Hyperbaric oxygenation for tumour sensitisation to radiotherapy.
Cancer is a common disease and radiotherapy is one well-established treatment for some solid tumours. Hyperbaric oxygenation therapy (HBOT) may improve the ability of radiotherapy to kill hypoxic cancer cells, so the administration of radiotherapy while breathing hyperbaric oxygen may result in a reduction in mortality and recurrence. To assess the benefits and harms of administering radiotherapy for the treatment of malignant tumours while breathing HBO. In September 2017 we searched the Cochrane Central Register of Controlled Trials (CENTRAL), the Cochrane Library Issue 8, 2017, MEDLINE, Embase, and the Database of Randomised Trials in Hyperbaric Medicine using the same strategies used in 2011 and 2015, and examined the reference lists of included articles. Randomised and quasi-randomised studies comparing the outcome of malignant tumours following radiation therapy while breathing HBO versus air or an alternative sensitising agent. Three review authors independently evaluated the quality of and extracted data from the included trials. We included 19 trials in this review (2286 participants: 1103 allocated to HBOT and 1153 to control).For head and neck cancer, there was an overall reduction in the risk of dying at both one year and five years after therapy (risk ratio (RR) 0.83, 95% confidence interval (CI) 0.70 to 0.98, number needed to treat for an additional beneficial outcome (NNTB) = 11 and RR 0.82, 95% CI 0.69 to 0.98, high-quality evidence), and some evidence of improved local tumour control immediately following irradiation (RR with HBOT 0.58, 95% CI 0.39 to 0.85, moderate-quality evidence due to imprecision). There was a lower incidence of local recurrence of tumour when using HBOT at both one and five years (RR at one year 0.66, 95% CI 0.56 to 0.78, high-quality evidence; RR at five years 0.77, 95% CI 0.62 to 0.95, moderate-quality evidence due to inconsistency between trials). There was also some evidence with regard to the chance of metastasis at five years (RR with HBOT 0.45 95% CI 0.09 to 2.30, single trial moderate quality evidence imprecision). No trials reported a quality of life assessment. Any benefits come at the cost of an increased risk of severe local radiation reactions with HBOT (severe radiation reaction RR 2.64, 95% CI 1.65 to 4.23, high-quality evidence). However, the available evidence failed to clearly demonstrate an increased risk of seizures from acute oxygen toxicity (RR 4.3, 95% CI 0.47 to 39.6, moderate-quality evidence).For carcinoma of the uterine cervix, there was no clear benefit in terms of mortality at either one year or five years (RR with HBOT at one year 0.88, 95% CI 0.69 to 1.11, high-quality evidence; RR at five years 0.95, 95% CI 0.80 to 1.14, moderate-quality evidence due to inconsistency between trials). Similarly, there was no clear evidence of a benefit of HBOT in the reported rate of local recurrence (RR with HBOT at one year 0.82, 95% CI 0.63 to 1.06, high-quality evidence; RR at five years 0.85, 95% CI 0.65 to 1.13, moderate-quality evidence due to inconsistency between trials). We also found no clear evidence for any effect of HBOT on the rate of development of metastases at both two years and five years (two years RR with HBOT 1.05, 95% CI 0.84 to 1.31, high quality evidence; five years RR 0.79, 95% CI 0.50 to 1.26, moderate-quality evidence due to inconsistency). There were, however, increased adverse effects with HBOT. The risk of a severe radiation injury at the time of treatment with HBOT was 2.05, 95% CI 1.22 to 3.46, high-quality evidence. No trials reported any failure of local tumour control, quality of life assessments, or the risk of seizures during treatment.With regard to the treatment of urinary bladder cancer, there was no clear evidence of a benefit in terms of mortality from HBOT at one year (RR 0.97, 95% CI 0.74 to 1.27, high-quality evidence), nor any benefit in the risk of developing metastases at two years (RR 2.0, 95% CI 0.58 to 6.91, moderate-quality evidence due to imprecision). No trial reported on failure of local control, local recurrence, quality of life, or adverse effects.When all cancer types were combined, there was evidence for an increased risk of severe radiation tissue injury during the course of radiotherapy with HBOT (RR 2.35, 95% CI 1.66 to 3.33, high-quality evidence) and of oxygen toxic seizures during treatment (RR with HBOT 6.76, 96% CI 1.16 to 39.31, moderate-quality evidence due to imprecision). We found evidence that HBOT improves local tumour control, mortality, and local tumour recurrence for cancers of the head and neck. These benefits may only occur with unusual fractionation schemes. Hyperbaric oxygenation therapy is associated with severe tissue radiation injury. Given the methodological and reporting inadequacies of the included studies, our results demand a cautious interpretation. More research is needed for head and neck cancer, but is probably not justified for uterine cervical or bladder cancer. There is little evidence available concerning malignancies at other anatomical sites.


Geochemical modeling of leaching from MSWI air-pollution-control residues.
This paper provides an improved understanding of the leaching behavior of waste incineration air-pollution-control (APC) residues in a long-term perspective. Leaching was investigated by a series of batch experiments reflecting leaching conditions after initial washout of highly soluble salts from residues. Leaching experiments were performed at a range of pH-values using carbonated and noncarbonated versions of two APC residues. The leaching data were evaluated by geochemical speciation modeling and discussed with respect to possible solubility control. The leaching of major elements as well as trace elements was generally found to be strongly dependent on pH. As leaching characterization was performed in the absence of high salt levels, the presented results represent long-term leaching after initial washout from a disposal site, that is, liquid-to-solid ratios above 1-2 L/kg. The leaching of Al, Ba, Ca, Cr, Pb, S, Si, V, and Zn was found influenced by solubility control from A12O3, Al(OH)3, Ba(S,Cr)04 solid solutions, BaSO4, Ca6Al2(SO4)3(OH)12 x 26H2O, CaAl2Si4O12 x 2H2O, Ca(OH)2, CaSiO3, CaSO4 x 2H2O, CaZn2(OH)6 x 2H2O, KAlSi2O6, PbCO3, PbCrO4, Pb2O3, Pb2V2O7, Pb3(VO4)2, ZnO, Zn2SiO4, and ZnSiO3. The presented dataset and modeling results form a thorough contribution to the assessment of long-term leaching behavior of APC residues under a wide range of conditions.


Oral health in renal transplant recipients administered cyclosporin A or tacrolimus.
The aim of this study was to determine the oral status of renal transplant recipients receiving cyclosporin A (CsA) or tacrolimus (FK-506) as immunosuppressant. A total of 88 renal transplant recipients receiving CsA (63 men and 25 women, mean age 51.4 years) and 67 receiving FK-506 (57 men and 10 women, mean age 33.5 years) were included in the study. Donor type, histocompatibility, cold ischemia time and prior delayed graft function were similar between the two groups. Demographics and pharmacological data were recorded for all subjects. The results demonstrated that CsA caused a greater number of oral diseases. A greater number of gingival overgrowth was present in patients treated with CsA. However, the combined use with calcium channel blockers increased the gingival overgrowth number. The occurrence of candida in saliva was observed in 80 renal recipients treated with CsA and 20 treated with FK-506. The presence of squamous oral carcinoma (n = 3) and herpes simplex (n = 10) was observed in patients treated with CsA. These alterations were not observed in renal recipients treated with FK-506. Renal recipients constitute a high-risk group for oral diseases, as they are immunocompromised. However, the FK-506 regime appears to ameliorate this effect, compared with CsA. Adequate pre- and post-transplant oral health care is recommended for these subjects, irrespective of the time interval for which the drug is administered.


Long-term Results After Ankle Syndesmosis Injuries.
Syndesmotic disruption occurs in more than 10% of ankle fractures. Operative treatment with syndesmosis screw fixation has been successfully performed for decades and is considered the gold standard of treatment. Few studies have reported the long-term outcomes of syndesmosis injuries. This study investigated long-term patient-reported, radiographic, and functional outcomes of syndesmosis injuries treated with screw fixation and subsequent timed screw removal. A retrospective cohort study was carried out at a Level I trauma center. The study group included 43 patients who were treated for ankle fractures with associated syndesmotic disruptions between December 2001 and May 2011. The study included case file reviews, self-reported questionnaires, radiologic reviews, and clinical assessments. At 5.1 (±1.76) years after injury, 60% of participants had pain, 26% had degenerative changes, 51% had loss of tibiofibular overlap, and 33% showed medial clear space widening. Retained syndesmotic positions on radiographs were linked to better self-reported outcomes. There is an inversely proportional relation between age at the time of injury and satisfaction with the outcome of the ankle fracture as well as a directly proportional relation between age at the time of injury and pain compared with the preinjury state. Optimal restoration and preservation of the syndesmosis is crucial. Syndesmotic disruption is associated with poor long-term outcomes after ankle fracture. Greater age is a risk factor for chronic pain and dissatisfaction with the outcome of ankle fracture and syndesmosis injury. Therefore, patient education to facilitate realistic expectations about recovery is vital, especially in older patients.


C-type cytochrome-initiated reduction of bacterial lytic polysaccharide monooxygenases.
The release of glucose from lignocellulosic waste for subsequent fermentation into biofuels holds promise for securing humankind's future energy needs. The discovery of a set of copper-dependent enzymes known as lytic polysaccharide monooxygenases (LPMOs) has galvanised new research in this area. LPMOs act by oxidatively introducing chain breaks into cellulose and other polysaccharides, boosting the ability of cellulases to act on the substrate. Although several proteins have been implicated as electron sources in fungal LPMO biochemistry, no equivalent bacterial LPMO electron donors have been previously identified, although the proteins Cbp2D and E from Cellvibrio japonicus have been implicated as potential candidates. Here we analyse a small c-type cytochrome (CjX183) present in Cellvibrio japonicus Cbp2D, and show that it can initiate bacterial CuII/I LPMO reduction and also activate LPMO-catalyzed cellulose-degradation. In the absence of cellulose, CjX183-driven reduction of the LPMO results in less H2O2 production from O2, and correspondingly less oxidative damage to the enzyme than when ascorbate is used as the reducing agent. Significantly, using CjX183 as the activator maintained similar cellulase boosting levels relative to the use of an equivalent amount of ascorbate. Our results therefore add further evidence to the impact that the choice of electron source can have on LPMO action. Furthermore, the study of Cbp2D and other similar proteins may yet reveal new insight into the redox processes governing polysaccharide degradation in bacteria.


Crowdsourcing novel childhood predictors of adult obesity.
Effective and simple screening tools are needed to detect behaviors that are established early in life and have a significant influence on weight gain later in life. Crowdsourcing could be a novel and potentially useful tool to assess childhood predictors of adult obesity. This exploratory study examined whether crowdsourcing could generate well-documented predictors in obesity research and, moreover, whether new directions for future research could be uncovered. Participants were recruited through social media to a question-generation website, on which they answered questions and were able to pose new questions that they thought could predict obesity. During the two weeks of data collection, 532 participants (62% female; age = 26.5±6.7; BMI = 29.0±7.0) registered on the website and suggested a total of 56 unique questions. Nineteen of these questions correlated with body mass index (BMI) and covered several themes identified by prior research, such as parenting styles and healthy lifestyle. More importantly, participants were able to identify potential determinants that were related to a lower BMI, but have not been the subject of extensive research, such as parents packing their children's lunch to school or talking to them about nutrition. The findings indicate that crowdsourcing can reproduce already existing hypotheses and also generate ideas that are less well documented. The crowdsourced predictors discovered in this study emphasize the importance of family interventions to fight obesity. The questions generated by participants also suggest new ways to express known predictors.


Thalamic sensitization transforms localized pain into widespread allodynia.
Focal somatic pain can evolve into widespread hypersensitivity to nonpainful and painful skin stimuli (allodynia and hyperalgesia, respectively). We hypothesized that transformation of headache into whole-body allodynia/hyperalgesia during a migraine attack is mediated by sensitization of thalamic neurons that process converging sensory impulses from the cranial meninges and extracephalic skin. Extracephalic allodynia was assessed using single unit recording of thalamic trigeminovascular neurons in rats and contrast analysis of blood oxygenation level-dependent (BOLD) signals registered in functional magnetic resonance imaging (fMRI) scans of patients exhibiting extracephalic allodynia. Sensory neurons in the rat posterior thalamus that were activated and sensitized by chemical stimulation of the cranial dura exhibited long-lasting hyperexcitability to innocuous (brush, pressure) and noxious (pinch, heat) stimulation of the paws. Innocuous, extracephalic skin stimuli that did not produce neuronal firing at baseline (eg, brush) became as effective as noxious stimuli (eg, pinch) in eliciting large bouts of neuronal firing after sensitization was established. In migraine patients, fMRI assessment of BOLD signals showed that brush and heat stimulation at the skin of the dorsum of the hand produced larger BOLD responses in the posterior thalamus of subjects undergoing a migraine attack with extracephalic allodynia than the corresponding responses registered when the same patients were free of migraine and allodynia. We propose that the spreading of multimodal allodynia and hyperalgesia beyond the locus of migraine headache is mediated by sensitized thalamic neurons that process nociceptive information from the cranial meninges together with sensory information from the skin of the scalp, face, body, and limbs.


Ischemia-related alteration of GABAA-operated chloride channel properties in gerbil hippocampus and cerebral cortex.
The properties of GABA-gated chloride (Cl-) channels in ischemia-reperfusion injury were studied by determination of the binding and dissociation kinetics of a specific Cl- channel ligand, tert-butylbicyclophosphoro[35S]thionate (TBPS) and by determination of 36CTl- uptake in the presence of the GABAA receptor agonist, muscimol. Four days after ischemia a small but insignificant decrease of [35S]TBPS binding to synaptic plasma membranes (SPM) was observed in the hippocampus and cerebral cortex as compared to control. The effect of ischemia was larger and statistically significant after the first and second month of reperfusion, constituting 20% inhibition of [35S]TBPS binding to SPM of sham-operated gerbils. On the other hand, the half-life of fast phase [35S]TBPS dissociation four days after ischemia was markedly diminished by about 40%-50% as compared to its control value and persisted during the first and second month of reperfusion in the hippocampal SPM. A similar but less potent reduction of the half-life of the fast phase of [35S]TBPS dissociation (about 30% versus control) appeared one and two months after ischemia in cerebral cortex SPM. One month after ischemia muscimol-stimulated 36Cl- uptake into cerebral cortex synaptoneurosomes was lowered as compared with control uptake, but remained statistically insignificant in the whole range of muscimol concentrations tested. Our results indicated that ischernia-reperfusion injury significantly decreases opening time of GABAA receptor-gated Cl- channels in the hippocampus and cerebral cortex, which may lower the hyperpolarization ability of this receptor complex leading to an imbalance between excitatory and inhibitory neurotransmitter pathways in these brain areas, and in consequence to neuronal dysfunction or degeneration.


An insight into the genetic variation of Schistosoma japonicum in mainland China using DNA microsatellite markers.
This study presents the first microsatellite investigation into the level of genetic variation among Schistosoma japonicum from different geographical origins. S. japonicum isolates were obtained from seven endemic provinces across mainland China: Zhejiang (Jiashan County), Anhui (Guichi County), Jiangxi (Yongxiu County), Hubei (Wuhan County), Hunan (Yueyang area), Sichuan 1 (Maoshan County), Sichuan 2 (Tianquan County), Yunnan (Dali County), and also one province in the Philippines (Sorsogon). DNA from 20 individuals from each origin were screened against 11 recently isolated and characterized S. japonicum microsatellites, and a set of nine loci were selected based on their polymorphic information content. High levels of polymorphism were obtained between and within population samples, with Chinese and Philippine strains appearing to follow different lineages, and with distinct branching between provinces. Moreover, across mainland China, genotype clustering appeared to be related to habitat type and/or intermediate host morph. These results highlight the suitability of microsatellites for population genetic studies of S. japonicum and suggest that there may be different strains of S. japonicum circulating in mainland China.


Assessing adherence to the 2010 antiretroviral guidelines at the antiretroviral rollout clinic in 1 military hospital, South Africa: a retrospective, cross sectional study.
HIV research is a therapeutic area for which well-defined population-specific treatment and prophylaxis guidelines exist. However, there are limited objective, evidence-based data for assessing adherence to these guidelines. To conduct a retrospective, cross sectional study of adult HIV-infected patients receiving treatment at the antiretroviral (ARV) roll-out clinic of the Infectious Diseases Clinic Pharmacy at 1 Military Hospital (1MH) over a period of 3 years to assess clinicians' adherence to the 2010 ARV guidelines. Pharmacy files from the pool of adult patients receiving treatment at the ARV roll-out clinic between 1 April 2009 and 31 March 2012 were selected. Variables used to establish adherence were assessed through evaluation of pharmacy scripts and laboratory tests. In accordance with the ARV guidelines, we found a switch in the first-line regimen from stavudine to tenofovir in the period following implementation. There was no difference in baseline blood tests conducted, suggesting that clinicians were recommending a standardised test panel. Notably, similar blood tests were routinely done during follow-up visits, despite no indication for doing so. While the number of blood tests was found to decrease over time, the type of blood tests requested for specific treatment regimens was not in accordance with ARV guidelines. We used an evidence-based approach to critically assess variations from the delineated ARV guidelines. Adherence to clinical guidelines at 1MH, while demonstrating improvement in patient outcomes, highlighted the need for increased vigilance in monitoring failure of prescribers to comply with ARV guidelines.


Molecular basis for vancomycin resistance in Enterococcus faecium BM4147: biosynthesis of a depsipeptide peptidoglycan precursor by vancomycin resistance proteins VanH and VanA.
Vancomycin resistance in Enterococcus faecium BM4147 is mediated by vancomycin resistance proteins VanA and VanH. VanA is a D-alanine:D-alanine ligase of altered substrate specificity [Bugg, T. D. H., Dutka-Malen, S., Arthur, M., Courvalin, P., & Walsh, C. T. (1991) Biochemistry 30, 2017-2021], while the sequence of VanH is related to those of alpha-keto acid dehydrogenases [Arthur, M., Molinas, C., Dutka-Malen, S., & Courvalin, P. (1991) Gene (submitted)]. We report purification of VanH to homogeneity, characterization as a D-specific alpha-keto acid dehydrogenase, and comparison with D-lactate dehydrogenases from Leuconostoc mesenteroides and Lactobacillus leichmanii. VanA was found to catalyze ester bond formation between D-alanine and the D-hydroxy acid products of VanH, the best substrate being D-2-hydroxybutyrate (Km = 0.60 mM). The VanA product D-alanyl-D-2-hydroxybutyrate could then be incorporated into the UDPMurNAc-pentapeptide peptidoglycan precursor by D-Ala-D-Ala adding enzyme from Escherichia coli or by crude extract from E. faecium BM4147. The vancomycin binding constant of a synthetic modified peptidoglycan analogue N-acetyl-D-alanyl-D-2-hydroxybutyrate (Kd greater than 73 mM) was greater than 1000-fold higher than the binding constant for N-acetyl-D-alanyl-D-alanine (Kd = 54 microM), partly due to the disruption of a hydrogen bond in the vancomycin-target complex, thus providing a molecular rationale for high-level vancomycin resistance.


Measurements of the radiographic cardiac silhouette of ospreys (Pandion haliaetus).
To evaluate and report measurements of the radiographic cardiac silhouette of healthy juvenile and adult ospreys (Pandion haliaetus). 54 ospreys (22 adults, 19 juveniles, and 13 birds of undetermined age) without clinical signs of cardiac disease and with adequate ventrodorsal radiographic images for cardiac silhouette assessment. Radiographs of ospreys were assessed to determine cardiac width at the widest point as well as sternal width and thoracic width at the same level. Two-way mixed-effects models were used to evaluate interrater reliability for mean rating. Multivariable linear regression analysis was used to create predictive models of cardiac width and to establish a theoretical reference range for healthy ospreys. Cardiac width of healthy ospreys was approximately 90% to 92% of sternal width and 67% to 69% of thoracic width. Both sternal width and thoracic width were significant predictors of cardiac width in independent predictive models as well as in a combined model after controlling for age. Thirty-four of 41 (83%) measured cardiac widths were within the theoretical reference range. Ospreys are sentinels used in monitoring environmental health. Environmental factors may have an impact on the cardiac health of ospreys, but reference values for healthy ospreys have not been established for use in assessing cardiomegaly in this species. The radiographic ratios and predictive model obtained in this study may be useful for objective evaluation of cardiomegaly in ospreys.


Bio-ethanol production from wet coffee processing waste in Ethiopia.
Large amounts of coffee residues are generated from coffee processing plants in Ethiopia. These residues are toxic and possess serious environmental problems following the direct discharge into the nearby water bodies which cause serious environmental and health problems. This study was aimed to quantify wet coffee processing waste and estimate its bio-ethanol production. The study showed that the wastes are potential environmental problems and cause water pollution due to high organic component and acidic nature. The waste was hydrolyzed by dilute H2SO4 (0.2, 0.4, 0.6, 0.8 and 1 M) and distilled water. Total sugar content of the sample was determined titrimetrically and refractometry. Maximum value (90%) was obtained from hydrolysis by 0.4 M H2SO4. Ethanol production was monitored by gas chromatography. The optimum yield of ethanol (78%) was obtained from the sample hydrolyzed by 0.4 M H2SO4 for 1 h at hydrolysis temperature of 100 °C and after fermentation for 24 h and initial pH of 4.5. Based on the data, it was concluded that reuse of the main coffee industry wastes is of significant importance from environmental and economical view points. In conclusion, this study has proposed to utilize the wet coffee processing waste to produce bio-ethanol which provides the alternative energy source from waste biomass and solves the environmental waste disposal as well as human health problem.


Ketamine self-administration in the rat: evidence for a critical role of setting.
The abuse of ketamine has been reported to be on the rise over the past 15 years, but its abuse appears to be limited almost exclusively to the context of music and dance settings, indicating a major role of context in modulating its reinforcing effects. We have previously reported that amphetamine, cocaine, and heroin self-administration (SA) in the rat are differentially influenced by the setting in which testing takes place. The aim of the present study is to extend this pre-clinical model to ketamine. Independent groups of rats with intravenous catheters were given the possibility to self-administer different doses of ketamine (125, 250, and 500 μg/kg per infusion) under two environmental conditions. Some animals were housed in the SA chambers (resident rats) whereas other rats were transported to the SA chambers only for the test sessions (non-resident rats). After training, within-subject dose effect curves (125, 250, 500, and 1,000 μg/kg per infusion) and break-point (during a progressive ratio session) were calculated. Non-resident rats readily acquired ketamine self-administration. In contrast, resident rats self-administered only the highest dose of ketamine (500 μg/kg), but still four times less than non-resident rats (11.0 ± 6.0 vs 44.4 ± 5.2 infusions during the last training session). No significant differences in break-point were found during the progressive ratio session. The present study confirms at a preclinical level the importance of setting for ketamine SA and further validates a previously described animal model of drug-environment interaction.


Glycosylation of the West Nile Virus envelope protein increases in vivo and in vitro viral multiplication in birds.
Many West Nile (WN) virus isolates associated with significant outbreaks possess a glycosylation site on the envelope (E) protein. E-protein glycosylated variants of New York (NY) strains of WN virus are more neuroinvasive in mice than the non-glycosylated variants. To determine how E protein glycosylation affects the interactions between WN virus and avian hosts, we inoculated young chicks with NY strains of WN virus containing either glycosylated or non-glycosylated variants of the E protein. The glycosylated variants were more virulent and had higher viremic levels than the non-glycosylated variants. The glycosylation status of the variant did not affect viral multiplication and dissemination in mosquitoes in vivo. Glycosylated variants showed more heat-stable propagation than non-glycosylated variants in mammalian (BHK) and avian (QT6) cells but not in mosquito (C6/36) cells. Thus, E-protein glycosylation may be a requirement for efficient transmission of WN virus from avian hosts to mosquito vectors.


Differential gene expression and protein abundance evince ontogenetic bias toward castes in a primitively eusocial wasp.
Polistes paper wasps are models for understanding conditions that may have characterized the origin of worker and queen castes and, therefore, the origin of paper wasp sociality. Polistes is "primitively eusocial" by virtue of having context-dependent caste determination and no morphological differences between castes. Even so, Polistes colonies have a temporal pattern in which most female larvae reared by the foundress become workers, and most reared by workers become future-reproductive gynes. This pattern is hypothesized to reflect development onto two pathways, which may utilize mechanisms that regulate diapause in other insects. Using expressed sequence tags (ESTs) for Polistes metricus we selected candidate genes differentially expressed in other insects in three categories: 1) diapause vs. non-diapause phenotypes and/or worker vs. queen differentiation, 2) behavioral subcastes of worker honey bees, and 3) no a priori expectation of a role in worker/gyne development. We also used a non-targeted proteomics screen to test for peptide/protein abundance differences that could reflect larval developmental divergence. We found that foundress-reared larvae (putative worker-destined) and worker-reared larvae (putative gyne-destined) differed in quantitative expression of sixteen genes, twelve of which were associated with caste and/or diapause in other insects, and they also differed in abundance of nine peptides/proteins. Some differentially-expressed genes are involved in diapause regulation in other insects, and other differentially-expressed genes and proteins are involved in the insulin signaling pathway, nutrient metabolism, and caste determination in highly social bees. Differential expression of a gene and a peptide encoding hexameric storage proteins is especially noteworthy. Although not conclusive, our results support hypotheses of 1) larval developmental pathway divergence that can lead to caste bias in adults and 2) nutritional differences as the foundation of the pathway divergence. Finally, the differential expression in Polistes larvae of genes and proteins also differentially expressed during queen vs. worker caste development in honey bees may indicate that regulatory mechanisms of caste outcomes share similarities between primitively eusocial and advanced eusocial Hymenoptera.


Dendritic cells mediate herpes simplex virus infection and transmission through the C-type lectin DC-SIGN.
Dendritic cells (DCs) are essential for the induction of specific immune responses against invading pathogens. Herpes simplex virus (HSV) is a common human pathogen that causes painful but mild infections of the skin and mucosa, and which results in latency and recurrent infections. Of the two HSV subtypes described, HSV-1 causes mainly oral-facial lesions, whilst HSV-2 is associated with genital herpes. DCs are involved in HSV-induced immune suppression, but little is known about the molecular interactions between DCs and HSV. This study demonstrated that HSV-1 and -2 both interact with the DC-specific C-type lectin DC-SIGN. Further analyses demonstrated that DC-SIGN interacts with the HSV glycoproteins gB and gC. Binding of HSV-1 to immature DCs depended on both DC-SIGN and heparan sulfate proteoglycans. Strikingly, HSV-1 infection of DCs was almost completely inhibited by blocking antibodies against DC-SIGN. Thus, DC-SIGN is an important attachment receptor for HSV-1 on immature DCs and enhances infection of DCs in cis. In addition, DC-SIGN captures HSV-1 for transmission to permissive target cells. These data strongly suggest that DC-SIGN is a potential target to prevent HSV infection and virus dissemination. Further studies will show whether these interactions are involved in HSV-induced immune suppression.


Evidence for auxiliary anaerobic metabolism in obligately aerobic Zetaproteobacteria.
Zetaproteobacteria are obligate chemolithoautotrophs that oxidize Fe(II) as an electron and energy source, and play significant roles in nutrient cycling and primary production in the marine biosphere. Zetaproteobacteria thrive under microoxic conditions near oxic-anoxic interfaces, where they catalyze Fe(II) oxidation faster than the abiotic reaction with oxygen. Neutrophilic Fe(II) oxidizing bacteria produce copious amounts of insoluble iron oxyhydroxides as a by-product of their metabolism. Oxygen consumption by aerobic respiration and formation of iron oxyhydroxides at oxic-anoxic interfaces can result in periods of oxygen limitation for bacterial cells. Under laboratory conditions, all Zetaproteobacteria isolates have been shown to strictly require oxygen as an electron acceptor for growth, and anaerobic metabolism has not been observed. However, genomic analyses indicate a range of potential anaerobic pathways present in Zetaproteobacteria. Heterologous expression of proteins from Mariprofundus ferrooxydans PV-1, including pyruvate formate lyase and acetate kinase, further support a capacity for anaerobic metabolism. Here we define auxiliary anaerobic metabolism as a mechanism to provide maintenance energy to cells and suggest that it provides a survival advantage to Zetaproteobacteria in environments with fluctuating oxygen availability.


Molecular and Translational Classifications of DAMPs in Immunogenic Cell Death.
The immunogenicity of malignant cells has recently been acknowledged as a critical determinant of efficacy in cancer therapy. Thus, besides developing direct immunostimulatory regimens, including dendritic cell-based vaccines, checkpoint-blocking therapies, and adoptive T-cell transfer, researchers have started to focus on the overall immunobiology of neoplastic cells. It is now clear that cancer cells can succumb to some anticancer therapies by undergoing a peculiar form of cell death that is characterized by an increased immunogenic potential, owing to the emission of the so-called "damage-associated molecular patterns" (DAMPs). The emission of DAMPs and other immunostimulatory factors by cells succumbing to immunogenic cell death (ICD) favors the establishment of a productive interface with the immune system. This results in the elicitation of tumor-targeting immune responses associated with the elimination of residual, treatment-resistant cancer cells, as well as with the establishment of immunological memory. Although ICD has been characterized with increased precision since its discovery, several questions remain to be addressed. Here, we summarize and tabulate the main molecular, immunological, preclinical, and clinical aspects of ICD, in an attempt to capture the essence of this phenomenon, and identify future challenges for this rapidly expanding field of investigation.


Pyruvate kinase variant of fission yeast tunes carbon metabolism, cell regulation, growth and stress resistance.
Cells balance glycolysis with respiration to support their metabolic needs in different environmental or physiological contexts. With abundant glucose, many cells prefer to grow by aerobic glycolysis or fermentation. Using 161 natural isolates of fission yeast, we investigated the genetic basis and phenotypic effects of the fermentation-respiration balance. The laboratory and a few other strains depended more on respiration. This trait was associated with a single nucleotide polymorphism in a conserved region of Pyk1, the sole pyruvate kinase in fission yeast. This variant reduced Pyk1 activity and glycolytic flux. Replacing the "low-activity" pyk1 allele in the laboratory strain with the "high-activity" allele was sufficient to increase fermentation and decrease respiration. This metabolic rebalancing triggered systems-level adjustments in the transcriptome and proteome and in cellular traits, including increased growth and chronological lifespan but decreased resistance to oxidative stress. Thus, low Pyk1 activity does not lead to a growth advantage but to stress tolerance. The genetic tuning of glycolytic flux may reflect an adaptive trade-off in a species lacking pyruvate kinase isoforms.


Immune tolerance against infused FVIII in hemophilia A is mediated by PD-L1+ Tregs.
A major complication of hemophilia A therapy is the development of alloantibodies (inhibitors) that neutralize intravenously administered coagulation factor VIII (FVIII). Immune tolerance induction therapy (ITI) by repetitive FVIII injection can eradicate inhibitors, and thereby reduce morbidity and treatment costs. However, ITI success is difficult to predict and the underlying immunological mechanisms are unknown. Here, we demonstrated that immune tolerance against FVIII under nonhemophilic conditions was maintained by programmed death (PD) ligand 1-expressing (PD-L1-expressing) regulatory T cells (Tregs) that ligated PD-1 on FVIII-specific B cells, causing them to undergo apoptosis. FVIII-deficient mice injected with FVIII lacked such Tregs and developed inhibitors. Using an ITI mouse model, we found that repetitive FVIII injection induced FVIII-specific PD-L1+ Tregs and reengaged removal of inhibitor-forming B cells. We also demonstrated the existence of FVIII-specific Tregs in humans and showed that such Tregs upregulated PD-L1 in patients with hemophilia after successful ITI. Simultaneously, FVIII-specific B cells upregulated PD-1 and became killable by Tregs. In summary, we showed that PD-1-mediated B cell tolerance against FVIII operated in healthy individuals and in patients with hemophilia A without inhibitors, and that ITI reengaged this mechanism. These findings may impact monitoring of ITI success and treatment of patients with hemophilia A.


Estimating the extent of Medicaid spend-down in nursing homes.
Although it is commonly thought that a very high proportion of private-pay nursing home patients become eligible for Medicaid during their stay, few national studies have been conducted to determine the extent of Medicaid "spend-down." The discharged resident survey of the 1985 National Nursing Home Survey was analyzed to determine the extent of spend-down among elderly nursing home patients during a single nursing home episode, the proportion of patients who remained Medicaid or private payers throughout their stay, and how the measures varied by patient characteristics. Overall, during a single nursing home episode only 10 percent of nursing home patients who entered as private payers received Medicaid at discharge. Roughly one-third of nursing home patients remained on Medicaid throughout their stay, and another third remained private payers throughout their stay.


Temporal and spatial dynamics of trypanosomes infecting the brush-tailed bettong (Bettongia penicillata): a cautionary note of disease-induced population decline.
The brush-tailed bettong or woylie (Bettongia penicillata) is on the brink of extinction. Its numbers have declined by 90% since 1999, with their current distribution occupying less than 1% of their former Australian range. Woylies are known to be infected with three different trypanosomes (Trypanosoma vegrandis, Trypanosoma copemani and Trypanosoma sp. H25) and two different strains of T. copemani that vary in virulence. However, the role that these haemoparasites have played during the recent decline of their host is unclear and is part of ongoing investigation. Woylies were sampled from five locations in southern Western Australia, including two neighbouring indigenous populations, two enclosed (fenced) populations and a captive colony. PCR was used to individually identify the three different trypanosomes from blood and tissues of the host, and to investigate the temporal and spatial dynamics of trypanosome infections. The spatial pattern of trypanosome infection varied among the five study sites, with a greater proportion of woylies from the Perup indigenous population being infected with T. copemani than from the neighbouring Kingston indigenous population. For an established infection, T. copemani detection was temporally inconsistent. The more virulent strain of T. copemani appeared to regress at a faster rate than the less virulent strain, with the infection possibly transitioning from the acute to chronic phase. Interspecific competition may also exist between T. copemani and T. vegrandis, where an existing T. vegrandis infection may moderate the sequential establishment of the more virulent T. copemani. In this study, we provide a possible temporal connection implicating T. copemani as the disease agent linked with the recent decline of the Kingston indigenous woylie population within the Upper Warren region of Western Australia. The chronic association of trypanosomes with the internal organs of its host may be potentially pathogenic and adversely affect their long term fitness and coordination, making the woylie more susceptible to predation.


Laser spectroscopy of trapped atomic ions.
Recent developments in laser spectroscopy of atomic ions stored in electromagnetic traps are reviewed with emphasis on techniques that appear to hold the greatest promise of attaining extremely high resolution. Among these techniques are laser cooling and the use of single, isolated ions as experimental samples. Doppler shifts and other perturbing influences can be largely eliminated. Atomic resonances with line widths of a few parts in 10(11) have been observed at frequencies ranging from the radio frequency to the ultraviolet. Experimental accuracies of one part in 10(18) appear to be attainable.


Management of thymic tumors: a survey of current practice among members of the European Society of Thoracic Surgeons.
Management of thymoma has largely been based on single-institution retrospective, observational studies. The European Society of Thoracic Surgeons (ESTS) Thymic Working Group has investigated the current practice among ESTS members. A questionnaire divided into seven sections with 24 questions was designed, and it was delivered to ESTS members. Forty-four centers replied to the questionnaire. The results indicate that there is a general agreement concerning (1) the value of computed tomography scan for preoperative assessment; (2) the uselessness of a routine histologic confirmation before surgery; (3) the role of the World Health Organization classification; (4) the importance of complete and extended resection; (5) the role of surgery for recurrent disease; and (6) the need of a multidisciplinary team, including thoracic surgeons, pathologists, medical, and radiation oncologists. On the other hand, there is still a considerable debate about (1) the role of positron emission tomography scan for preoperative assessment; (2) a consistent and reliable staging system; (3) the usefulness of postoperative treatments for stages II and III diseases; (4) the management of type C thymoma; and (5) the role of extrapleural pneumonectomy for stage IVA thymoma. The survey provides a large, multiinstitutional overview of the clinical practice in the management of thymic tumors by ESTS members. Responses show some areas of agreement along with several areas of controversy. It is conceivable that a consequent step forward will be the creation of a collaborative effort within the ESTS and with other organizations for the creation of standard recommendations and guidelines for the management of thymic malignancies.


Soccer training: high-intensity interval training is mood disturbing while small sided games ensure mood balance.
The aim of the study was to compare the effects of high-intensity intermittent training (HIIT) versus small-sided games (SSG) in soccer on both the physiological responses and the mood state of players. Sixteen professional soccer players took part in the study (age: 24.1±0.9 years). Testing of players was conducted on separate days in a randomized and counter-balanced order (each training session: 28 minutes: 4x4 minutes work with 3 minutes of passive recovery in-between). Effort: HIIT: intermittent 15-s runs at 110% maximal aerobic speed with 15 seconds of passive recovery in-between. SSG: 4 versus 4 players on a 25x35 meter pitch size with full-involvement play. Psychological responses before and after each training-session were assessed using the profile of mood-state (POMS: Tension, Depression, Anger, Vigor, Fatigue, and Confusion). The players' heart rate (HR) was continuously measured, whereas ratings of perceived exertion (RPE) and blood lactate concentration ([La]) were collected ~3-min after each training-session. HIIT and SSG showed no significant difference in HR, RPE and [La] responses. The HIIT compared with SSG resulted in: an increased total mood disturbance (P<0.001), tension (P<0.05), fatigue (P<0.01) and a decreased vigor (P<0.001). Both HIIT and SSG sessions induced similar physiological responses, in contrast, HIIT produced a mood disturbance while SSG ensured mood balance. Practitioners could choose between these two exercises according to the objective of their training, keeping in mind the mood-related advantages of the SSG shown in the present study.


Analytical expressions for the angle of arrival fluctuations for optical waves' propagation through moderate-to-strong non-Kolmogorov refractive turbulence.
The effects of moderate-to-strong non-Kolmogorov turbulence on the angle of arrival (AOA) fluctuations for plane and spherical waves are investigated in detail both analytically and numerically. New analytical expressions for the variance of AOA fluctuations are derived for moderate-to-strong non-Kolmogorov turbulence. The new expressions cover a wider range of non-Kolmogorov turbulence strength and reduce correctly to previously published analytic expressions for the cases of plane and spherical wave propagation through both weak non-Kolmogorov turbulence and moderate-to-strong Kolmogorov turbulence cases. The final results indicate that, as turbulence strength becomes greater, the expressions developed with the Rytov theory deviate from those given in this work. This deviation becomes greater with stronger turbulence, up to moderate-to-strong turbulence strengths. Furthermore, general spectral power law has significant influence on the variance of AOA fluctuations in non-Kolmogorov turbulence. These results are useful for understanding the potential impact of deviations from the standard Kolmogorv spectrum.


Circulating intercellular adhesion molecule-1 (ICAM-1) in patients with hepatolithiasis.
We evaluated the role of circulating intercellular adhesion molecule-1 in the pathogenesis of hepatolithiasis. From December 1994 to May 1995, 40 patients with hepatolithiasis were included. All the patients met the following criteria: (1) presence of hepatolithiasis, (2) no obvious clinical evidence of an associated intrahepatic cholangiocarcinoma, (3) no clinical manifestation of cholangitis for at least 72 hr, (4) no immunomodulatory agents in the last three weeks, and (5) no blood transfusion in the last three weeks. Venous blood samples were collected both before surgery and at least three months after complete clearance of the stones, and the serum concentrations of circulating intercellular adhesion molecule-1 were measured with a sandwich enzyme immunoassay method. Fifteen healthy subjects were used as a control group. Bile specimens routinely obtained during surgery were cultured for aerobes and anaerobes. The x-ray films of cholangiography were all reviewed in detail. The mean value (834 +/- 128 ng/ml) of circulating intercellular adhesion molecule-1 (ICAM-1) in the patient group before surgery was significantly higher than that (346 +/- 68 ng/ml) of the control group (P < 0.01). The mean value (677 +/- 139 ng/ml) of circulating ICAM-1 in the patient group at least three months after complete clearance of the stones was significantly lower than that (834 +/- 128 ng/ml) of the patients before surgery (P < 0.01), but this mean value (677 +/- 139 ng/ml) was still significantly higher than that (346 +/- 68 ng/ml) of the control group (P < 0.01). Bacteria was present in the bile of all patients. The total number of bacterial species was 135, and there were an average of 3.4 bacterial species cultured per patient. Intrahepatic stricture was demonstrated in cholangiography in 33 patients (82.5%). In addition to the high incidence of intrahepatic bile duct strictures and bile infection, a significant elevation in circulating intercellular adhesion molecule-1 (sICAM-1) was shown in patients with hepatolithiasis. Our preliminary results seem to be promising and the real role of sICAM-1 deserves further investigation and elucidation.


Chordoid glioma of the third ventricle: a patient presenting with SIADH and a review of this rare tumor.
Chordoid glioma of the third ventricle is a rare and recently described tumor characterized by a unique histomorphology and exclusive association with the suprasellar/third ventricular compartment. Its clinical, radiological and histological features may vary. Despite the fact that chordoid glioma is a low-grade tumor, its prognosis has been relatively poor because of its insidious presentation and the difficulty in obtaining complete surgical resection. Here, we report on a new case of chordoid glioma occurring in a 48-year-old woman, presented with hyponatremia, and on the initial work-up with a diagnosis of hyponatremia due at least in part to SIADH. We review the current literature on this rare pathology, discuss the radiological and histopathologic findings, and discuss the optimal management of chordoid glioma in general. Based on this new case and the previous literature reports, we suggest that chordoid glioma should be included in the differential diagnosis of uncommon masses of the third ventricle, especially in middle-aged women, and we emphasize current management guidelines.


Long-range effects of retroviral insertion on c-myb: overexpression may be obscured by silencing during tumor growth in vitro.
The c-myb oncogene is a frequent target for retroviral activation in hemopoietic tumors of avian and mammalian species. While insertions can target the gene directly, numerous clusters of retroviral insertion sites have been identified which map close to c-myb and outside the transcription unit in T-lymphomas (Ahi-1, fit-1, and Mis-2) and monocytic and myeloid leukemias (Mml1, Mml2, Mml3, and Epi-1). Previous analyses showed no consistent effect of these insertions on c-myb expression, raising the possibility that other nearby genes were the true targets. In contrast, our analysis of four cell lines established from lymphomas bearing insertions at fit-1 (fti-1) (feline leukemia virus) and Ahi-1 (Moloney murine leukemia virus) shows that these display higher expression levels of c-myb RNA and protein compared to a panel of phenotypically similar cell lines lacking such insertions. An interesting feature of the cell lines with long-range c-myb insertions was that each also carried an activated Myc allele. The potential for oncogenic synergy between Myb and Myc in T-cell lymphoma was confirmed in transgenic mice overexpressing alleles of both genes in the T-cell compartment, lending further credence to the case for c-myb as the major target for long-range activation. In contrast, mapping and analysis of c-myb neighboring genes (HBS1 and FLJ20069) showed that the expression of these genes did not correlate well with the presence of proviral insertions. A possible explanation for the paradoxical behavior of c-myb was provided by one of the murine T-lymphoma lines bearing an insertion at Ahi-1 (p/m16i) that reproducibly down-regulated c-myb RNA and protein to very low levels or undetectable levels on prolonged culture. Our observations implicate c-myb as a key target of upstream and downstream retroviral insertions. However, overexpression may become dispensable during outgrowth in vitro, and perhaps during tumor progression in vivo, providing a potential rationale for the previously observed discordance between retroviral insertion and c-myb expression levels.


A theory of social thermoregulation in human primates.
Beyond breathing, the regulation of body temperature-thermoregulation-is one of the most pressing concerns for many animals. A dysregulated body temperature has dire consequences for survival and development. Despite the high frequency of social thermoregulation occurring across many species, little is known about the role of social thermoregulation in human (social) psychological functioning. We outline a theory of social thermoregulation and reconsider earlier research on people's expectations of their social world (i.e., attachment) and their prediction of the social world. We provide support and outline a research agenda that includes consequences for individual variation in self-regulatory strategies and capabilities. In our paper, we discuss physiological, neural, and social processes surrounding thermoregulation. Emphasizing social thermoregulation in particular, we appeal to the economy of action principle and the hierarchical organization of human thermoregulatory systems. We close with future directions of a crucial aspect of human functioning: the social regulation of body temperature.


Sex differences in stress-induced social withdrawal: role of brain derived neurotrophic factor in the bed nucleus of the stria terminalis.
Depression and anxiety disorders are more common in women than men, and little is known about the neurobiological mechanisms that contribute to this disparity. Recent data suggest that stress-induced changes in neurotrophins have opposing effects on behavior by acting in different brain networks. Social defeat has been an important approach for understanding neurotrophin action, but low female aggression levels in rats and mice have limited the application of these methods primarily to males. We examined the effects of social defeat in monogamous California mice (Peromyscus californicus), a species in which both males and females defend territories. We demonstrate that defeat stress increases mature brain-derived neurotrophic factor (BDNF) protein but not mRNA in the bed nucleus of the stria terminalis (BNST) in females but not males. Changes in BDNF protein were limited to anterior subregions of the BNST, and there were no changes in the adjacent nucleus accumbens (NAc). The effects of defeat on social withdrawal behavior and BDNF were reversed by chronic, low doses of the antidepressant sertraline. However, higher doses of sertraline restored social withdrawal and elevated BDNF levels. Acute treatment with a low dose of sertraline failed to reverse the effects of defeat. Infusions of the selective tyrosine-related kinase B receptor (TrkB) antagonist ANA-12 into the anterior BNST specifically increased social interaction in stressed females but had no effect on behavior in females naïve to defeat. These results suggest that stress-induced increases in BDNF in the anterior BNST contribute to the exaggerated social withdrawal phenotype observed in females.


The phytotoxin coronatine contributes to pathogen fitness and is required for suppression of salicylic acid accumulation in tomato inoculated with Pseudomonas syringae pv. tomato DC3000.
The roles of the phytotoxin coronatine (COR) and salicylic acid (SA)-mediated defenses in the interaction of Pseudomonas syringae pv. tomato DC3000 and tomato (Solanum lycopersicum) were investigated. Unlike findings reported for Arabidopsis thaliana, DC3000 mutants impaired for production of COR or one of its components, coronafacic acid (CFA) or coronamic acid (CMA), induced distinctly different disease lesion phenotypes in tomato. Tomato plants inoculated with the CFA- CMA- mutant DB29 showed elevated transcript levels of SlICS, which encodes isochorismate synthase, an enzyme involved in SA biosynthesis in S. lycopersicum. Furthermore, expression of genes encoding SA-mediated defense proteins were elevated in DB29-inoculated plants compared with plants inoculated with DC3000, suggesting that COR suppresses SlICS-mediated SA responses. Sequence analysis of SlICS revealed that it encodes a protein that is 55 and 59.6% identical to the A. thaliana ICS-encoded proteins AtICS1 and AtICS2, respectively. Tomato plants silenced for SlICS were hypersusceptible to DC3000 and accumulated lower levels of SA after infection with DC3000 compared with inoculated wild-type tomato plants. Unlike what has been shown for A. thaliana, the COR- mutant DB29 was impaired for persistence in SlICS-silenced tomato plants; thus, COR has additional roles in virulence that are SA independent and important in the latter stages of disease development. In summary, the infection assays, metabolic profiling, and gene expression results described in this study indicate that the intact COR molecule is required for both suppression of SA-mediated defense responses and full disease symptom development in tomato.


COVID-19: Review of a 21st Century Pandemic from Etiology to Neuro-psychiatric Implications.
COVID-19 is a severe infectious disease that has claimed >150,000 lives and infected millions in the United States thus far, especially the elderly population. Emerging evidence has shown the virus to cause hemorrhagic and immunologic responses, which impact all organs, including lungs, kidneys, and the brain, as well as extremities. SARS-CoV-2 also affects patients', families', and society's mental health at large. There is growing evidence of re-infection in some patients. The goal of this paper is to provide a comprehensive review of SARS-CoV-2-induced disease, its mechanism of infection, diagnostics, therapeutics, and treatment strategies, while also focusing on less attended aspects by previous studies, including nutritional support, psychological, and rehabilitation of the pandemic and its management. We performed a systematic review of >1,000 articles and included 425 references from online databases, including, PubMed, Google Scholar, and California Baptist University's library. COVID-19 patients go through acute respiratory distress syndrome, cytokine storm, acute hypercoagulable state, and autonomic dysfunction, which must be managed by a multidisciplinary team including nursing, nutrition, and rehabilitation. The elderly population and those who are suffering from Alzheimer's disease and dementia related illnesses seem to be at the higher risk. There are 28 vaccines under development, and new treatment strategies/protocols are being investigated. The future management for COVID-19 should include B-cell and T-cell immunotherapy in combination with emerging prophylaxis. The mental health and illness aspect of COVID-19 are among the most important side effects of this pandemic which requires a national plan for prevention, diagnosis and treatment.


Separation of isoclinics and isochromatics from photoelastic data with a regularized phase-tracking technique.
We present the application of a regularization algorithm to the processing of photoelastic fringe patterns. The method used is a modified regularized phase-tracking (RPT) algorithm applied to phase-shifted images. In particular, we present an algorithm for isoclinic-isochromatic separation that uses only five images. In the case of isoclinics the method can deal with problems associated with modulation of isochromatics and with isotropic points by means of a modified cost functional. With respect to the isochromatics the problems associated with regions of high fringe density are solved in a robust way by the unmodified RPT algorithm by use of the modulation information. The performance of the method is discussed, and experimental results are presented.


Cell cycle progression dictates the requirement for BCL2 in natural killer cell survival.
Natural killer (NK) cells are innate lymphoid cells with antitumor functions. Using an N-ethyl-N-nitrosourea (ENU)-induced mutagenesis screen in mice, we identified a strain with an NK cell deficiency caused by a hypomorphic mutation in the Bcl2 (B cell lymphoma 2) gene. Analysis of these mice and the conditional deletion of Bcl2 in NK cells revealed a nonredundant intrinsic requirement for BCL2 in NK cell survival. In these mice, NK cells in cycle were protected against apoptosis, and NK cell counts were restored in inflammatory conditions, suggesting a redundant role for BCL2 in proliferating NK cells. Consistent with this, cycling NK cells expressed higher MCL1 (myeloid cell leukemia 1) levels in both control and BCL2-null mice. Finally, we showed that deletion of BIM restored survival in BCL2-deficient but not MCL1-deficient NK cells. Overall, these data demonstrate an essential role for the binding of BCL2 to BIM in the survival of noncycling NK cells. They also favor a model in which MCL1 is the dominant survival protein in proliferating NK cells.


Minimizing off-target effects by using diced siRNAs for RNA interference.
Microarray studies have shown that individual synthetic small interfering RNAs (siRNAs) can have substantial off-target effects. Pools of siRNAs, produced by incubation of dsRNAs with recombinant Dicer or RNase III, can also be used to silence genes. Here we show that diced siRNA pools are highly complex, containing hundreds of different individual siRNAs. This high complexity could either compound the problem of off-target effects, since the number of potentially problematic siRNAs is high, or it could diminish the problem, since the concentration of any individual problematic siRNA is low. We therefore compared the off-target effects of diced siRNAs to chemically synthesized siRNAs. In agreement with previous reports, we found that two chemically synthesized siRNAs targeted against p38alpha MAPK (MAPK14) induced off-target changes in the abundance of hundreds of mRNAs. In contrast, three diced siRNA pools against p38alpha MAPK had almost no off-target effects. The off-target effects of a synthetic siRNA were reduced when the siRNA was diluted 3-fold in a diced pool and completely alleviated when it was diluted 30- or 300-fold, suggesting that when problematic siRNAs are present within a diced pool, their absolute concentration is too low to result in significant off-target effects. These data rationalize the observed high specificity of RNA interference in C. elegans and D. melanogaster, where gene suppression is mediated by endogenously-generated diced siRNA pools, and provide a strategy for improving the specificity of RNA interference experiments and screens in mammalian cells.


Approach to critical illness polyneuropathy and myopathy.
A newly acquired neuromuscular cause of weakness has been found in 25-85% of critically ill patients. Three distinct entities have been identified: (1) critical illness polyneuropathy (CIP); (2) acute myopathy of intensive care (itself with three subtypes); and (3) a syndrome with features of both 1 and 2 (called critical illness myopathy and/or neuropathy or CRIMYNE). CIP is primarily a distal axonopathy involving both sensory and motor nerves. Electroneurography and electromyography (ENG-EMG) is the gold standard for diagnosis. CIM is a proximal as well as distal muscle weakness affecting both types of muscle fibres. It is associated with high use of non-depolarising muscle blockers and corticosteroids. Avoidance of systemic inflammatory response syndrome (SIRS) is the most effective way to reduce the likelihood of developing CIP or CIM. Outcome is variable and depends largely on the underlying illness. Detailed history, careful physical examination, review of medication chart and analysis of initial investigations provides invaluable clues towards the diagnosis.


The shaken baby syndrome. A clinical, pathological, and biomechanical study.
Because a history of shaking is often lacking in the so-called "shaken baby syndrome," diagnosis is usually based on a constellation of clinical and radiographic findings. Forty-eight cases of infants and young children with this diagnosis seen between 1978 and 1985 at the Children's Hospital of Philadelphia were reviewed. All patients had a presenting history thought to be suspicious for child abuse, and either retinal hemorrhages with subdural or subarachnoid hemorrhages or a computerized tomography scan showing subdural or subarachnoid hemorrhages with interhemispheric blood. The physical examination and presence of associated trauma were analyzed; autopsy findings for the 13 fatalities were reviewed. All fatal cases had signs of blunt impact to the head, although in more than half of them these findings were noted only at autopsy. All deaths were associated with uncontrollably increased intracranial pressure. Models of 1-month-old infants with various neck and skull parameters were instrumented with accelerometers and shaken and impacted against padded or unpadded surfaces. Angular accelerations for shakes were smaller than those for impacts by a factor of 50. All shakes fell below injury thresholds established for subhuman primates scaled for the same brain mass, while impacts spanned concussion, subdural hematoma, and diffuse axonal injury ranges. It was concluded that severe head injuries commonly diagnosed as shaking injuries require impact to occur and that shaking alone in an otherwise normal baby is unlikely to cause the shaken baby syndrome.


Phospho-specific flow cytometry identifies aberrant signaling in indolent B-cell lymphoma.
Knowledge about signaling pathways in malignant cells may provide prognostic and diagnostic information in addition to identify potential molecular targets for therapy. B-cell receptor (BCR) and co-receptor CD40 signaling is essential for normal B cells, and there is increasing evidence that signaling via BCR and CD40 plays an important role in the pathogenesis of B-cell lymphoma. The aim of this study was to investigate basal and induced signaling in lymphoma B cells and infiltrating T cells in single-cell suspensions of biopsies from small cell lymphocytic lymphoma/chronic lymphocytic leukemia (SLL/CLL) and marginal zone lymphoma (MZL) patients. Samples from untreated SLL/CLL and MZL patients were examined for basal and activation induced signaling by phospho-specific flow cytometry. A panel of 9 stimulation conditions targeting B and T cells, including crosslinking of the B cell receptor (BCR), CD40 ligand and interleukins in combination with 12 matching phospho-protein readouts was used to study signaling. Malignant B cells from SLL/CLL patients had higher basal levels of phosphorylated (p)-SFKs, p-PLCγ, p-ERK, p-p38, p-p65 (NF-κB), p-STAT5 and p-STAT6, compared to healthy donor B cells. In contrast, anti-BCR induced signaling was highly impaired in SLL/CLL and MZL B cells as determined by low p-SFK, p-SYK and p-PLCγ levels. Impaired anti-BCR-induced p-PLCγ was associated with reduced surface expression of IgM and CD79b. Similarly, CD40L-induced p-ERK and p-p38 were also significantly reduced in lymphoma B cells, whereas p-p65 (NF-κB) was equal to that of normal B cells. In contrast, IL-2, IL-7 and IL-15 induced p-STAT5 in tumor-infiltrating T cells were not different from normal T cells. BCR signaling and CD40L-induced p-p38 was suppressed in malignant B cells from SLL/CLL and MZL patients. Single-cell phospho-specific flow cytometry for detection of basal as well as activation-induced phosphorylation of signaling proteins in distinct cell populations can be used to identify aberrant signaling pathways.


Predicting difficult-to-treat chronic rhinosinusitis by noninvasive biological markers.
Previous studies have been limited to the utility of clinical features and invasive nasal mucosal biomarkers in the prediction of chronic rhinosinusitis (CRS) outcomes. This study aimed to identify noninvasive biomarkers associated with difficult- to-treat CRS, enabling physicians to subgroup patients into risk groups for poor outcome before surgery. Three hundred and nine CRS patients undergoing endoscopic sinus surgery were finally enrolled. Patients treated with oral or intranasal glucocorticoids within 3 months or 1 month before surgery, respectively, were excluded. Baseline clinical charac- teristics, nasal secretions and peripheral blood samples were collected before surgery. The protein levels of 39 biological mar- kers were detected by the Bio-Plex suspension chip method. Classification and regression tree analysis was applied to establish prediction model for difficult-to-treat CRS determined one year after surgery. A random forest algorithm was used to confirm the discriminating factors that formed the classification tree. In the cohort with nasal secretion sample (n = 189), 21% of CRS patients were diagnosed as difficult-to-treat after 1 year of follow-up. Nasal secretion CCL17 level, hyposmia score, allergic rhinitis comorbidity, and nasal secretion MIP-1β level were found important predictors of difficult-to-treat CRS. A classification tree separated patients into 5 subgroups leading to an overall predictive accuracy of 94%. However, none of the plasma biological markers were associated with difficult-to-treat CRS in the cohort with blood sample (n = 128). Patients with difficult-to-treat-CRS were characterized by higher nasal secretion levels of CCL17 and MIP-1β severe hyposmia and concomitant allergic rhinitis. The classification tree could be useful to identify patients with high risk of poor outcome prior to surgery and offer more personalized interventions. However, since only patients without preoperative steroid treatments were included in this study, the generalization of our predictive model in other patient populations should be conside- red with caution.


Upper gastrointestinal adverse events and cyclical etidronate.
Recently, there have been several published case reports of esophagitis associated with the use of aminobisphosphonates. The objective of this study was to evaluate the upper gastrointestinal (GI) safety of cyclical etidronate, an alkylbisphosphonate, in routine clinical practice. Information was obtained from 550 general practices in the United Kingdom that provide the medical records to the General Practice Research Database. A group of 7977 cyclical etidronate takers and 2 age-, gender-, and practice-matched control groups (1 with osteoporosis and 1 without) were analyzed. For cyclical etidronate takers, the average age was 71.6 years and total follow-up was 10,328 person-years. The risk of upper GI events (inflammation or ulcer of esophagus, stomach, or duodenum) was comparable between patients taking etidronate and the two control groups. The adjusted relative risk of upper GI events was 0.92 (95% confidence interval [CI] 0.78 to 1.09) for etidronate takers compared with osteoporosis controls and 1.12 (CI 0.91 to 1.37) compared to nonosteoporosis controls. For esophagitis and esophageal ulcers, the relative risks were 0.83 (CI 0.64 to 1.08) and 0.97 (CI 0.71 to 1.31) respectively. The incidence of upper GI events during nonsteroidal anti-inflammatory drug (NSAID), aspirin, or corticosteroid use was similar across the three groups. The upper GI risks of etidronate NSAID users were 0.71 (CI 0.45 to 1.11) and 2.06 (CI 0.98 to 4.35) compared with NSAID users in the two control groups. These results support the GI tolerability and safety profile of cyclical etidronate in routine clinical practice. Concomitant use of cyclical etidronate with NSAIDs, aspirin, or corticosteroids did not increase the incidence of upper GI events.


N. elongata produces type IV pili that mediate interspecies gene transfer with N. gonorrhoeae.
The genus Neisseria contains at least eight commensal and two pathogenic species. According to the Neisseria phylogenetic tree, commensals are basal to the pathogens. N. elongata, which is at the opposite end of the tree from N. gonorrhoeae, has been observed to be fimbriated, and these fimbriae are correlated with genetic competence in this organism. We tested the hypothesis that the fimbriae of N. elongata are Type IV pili (Tfp), and that Tfp functions in genetic competence. We provide evidence that the N. elongata fimbriae are indeed Tfp. Tfp, as well as the DNA Uptake Sequence (DUS), greatly enhance N. elongata DNA transformation. Tfp allows N. elongata to make intimate contact with N. gonorrhoeae and to mediate the transfer of antibiotic resistance markers between these two species. We conclude that Tfp functional for genetic competence is a trait of a commensal member of the Neisseria genus. Our findings provide a mechanism for the horizontal gene transfer that has been observed among Neisseria species.


Body fat and risk of colorectal cancer among postmenopausal women.
Studies of the relationship between anthropometric indices of obesity and colorectal cancer risk in women have shown only weak and inconsistent associations. Given the limitations of such indices, we used dual-energy X-ray absorptiometry (DXA)-derived measures of body fat obtained in the Women's Health Initiative to examine the association between body fat and risk of incident colorectal cancer. We compared these risk estimates with those obtained using conventional anthropometric measurements (body mass index and waist circumference). After exclusions, the study population consisted of 11,124 postmenopausal women with DXA measurements at baseline and no history of colorectal cancer. After a median follow-up period of 12.9 years, 169 incident colorectal cancer cases were ascertained. Cox's proportional hazards models were used to estimate hazard ratios and 95 % confidence intervals for the exposures of interest. Neither DXA-derived body fat measures nor anthropometric measures showed significant associations with risk. In view of the limited number of cases, we cannot rule out the existence of weak associations of these measures with risk of colorectal cancer.


Thermal stability and reactivity of metal halide filled single-walled carbon nanotubes.
Thermal stability and reactivity to oxidation of several nanocomposite systems obtained by encapsulation of metal halides in single-walled carbon nanotubes are studied. Thermogravimetric analysis coupled with Raman spectroscopy allows insight into the various contributing factors, such as charge transfer, strain, and defect formation, and establishing a hierarchy of reactivity for the systems studied (AgX@SWCNTs, with X = Br, I; SWCNTs = arc discharge and HiPCO). The activation energy for oxidation decreases considerably after filling, indicating that filled nanotubes are more amenable to controlled modifications based on chemical reactivity than the originating empty nanotubes. The complete removal of the carbon shell at high temperatures does not preserve the nanowire morphology of the encapsulated halides; these are freed on surfaces in the form of nanoparticles arranged in 1D patterns. Metallic nanoparticles were obtained after hydrogen reduction of the halides, and growth of silicon nanowires in the footprint of the originating nanocomposites was demonstrated from such Co seeds. MX@SWCNTs (M = Ag, Co) can thus be used as environmentally stable nanoscale containers that allow the deliverance of catalytic nanoparticles in a prepatterned and aligned way.


Genome-Wide and Abdominal MRI Data Provide Evidence That a Genetically Determined Favorable Adiposity Phenotype Is Characterized by Lower Ectopic Liver Fat and Lower Risk of Type 2 Diabetes, Heart Disease, and Hypertension.
Recent genetic studies have identified alleles associated with opposite effects on adiposity and risk of type 2 diabetes. We aimed to identify more of these variants and test the hypothesis that such favorable adiposity alleles are associated with higher subcutaneous fat and lower ectopic fat. We combined MRI data with genome-wide association studies of body fat percentage (%) and metabolic traits. We report 14 alleles, including 7 newly characterized alleles, associated with higher adiposity but a favorable metabolic profile. Consistent with previous studies, individuals carrying more favorable adiposity alleles had higher body fat % and higher BMI but lower risk of type 2 diabetes, heart disease, and hypertension. These individuals also had higher subcutaneous fat but lower liver fat and a lower visceral-to-subcutaneous adipose tissue ratio. Individual alleles associated with higher body fat % but lower liver fat and lower risk of type 2 diabetes included those in PPARG, GRB14, and IRS1, whereas the allele in ANKRD55 was paradoxically associated with higher visceral fat but lower risk of type 2 diabetes. Most identified favorable adiposity alleles are associated with higher subcutaneous and lower liver fat, a mechanism consistent with the beneficial effects of storing excess triglycerides in metabolically low-risk depots.


Low consumption of fruits and dairy foods is associated with metabolic syndrome in Korean adults from outpatient clinics in and near Seoul.
The aim of this study was to examine differences in nutrient intake and food consumption by the presence of metabolic syndrome in Korean adults. Study subjects in this cross-sectional study were recruited from four outpatient clinics in and near the Seoul metropolitan area of South Korea between 2006 and 2012. A total of 668 subjects (413 men and 255 women) aged ≥ 30 y were included in the final data analyses. For each subject, daily nutrient intake and food consumption were calculated using three days of dietary intake data obtained from a combination of 24-hour recalls and dietary records. To evaluate food consumption, mean number of servings consumed per day and percentages of recommended number of servings for six food groups were calculated. Metabolic syndrome was defined using the National Cholesterol Education Program Adult Treatment Panel III criteria. The general linear model was performed to examine differences in nutrient intake and food consumption by sex and the presence of metabolic syndrome after adjustment for potential confounding variables. Nutrient intake did not differ by the presence of metabolic syndrome in both men and women. Men with metabolic syndrome had lower consumption and percentage of the recommendation for fruits compared with those without metabolic syndrome (1.6 vs. 1.1 servings/day, P-value = 0.001; 63.5 vs. 49.5%, P-value = 0.013). Women with metabolic syndrome showed lower consumption and percentage of the recommendation for dairy foods than those without metabolic syndrome (0.8 vs. 0.5 servings/day, P-value = 0.001; 78.6 vs. 48.9%, P-value = 0.001). Low intakes of fruits and dairy foods might be associated with the risk of having metabolic syndrome among Korean adults. Dietary advice on increasing consumption of these foods is needed to prevent and attenuate the risk of metabolic syndrome.


Efficacy and safety of retapamulin ointment as treatment of impetigo: randomized double-blind multicentre placebo-controlled trial.
Impetigo is a common skin infection, primarily caused by Staphylococcus aureus and mainly occurring in children. It is usually treated topically with antibiotics to achieve a quick cure and prevent spread of the infection. Worldwide, resistance rates of S. aureus against commonly used antibiotics are rising. Retapamulin belongs to a newly developed class of antibiotics for the treatment of uncomplicated skin infections. Our aim was to compare the efficacy and safety of topical application of retapamulin ointment with topical placebo ointment in the treatment of primary impetigo. In a randomized, double-blind, multicentre study, patients received either topical retapamulin ointment 1% twice daily for 5 days or topical placebo. Patients were enrolled into the study for 14 days and attended the clinic for three visits during which clinical and laboratory evaluations were performed. Two hundred and thirteen patients were randomized, with 139 evaluable patients in the retapamulin group and 71 in the placebo group. Based on the primary efficacy endpoint of clinical response after 7 days (intention to treat), retapamulin ointment was superior to placebo (success rate 85.6% vs. 52.1%; P<0.0001). Similar results were found in the per protocol analysis and in the subgroup of patients who had a pathogen isolated at baseline. The most common adverse effect, pruritus at the application site, was reported by 6% and 1% of patients in the retapamulin and placebo groups, respectively. This study shows that topical retapamulin is effective and safe in the treatment of primary impetigo, offering a new treatment option.


Assessing explicit strategies in force field adaptation.
In recent years, it has become increasingly clear that a number of learning processes are at play in visuomotor adaptation tasks. In addition to implicitly adapting to a perturbation, learners can develop explicit knowledge allowing them to select better actions in responding to it. Advances in visuomotor rotation experiments have underscored the important role of such "explicit learning" in shaping adaptation to kinematic perturbations. Yet, in adaptation to dynamic perturbations, its contribution has been largely overlooked. We therefore sought to approach the assessment of explicit learning in adaptation to dynamic perturbations, by developing two novel modifications of a force field experiment. First, we asked learners to abandon any cognitive strategy before selected force channel trials to expose consciously accessible parts of overall learning. Here, learners indeed reduced compensatory force compared with standard Catch channels. Second, we instructed a group of learners to mimic their right hand's adaptation by moving with their naïve left hand. While a control group displayed negligible left hand force compensation, the mimicking group reported forces that approximated right hand adaptation but appeared to under-report the velocity component of the force field in favor of a more position-based component. Our results highlight the viability of explicit learning as a potential contributor to force field adaptation, though the fraction of learning under participants' deliberate control on average remained considerably smaller than that of implicit learning, despite task conditions favoring explicit learning. The methods we employed provide a starting point for investigating the contribution of explicit strategies to force field adaptation.NEW & NOTEWORTHY While the contribution of explicit learning has been increasingly studied in visuomotor adaptation, its contribution to force field adaptation has not been studied extensively. We employed two novel methods to assay explicit learning in a force field adaptation task and found that learners can voluntarily control aspects of compensatory force production and manually report it with their untrained limb. This supports the general viability of the contribution of explicit learning also in force field adaptation.


Mesh diagram analysis: developing a norm for African Americans.
The mesh analysis is a proportionate cephalometric analysis that enables a patient to serve as his or her own control to create a template from which to assess skeletal, dental, and craniofacial disharmonies. The norms for this analysis were originally created from a white, European American sample. The purposes of this study were: (1) to create a normal mesh diagram from a black, African American population; (2) to compare mesh diagrams from black and white Americans; and (3) to evaluate the usefulness of a panel of diverse members in selecting subjects. All subjects were black Americans of African descent, had no prior orthodontic treatment, and had Class I dental occlusion with minimal crowding (4 mm or less). The panel selected as esthetically pleasing 18 males and 25 females from a group of 77 patients meeting the study criteria. Male and female normal mesh diagrams were created from the cephalographs and compared with Caucasian normal diagrams developed by Moorrees in 1976. Linear and angular components of the hard and soft tissues were compared. Comparisons were also made with previously published normal values. Differences between the African American and Caucasian samples were more notable close to the dentoalveolar complex. Similarities were limited to the soft tissue of the upper face, the cranial base, and the midface. Analysis of the panel selection results did not suggest any trends between or within race, sex, or occupation of the panel members. However, agreement between the races was good.


Beyond leopards: tooth marks and the contribution of multiple carnivore taxa to the accumulation of the Swartkrans Member 3 fossil assemblage.
The ca. 1.0 myr old fauna from Swartkrans Member 3 (South Africa) preserves abundant indication of carnivore activity in the form of tooth marks (including pits) on many bone surfaces. This direct paleontological evidence is used to test a recent suggestion that leopards, regardless of prey body size, may have been almost solely responsible for the accumulation of the majority of bones in multiple deposits (including Swartkrans Member 3) from various Sterkfontein Valley cave sites. Our results falsify that hypothesis and corroborate an earlier hypothesis that, while the carcasses of smaller animals may have been deposited in Swartkrans by leopards, other kinds of carnivores (and hominids) were mostly responsible for the deposition of large animal remains. These results demonstrate the importance of choosing appropriate classes of actualistic data for constructing taphonomic inferences of assemblage formation. In addition, they stress that an all-encompassing model of assemblage formation for the hominid-bearing deposits of the Sterkfontein Valley is inadequate and that each must be evaluated individually using not just analogical reasoning but also incorporating empirical data generated in the preserved fossil samples.


Glyoxalase 1 increases anxiety by reducing GABAA receptor agonist methylglyoxal.
Glyoxalase 1 (Glo1) expression has previously been associated with anxiety in mice; however, its role in anxiety is controversial, and the underlying mechanism is unknown. Here, we demonstrate that GLO1 increases anxiety by reducing levels of methylglyoxal (MG), a GABAA receptor agonist. Mice overexpressing Glo1 on a Tg bacterial artificial chromosome displayed increased anxiety-like behavior and reduced brain MG concentrations. Treatment with low doses of MG reduced anxiety-like behavior, while higher doses caused locomotor depression, ataxia, and hypothermia, which are characteristic effects of GABAA receptor activation. Consistent with these data, we found that physiological concentrations of MG selectively activated GABAA receptors in primary neurons. These data indicate that GLO1 increases anxiety by reducing levels of MG, thereby decreasing GABAA receptor activation. More broadly, our findings potentially link metabolic state, neuronal inhibitory tone, and behavior. Finally, we demonstrated that pharmacological inhibition of GLO1 reduced anxiety, suggesting that GLO1 is a possible target for the treatment of anxiety disorders.


Trends in antibiotic utilization in eight Latin American countries, 1997-2007.
To describe the trends in antibiotic utilization in eight Latin American countries between 1997-2007 We analyzed retail sales data of oral and injectable antibiotics (World Health Organization (WHO) Anatomic Therapeutic Chemical (ATC) code J01) between 1997 and 2007 for Argentina, Brazil, Chile, Colombia, Mexico, Peru, Uruguay, and Venezuela. Antibiotics were aggregated and utilization was calculated for all antibiotics (J01); for macrolides, lincosamindes, and streptogramins (J01 F); and for quinolones (J01 M). The kilogram sales of each antibiotic were converted into defined daily dose per 1 000 inhabitants per day (DID) according to the WHO ATC classification system. We calculated the absolute change in DID and relative change expressed in percent of DID variation, using 1997 as a reference Total antibiotic utilization has increased in Peru, Venezuela, Uruguay, and Brazil, with the largest relative increases observed in Peru (5.58 DID, +70.6%) and Venezuela (4.81 DID, +43.0%). For Mexico (-2.43 DID; -15.5%) and Colombia (-4.10; -33.7%), utilization decreased. Argentina and Chile showed major reductions in antibiotic utilization during the middle of this period. In all countries, quinolone use increased, particularly sharply in Venezuela (1.86 DID, +282%). The increase in macrolide, lincosaminde, and streptogramin use was greatest in Peru (0.76 DID, +82.1%), followed by Brazil, Argentina, and Chile Analyzing antibiotic utilization in Latin America presents a series of challenges. Creating policy-relevant evidence based on antimicrobial consumption patterns is needed in order to foster policies aimed at improving appropriate use of antibiotics in the region.


Crosslink Density Is Reduced and Oxidation Is Increased in Retrieved Highly Crosslinked Polyethylene TKA Tibial Inserts.
The wear resistance of highly crosslinked polyethylene depends on crosslink density, which may decrease with in vivo loading, leading to more wear and increased oxidation. The relationship among large and complex in vivo mechanical stresses, breakdown of the polyethylene crosslinks, and oxidative degradation is not fully understood in total knee arthroplasty (TKA). We wished to determine whether crosslink density is reduced at the articular surfaces of retrieved tibial inserts in contact areas exposed to in vivo mechanical stress. (1) Does polyethylene crosslink density decrease preferentially in regions of the articular surface of thermally stabilized crosslinked polyethylene tibial components exposed to mechanical stress in vivo; and (2) what is the ramification of decreased crosslink density in TKA in terms of accompanying oxidation of the polyethylene? From May 2011 to January 2014, 90 crosslinked polyethylene tibial components were retrieved during revision surgery as a part of a long-standing implant retrieval program. Forty highly crosslinked polyethylene tibial inserts (27 posterior-stabilized designs and 13 cruciate-retaining designs) retrieved for instability (15 cases), stiffness (11), infection (six), aseptic loosening (four), pain (two), and malposition (two) after a mean time of 18 months were inspected microscopically to identify loaded (burnished) and unloaded (unburnished) regions on the articular surfaces. Swell ratio testing was done according to ASTM F2214 to calculate crosslink density and infrared spectroscopy was used according to ASTM F2102 to measure oxidation. The region of the tibial insert influenced crosslink density. Loaded surface regions had a mean crosslink density of 0.19 (95% confidence interval [CI], 0.18-0.19) mol/dm3, lower than the other three regions (loaded subsurface, unloaded surface, and unloaded subsurface), which had crosslink densities of 0.21 (95% CI, 0.21-0.22; p < 0.01) mol/dm3. Peak oxidation levels were higher in loaded regions with a mean oxidation index (OI) of 0.67 (95% CI, 0.56-0.78) versus unloaded regions with a mean OI of 0.36 (95% CI, 0.27-0.45; p < 0.01). Peak oxidation levels were higher in annealed samples with a mean OI of 0.66 (95% CI, 0.52-0.81) versus remelted samples with a mean OI of 0.40 (95% CI, 0.34-0.47; p < 0.01). The results suggest that the crosslink density decreases and accompanying oxidation is driven predominantly by contact stress conditions. If crosslink density continues to decrease with continued loading over time, crosslinked polyethylene may not provide a clinical advantage over conventional polyethylene in the long term for TKA. Therefore, we will continue to collect longer term retrievals to evaluate mechanical property changes in crosslinked polyethylenes. Although we found a decrease in crosslink density and increase in oxidation in the tibial inserts, the degree of oxidation does not suggest, for now, a reason for concern in these early retrievals. The OI values of the tibial inserts in this study were lower than the critical oxidation level (OI > 3) reported in the literature where polyethylene may lose mechanical properties and have the compromised ability to withstand mechanical loading.


The dynamic behaviour of metabolic syndrome and its components in an eight-year population-based cohort from the Mediterranean.
The significant rise in the prevalence of obesity coincides with the considerable increase in the prevalence of metabolic syndrome (MS) currently being observed worldwide. The components of MS are not static and their dynamics, such as the order of their occurrence, or the time of exposure to them are, as yet, unknown but could well be clinically relevant. Our objective was to study the dynamic behaviour of MS and its components in a large population-based cohort from a Mediterranean region. Our study employed a retrospective cohort (between January 1, 2005 and December 31, 2012) made up of individuals from the general population in a region in the northeast of Catalonia, Spain. Given that most of the explicative variables of the risk of having MS were time dependent and, therefore, the risk was not proportional, we used the Andersen-Gill (AG) model to perform a multivariate survival analysis and inferences were performed using a Bayesian framework. Thirty-nine percent of the participants developed MS; 44.6% of them with a single limited episode. Triglycerides and low HDL cholesterol, together with obesity, are components associated with the first occurrence of MS. Components related to the metabolism of glucose are associated with a medium risk of having a first episode of MS, and those related to blood pressure are associated with a lower risk. When the components related to blood pressure and the metabolism of glucose appear first, they determine the appearance of the first episode of MS. The variables concerning the persistence of MS are those that correspond to clinical conditions that do not have well-established drug treatment criteria. Our results suggest that the components related to the metabolism of glucose and to high blood pressure appear early on and act as biomarkers for predicting MS, while the components related to obesity and dyslipidaemia, although essential for the development of MS, appear later. Making lifestyle changes reduces the conditions associated with the persistence of MS.


Is hepatitis E virus an emerging problem in industrialized countries?
Hepatitis E virus (HEV) is yearly responsible for approximately 20 million infections worldwide. Although most infections occur in developing countries, HEV appears to be an emerging problem in several industrialized countries, where it is mostly associated with either traveling to an HEV endemic area or contact with pigs, which represent a major reservoir of HEV. The major risk groups for HEV infection and its ensuing complications are elderly men, pregnant women, young children, immunocompromised patients, patients with preexisting liver disease, and workers that come into close contact with HEV-infected animals. Whereas HEV mainly causes acute self-limiting infections, chronic infections may occur among immunocompromised patients (e.g., transplant recipients and human immunodeficiency virus [HIV]-infected patients). Accordingly, HEV-HIV coinfection leads to accelerated liver cirrhosis and increased mortality rates compared to HEV infection alone, which is, except during pregnancy, usually associated with only low mortality. In the Western world, the most common genotype (gt) causing HEV infection is gt 3. Ribavirin (RBV) and interferon have been used successfully for treatment of HEV, but this treatment is contraindicated in certain patient groups. Therefore, novel antiviral compounds are highly needed, especially given that viral isolates with RBV resistance have been recently identified. Moreover, eradication of HEV is hampered by long-term environmental persistence of the virus, which represents a continuous source of the virus. In 2011, the first prophylactic HEV vaccine, Hecolin, was approved in China, but it is not yet globally available. In this review, we will discuss the molecular virology of HEV, mode of transmission in industrialized countries, and potential implications for different specific patient populations.


Methylene blue fails to inhibit Tau and polyglutamine protein dependent toxicity in zebrafish.
Methylene blue is an FDA approved compound with a variety of pharmacologic activities. It inhibits aggregation of several amyloidogenic proteins known to be deposited in neurodegenerative diseases. Recently, it has been proposed that methylene blue shows significant beneficial effects in a phase 2 clinical trial by slowing cognitive decline in Alzheimer's disease patients. To analyze its therapeutic potential, we investigated the effect of methylene blue on neurotoxicity in a zebrafish model for tauopathies. Transgenic expression of the frontotemporal dementia associated Tau-P301L mutation recapitulates a number of the pathological features observed in humans including abnormal phosphorylation and folding of Tau, tangle formation and Tau dependent neuronal loss. Upon incubation of zebrafish larvae with methylene blue, neither abnormal phosphorylation nor neuronal cell loss, reduced neurite outgrowth or a swimming defect were rescued. Methylene blue is biologically active in zebrafish since it reduced aggregation of a huntingtin variant containing a stretch of 102 glutamine residues. However, although huntingtin aggregation was largely prevented by methylene blue, huntingtin-dependent toxicity was unaffected. Our findings are consistent with the hypothesis that toxicity is not necessarily associated with deposition of insoluble amyloid proteins.


Severe FOXP3+ and naïve T lymphopenia in a non-IPEX form of autoimmune enteropathy combined with an immunodeficiency.
Immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome is the best-characterized form of a rare entity called autoimmune enteropathy (AIE). IPEX syndrome is due to mutations in the FOXP3 gene, a transcription factor essential for the development and function of the natural regulatory CD25(+)CD4(+) T cells. We studied a female patient with a polyautoimmune AIE syndrome resembling a mild form of IPEX syndrome but associated with recurrent bacterial infections and mild hypogammaglobulinemia. We hypothesized that this syndrome combined a deficit of FOXP3(+) cells and other lymphocyte populations. We analyzed the major lymphocyte subsets and the FOXP3(+) regulatory system in blood samples obtained during the 2-year period that followed the last autoimmune manifestation. The patient had severe naïve T lymphopenia and a major deficit of FOXP3(+)CD4(+) T cells, both in circulation and in the highly inflamed intestinal mucosa, but mutations in the FOXP3 locus were excluded. The blood FOXP3(+) pool was devoid of CD25(high) cells, but the few regulatory CD25(+) cells were functional. Intrinsic defects in the expression of CD25, FOXP3, and interleukin 2 were excluded. Upon activation, a small subset of cells, presumably committed to regulatory function, sustained expression of CD25 and FOXP3. Peripheral T lymphopenia of both naïve and natural regulatory T cells might be the consequence of defective thymic production or the short life span of exported T cells. This case sheds new light in the etiology of autoimmune manifestations in T-cell immunodeficiencies and in the heterogeneity of AIE.