Although the etiology of squamous cell carcinomas of the oral mucosa is well understood, the cellular origin and the exact molecular mechanisms leading to their formation are not. Previously, we observed the coordinated loss of E-cadherin (CDH1) and transforming growth factor beta receptor II (TGFBR2) in esophageal squamous tumors. To investigate if the coordinated loss of Cdh1 and Tgfbr2 is sufficient to induce tumorigenesis in vivo, we developed two mouse models targeting ablation of both genes constitutively or inducibly in the oral-esophageal epithelium. We show that the loss of both Cdh1 and Tgfbr2 in both models is sufficient to induce squamous cell carcinomas with animals succumbing to the invasive disease by 18 months of age. Advanced tumors have the ability to invade regional lymph nodes and to establish distant pulmonary metastasis. The mouse tumors showed molecular characteristics of human tumors such as overexpression of Cyclin D1. We addressed the question whether TGFβ signaling may target known stem cell markers and thereby influence tumorigenesis. From our mouse and human models, we conclude that TGFβ signaling regulates key aspects of stemness and quiescence in vitro and in vivo. This provides a new explanation for the importance of TGFβ in mucosal homeostasis.


Adaptive gene amplification as an intermediate step in the expansion of virus host range.
The majority of recently emerging infectious diseases in humans is due to cross-species pathogen transmissions from animals. To establish a productive infection in new host species, viruses must overcome barriers to replication mediated by diverse and rapidly evolving host restriction factors such as protein kinase R (PKR). Many viral antagonists of these restriction factors are species specific. For example, the rhesus cytomegalovirus PKR antagonist, RhTRS1, inhibits PKR in some African green monkey (AGM) cells, but does not inhibit human or rhesus macaque PKR. To model the evolutionary changes necessary for cross-species transmission, we generated a recombinant vaccinia virus that expresses RhTRS1 in a strain that lacks PKR inhibitors E3L and K3L (VVΔEΔK+RhTRS1). Serially passaging VVΔEΔK+RhTRS1 in minimally-permissive AGM cells increased viral replication 10- to 100-fold. Notably, adaptation in these AGM cells also improved virus replication 1000- to 10,000-fold in human and rhesus cells. Genetic analyses including deep sequencing revealed amplification of the rhtrs1 locus in the adapted viruses. Supplying additional rhtrs1 in trans confirmed that amplification alone was sufficient to improve VVΔEΔK+RhTRS1 replication. Viruses with amplified rhtrs1 completely blocked AGM PKR, but only partially blocked human PKR, consistent with the replication properties of these viruses in AGM and human cells. Finally, in contrast to AGM-adapted viruses, which could be serially propagated in human cells, VVΔEΔK+RhTRS1 yielded no progeny virus after only three passages in human cells. Thus, rhtrs1 amplification in a minimally permissive intermediate host was a necessary step, enabling expansion of the virus range to previously nonpermissive hosts. These data support the hypothesis that amplification of a weak viral antagonist may be a general evolutionary mechanism to permit replication in otherwise resistant host species, providing a molecular foothold that could enable further adaptations necessary for efficient replication in the new host.


Tracking the dynamic interplay between bacterial and host factors during pathogen-induced vacuole rupture in real time.
Escape into the host cell cytosol following invasion of mammalian cells is a common strategy used by invasive pathogens. This requires membrane rupture of the vesicular or vacuolar compartment formed around the bacteria after uptake into the host cell. The mechanism of pathogen-induced disassembly of the vacuolar membrane is poorly understood. We established a novel, robust and sensitive fluorescence microscopy method that tracks the precise time point of vacuole rupture upon uptake of Gram-negative bacteria. This revealed that the enteroinvasive pathogen Shigella flexneri escapes rapidly, in less than 10 min, from the vacuole. Our method demonstrated the recruitment of host factors, such as RhoA, to the bacterial entry site and their continued presence at the point of vacuole rupture. We found a novel host marker for ruptured vacuoles, galectin-3, which appears instantly in the proximity of bacteria after escape into the cytosol. Furthermore, we show that the Salmonella effector proteins, SifA and PipB2, stabilize the vacuole membrane inhibiting bacterial escape from the vacuole. Our novel approach to track vacuole rupture is ideally suited for high-content and high-throughput approaches to identify the molecular and cellular mechanisms of membrane rupture during invasion by pathogens such as viruses, bacteria and parasites.


Botulinum toxin management of spasmodic dysphonia (laryngeal dystonia): a 12-year experience in more than 900 patients.
This paper reviews a 12-year experience in more than 900 patients with spasmodic dysphonia who have been treated with botulinum toxin. This is a retrospective analysis of patients with adductor spasmodic dysphonia (strain-strangled voice), abductor spasmodic dysphonia (whispering voice), and adductor breathing dystonia (paradoxical vocal fold motion), all of whom have been treated with botulinum toxin injections for relief of symptom. All of the patients were studied with a complete head and neck and neurologic examination; fiberoptic laryngostroboscopy; acoustic and aerodynamic measures; and a speech evaluation including the Universal spasmodic dysphonia rating scale. Some were given electromyography. All patients received botulinum toxin injections into the affected muscles under electromyographic guidance. The adductor patients had an average benefit of 90% of normal function lasting an average of 15.1 weeks. The abductor patients had an average benefit of 66.7% of normal function lasting an average of 10.5 weeks. Adverse effects included mild breathiness and coughing on fluids in the adductor patients, and mild stridor in a few of the abductor patients. Botulinum toxin A injection of the laryngeal hyperfunctional muscles has been found over the past 12 years to be the treatment of choice to control the dystonic symptoms in most patients with spasmodic dysphonia.


Folate and MTHFR: risk of adenoma recurrence in the Polyp Prevention Trial.
Low dietary folate intake has been associated with colorectal cancer risk and adenoma recurrence. A C/T transition at position 677 in the gene encoding methlylenetetrahydrofolate reductase (MTHFR C677T) has been reported to interact with folate intake to modulate colorectal adenoma recurrence or cancer risk. We investigated the association between MTHFR, total folate, and the risk of colorectal adenoma recurrence in the Polyp Prevention Trial. We compared 625 individuals with any adenoma recurrence after 4 years (266 individuals with multiple (> or =2) recurrent adenomas and 101 individuals with advanced adenoma recurrence) to 978 individuals with no adenoma recurrence. Odds ratios (OR) and 95% confidence intervals (CI) for risk of adenoma recurrence were calculated using unconditional logistic regression. We also investigated effect modification of the MTHFR genotype associations by total folate intake. In general, no statistically significant associations were found between quartile of folate intake (dietary or total) and adenoma recurrence. The MTHFR CT genotype was associated with a significantly increased risk of multiple adenoma recurrence (OR: 1.34, 95% CI: 1.00, 1.81). No significant interaction was noted for total folate and MTHFR genotype, though an increased risk of recurrence noted for the MTHFR CT genotype was statistically significant only for those individuals with below median intake of total folate. We report that the MTHFR 677 CT genotype was associated with increased risk of adenoma recurrence (specifically multiple adenoma recurrence) 4 years after polypectomy.


The promise of evolutionary systems biology: lessons from bacterial chemotaxis.
Bacterial chemotaxis and the signaling networks underlying it provide us with a model system for studying the molecular basis of behavior and information processing. Although chemotaxis is well characterized at both the phenotype and genotype levels in the model organism Escherichia coli, it is not yet possible to predict chemotaxis behavior in diverse bacteria on the basis of their environment or genome sequence. Moreover, we still cannot propose a plausible evolutionary trajectory from minimal systems to present-day chemotaxis networks. The analysis of all sequenced bacterial genomes provides a prediction of their chemotaxis networks and reveals substantial structural diversity. Additionally, it uncovers a set of previously unknown proteins that could be the "missing" link between complex present-day chemotaxis networks and simpler, ancestral systems composed of a few proteins. Further evaluation of these findings with experimental and modeling studies will allow us to distill evolutionary design principles in chemotaxis signaling networks.


Registry of emergency airways arriving at combat hospitals.
Prehospital Emergency Medical Services have demonstrated variable success with regards to prehospital airway management in U.S. civilian settings. We attempted to identify the incidence of successful prehospital endotracheal intubations in the modern combat environment. This was a prospective, observational study. Data collection occurred at Combat Support Hospitals (CSH) within Operation Iraqi Freedom locations between January 2005 and March 2007. Military trauma physicians systematically examined casualties presenting to the CSH that received advanced prehospital airway management. Correct endotracheal tube (ETT) positioning was verified using an explicit combination of clinical findings and colorimetric end-tidal carbon dioxide detection. The primary outcome was correct placement of the ETTs by combat prehospital providers. A total of 6,875 combat casualties presented to participating CSHs during the study period, of which there were 293 (4.2%) advanced prehospital airways, of which 282 (97.3%) were trauma patients. Prehospital airway management included: 253 endotracheal intubations (86.6%); 23 supraglottic airways (7.5%), and 17 cricothyrotomies (5.8%). Of the ETTs, upon arrival to the CSH, 242 (95.7%) were determined to be correctly placed. There were 11 incorrectly placed ETTs: 10 were in the right mainstem bronchus, and 1 was found to be dislodged in the hypopharynx. There were no unrecognized battlefield esophageal intubations. Under combat conditions, the overall rate of correctly placed ETTs performed by military prehospital providers was comparable with that of published U.S. civilian paramedic data.


Invasive lobular carcinoma cell lines are characterized by unique estrogen-mediated gene expression patterns and altered tamoxifen response.
Invasive lobular carcinoma (ILC) is a histologic subtype of breast cancer that is frequently associated with favorable outcomes, as approximately 90% of ILC express the estrogen receptor (ER). However, recent retrospective analyses suggest that patients with ILC receiving adjuvant endocrine therapy may not benefit as much as patients with invasive ductal carcinoma. On the basis of these observations, we characterized ER function and endocrine response in ILC models. The ER-positive ILC cell lines MDA MB 134VI (MM134) and SUM44PE were used to examine the ER-regulated transcriptome via gene expression microarray analyses and ER ChIP-Seq, and to examine response to endocrine therapy. In parallel, estrogen response was assessed in vivo in the patient-derived ILC xenograft HCI-013. We identified 915 genes that were uniquely E2 regulated in ILC cell lines versus other breast cancer cell lines, and a subset of these genes were also E2 regulated in vivo in HCI-013. MM134 cells were de novo tamoxifen resistant and were induced to grow by 4-hydroxytamoxifen, as well as other antiestrogens, as partial agonists. Growth was accompanied by agonist activity of tamoxifen on ER-mediated gene expression. Though tamoxifen induced cell growth, MM134 cells required fibroblast growth factor receptor (FGFR)-1 signaling to maintain viability and were sensitive to combined endocrine therapy and FGFR1 inhibition. Our observation that ER drives a unique program of gene expression in ILC cells correlates with the ability of tamoxifen to induce growth in these cells. Targeting growth factors using FGFR1 inhibitors may block survival pathways required by ILC and reverse tamoxifen resistance.


Coherent-form energy conservation relation for the elastic scattering of a guided mode in a symmetric scattering system.
We propose a coherent-form energy conservation relation (ECR) that is generally valid for the elastic transmission and reflection of a guided mode in a symmetric scattering system. In contrast with the classical incoherent-form ECR, |τ|2 + |ρ|2≤1 with τ and ρ denoting the elastic transmission and reflection coefficients of a guided mode, the coherent-form ECR is expressed as |τ + ρ|≤1, which imposes a constraint on a coherent superposition of the transmitted and reflected modes. The coherent-form ECR is rigorously demonstrated and is numerically tested by considering different types of modes in various scattering systems. Further discussions with the scattering matrix formalism indicate that two coherent-form ECRs, |τ + ρ|≤1 and |τ-ρ|≤1, along with the classical ECR |τ|2 + |ρ|2≤1 constitute a complete description of the energy conservation for the elastic scattering of a guided mode in a symmetric scattering system. The coherent-form ECR provides a common tool in terms of energy transfer for understanding and analyzing the scattering dynamics in currently interested scattering systems.


Preemptive analgesia for postoperative hysterectomy pain control: systematic review and clinical practice guidelines.
The objective of the study was to investigate the effectiveness of preemptive analgesia at pain control in women undergoing total abdominal hysterectomy. Eligible studies, published through May 31, 2016, were retrieved through Medline, Cochrane Central Register for Controlled Trials, and Cochrane Database of Systematic Reviews. We included randomized controlled trials with the primary outcome of pain control in women receiving a preemptive medication prior to total abdominal hysterectomy. Comparators were placebo, different doses of the same medication as intervention, or other nonnarcotic or narcotic medication. Study data were extracted by one reviewer and confirmed by a second reviewer. For each outcome we graded the quality of the evidence. Studies were classified by the type of medication used and by outcome type. Eighty-four trials met eligibility, with 69 included. Among nonnarcotic medications, paracetamol, gabapentin, and rofecoxib combined with gabapentin resulted in improvements in pain assessment compared with placebo and other nonnarcotic medications. Patient satisfaction was higher in patients who were given gabapentin combined with paracetamol compared with gabapentin alone. Use of preemptive paracetamol, gabapentin, bupivacaine, and phenothiazine resulted in less narcotic usage than placebo. All narcotics (ketamine, morphine, fentanyl) resulted in improved pain control compared with placebo. Narcotics had a greater reduction in pain assessment scores compared with nonnarcotics, and their use resulted in lower total narcotic usage. Preemptive nonnarcotic and narcotic medications prior to abdominal hysterectomy decrease total narcotic requirements and improve patient postoperative pain assessment and satisfaction scores.


Structural and energetic consequences of mutations in a solvated hydrophobic cavity.
The structural and energetic consequences of modifications to the hydrophobic cavity of interleukin 1-beta (IL-1beta) are described. Previous reports demonstrated that the entirely hydrophobic cavity of IL-1beta contains positionally disordered water. To gain a better understanding of the nature of this cavity and the water therein, a number of mutant proteins were constructed by site-directed mutagenesis, designed to result in altered hydrophobicity of the cavity. These mutations involve the replacement of specific phenylalanine residues, which circumscribe the cavity, with tyrosine, tryptophan, leucine and isoleucine. Using differential scanning calorimetry to determine the relative stabilities of the wild-type and mutant proteins, we found all of the mutants to be destabilizing. X-ray crystallography was used to identify the structural consequences of the mutations. No clear correlation between the hydrophobicities of the specific side-chains introduced and the resulting stabilities was found.


Distributed fiber optic sensing based on counterpropagating waves.
The spatially and temporally resolved birefringence of a single-mode optical fiber can be ascertained using backward stimulated Raman scattering. The magnitude of the birefringence is determined from the optical power exchanged between two counterpropagating light pulses. The degree to which a signal pulse is amplified by a pump pulse is governed by their relative states of polarization when they overlap. A novel normalization procedure is proposed that eliminates many of the unknowns. An example of how this technique could be used to evaluate a distributed strain field is provided.


Synergistic neuroprotective effects by combining an NMDA or AMPA receptor antagonist with nitric oxide synthase inhibitors in global cerebral ischaemia.
We have investigated the neuroprotective effects of combining an NMDA or AMPA receptor antagonist with a nitric oxide synthase (NOS) inhibitor in the gerbil model of global cerebral ischaemia. Ischaemia was induced by occlusion of the common carotid arteries for 5 min. (5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,1 0-imine (MK-801, 2.5 mg/kg i.p.) or (3S,4aR,6R,8aR)-6-[2-(1(2)H-tetrazole-5-yl)]decahydroisoq uinoline-3-carboxylic acid (LY293558, 20 mg/kg i.p.) and 7-nitroindazole (25 mg/kg i.p.) or N-[4-(2-[[(3-chlorophenyl)methyl]amino]ethyl) phenyl]-2-thiophenecarboximidamide dihydrochloride (ARL17477, 25 mg/kg i.p.) were administered alone or in combination (i.e., MK-801 with 7-nitroindazole or ARL17477 or LY293558 with 7-nitroindazole or ARL17477). In the present studies, both MK-801 and LY293558 provided significant degree of neuroprotection, while 7-nitroindazole and ARL17477 also provided some neuroprotection, which failed to reach significance in every case. However, the combination of MK-801 with 7-nitroindazole or ARL17477 provided 21% or 44% greater protection than the total protection or either alone. Likewise, the combination of LY293558 with 7-nitroindazole or ARL17477 provided 14.5% and 35% greater protection than total protection of either compound alone. These results indicate that several pathways contribute to ischaemic cell death and combining excitatory amino antagonists and NOS inhibitors provides greater protection than either alone. Therefore, combination therapy should be considered as an approach for treating ischaemic conditions.


The reality of pediatric emergency mass critical care in the developing world.
Public health emergencies resulting from major man-made crises and large-scale natural disasters severely impact developing countries, causing unprecedented rates of indirect mortality and morbidity, especially in children and women. Concomitantly, the state of children's health in the least-developed countries is the worst since the 1950s before the Declaration of Alma Ata. Worldwide decline in public health protections, infrastructures, and systems, and a health worker crisis primarily in Africa and Asia, limit the delivery of intensive and critical care services. In May 2008, the Task Force for Mass Critical Care published guidance on provision of mass critical care to adults. Acknowledging that the critical care needs of children during disasters were unaddressed by this effort, a 17-member Steering Committee, assembled by the Oak Ridge Institute for Science and Education with guidance from members of the American Academy of Pediatrics, convened in April 2009 to determine priority topic areas for pediatric emergency mass critical care recommendations.Steering Committee members established subgroups by topic area and performed literature reviews of MEDLINE and Ovid databases. The Steering Committee produced draft outlines through consensus-based study of the literature and convened October 6-7, 2009, in New York, NY, to review and revise each outline. Eight draft documents were subsequently developed from the revised outlines as well as through searches of MEDLINE updated through March 2010.The Pediatric Emergency Mass Critical Care Task Force, composed of 36 experts from diverse public health, medical, and disaster response fields, convened in Atlanta, GA, on March 29-30, 2010. Feedback on each manuscript was compiled and the Steering Committee revised each document to reflect expert input in addition to the most current medical literature. Using pandemics as a model of public health emergencies, steps to improve care to the most vulnerable of populations are outlined, including mandates under the International Health Regulations Treaty of 2007 and World Health Organization guidelines. Recommendations include an emphasis on first improving primary care, prevention, and basic emergency care, where possible. Advances in care should move incrementally without compromising primary care resources. A first step in preparing for a pandemic in developing countries involves building capacity in public health surveillance and proven community containment and mitigation strategies. Given the severe lack of healthcare workers in at least 57 countries, the Task Force also supports World Health Organization's recommendations that planning for a public health emergency include means for health workers to collaborate with staff in the military, transport, and education sectors as well as international healthcare workers to maximize the efficiency of scarce human resources. Rapid response teams can be augmented by international subject matter experts if these do not exist at the country level.


Sleep enhances exposure therapy.
Sleep benefits memory consolidation. Here, we tested the beneficial effect of sleep on memory consolidation following exposure psychotherapy of phobic anxiety. A total of 40 individuals afflicted with spider phobia according to DSM-IV underwent a one-session virtual reality exposure treatment and either slept for 90 min or stayed awake afterwards. Sleep following exposure therapy compared with wakefulness led to better reductions in self-reported fear (p = 0.045, d = 0.47) and catastrophic spider-related cognitions (p = 0.026, d = 0.53) during approaching a live spider, both tested after 1 week. Both reductions were associated with greater percentages of stage 2 sleep. Our results indicate that sleep following successful psychotherapy, such as exposure therapy, improves therapeutic effectiveness, possibly by strengthening new non-fearful memory traces established during therapy. These findings offer an important non-invasive alternative to recent attempts to facilitate therapeutic memory extinction and consolidation processes with pharmacological or behavioral interventions.


Nitrogen and energy relationships in malnourished patients with emphysema.
The purpose of this study was to examine the impact of nutritional support on nitrogen-energy relationships and functional parameters in malnourished patients with emphysema. Malnourished patients without lung disease served as the control group. Ten ambulatory, stable patients with emphysema and six patients without lung disease received an infusion of 5% dextrose (baseline) plus electrolytes (D5W) for two days, which was followed by an enteral or a parenteral infusion of either a carbohydrate-based (CB, 53% carbohydrate) or a fat-based diet (FB, 55% fat) for 1 wk each, in a randomized cross-over design. All patients had greater than 10% weight loss. Caloric intake was set at 1.7 times the resting energy expenditure (REE) as measured during the baseline period. The REE of patients with emphysema was 23 and 27% above that of the control group during baseline and refeeding periods, respectively. The increased REE was met primarily by an increased carbohydrate oxidation. During the infusion of D5W, N balance was lower in patients with emphysema, but during repletion N balance was similar in both groups of patients. Two weeks of nutritional support with either a CB or a FB diet increased body weight, N balance, and arm muscle area and improved maximal inspiratory pressure, skeletal muscle strength, and endurance-strength (using quadriceps, hamstring, and handgrip) to a similar degree in malnourished patients with and without lung disease. In other stress states, such as infection, it has been shown that hypermetabolism, hypercatabolism, and preferential fat oxidation occur concomitantly. Patients with emphysema are unusual because, although they are hypermetabolic, they are not hypercatabolic and do not demonstrate preferential fat oxidation.


Intrinsically disordered p53 and its complexes populate compact conformations in the gas phase.
Spontaneous shrinking: the intrinsically disordered tumor suppressor protein p53 was analyzed by using a combination of ion mobility mass spectrometry and molecular dynamics simulations. Structured p53 subdomains retain their overall topology upon transfer into the gas phase. When intrinsically disordered segments are introduced into the protein sequence, however, the complex spontaneously collapses in the gas phase to a compact conformation.


Glass formation of a DMSO-water mixture probed with a photosynthetic pigment.
Despite their extensive industrial usage, glass-forming liquids are not fully understood, and methods to investigate their dynamical heterogeneity are sought after. Here we show how the appearance of a second component in the visible absorption spectrum of a photosynthetic pigment upon cooling can be used to probe the glass transition of a dimethylsulfoxide-water mixture. The changes in the relative ratio of the two components with respect to temperature follow a sigmoid curve, and we show that the second component arises due to protonation of the pigment at low temperatures. Furthermore, from visible transient absorption spectra we show that, unlike the first component, the dynamics of the second component slows down significantly at lower temperatures, suggesting that there are two distinct environments with fast and slow fluctuations. Our results therefore enable a new method to characterize the dynamical heterogeneity of glass-forming liquids.


A comparison of the effects of incubation on cells containing haemoglobin S or haemoglobin A.
1. A centrifugal packing technique has been used to investigate the effect of incubation on the flexibility of erythrocytes containing normal (Hb A) or sickle-cell (Hb S) haemoglobin. The effects on cell morphology and on the reversibility of morphological changes were also investigated. 2. The two types of cell reacted similarly to incubation. Sickling of the Hb-S was shown not to be responsible for the production of irreversibly deformed cells in these circumstances. 3. It is concluded that for both cell types the membrane inflexibility, cellular deformation and morphological irreversibility induced by the incubation were due to metabolic depletion.


Increased concentrations of presynaptic proteins in the cingulate cortex of subjects with schizophrenia.
Cytoarchitectural and neurochemical studies demonstrate disorganization in the cerebral cortex in schizophrenia, which perhaps underlies the severe behavioral disturbances of the disease. This neuronal disarray should be accompanied by synaptic abnormalities. As such, presynaptic proteins have proved valuable indexes of synaptic density and their concentrations have correlated markedly with synaptic loss. Our study sought to determine whether abnormalities exist in the concentrations of presynaptic proteins in the postmortem cerebral cortex of subjects with schizophrenia. Presynaptic protein immunoreactivities were assessed in 4 different cerebrocortical regions derived from 16 elderly controls, 19 elderly subjects with schizophrenia, and 24 subjects with Alzheimer's disease. Tissues were assayed with the monoclonal antibodies EP10 and SP4, which recognize synaptophysin, and the monoclonal antibodies SP6 and SP14, which detect syntaxin and synaptosomal-associated protein-25-kd immunoreactivities, respectively. In subjects with schizophrenia relative to controls, presynaptic proteins were increased in the cingulate cortex, but were unchanged in the temporal, frontal, and parietal cortices. In contrast, when cases with Alzheimer's disease were compared with controls, presynaptic proteins were decreased in the frontal, temporal, and parietal samples. These findings reveal changes in the synaptic organization of the cingulate cortex in schizophrenia relative to other areas examined. These changes are distinct from the deficits in presynaptic proteins observed in Alzheimer's disease.


Comparative Study of Endothelial Function and Uterine Artery Doppler Velocimetry between Pregnant Women with or without Preeclampsia Development.
Background. Poor placentation and systemic endothelial dysfunction have been identified as main events in Preeclampsia (PE). The relationship and chronology of these phenomena are important if we are to understand the pathophysiological mechanisms underlying this major clinical problem. Objectives. To compare the evolution of placentation and endothelial function in normotensive and preeclamptic pregnancies. Patients and methods. In a prospective cohort study, 59 pregnant women with a high risk of developing PE were subjected to flow-mediated dilation (FMD) and to Doppler velocimetry of uterine arteries in order to obtain their Pulsatility Index (UtA-PI). The variations in the FMD and UtA-PI values, between 16(+0) and 19(+6) and 24(+0) and 27(+6) weeks of gestation, were compared, taking PE development into consideration. Results. Nine patients developed PE and the other 50 women remained normotensive. At 16(+0) to 19(+6) weeks of pregnancy, patients that developed PE presented higher values of UtA-PI than the normotensive group, but there was no difference in FMD results between them. At 24(+0) to 27(+6) weeks, the patients that developed PE presented higher values of UtA-PI and lower values of FMD than the women that remained normotensive. Conclusions. These results corroborate the evidence that endothelial injury is secondary to poor placentation.


Users' guide to the surgical literature: how to assess an article about harm in surgery.
You are a new plastic surgeon in the community and you are referred a patient interested in breast reconstruction. The patient is a 35-year-old female school teacher who had a bilateral prophylactic mastectomy 2 years earlier, as she was a BRCA gene carrier. Since she is of a petite build with very little subcutaneous tissue or extra skin in the lower abdomen, you decide that she is not a suitable candidate for an abdomen-based autologous tissue reconstruction. You recommend the technique of tissue expansion and silicone gel implants. She is concerned, however, about the possibility of anaplastic large cell lymphoma (ALCL) developing in her breasts. She read in a magazine recently that ALCL, an unusual form of breast cancer, has been occurring in patients who have breast implants. She is very concerned that she might be at risk and asks for your opinion as to whether she should proceed with the procedure or not.


State-Level Community Benefit Regulation and Nonprofit Hospitals' Provision of Community Benefits.
Do nonprofit hospitals provide enough community benefits to justify their tax exemptions? States have sought to enhance nonprofit hospitals' accountability and oversight through regulation, including requirements to report community benefits, conduct community health needs assessments, provide minimum levels of community benefits, and adhere to minimum income eligibility standards for charity care. However, little research has assessed these regulations' impact on community benefits. Using 2009-11 Internal Revenue Service data on community benefit spending for more than eighteen hundred hospitals and the Hilltop Institute's data on community benefit regulation, we investigated the relationship between these four types of regulation and the level and types of hospital-provided community benefits. Our multivariate regression analyses showed that only community health needs assessments were consistently associated with greater community benefit spending. The results for reporting and minimum spending requirements were mixed, while minimum income eligibility standards for charity care were unrelated to community benefit spending. State adoption of multiple types of regulation was consistently associated with higher levels of hospital-provided community benefits, possibly because regulatory intensity conveys a strong signal to the hospital community that more spending is expected. This study can inform efforts to design regulations that will encourage hospitals to provide community benefits consistent with policy makers' goals.


Parent attitudes toward integrating parent involvement into teenage driver education courses.
The widespread adoption of graduated driver licensing (GDL) policies has effectively reduced crash risk for young drivers; however, parents must support, reinforce, and enforce GDL for it to be effective, and research indicates that parents need better information and instruction for adhering to GDL requirements, conducting supervised practice driving, and restricting independent teenage driving. Because teenagers in most states must take driver education to enter the licensing process prior to age 18, integrating parent involvement into driver education may be an effective way to inform and instruct parents on a large scale about teen driver safety. This study assessed parent attitudes (overall and by rural status, minority status, and income level) toward integrating parent involvement into teenage driver education classes. In this study, 321 parents of teenagers enrolled in driver education classes across the state of Montana completed surveys about current involvement in driver education and attitudes toward required involvement. The results indicated that parents were not very involved currently in their teenagers' driver education classes, but 76% reported that parents should be required to be involved. If involvement were required, parents would prefer having written materials sent home, access to information over the Internet, or discussions in person with the instructor; far fewer would prefer to attend classes or behind-the-wheel driving instruction. There were few differences in parent attitudes by rural or minority status but many by income level. Compared to higher income parents, lower income parents were more likely to endorse required parent involvement in teenage driver education classes and to want parent information from driver education about many teen driving issues. That the majority of parents are open to required involvement in their teenagers' driver education classes is promising because doing so could better prepare parents to understand and adhere to GDL policies, supervise teenagers' practice driving, and manage teen independent driving, all of which could further increase teen driver safety.


Dry Eye Disease: Present Challenges in the Management and Future Trends.
Dry eye disease (DED) is a tear film disorder resulting in hyperosmolarity of the tear film and inflammation of the ocular surface. DED is also referred to as keratoconjunctivitis sicca (KCS) and dry eye syndrome. DED represents a significant public health issue, particularly in older adults, and needs more research and attention. Despite the urgent need for safe and effective pharmacotherapies, there is currently only one approved medication, Restasis®, to tackle DED. In this review article, we present an overview of DED, classification, epidemiology, pathophysiology, diagnosis, and risk factors of DED. Special emphasis is placed on current treatment options for DED such as artificial tears, lipid-containing lubricants, liposomal spray, inserts, anti-inflammatory or immunosuppressant drops, antibiotics, dietary omega-3 essential fatty acids, autologous serum, intense-pulsed-light (IPL), punctual plugs, moisture-retaining eyeglasses, hydrophilic bandage contact lenses and secretagogues. The review also summarizes trends in DED treatment that are patented and are currently under investigation in clinical trials.


Shared Decision Making and Autonomy Among US Participants with Multiple Sclerosis in the NARCOMS Registry.
Treatment decisions in multiple sclerosis (MS) are affected by many factors and are made by the patient, doctor, or both. With new disease-modifying therapies (DMTs) emerging, the complexity surrounding treatment decisions is increasing, further emphasizing the importance of understanding decision-making preferences. North American Research Committee on Multiple Sclerosis (NARCOMS) Registry participants completed the Fall 2014 Update survey, which included the Control Preferences Scale (CPS). The CPS consists of five images showing different patient/doctor roles in treatment decision making. The images were collapsed to three categories: patient-centered, shared, and physician-centered decision-making preferences. Associations between decision-making preferences and demographic and clinical factors were evaluated using multivariable logistic regression. Of 7009 participants, 79.3% were women and 93.5% were white (mean [SD] age, 57.6 [10.3] years); 56.7% reported a history of relapses. Patient-centered decision making was most commonly preferred by participants (47.9%), followed by shared decision making (SDM; 42.8%). SDM preference was higher for women and those taking DMTs and increased with age and disease duration (all P < .05). Patient-centered decisions were most common for respondents not taking a DMT at the time of the survey and were preferred by those who had no DMT history compared with those who had previously taken a DMT (P < .0001). There was no difference in SDM preference by current MS disease course after adjusting for other disease-related factors. Responders reported most commonly considering their doctor's opinion before making a treatment decision and making decisions jointly with their doctor. DMT use, gender, and age were associated with decision-making preference.


Speaker sex influences processing of grammatical gender.
Spoken words carry linguistic and indexical information to listeners. Abstractionist models of spoken word recognition suggest that indexical information is stripped away in a process called normalization to allow processing of the linguistic message to proceed. In contrast, exemplar models of the lexicon suggest that indexical information is retained in memory, and influences the process of spoken word recognition. In the present study native Spanish listeners heard Spanish words that varied in grammatical gender (masculine, ending in -o, or feminine, ending in -a) produced by either a male or a female speaker. When asked to indicate the grammatical gender of the words, listeners were faster and more accurate when the sex of the speaker "matched" the grammatical gender than when the sex of the speaker and the grammatical gender "mismatched." No such interference was observed when listeners heard the same stimuli, but identified whether the speaker was male or female. This finding suggests that indexical information, in this case the sex of the speaker, influences not just processes associated with word recognition, but also higher-level processes associated with grammatical processing. This result also raises questions regarding the widespread assumption about the cognitive independence and automatic nature of grammatical processes.


Interventional optical molecular imaging guidance during percutaneous biopsy.
To investigate indocyanine green (ICG) as a molecular beacon for malignant lesions within the liver and evaluate the ability of a developed handheld imaging system to allow measurement of ICG fluorescence within focal hepatic lesions with high target-to-background ratios in a mouse model. All animal experiments were approved by the institutional animal care committee. A handheld optical molecular imaging device was constructed to pass through the introducer needle of a standard percutaneous biopsy kit. An ex vivo phantom system was constructed to quantify tissue attenuation properties of ICG in liver parenchyma. Subsequently, intrahepatic colorectal cancer metastases were generated in nude mice, and epifluorescence imaging of ICG, as well as histologic analysis of the explanted livers, was performed at 3 weeks after implantation (n = 6). Epifluorescence imaging with the handheld imaging device was then performed on intrahepatic colorectal metastases after the administration of ICG (n = 15) at 3, 6, and 24 hours after injection. Target-to-background ratios were calculated for each time point. Subsequently, a core biopsy of intrahepatic colorectal metastases was performed by using a standard clinical 18-gauge biopsy needle. There was avid localization of ICG to the focal lesions at all time points. Similarly, fluorescence within the tumors was greater than that within normal liver, as detected with the handheld imaging system (mean target-to-background ratio ± standard deviation, 3.9 ± 0.2 at 24 hours). A core biopsy of tumor and normal adjacent liver by using a standard biopsy needle demonstrated a sharp margin of fluorescence intensity at the tumor-liver interface. The custom-designed molecular imaging device, in combination with ICG, readily allowed differentiation between normal and malignant tissue in a murine model of intrahepatic colorectal metastasis.


Young children's preference for solitary play: Implications for socio-emotional and school adjustment.
The purpose of this study was to provide additional psychometric support for the Preference for Solitary Play Interview (PSPI) and to examine the associations between self-reported preference for solitary play and indices of adjustment in early childhood. Participants were N = 340 children attending kindergarten and grade 1. Children completed the PSPI, and teachers provided assessments of children's socio-emotional and school adjustment. In support of the validity of the PSPI, preference for solitary play was positively associated with asocial behaviours. Further, preference for solitary play displayed an indirect (but not direct) association with peer exclusion via asocial behaviours. Findings are discussed in terms of the social and behavioural implications of preference for solitary play in early childhood. Statement of contribution What is already known on this subject? Children who spend more time alone are at increased risk of adjustment difficulties. However, some individuals desire to spend time alone because of an appreciation for solitude. A preference for solitude is not associated with negative adjustment in adults and older youth. What does this study add? This study is among the first to examine self-reported preference for solitary in early childhood. Preference for solitude may not be related to emotional or school difficulties in young children. However, a heightened display of solitary behaviours may still evoke negative responses from peers.


A self management plan in the treatment of adult asthma.
To determine whether routine assessment of peak expiratory flow (PEF) in association with a self management plan based on inhaled corticosteroid use is effective in the management of chronic asthma, 36 consecutive adult patients with asthma attending an outpatient chest clinic were admitted to an open prospective study. Patients were treated with inhaled salbutamol and beclomethasone dipropionate in an attempt to achieve normal lung function. Each patient had a "potential normal value," which was either the predicted normal or the maximum PEF value achieved by the patient, whichever was the higher. Patients measured PEF at home and if it fell by more than 30% from the potential normal value the dose of beclomethasone was doubled until PEF returned to the potential normal value, then continued at 20 mg/day for the same number of days. If PEF fell to below 150-200 l/min patients were asked to obtain emergency medical assistance. In the 30 patients who completed the trial the six months before and the six months after intervention with the self management plan were compared. There was a substantial improvement in both subjective and objective measurements of asthma severity, with a significant reduction in nights woken, days lost from work, and requirement for oral corticosteroids and a significant increase in baseline lung function. Routine measurement of PEF in association with a self management plan appears to be effective in reducing symptoms of asthma and improving lung function.


COVID-19: Technology, Social Connections, Loneliness, and Leisure Activities: An International Study Protocol.
Drawn from the stress process model, the pandemic has imposed substantial stress to individual economic and mental well-being and has brought unprecedented disruptions to social life. In light of social distancing measures, and in particular physical distancing because of lockdown policies, the use of digital technologies has been regarded as the alternative to maintain economic and social activities. This paper aims to describe the design and implementation of an online survey created as an urgent, international response to the COVID-19 pandemic. The online survey described here responds to the need of understanding the effects of the pandemic on social interactions/relations and to provide findings on the extent to which digital technology is being utilized by citizens across different communities and countries around the world. It also aims to analyze the association of use of digital technologies with psychological well-being and levels of loneliness. The data will be based on the ongoing survey (comprised of several existing and validated instruments on digital use, psychological well-being and loneliness), open for 3 months after roll out (ends September) across 11 countries (Austria, France, Germany, India, Malta, Portugal, Romania, Spain, Turkey, and UK). Participants include residents aged 18 years and older in the countries and snowball sampling is employed via social media platforms. We anticipate that the findings of the survey will provide useful and much needed information on the prevalence of use and intensities of digital technologies among different age groups, gender, socioeconomic groups in a comparative perspective. Moreover, we expect that the future analysis of the data collected will show that different types of digital technologies and intensities of use are associated with psychological well-being and loneliness. To conclude, these findings from the study are expected to bring in our understanding the role of digital technologies in affecting individual social and emotional connections during a crisis.


Quantitative phase imaging of weakly scattering objects using partially coherent illumination.
In this paper, we extend our recent work on partially coherent quantitative phase imaging (pcQPI) from two-dimensional (2D) to three-dimensional (3D) imaging of weakly scattering samples. Due to the mathematical complexity, most theoretical modeling of quantitative phase image formation under partial coherence has focused on thin, well-focused samples. It is unclear how these abberations are affected by defocusing. Also, as 3D QPI techniques continue to develop, a better model needs to be developed to understand and quantify these aberrations when imaging thicker samples. Here, using the first order Born's approximation, we derived a mathematical framework that provides an intuitive model of image formation under varying degrees of coherence. Our description provides a clear connection between the halo effect and phase underestimation, defocusing and the 3D structure of the sample under investigation. Our results agree very well with the experiments and show that the microscope objective defines the sectioning ability of the imaging system while the condenser lens is responsible for the halo effect.


Professional doctorate supervision: exploring student and supervisor experiences.
This paper discusses research supervision within one professional doctorate programme; focusing on the processes and issues students and supervisors gave priority. An exploratory, descriptive approach was used to develop a pilot study. Data was collected from three sources. First student perceptions and experiences of supervision were obtained during a group workshop. Observations and field notes were gathered during this event. A second, similar event was undertaken with professional doctorate research supervisors. Finally a convenience sample of students and supervisors (two of each) participated in open ended one to one discussions regarding supervision. The discussions were tape recorded, transcribed and analysed, along with the workshop field notes and observations. There are implied differences between professional doctorates and the PhD relating to process, purpose and outcome [Yam, B., 2005. Professional doctorate and nursing practice. Nurse Education Today 25 (7), 564-572; Laing, S., 2000. Linking research to practice. Physiotherapy 86 (7), 371]. Given such implications professional doctorate research supervision could, or should, embrace critical engagement with issues related to the leadership of research in professional practice; moving beyond research and methodological issues. Indeed it had been presupposed such issues would emerge in this study. For example it had been anticipated that students would want a supervisor or adviser from within their practice setting, to provide insight and support in relation to leading research in practice and to complement the role of their research supervisor. Such complementary supervisory roles within the practice context are given credence in the literature [Kemp, S., 2004. Professional doctorates and doctoral education. International Journal of Organisational Behaviour 7 (4), 401-410]. However students preferred to utilise support networks within the professional doctorate itself to address professional issues.


Novel Fluorescence-Based High-Throughput FLIPR Assay Utilizing Membrane-Tethered Genetic Calcium Sensors to Identify T-Type Calcium Channel Modulators.
T-type voltage-gated Ca2+ channels have been implicated in many human disorders, and there has been increasing interest in developing highly selective and potent T-type Ca2+ channel modulators for potential clinical use. However, the unique biophysical properties of T-type Ca2+ channels are not conducive for developing high-throughput screening (HTS) assays to identify modulators, particularly potentiators. To illustrate, T-type Ca2+ channels are largely inactivated and unable to open to allow Ca2+ influx at -25 mV, the typical resting membrane potential of the cell lines commonly used in cellular screening assays. To address this issue, we developed cell lines that express Kir2.3 channels to hyperpolarize the membrane potential to -70 mV, thus allowing T-type channels to return to their resting state where they can be subsequently activated by membrane depolarization in the presence of extracellular KCl. Furthermore, to simplify the HTS assay and to reduce reagent cost, we stably expressed a membrane-tethered genetic calcium sensor, GCaMP6s-CAAX, that displays superior signal to the background compared to the untethered GCaMP6s or the synthetic Ca2+ sensor Fluo-4AM. Here, we describe a novel GCaMP6s-CAAX-based calcium assay utilizing a high-throughput fluorometric imaging plate reader (Molecular Devices, Sunnyvale, CA) format that can identify both activators and inhibitors of T-type Ca2+ channels. Lastly, we demonstrate the utility of this novel fluorescence-based assay to evaluate the activities of two distinct G-protein-coupled receptors, thus expanding the use of GCaMP6s-CAAX to a wide range of applications relevant for developing cellular assays in drug discovery.


Host genes associated with HIV-1 replication in lymphatic tissue.
Much effort has been spent recently in identifying host factors required for HIV-1 to effectively replicate in cultured human cells. However, much less is known about the genetic factors in vivo that impact viral replication in lymphatic tissue, the primary anatomical site of virus-host interactions where the bulk of viral replication and pathogenesis occurs. To identify genetic determinants in lymphatic tissue that critically affect HIV-1 replication, we used microarrays to transcriptionally profile and identify host genes expressed in inguinal lymph nodes that were associated determinants of viral load. Strikingly, ∼95% of the transcripts (558) in this data set (592 transcripts total) were negatively associated with HIV-1 replication. Genes in this subset 1) inhibit cellular activation/proliferation (e.g., TCFL5, SOCS5 and SCOS7, KLF10), 2) promote heterochromatin formation (e.g., HIC2, CREBZF, ZNF148/ZBP-89), 3) increase collagen synthesis (e.g., PLOD2, POSTN, CRTAP), and 4) reduce cellular transcription and translation. Potential anti-HIV-1 restriction factors were also identified (e.g., NR3C1, HNRNPU, PACT). Only ∼5% of the transcripts (34) were positively associated with HIV-1 replication. Paradoxically, nearly all of these genes function in innate and adaptive immunity, particularly highlighting heightened gene expression in the IFN system. We conclude that this conventional host response cannot contain HIV-1 replication and, in fact, could well contribute to increased replication through immune activation. More importantly, genes that have a negative association with virus replication point to target cell availability and potentially new viral restriction factors as principal determinants of viral load.


Sex-trafficking, violence, negotiating skill, and HIV infection in brothel-based sex workers of eastern India, adjoining Nepal, Bhutan, and Bangladesh.
A community-based cross-sectional study was conducted among brothel-based sex workers of West Bengal, eastern India, to understand sex-trafficking, violence, negotiating skills, and HIV infection in them. In total, 580 sex workers from brothels of four districts participated in the study. A pretested questionnaire was introduced to study their sociodemography, sex-trafficking, violence, and negotiating skills. Blood sample of 4-5 mL was collected from each sex worker using an unlinked anonymous method to study their HIV status. Data were edited and entered into a computer using the Epi Info software (6.04d version). Both univariate and multivariate analyses were done to find out any association between HIV and relevant risk factors. Results of the study revealed that a sizeable number of the participants were from Nepal (9%) and Bangladesh (7%). The seroprevalence of HIV was strikingly higher among Nepalese (43%) than among Bangladeshis (7%) and Indians (9%). Almost one in every four sex workers (24%) had joined the profession by being trafficked. Violence at the beginning of this profession was more among the trafficked victims, including those sold by their family members (57%) compared to those who joined the profession voluntarily (15%). The overall condom negotiation rate with most recent two clients was 38%. By multivariate analysis, HIV was significantly associated with sexual violence (odds ratio=2.3; 95% confidence interval 1.2-4.5). The study has documented that the trafficked victims faced violence, including sexual violence, to a greater magnitude, and sexual violence was associated with acquiring HIV in them. There is a need for an in-depth study to understand the problem of trafficking and its consequences.


Importance of extended protease substrate recognition motifs in steering BNIP-2 cleavage by human and mouse granzymes B.
Previous screening of the substrate repertoires and substrate specificity profiles of granzymes resulted in long substrate lists highly likely containing bystander substrates. Here, a recently developed degradomics technology that allows distinguishing efficiently from less efficiently cleaved substrates was applied to study the degradome of mouse granzyme B (mGrB). In vitro kinetic degradome analysis resulted in the identification of 37 mGrB cleavage events, 9 of which could be assigned as efficiently targeted ones. Previously, cleavage at the IEAD75 tetrapeptide motif of Bid was shown to be efficiently and exclusively targeted by human granzyme B (hGrB) and thus not by mGrB. Strikingly, and despite holding an identical P4-P1 human Bid (hBid) cleavage motif, mGrB was shown to efficiently cleave the BCL2/adenovirus E1B 19 kDa protein-interacting protein 2 or BNIP-2 at IEAD28. Like Bid, BNIP-2 represents a pro-apoptotic Bcl-2 protein family member and a potential regulator of GrB induced cell death. Next, in vitro analyses demonstrated the increased efficiency of human and mouse BNIP-2 cleavage by mGrB as compared to hGrB indicative for differing Bid/BNIP-2 substrate traits beyond the P4-P1 IEAD cleavage motif influencing cleavage efficiency. Murinisation of differential primed site residues in hBNIP-2 revealed that, although all contributing, a single mutation at the P3' position was found to significantly increase the mGrB/hGrB cleavage ratio, whereas mutating the P1' position from I29 > T yielded a 4-fold increase in mGrB cleavage efficiency. Finally, mutagenesis analyses revealed the composite BNIP-2 precursor patterns to be the result of alternative translation initiation at near-cognate start sites within the 5' leader sequence (5'UTR) of BNIP-2. Despite their high sequence similarity, and previously explained by their distinct tetrapeptide specificities observed, the substrate repertoires of mouse and human granzymes B only partially overlap. Here, we show that the substrate sequence context beyond the P4-P1 positions can influence orthologous granzyme B cleavage efficiencies to an unmatched extent. More specifically, in BNIP-2, the identical and hGrB optimal IEAD tetrapeptide substrate motif is targeted highly efficiently by mGrB, while this tetrapeptide motif is refractory towards mGrB cleavage in Bid.


Co-proxamol and suicide: preventing the continuing toll of overdose deaths.
Restricting means for suicide is a key element in suicide prevention strategies of all countries where these have been introduced. Preventing deaths from analgesic overdoses is highlighted in the National Suicide Prevention Strategy for England. The problem of self-poisoning with the prescription-only drug co-proxamol (dextropropoxyphene plus paracetamol) has received attention in several countries. We have conducted a review of the international literature related to possible strategies to tackle this problem. In England and Wales in 1997-1999, 18% of drug-related suicides involved co-proxamol; these constituted 5% of all suicides. Death usually results from the toxic effects of dextropropoxyphene on respiration or cardiac function. Death from co-proxamol overdose may occur rapidly, the lethal dose can be relatively low, and the effects are potentiated by alcohol and other CNS depressants. The majority of co-proxamol overdose deaths occur before hospital treatment can be received. The risk can extend to others in the household of the person for whom the drug is prescribed. While there is limited evidence that educational strategies have been effective in reducing deaths from co-proxamol poisoning, initiatives in Scandinavia, Australia and the UK to restrict availability of co-proxamol have produced promising results. Given the paucity of evidence for superior therapeutic efficacy of co-proxamol over other less toxic analgesics, there are good reasons to question whether it should continue to be prescribed.


Antimicrobial Peptides from Rat-Tailed Maggots of the Drone Fly Eristalis tenax Show Potent Activity against Multidrug-Resistant Gram-Negative Bacteria.
The spread of multidrug-resistant Gram-negative bacteria is an increasing threat to human health, because novel compound classes for the development of antibiotics have not been discovered for decades. Antimicrobial peptides (AMPs) may provide a much-needed breakthrough because these immunity-related defense molecules protect many eukaryotes against Gram-negative pathogens. Recent concepts in evolutionary immunology predict the presence of potent AMPs in insects that have adapted to survive in habitats with extreme microbial contamination. For example, the saprophagous and coprophagous maggots of the drone fly Eristalis tenax (Diptera) can flourish in polluted aquatic habitats, such as sewage tanks and farmyard liquid manure storage pits. We used next-generation sequencing to screen the E. tenax immunity-related transcriptome for AMPs that are synthesized in response to the injection of bacterial lipopolysaccharide. We identified 22 AMPs and selected nine for larger-scale synthesis to test their activity against a broad spectrum of pathogens, including multidrug-resistant Gram-negative bacteria. Two cecropin-like peptides (EtCec1-a and EtCec2-a) and a diptericin-like peptide (EtDip) displayed strong activity against the pathogens, even under simulated physiological conditions, and also achieved a good therapeutic window. Therefore, these AMPs could be used as leads for the development of novel antibiotics.


A novel dehydrin-like protein from Aspergillus fumigatus regulates freezing tolerance.
We recently reported on DprA and DprB, dehydrin-like proteins involved in the protection against oxidative, osmotic and pH stress in Aspergillus fumigatus. A third dehydrin-like gene, DprC, was detected in the genome of the filamentous fungus and repressed during conidial germination. The deletion mutant was impaired in freezing tolerance. The DprC-mediated response was dependent on the SakA MAP kinase pathway, and accordingly, SakA phosphorylation was induced by cold shock. When fused to eGFP, the protein was associated with the vacuoles.


Enhanced 5-methylcytosine detection in single-molecule, real-time sequencing via Tet1 oxidation.
DNA methylation serves as an important epigenetic mark in both eukaryotic and prokaryotic organisms. In eukaryotes, the most common epigenetic mark is 5-methylcytosine, whereas prokaryotes can have 6-methyladenine, 4-methylcytosine, or 5-methylcytosine. Single-molecule, real-time sequencing is capable of directly detecting all three types of modified bases. However, the kinetic signature of 5-methylcytosine is subtle, which presents a challenge for detection. We investigated whether conversion of 5-methylcytosine to 5-carboxylcytosine using the enzyme Tet1 would enhance the kinetic signature, thereby improving detection. We characterized the kinetic signatures of various cytosine modifications, demonstrating that 5-carboxylcytosine has a larger impact on the local polymerase rate than 5-methylcytosine. Using Tet1-mediated conversion, we show improved detection of 5-methylcytosine using in vitro methylated templates and apply the method to the characterization of 5-methylcytosine sites in the genomes of Escherichia coli MG1655 and Bacillus halodurans C-125. We have developed a method for the enhancement of directly detecting 5-methylcytosine during single-molecule, real-time sequencing. Using Tet1 to convert 5-methylcytosine to 5-carboxylcytosine improves the detection rate of this important epigenetic marker, thereby complementing the set of readily detectable microbial base modifications, and enhancing the ability to interrogate eukaryotic epigenetic markers.


Longitudinal analysis of the lung microbiota of cynomolgous macaques during long-term SHIV infection.
Longitudinal studies of the lung microbiome are challenging due to the invasive nature of sample collection. In addition, studies of the lung microbiome in human disease are usually performed after disease onset, limiting the ability to determine early events in the lung. We used a non-human primate model to assess lung microbiome alterations over time in response to an HIV-like immunosuppression and determined impact of the lung microbiome on development of obstructive lung disease. Cynomolgous macaques were infected with the SIV-HIV chimeric virus SHIV89.6P. Bronchoalveolar lavage fluid samples were collected pre-infection and every 4 weeks for 53 weeks post-infection. The microbiota was characterized at each time point by 16S ribosomal RNA (rRNA) sequencing. We observed individual variation in the composition of the lung microbiota with a proportion of the macaques having Tropheryma whipplei as the dominant organism in their lungs. Bacterial communities varied over time both within and between animals, but there did not appear to be a systematic alteration due to SHIV infection. Development of obstructive lung disease in the SHIV-infected animals was characterized by a relative increase in abundance of oral anaerobes. Network analysis further identified a difference in community composition that accompanied the development of obstructive disease with negative correlations between members of the obstructed and non-obstructed groups. This emphasizes how species shifts can impact multiple other species, potentially resulting in disease. This study is the first to investigate the dynamics of the lung microbiota over time and in response to immunosuppression in a non-human primate model. The persistence of oral bacteria in the lung and their association with obstruction suggest a potential role in pathogenesis. The lung microbiome in the non-human primate is a valuable tool for examining the impact of the lung microbiome in human health and disease.


Antibacterial action of several tannins against Staphylococcus aureus.
We examined the antibacterial action of several tannins on plasma coagulation by Staphylococcus aureus and the effect of conventional chemotherapy combined with tannic acid below the MIC. Coagulation was inhibited in plasma containing tannic acid (100 mg/L), gallic acid (5000 mg/L), ellagic acid (5000 mg/L), (-)-epicatechin (1500 mg/L), (-)-epicatechin gallate (500 mg/L) or (-)-epigallocatechin gallate (200 mg/L) after incubation for 24 h. All tannins inhibited coagulation at a concentration below the MIC. The MICs of oxacillin and cefdinir for S. aureus were reduced to < or = 0.06 mg/L in Mueller-Hinton agar plates with tannic acid (100 mg/L) at a concentration below the MIC. The antistaphylococcal activity of tannic acid was reduced in plates with 10% rabbit blood, but not in those with 10% rabbit plasma. Membranous structures formed in a culture medium containing equal proportions of plasma and tryptic soy broth after incubation for 24 h. The colony counts of S. aureus in membranous structures in the medium containing oxacillin (40 mg/L) and tannic acid (100 mg/L) were c. 10-fold lower than those in medium containing oxacillin (40 mg/L) alone (P < 0.01). Tannic acid merits further investigation as a possible adjuvant agent against S. aureus skin infections treated with beta-lactam antibiotics.


Phospho-sulindac (OXT-922) inhibits the growth of human colon cancer cell lines: a redox/polyamine-dependent effect.
Non-steroidal anti-inflammatory drugs such as sulindac are promising chemoprevention agents against colon cancer, but their weak potency and side effects limit their use for both chemoprevention and chemotherapy. Here, we evaluated the effect of a new sulindac derivative, phospho-sulindac or OXT-922, on the growth of human cancer cell lines and its mechanism of action. OXT-922 inhibited the growth of human cancer cell lines originating from colon, pancreas and breast ~11- to 30-fold more potently than sulindac. This effect was mediated by a strong cytokinetic effect. Compared with control, OXT-922 inhibited cell proliferation by up to 67%, induced apoptosis 4.1-fold over control and blocked the G(1) to S cell cycle phase transition. OXT-922 suppressed the levels of cell cycle regulating proteins, including cyclins D(1) and D(3) and Cyclin-dependent kinases (CDK) 4 and 6. The levels of intracellular reactive oxygen species (ROS), especially those of mitochondrial O₂ⁱ⁻, were markedly elevated (5.5-fold) in response to OXT-922. ROS collapsed the mitochondrial membrane potential and triggered apoptosis, which was largely abrogated by antioxidants. OXT-922 suppressed nuclear factor-kappaB activation and downregulated thioredoxin-1 expression. It also suppressed the production of prostaglandin E(2) and decreased cyclooxygenase-1 expression. Similar to sulindac, OXT-922 enhanced spermidine/spermine N(1)-acetyltransferase activity, reduced the cellular polyamine content and synergized with difluoromethylornithine to inhibit cancer cell proliferation and induce apoptosis. Our results suggest that OXT-922 possesses promising anticancer properties and deserves further evaluation.


Determinants of exercise adherence and maintenance among cancer survivors: a systematic review.
For an exercise intervention to be successful, it is important that cancer survivors adhere to the prescribed program. To be able to improve adherence and to preserve achieved beneficial effects, insights into the relevant and modifiable determinants is important. Therefore, we aimed to systematically review determinants of exercise adherence and maintenance in cancer survivors using a socio-ecological approach.Studies were identified in PubMed, Embase, PsycINFO and SPORTDiscus up to July 2013. We included full-text articles that: 1) were conducted among adult cancer survivors; 2) quantitatively assessed factors associated with intervention adherence and maintenance, and 3) were published in English. The methodological quality of the selected studies was examined. A best evidence synthesis was applied. Eighteen studies were included. Median methodological quality was 53% and ranged from 21-78% of maximum score. Twelve studies focused on determinants of exercise adherence and evaluated 71 potential determinants: 29 demographic and clinical, 27 psychological, ten physical, four social factors, and one environmental factor. Six studies focused on determinants of exercise maintenance after completion of an intervention, and investigated 63 factors: 22 demographic and clinical, 28 psychosocial, nine physical, three social and one environmental factor. We found moderate evidence for a positive association between exercise history and exercise adherence. Inconsistent findings were found for age, gender and education as well as for psychological factors such as stage of change, perceived behavioral control, self-efficacy, extraversion, attitude, intention, fatigue, and quality of life, and physical factors including cardiovascular fitness, body mass index, and baseline physical activity.Exercise history is positively associated with exercise adherence. Future trials should further study the influence of social and environmental determinants on exercise adherence and maintenance in addition to demographic, psychological and physical determinants.


HLA-DR3 transgenic mice immunized with adenovirus encoding the thyrotropin receptor: T cell epitopes and functional analysis of the CD40 Graves' polymorphism.
The major histocompatibility (MHC) molecule HLA-DR3 is a susceptibility gene for Graves' disease (GD) in Caucasians. Mice lacking murine MHC and expressing human HLA-DR3 develop thyrotropin receptor (TSHR) antibodies and sometimes hyperthyroidism after vaccination with TSHR-DNA. MHC molecules present peptides processed from antigens to T cells. Therefore, we used DR3-transgenic mice to investigate recognition of TSHR ectodomain peptides. After immunization with TSHR A-subunit adenovirus (A-subunit-Ad) but not control-adenovirus (Control-Ad), splenocytes from DR3 mice responded to A-subunit protein in culture by producing interferon-gamma (IFN-gamma). When challenged with 29 overlapping TSHR peptides, splenocytes from A-subunit-Ad- or Control-Ad-immunized mice responded to several peptides. However, in splenocytes from A-subunit-Ad but not Control-Ad mice, a peptide containing TSHR residues 142-161 induced significantly more IFN-gamma than the same splenocytes in medium alone. Immunized DR3 mice also permitted testing the TSHR-specific function of the CD40 single nucleotide polymorphism (C vs. T) associated with GD. Of three human DR3 human Epstein-Barr virus lines (EBVL), two had C in both alleles (CC) and one was CT. However, these EBVL presented peptides poorly and there was no difference between CC vs. CT EBVL in peptide presentation to splenocytes from immunized mice. A peptide corresponding to residues 145-163 is one of seven suggested to be important in GD based on HLA-binding affinities, T-epitope algorithms, and human studies. Consequently, as in human GD, TSHR amino acids 142-161 appear to include a major T cell epitope in HLA-DR3 transgenic mice immunized with A-subunit-Ad.


Determinants of drug absorption in different ECMO circuits.
The aim of this in vitro study was to evaluate potential determinants of drug loss in different ECMO circuits. Midazolam, morphine, fentanyl, paracetamol, cefazolin, meropenem and vancomycin were injected into three neonatal roller pump, two paediatric roller pump and two clinically used neonatal roller pump circuits, all with a silicone membrane, and two neonatal centrifugal pump circuits with polypropylene hollow-fibre membranes. Serial blood samples were taken from a post-oxygenator site. Drug recovery was calculated as the ratio between the determined and the theoretical maximum concentration. The latter was obtained by dividing dose by theoretical circuit volume. Average drug recoveries at 180 min in three neonatal silicone membrane roller pump circuits were midazolam 0.62%, morphine 23.9%, fentanyl 0.35%, paracetamol 34.0%, cefazolin 84.3%, meropenem 82.9% and vancomycin 67.8%. There was a significant correlation between the lipophilicity of the drug expressed as log P and the extent of drug absorption, p < 0.001. The recovery of midazolam and fentanyl in centrifugal pump circuits with hollow-fibre membrane oxygenator was significantly higher compared to neonatal roller pump circuits with silicone membranes: midazolam 63.4 versus 0.62%, fentanyl 33.8 versus 0.35%, p < 0.001. Oxygenator size and used circuits do not significantly affect drug losses. Significant absorption of drugs occurs in the ECMO circuit, correlating with increased lipophilicity of the drug. Centrifugal pump circuits with hollow-fibre membrane oxygenators show less absorption for all drugs, most pronounced for lipophilic drugs. These results suggest that pharmacokinetics and hence optimal doses of these drugs may be altered during ECMO.


An evaluation of a hybrid occupational therapy and supported employment program in Japan for persons with schizophrenia.
A vocational rehabilitation program (occupational therapy and supported employment) for promoting the return to the community of long-stay persons with schizophrenia was established at a psychiatric hospital in Japan. The purpose of the study was to evaluate the program in terms of hospitalization rates, community tenure, and social functioning with each individual serving as his or her control. Fifty-two participants, averaging 8.9 years of hospitalization, participated in the vocational rehabilitation program consisting of 2 to 6 hours of in-hospital occupational therapy for 6 days per week and a post-discharge supported employment component. Seventeen years after the program was established, a retrospective study was conducted to evaluate the impact of the program on hospitalizations, community tenure, and social functioning after participants' discharge from hospital, using an interrupted time-series analysis. The postdischarge period was compared with the period from onset of illness to the index discharge on the three outcome variables. After discharge from the hospital, the length of time spent by participants out of the hospital increased, social functioning improved, and risk of hospitalization diminished by 50%. Female participants and those with supportive families spent more time out of the hospital than participants who were male or came from nonsupportive families. A combined program of occupational therapy and supported employment was successful in a Japanese psychiatric hospital when implemented with the continuing involvement of a clinical team. Interventions that improve the emotional and housing supports provided to persons with schizophrenia by their families are likely to enhance the outcome of vocational services.


Neurocognitive outcomes after cardiac surgery.
To summarize recent studies of neurocognitive dysfunction after cardiac surgery, as well as to outline efforts and approaches toward advancing the field. Observational studies have improved our understanding of the incidence and the trajectory of cognitive decline after cardiac surgery; however, the magnitude of this neurocognitive change remains controversial because of the inconsistent definitions and the lack of a gold-standard diagnostic modality. Nonetheless, physicians commonly see patients with functional and cognitive impairments after cardiac surgery, which utilize healthcare resources and impact quality of life. Novel approaches have utilized advanced neuroimaging techniques as well as innovative monitoring modalities to improve the efficiency of neuroprotective strategies during cardiac surgery. Adverse cognitive and neurologic outcomes following cardiac surgery range from discrete neurocognitive deficits to severe neurologic injury such as stroke and even death. The elderly are at higher risk of suffering these outcomes and the public health dimension of this problem is expected to accelerate. Future studies should combine advanced neuroimaging with genomic, transcriptional, proteomic, and metabolomic profiling to improve our understanding of the pathophysiologic mechanisms and optimize the diagnosis, prevention, and treatment of neurocognitive injury.


Staphylococcus aureus protein A causes osteoblasts to hyper-mineralise in a 3D extra-cellular matrix environment.
Osteomyelitis is an inflammatory bone infection that is caused most commonly by the opportunistic pathogen Staphylococcus aureus. Research into staphylococcal induced bone infection is typically conducted using traditional 2D in vitro culture settings, which is not fully representative of the dynamic in vivo environment. In this study we utilised a collagen glycosaminoglycan scaffold, previously developed for bone tissue engineering, as a representative 3D model of infection. The scaffold resisted degradation and retained its pore structure, which is important for cellular function and survival, when seeded with both cells and bacteria. Using this model, we showed that in the presence of S. aureus, osteoblast proliferation was reduced over 21 days. Interestingly however these cells were more metabolically active compared to the uninfected cells and demonstrated increased mineralisation. Protein A (SpA) is a virulence factor found on the surface of S. aureus and has been shown to interact with osteoblasts. When SpA was removed from the surface of S. aureus, the osteoblasts show comparable activity with the uninfected cells-demonstrating the importance of SpA in the interaction between bone cells and S. aureus. Our results suggest that infected osteoblasts are capable of over-compensating for bone loss and bone destruction by increasing mineralisation in a 3D environment, key elements required for ensuring bone strength. It also reinforces our previously established result that S. aureus SpA is a critical mediator in osteomyelitis and might be a potential novel drug target to treat osteomyelitis by preventing the interaction between S. aureus and osteoblasts.


Efficient bipedal robots based on passive-dynamic walkers.
Passive-dynamic walkers are simple mechanical devices, composed of solid parts connected by joints, that walk stably down a slope. They have no motors or controllers, yet can have remarkably humanlike motions. This suggests that these machines are useful models of human locomotion; however, they cannot walk on level ground. Here we present three robots based on passive-dynamics, with small active power sources substituted for gravity, which can walk on level ground. These robots use less control and less energy than other powered robots, yet walk more naturally, further suggesting the importance of passive-dynamics in human locomotion.


Mast cells in innate immunity.
Mast cells are known to be the main effector cells in the elicitation of the IgE-mediated allergic response. The specific location of mast cells within tissues that interface the external environment, and the extent of their functional capacity, including the ability to phagocytose and to produce and secrete a wide spectrum of mediators, have led investigators to propose a potential role for mast cells in innate immune responses. Certain microorganisms have been found to interact either directly or indirectly with mast cells. This interaction results in mast cell activation and mediator release which elicit an inflammatory response or direct killing leading to bacterial clearance. The in vivo relevance of these in vitro observations has been demonstrated by the use of complement-deficient and/or mast cell-deficient and mast cell-reconstituted mice. In thus has been shown that both C3 and mast cell- and tumor necrosis factor-alpha-dependent recruitment of circulating leukocytes with bactericidal properties are crucial to a full response in certain models of acute infection. Modulation of mast cell numbers in vivo was also found to affect the host response against bacterial infection. Thus, mast cells do have a role in innate immunity in defined animal models of bacterial infection. Whether mast cells participate in innate immune responses in the protection of the human host against bacteria remains to be determined.


Personality factors in recently diagnosed multiple sclerosis patients: a preliminary investigation with the NEO-FFI scale.
This article describes some prevalent personality dimensions of recently diagnosed multiple sclerosis patients. A sample of 33 female recently diagnosed with relapsing-remitting multiple sclerosis (RRMS) was assessed with the NEO-FFI personality scale. Beck depression (BDI) and anxiety (BAI) scales were also used. No significant levels of anxiety or depression were identified in this group. As for personality factors, conscientiousness was the most common factor found, whereas openness to experience was the least observed. Literature on the relationship between personality and MS is scarce and there are no Brazilian studies on this subject. Some personality traits might complicate or facilitate the experience of living with a chronic, disabling and uncertain neurological condition such as MS.


White matter integrity in polydrug users in relation to attachment and personality: a controlled diffusion tensor imaging study.
The relationship between substance use disorders (SUD) and brain deficits has been studied extensively. However, there is still a lack of research focusing on the structural neural connectivity in long-term polydrug use disorder (PUD). Since a deficiency in white matter integrity has been reported as being related to various parameters of increased psychopathology, it might be considered an aggravating factor in the treatment of SUD. In this study we compared two groups of PUD inpatients (abstinent: n = 18, in maintenance treatment: n = 15) to healthy controls (n = 16) with respect to neural connectivity in white matter, and their relation to behavioral parameters of personality factors/organization and attachment styles. Diffusion Tensor Imaging was used to investigate white matter structure. Compared with healthy controls, the PUD patients showed reduced fractional anisotropy (FA) and increased radial diffusivity (RD) mainly in the superior fasciculus longitudinalis and the superior corona radiata. These findings suggest diminished neural connectivity as a result of myelin pathology in PUD patients. In line with our assumptions, we observed FA in the biggest cluster as negatively correlated with anxious attachment (r = 0.36, p < 0.05), personality dysfunctioning (r = -0.41; p < 0.01) as well positively correlated with personality factors Openness (r = 0.34; p < 0.05) and Agreeableness (r = 0.28; p < 0.05). Correspondingly these findings were inversely mirrored by RD. Further research employing enhanced samples and addressing longitudinally neuronal plastic effects of SUD treatment in relation to changes in personality and attachment is recommended.


Formation of phase gratings on the end of gradient-index lenses with ultraviolet ablation at 193 nm.
We present a simple, cost-effective method for creating diffractive optical elements on the surfaces of optical fibers and fiber-optic components by use of 193-nm ablation techniques. It is an outgrowth of a more fundamental investigation of the effects of intense UV radiation fields on SiO2- and Ge-SiO2-based structures (specifically optical fibers and preforms) and allows the inexpensive fabrication of structures such as the suggested evanescent-field-based sensing device.


Improved sensitive high performance liquid chromatography assay for glucosamine in human and rat biological samples with fluorescence detection.
An improved HPLC method with fluorescence detection was developed and validated for determination of glucosamine in human and rat biological samples. Aliquot of 0.1 mL plasma was spiked with mannosamine HCl as the internal standard (IS); proteins were precipitated with acetonitrile; the clear layer was derivatized with 9-fluorenylmethyl chloroformate (8 mM/acetonitrile) in presence of borate 0.2 M buffer at 30 degrees C for 30 min. The excess derivatizing agent was removed with 1-aminoadamantane HCl (300 mM in acetonitrile-water 1:1). Chromatographic separation was achieved on a C18 (100 mm X 4.6 mm, id 3 microm) reversed phase column using 0.1% acetic acid/acetoniltrile gradient mobile phase at 1 mL/min flow rate. Glucosamine was determined in the plasma of a human and rats and also in rat urine. The analytes were detected at excitation and emission wavelengths of 263 and 315 nm, respectively. The assay was linear over the range of 0.05-20 microg/mL with a typical correlation coefficient of 0.999 and intra-day and inter-day coefficient of variation of <15%. The lowest limit of quantification was set at 50 ng/mL. The recovery for glucosamine and mannosamine was 98 and 96%, respectively. We were able to improve glucosamine assay suitable to quantify glucosamine in both human and rat plasma and rat urine.


Arterial territory-specific phosphorylated retinoblastoma protein species and CDK2 promote differences in the vascular smooth muscle cell response to mitogens.
Despite recent advances in medical procedures, cardiovascular disease remains a clinical challenge and the leading cause of mortality in the western world. The condition causes progressive smooth muscle cell (SMC) dedifferentiation, proliferation, and migration that contribute to vascular restenosis. The incidence of disease of the internal mammary artery (IMA), however, is much lower than in nearly all other arteries. The etiology of this IMA disease resistance is not well understood. Here, using paired primary IMA and coronary artery SMCs, serum stimulation, siRNA knockdowns, and verifications in porcine vessels in vivo, we investigate the molecular mechanisms that could account for this increased disease resistance of internal mammary SMCs. We show that the residue-specific phosphorylation profile of the retinoblastoma tumor suppressor protein (Rb) appears to differ significantly between IMA and coronary artery SMCs in cultured human cells. We also report that the differential profile of Rb phosphorylation may follow as a consequence of differences in the content of cyclin-dependent kinase 2 (CDK2) and the CDK4 phosphorylation inhibitor p15. Finally, we present evidence that siRNA-mediated CDK2 knockdown alters the profile of Rb phosphorylation in coronary artery SMCs, as well as the proliferative response of these cells to mitogenic stimulation. The intrinsic functional and protein composition specificity of the SMCs population in the coronary artery may contribute to the increased prevalence of restenosis and atherosclerosis in the coronary arteries as compared with the internal mammary arteries.