Paramuscular perforators in DIEAP flap for breast reconstruction.
One of the main steps in perforator flap surgery is to identify the dominant perforator. Using multidetector row computed tomography (MDCT) for the preoperative planning of deep inferior epigastric artery perforator (DIEAP) flap surgery, we identified a perforator with a large caliber, an excellent location in the middle abdominal region, and a totally extramuscular trajectory in a significant number of patients. We describe the frequency of this perforator and determine its characteristics. We conducted a retrospective study of 482 patients who underwent 526 DIEAP flaps for breast reconstruction from October 2003 to October 2011. Mean age at surgery was 51.3 years old. A preoperative MDCT of abdominal vascularization was performed in all patients. MDCT identified a dominant perforator with a paramuscular course in 12.4% of abdominal walls. In all cases, it was located in the midline and emerged directly from the deep inferior epigastric system. Its mean caliber was 1.9 mm. The flap was harvested based on this perforator in all these patients, and mean harvest time was 51 minutes. The characteristics of this perforator made dissection easier and reduced morbidity at the donor site. There were no flap losses and the only complications were minor. We located a paramuscular perforator in 12.4% of patients undergoing breast reconstruction with abdominal perforator flaps. Its morphological features and extramuscular course make it the perforator of choice in DIEAP flap surgery.


Phenytoin versus levetiracetam as prophylaxis for postcraniotomy seizure in patients with no history of seizures: systematic review and meta-analysis.
OBJECTIVEDe novo seizure following craniotomy (DSC) for nontraumatic pathology may adversely affect medical and neurological outcomes in patients with no history of seizures who have undergone craniotomies. Antiepileptic drugs (AEDs) are commonly used prophylactically in patients undergoing craniotomy; however, evidence supporting this practice is limited and mixed. The authors aimed to collate the available evidence on the efficacy and tolerability of levetiracetam monotherapy and compare it with that of the classic AED, phenytoin, for DSC.METHODSPubMed, Embase, Web of Science, and the Cochrane Library were searched for studies that compared levetiracetam with phenytoin for DSC prevention. Inclusion criteria were adult patients with no history of epilepsy who underwent craniotomy with prophylactic usage of phenytoin, a comparator group with levetiracetam treatment as the main treatment difference between the two groups, and availability of data on the numbers of patients and seizures for each group. Patients with brain injury and previous seizure history were excluded. DSC occurrence and adverse drug reaction (ADR) were evaluated. Seizure occurrence was calculated using the Peto odds ratio (POR), which is the relative effect estimation method of choice for binary data with rare events.RESULTSData from 7 studies involving 803 patients were included. The DSC occurrence rate was 1.26% (4/318) in the levetiracetam cohort and 6.60% (32/485) in the phenytoin cohort. Meta-analysis showed that levetiracetam is significantly superior to phenytoin for DSC prevention (POR 0.233, 95% confidence interval [CI] 0.117-0.462, p < 0.001). Subgroup analysis demonstrated that levetiracetam is superior to phenytoin for DSC due to all brain diseases (POR 0.129, 95% CI 0.039-0.423, p = 0.001) and tumor (POR 0.282, 95% CI 0.117-0.678, p = 0.005). ADRs in the levetiracetam group were cognitive disturbance, thrombophlebitis, irritability, lethargy, tiredness, and asthenia, whereas rash, anaphylaxis, arrhythmia, and hyponatremia were more common in the phenytoin group. The overall occurrence of ADR in the phenytoin (34/466) and levetiracetam (26/432) groups (p = 0.44) demonstrated no statistically significant difference in ADR occurrence. However, the discontinuation rate of AEDs due to ADR was 53/297 in the phenytoin group and 6/196 in the levetiracetam group (POR 0.266, 95% CI 0.137-0.518, p < 0.001).CONCLUSIONSLevetiracetam is superior to phenytoin for DSC prevention for nontraumatic pathology and has fewer serious ADRs that lead to discontinuation. Further high-quality studies that compare levetiracetam with placebo are necessary to provide evidence for establishing AED guidelines.


Live or computerized simulation of clinical encounters: do clinicians work up patient cases differently?
Computer simulation of clinical encounters is increasingly used in clinical settings to train patient work-up. The aim of this prospective, controlled study was to compare the characteristics of data collection and diagnostic exploration of physicians working up cases with a standardized patient and in a computerized simulation. Six clinicians of different clinical experience in internal medicine worked up three cases with a standardized patient and through a computer simulation allowing free inquiry. After each encounter, we asked the subjects to justify the information collected and to comment on their working diagnoses. The characteristics of data collected and working diagnoses generated were assessed and compared, according to the simulation method used. In the computer simulation, physicians limited their data collection and focused earlier and more specifically on information and working diagnoses with high levels of relevance. They reached a similar diagnostic accuracy and made decisions of a similar relevance. Computer simulation with a free-inquiry approach reproduces the data collection and the diagnostic exploration observed in a standardized-patient simulation and promotes an early collection of relevant data. Its contribution to extend the competence of learners in clinical settings should be further evaluated.


Hydrogenation of ethylene over palladium: evolution of the catalyst structure by operando synchrotron-based techniques.
Palladium-based catalysts are exploited on an industrial scale for the selective hydrogenation of hydrocarbons. The formation of palladium carbide and hydride phases under reaction conditions changes the catalytic properties of the material, which points to the importance of operando characterization for determining the relation between the relative fractions of the two phases and the catalyst performance. We present a combined time-resolved characterization by X-ray absorption spectroscopy (in both near-edge and extended regions) and X-ray diffraction of a working palladium-based catalyst during the hydrogenation of ethylene in a wide range of partial pressures of ethylene and hydrogen. Synergistic coupling of multiple techniques allowed us to follow the structural evolution of the palladium lattice as well as the transitions between the metallic, hydride and carbide phases of palladium. The nanometric dimensions of the particles resulted in the considerable contribution of both surface and bulk carbides to the X-ray absorption spectra. During the reaction, palladium carbide is formed, which does not lead to a loss of activity. Unusual contraction of the unit cell parameter of the palladium lattice in the spent catalyst was observed upon increasing hydrogen partial pressure.


Merkel Cell Polyomavirus in Merkel Cell Carcinoma: Integration Sites and Involvement of the KMT2D Tumor Suppressor Gene.
Merkel cell carcinoma (MCC) is an uncommon, lethal cancer of the skin caused by either Merkel cell polyomavirus (MCPyV) or UV-linked mutations. MCPyV is found integrated into MCC tumor genomes, accompanied by truncation mutations that render the MCPyV large T antigen replication incompetent. We used the open access HPV Detector/Cancer-virus Detector tool to determine MCPyV integration sites in whole-exome sequencing data from five MCC cases, thereby adding to the limited published MCPyV integration site junction data. We also systematically reviewed published data on integration for MCPyV in the human genome, presenting a collation of 123 MCC cases and their linked chromosomal sites. We confirmed that there were no highly recurrent specific sites of integration. We found that chromosome 5 was most frequently involved in MCPyV integration and that integration sites were significantly enriched for genes with binding sites for oncogenic transcription factors such as LEF1 and ZEB1, suggesting the possibility of increased open chromatin in these gene sets. Additionally, in one case we found, for the first time, integration involving the tumor suppressor gene KMT2D, adding to previous reports of rare MCPyV integration into host tumor suppressor genes in MCC.


[Intermittent fasting : A solution for metabolic disorders?]
The management of obesity comprises lifestyle changes targeting nutrient content, eating behavior and regular physical activity. Medication (orlistat, liraglutide) and bariatric surgery can later be used, but they require a clear indication and a close follow-up. Studies in chronobiology are now exploring the metabolic benefits of intermittent fasting, which restricts food intake and calorie-containing beverages to a certain window of the 24h cycle, or to certain days of the week/month, thus reinstating the alternance between anabolism and catabolism. However, the current scientific evidence is limited by the sample size and duration of the studies. It is therefore too early for a blanket strategy based on intermittent fasting in all patients with metabolic disorders.


Double-labeled rabies virus: live tracking of enveloped virus transport.
Here we describe a strategy to fluorescently label the envelope of rabies virus (RV), of the Rhabdoviridae family, in order to track the transport of single enveloped viruses in living cells. Red fluorescent proteins (tm-RFP) were engineered to comprise the N-terminal signal sequence and C-terminal transmembrane spanning and cytoplasmic domain sequences of the RV glycoprotein (G). Two variants of tm-RFP were transported to and anchored in the cell surface membrane, independent of glycosylation. As shown by confocal microscopy, tm-RFP colocalized at the cell surface with the RV matrix and G protein and was incorporated into G gene-deficient virus particles. Recombinant RV expressing the membrane-anchored tm-RFP in addition to G yielded infectious viruses with mosaic envelopes containing both tm-RFP and G. Viable double-labeled virus particles comprising a red fluorescent envelope and a green fluorescent ribonucleoprotein were generated by expressing in addition an enhanced green fluorescent protein-phosphoprotein fusion construct (S. Finke, K. Brzozka, and K. K. Conzelmann, J. Virol. 78:12333-12343, 2004). Individual enveloped virus particles were observed under live cell conditions as extracellular particles and inside endosomal vesicles. Importantly, double-labeled RVs were transported in the retrograde direction over long distances in neurites of in vitro-differentiated NS20Y neuroblastoma cells. This indicates that the typical retrograde axonal transport of RV to the central nervous system involves neuronal transport vesicles in which complete enveloped RV particles are carried as a cargo.


A core group model for disease transmission.
Models for sexually transmitted diseases generally assume that the size of the core group is fixed. Publicly available information on disease prevalence may influence the recruitment of new susceptibles into highly sexually active populations. It is assumed that the recruitment rate into the core population is low while disease prevalence is high, core group members mix only with each other, disease levels outside the core are negligible, and some core group members reduce their risk through the use of a partially effective vaccine or prophylactics. A demographic-epidemic model is formulated in which the combined size of the core and non-core population is constant. A simpler version models the epidemic in an isolated core population of constant size under the influence of educational programs and measures that reduce susceptibility. The threshold condition for an endemic infection is determined. Backward bifurcations, multiple infective stationary states, and hysteresis phenomena can be observed even in the simplified version. Abrupt changes in disease prevalence levels may result from small changes in the disease management parameters and do not occur in the absence of such a program. The general conclusion is that partially effective vaccination or education programs may increase the total number of cases while decreasing the relative frequency of cases in the core group. The study throws some new light on the role of the reproduction number in connection with elimination attempts. It shows that although the reproduction number defines the threshold for the spread of the disease in a susceptible population, it is of limited value when elimination of an existing epidemic is planned.


Stressful life events and daily stressors affect awakening cortisol level in midlife mothers of individuals with autism spectrum disorders.
The current study examines the awakening cortisol level in midlife mothers (M = 51.4 years old, SD = 8.4) of individuals (M = 22.1 years old, SD = 7.1) with autism spectrum disorders (ASD) under stressful conditions that are not specific to their son or daughter's ASD symptoms. In addition to completing a set of questionnaires and in-home interviews, 82 mothers from the Adolescents and Adults with Autism Study (AAA) participated in a Daily Diary Study. Findings from the multilevel models indicated that mothers who previously were exposed to no negative life events in the previous period had an increased awakening cortisol level on days following a greater number and more severe stressors, a normative stress response. In contrast, we observed a flatter cortisol level of daily stressors in mothers who experienced a greater number of negative life events in the previous period. These findings highlight the sustained toll that global and everyday stressors have on awakening cortisol level of midlife and aging mothers of individuals with ASD.


Scented Sugar Baits Enhance Detection of St. Louis Encephalitis and West Nile Viruses in Mosquitoes in Suburban California.
Scented sugar baits deployed in California deserts detected early West Nile virus (WNV) transmission by mosquitoes, representing a potential improvement to conventional arbovirus surveillance that relies heavily on infection rates in mosquito pools. In this study, we expanded deployment of scented sugar baits into suburban Sacramento and Yolo (2015, 2016) and Riverside Counties (2016), California. The goal of the study was to determine whether scented sugar baits detect WNV and St. Louis encephalitis virus (SLEV) concurrent with mosquito infections in trapped pools in areas of high human density. Between 8 and 10% of sugar baits were WNV RNA positive in both study years across the three counties. In Riverside County, where SLEV re-emerged in 2015, 1% of sugar baits were SLEV positive in 2016. Rates of sugar bait positives were at least 100 times higher than infection rates in trapped mosquitoes in the same districts. The prevalence of sugar bait positives varied temporally and did not coincide with infections in mosquitoes collected at the same sites each week. WNV RNA positive sugar baits were detected up to 2 wk before and after concurrent surveillance detected infection in mosquito pools at the same sites. Sugar baits also detected WNV in Riverside County at locations where no WNV activity was detected in mosquito pools. Sugar baits generated between 0.8 and 1.2 WNV positives per $1,000 and can be more economical than carbon dioxide baited traps that produce 0.8 positives per $1,000. These results indicate that the sugar bait approach enhances conventional arbovirus surveillance in mosquitoes in suburban California.


Intestinal barrier dysfunction orchestrates the onset of inflammatory host-microbiome cross-talk in a human gut inflammation-on-a-chip.
The initiation of intestinal inflammation involves complex intercellular cross-talk of inflammatory cells, including the epithelial and immune cells, and the gut microbiome. This multicellular complexity has hampered the identification of the trigger that orchestrates the onset of intestinal inflammation. To identify the initiator of inflammatory host-microbiome cross-talk, we leveraged a pathomimetic "gut inflammation-on-a-chip" undergoing physiological flow and motions that recapitulates the pathophysiology of dextran sodium sulfate (DSS)-induced inflammation in murine models. DSS treatment significantly impaired, without cytotoxic damage, epithelial barrier integrity, villous microarchitecture, and mucus production, which were rapidly recovered after cessation of DSS treatment. We found that the direct contact of DSS-sensitized epithelium and immune cells elevates oxidative stress, in which the luminal microbial stimulation elicited the production of inflammatory cytokines and immune cell recruitment. In contrast, an intact intestinal barrier successfully suppressed oxidative stress and inflammatory cytokine production against the physiological level of lipopolysaccharide or nonpathogenic Escherichia coli in the presence of immune elements. Probiotic treatment effectively reduced the oxidative stress, but it failed to ameliorate the epithelial barrier dysfunction and proinflammatory response when the probiotic administration happened after the DSS-induced barrier disruption. Maintenance of epithelial barrier function was necessary and sufficient to control the physiological oxidative stress and proinflammatory cascades, suggesting that "good fences make good neighbors." Thus, the modular gut inflammation-on-a-chip identifies the mechanistic contribution of barrier dysfunction mediated by intercellular host-microbiome cross-talk to the onset of intestinal inflammation.


Cytomegalovirus vaccine development.
Although infection with human cytomegalovirus (HCMV) is ubiquitous and usually asymptomatic, there are individuals at high risk for serious HCMV disease. These include solid organ and hematopoietic stem cell (HSC) transplant patients, individuals with HIV infection, and the fetus. Since immunity to HCMV ameliorates the severity of disease, there have been efforts made for over 30 years to develop vaccines for use in these high-risk settings. However, in spite of these efforts, no HCMV vaccine appears to be approaching imminent licensure. The reasons for the failure to achieve the goal of a licensed HCMV vaccine are complex, but several key problems stand out. First, the host immune correlates of protective immunity are not yet clear. Secondly, the viral proteins that should be included in a HCMV vaccine are uncertain. Third, clinical trials have largely focused on immunocompromised patients, a population that may not be relevant to the problem of protection of the fetus against congenital infection. Fourth, the ultimate target population for HCMV vaccination remains unclear. Finally, and most importantly, there has been insufficient education about the problem of HCMV infection, particularly among women of child-bearing age and in the lay public. This review considers the strategies that have been explored to date in development of HCMV vaccines, and summarizes both active clinical trials as well as novel technologies that merit future consideration toward the goal of prevention of this significant public health problem.


Mismatch repair mutations of Escherichia coli K12 enhance transposon excision.
Excision of the prokaryotic transposon Tn10 is a host-mediated process that occurs in the absence of recA function or any transposon-encoded functions. To determine which host functions might play a role in transposon excision, we have isolated 40 mutants of E. coli K12, designated tex, which increase the frequency of Tn10 precise excision. Three of these mutations (texA) have been shown to qualitatively alter RecBC function. We show that 21 additional tex mutations with a mutator phenotype map to five genes previously identified as components of a methylation-directed pathway for repair of base pair mismatches: uvrD, mutH, mutL, mutS and dam. Previously identified alleles of these genes also have a Tex phenotype.--Several other E. coli mutations affecting related functions have been analyzed for their effects on Tn10 excision. Other mutations affecting the frequency of spontaneous mutations (mutT, polA, ung), different excision repair pathways (uvrA, uvrB) or the state of DNA methylation (dcm) have no effect on Tn10 excision. Mutations ssb-113 and mutD5, however, do increase Tn10 excision.--The products of the mismatch correction genes probably function in a coordinated way during DNA repair in vivo. Thus, mutations in these genes might also enhance transposon excision by a single general mechanism. Alternatively, since mutations in each gene have qualitatively and quantitatively different effects on transposon excision, defects in different mismatch repair genes may enhance excision by different mechanisms.


Early and late benefits of prasugrel in patients with acute coronary syndromes undergoing percutaneous coronary intervention: a TRITON-TIMI 38 (TRial to Assess Improvement in Therapeutic Outcomes by Optimizing Platelet InhibitioN with Prasugrel-Thrombolysis In Myocardial Infarction) analysis.
We evaluated the relative contributions of the loading and maintenance doses of prasugrel on events in a TRITON-TIMI 38 (TRial to Assess Improvement in Therapeutic Outcomes by Optimizing Platelet InhibitioN with Prasugrel-Thrombolysis In Myocardial Infarction) analysis. Prasugrel is superior to clopidogrel in preventing ischemic events in patients with an acute coronary syndrome who are undergoing percutaneous coronary intervention, but it is associated with an increased risk of major bleeding. Landmark analyses for efficacy, safety, and net clinical benefit were performed from randomization to day 3 and from day 3 to the end of the trial. Significant reductions in ischemic events, including myocardial infarction, stent thrombosis, and urgent target vessel revascularization, were observed with the use of prasugrel both during the first 3 days and from 3 days to the end of the trial. Thrombolysis In Myocardial Infarction major non-coronary artery bypass graft bleeding was similar to clopidogrel during the first 3 days but was significantly greater with the use of prasugrel from 3 days to the end of the study. Net clinical benefit significantly favored prasugrel both early and late in the trial. Both the loading dose and maintenance dose of prasugrel were superior to clopidogrel for the reduction of ischemic events. This result emphasizes the importance of maintaining high levels of inhibition of platelet aggregation via P2Y(12) receptor inhibition, not only for the prevention of periprocedural ischemic events but also during long-term follow-up. The excess major bleeding observed with the use of prasugrel occurred predominantly during the maintenance phase. Approaches to reduce the relative excess of bleeding with prasugrel should focus on the maintenance dose (e.g., reduction in maintenance dose in previously reported high-risk subgroups, such as the elderly and those patients with low body weight). (A Comparison of CS-747 and Clopidogrel in Acute Coronary Syndrome Subjects Who Are to Undergo Percutaneous Coronary Intervention; NCT00097591).


Local flaps in scar revision.
Local and regional skin flaps provide the foundation for reconstructive surgery of the face. When planned and executed properly, local skin flaps provide rapid reconstruction using donor tissue with excellent color and texture match, and a reliable, adequate blood supply. The majority of local skin flaps are utilized for reconstruction after superficial cancer extirpation, however, unfavorable scars from previous trauma, burns, or reconstruction efforts provide the motivated patient and surgeon another opportunity to restore form and function. This article reviews techniques commonly used to reconstruct the face and provides several cases where these techniques were successfully employed.


Sex differences in facial skin temperature when exposed to darkness with and without warning.
The changes in skin temperature under anxiety were investigated by noninvasive measurements. The nasal skin temperature of 20 subjects was assessed by thermography. The measurement of the skin temperature by thermistor causes stress in subjects. The primary advantages of a thermography over use of a thermistor are noninvasive and accurate. Each subject was made anxious under the 2 conditions of sudden darkness (Sudden darkness condition) or experienced darkness (Experienced darkness condition). Under former, after a baseline facial skin temperature was established, the room light was suddenly turned off. The subject remained seated in darkness for 2 min. The time-series changes were significant for women, but not for men. For women, skin temperature significantly declined in sudden darkness. Experiments on experienced darkness were performed a week later. After a baseline facial skin temperature was established, the subject was told; "The room light would be turned off from now on, but please remain seated without moving". After the instruction, the room light was turned off. The subject remained seated in darkness for 2 min. The resultant time-series changes in skin temperatures were significant in neither men nor women.


Placental Exosomes as Biomarkers for Maternal Diseases: Current Advances in Isolation, Characterization, and Detection.
Serving as the interface between fetal and maternal circulation, the placenta plays a critical role in fetal growth and development. Placental exosomes are small membrane-bound extracellular vesicles released by the placenta during pregnancy. They contain a variety of biomolecules, including lipids, proteins, and nucleic acids, which can potentially be biomarkers of maternal diseases. An increasing number of studies have demonstrated the utility of placental exosomes for the diagnosis and monitoring of pathological conditions such as pre-eclampsia and gestational diabetes. This suggests that placental exosomes may serve as new biomarkers in liquid biopsy analysis. This review provides an overview of the current understanding of the biological function of placental exosomes and their potential as biomarkers of maternal diseases. Additionally, this review highlights current barriers and the way forward for standardization and validation of known techniques for exosome isolation, characterization, and detection. Finally, microfluidic devices for exosome research are discussed.


Engineering Pyrrolysyl-tRNA Synthetase for the Incorporation of Non-Canonical Amino Acids with Smaller Side Chains.
Site-specific incorporation of non-canonical amino acids (ncAAs) into proteins has emerged as a universal tool for systems bioengineering at the interface of chemistry, biology, and technology. The diversification of the repertoire of the genetic code has been achieved for amino acids with long and/or bulky side chains equipped with various bioorthogonal tags and useful spectral probes. Although ncAAs with relatively small side chains and similar properties are of great interest to biophysics, cell biology, and biomaterial science, they can rarely be incorporated into proteins. To address this gap, we report the engineering of PylRS variants capable of incorporating an entire library of aliphatic "small-tag" ncAAs. In particular, we performed mutational studies of a specific PylRS, designed to incorporate the shortest non-bulky ncAA (S-allyl-l-cysteine) possible to date and based on this knowledge incorporated aliphatic ncAA derivatives. In this way, we have not only increased the number of translationally active "small-tag" ncAAs, but also determined key residues responsible for maintaining orthogonality, while engineering the PylRS for these interesting substrates. Based on the known plasticity of PylRS toward different substrates, our approach further expands the reassignment capacities of this enzyme toward aliphatic amino acids with smaller side chains endowed with valuable functionalities.


Close Relationship between cIAP2 and Human ARDS Induced by Severe H7N9 Infection.
cIAP2 is involved in necroptosis as a key upstream regulation factor. We aimed to investigate the role of cIAP2 in ARDS/ALI induced by H7N9 virus through regulating the RIPK1/3 necroptosis pathway. Lung tissues of 11 patients who died from ARDS-complicated H7N9 infection between 2013 and 2016 were obtained as the H7N9-ARDS group. Lung tissues near benign lung nodules were acquired as the control group. Histological changes were evaluated by H&E staining. Protein levels of cIAP2, RIPK1, RIPK3, p-RIPK3, MLKL, and p-MLKL in the lung tissues were detected by Western Blot. The mRNA levels of cIAP2, RIPK1, and RIPK3 were detected by real-time PCR. H7N9 virus infection had a high mortality, with ARDS being the leading cause of death. The protein level of cIAP2 in the experimental group was lower than that in the control group (P<0.05). However, the experimental group showed higher RIPK1, RIPK3, and p-RIPK3 protein levels than the control group (P<0.05), as well as the expression level of MLKL and p-MLKL protein, which is a key downstream protein in necroptosis (P<0.05). In tissues from patients with fatal H7N9, downregulation of cIAP2 and induction of necroptosis was observed. We could speculate that necroptosis of the pulmonary epithelium is associated with severe H7N9 infection leading to ARDS. Thus, necroptosis inhibition may be a novel therapy for H7N9 influenza virus.


A resistance exercise program improves functional capacity of patients with psoriatic arthritis: a randomized controlled trial.
The objective of this study is to assess the effectiveness of resistance training in patients with psoriatic arthritis (PsA). The study is a randomized controlled trial with 41 patients aged between 18 and 65 years with diagnosis of psoriatic arthritis (PsA). The patients were randomized into the following: intervention group (IG) and control group (CG). The IG underwent resistance exercise twice a week, for 12 weeks. The CG remained with the conventional drug therapy. The outcome measurements were the following: BASFI and HAQ-S for functional capacity, one maximum repetition test (1RM) for muscle strength, SF-36 questionnaire for general quality of life, and BASDAI and DAS-28 for disease activity. The evaluations were done by a blinded evaluator at baseline (T0) after 6 (T6) and 12 weeks (T12). At baseline, the groups were homogeneous regarding clinical and demographic characteristics. The IG significantly improved functional capacity measured by HAQ-S and disease activity measured by BASDAI, compared to CG, at week 12. Regarding quality of life, the IG improved the domains "pain" and "general health status" compared to CG (p < 0.05). There was improvement in muscular strength in almost all exercises in IG, except in the exercise for biceps. However, there were statistical differences between groups only on exercise "leg extension" in IG compared to CG. Resistance training is effective in improving functional capacity, disease activity, and quality of life of patients with psoriatic arthritis. The clinical improvements were not coupled to significant changes in muscular strength.


Global burden of disease: acute-on-chronic liver failure, a systematic review and meta-analysis.
Acute-on-chronic liver failure (ACLF) is characterised by acute decompensation of cirrhosis associated with organ failures. We systematically evaluated the geographical variations of ACLF across the world in terms of prevalence, mortality, aetiology of chronic liver disease (CLD), triggers and organ failures. We searched EMBASE and PubMed from 3/1/2013 to 7/3/2020 using the ACLF-EASL-CLIF (European Association for the Study of the Liver-Chronic Liver Failure) criteria. Two investigators independently conducted the abstract selection/abstraction of the aetiology of CLD, triggers, organ failures and prevalence/mortality by presence/grade of ACLF. We grouped countries into Europe, East/South Asia and North/South America. We calculated the pooled proportions, evaluated the methodological quality using the Newcastle-Ottawa Scale and statistical heterogeneity, and performed sensitivity analyses. We identified 2369 studies; 30 cohort studies met our inclusion criteria (43 206 patients with ACLF and 140 835 without ACLF). The global prevalence of ACLF among patients admitted with decompensated cirrhosis was 35% (95% CI 33% to 38%), highest in South Asia at 65%. The global 90-day mortality was 58% (95% CI 51% to 64%), highest in South America at 73%. Alcohol was the most frequently reported aetiology of underlying CLD (45%, 95% CI 41 to 50). Infection was the most frequent trigger (35%) and kidney dysfunction the most common organ failure (49%). Sensitivity analyses showed regional estimates grossly unchanged for high-quality studies. Type of design, country health index, underlying CLD and triggers explained the variation in estimates. The global prevalence and mortality of ACLF are high. Region-specific variations could be explained by the type of triggers/aetiology of CLD or grade. Health systems will need to tailor early recognition and treatment of ACLF based on region-specific data.


Declining fertility and economic well-being: do education and health ride to the rescue?
It is widely argued that declining fertility slows the pace of economic growth in industrialized countries through its negative effect on labor supply. There are, however, theoretical arguments suggesting that the effect of falling fertility on effective labor supply can be offset by associated behavioral changes. We formalize these arguments by setting forth a dynamic consumer optimization model that incorporates endogenous fertility as well as endogenous education and health investments. The model shows that a fertility decline induces higher education and health investments that are able to compensate for declining fertility under certain circumstances. We assess the theoretical implications by investigating panel data for 118 countries over the period 1980 to 2005 and show that behavioral changes partly mitigate the negative impact of declining fertility on effective labor supply.


Ultracentrifugation deforms unfixed influenza A virions.
Negatively stained influenza virions sometimes show irregular morphology and are often referred to as pleomorphic. However, this irregular morphology has not been visualized when ultrathin-section transmission and scanning electron microscopies are used. This study focused on the effects of ultracentrifugation on influenza A virion morphology, as negative staining often involves ultracentrifugation to concentrate or purify virions. The morphologies of unfixed, glutaraldehyde-fixed and osmium tetroxide-fixed virions were quantitatively compared before and after ultracentrifugation, and it was found that, without chemical fixation, approximately 30% of virions were altered from oval to irregular shapes following ultracentrifugation. By contrast, most glutaraldehyde-fixed virions remained uniformly elliptical, even after ultracentrifugation. When a virus with an 11 aa deletion at the C terminus of its M2 cytoplasmic tail was ultracentrifuged, its morphology was appreciably deformed compared with that of the wild-type virus. These results demonstrate that the native morphology of influenza A virions is regular but is disrupted by ultracentrifugation, and that the cytoplasmic tail of M2 is important for virion integrity.


Hepatic stellate cells increase in Toxoplasma gondii infection in mice.
Toxoplasma gondii is a ubiquitous protozoan parasite that can infect humans and animals. The severity of toxoplasmosis varies according to the immune status of the individual, parasite strain, and host species. In mammalian species, it has been observed that severe lesions of acute toxoplasmosis form in visceral organs such as the liver, lung, and spleen. Some epidemiological studies have reported an association of T. gondii infection with liver cirrhosis. Acute infection was induced in fifteen 30-day-old normal Swiss albino mice. The mice were infected by intraperitoneal inoculation of 5000 T. gondii RH strain tachyzoites. The mice were sacrificed in groups of 5 at 2, 4, and 6 days after inoculation. Another group of 5 mice were used as the controls. Anti-glial fibrillary acidic protein (GFAP) and anti-T. gondii antibodies were used to compare GFAP-immunoreactive cells and anti-T. gondii-immunopositive areas in the liver between the T. gondii-infected groups and the healthy controls, respectively. There was a significant correlation between the numbers of GFAP-positive hepatic stellate cells (HSCs) when they were compared with T. gondii antigen immunostaining (p < 0.05). The amount of T. gondii immunostaining increased significantly with the increase in the number of HSCs. There is a significant relationship between the number of HSCs and T. gondii antigens, which may represent an active role of HSCs in liver pathology and the pathobiology of T. gondii-related hepatitis.


Cognition and connectomes in nondementia idiopathic Parkinson's disease.
In this study, we investigate the organization of the structural connectome in cognitively well participants with Parkinson's disease (PD-Well; n = 31) and a subgroup of participants with Parkinson's disease who have amnestic disturbances (PD-MI; n = 9). We explore correlations between connectome topology and vulnerable cognitive domains in Parkinson's disease relative to non-Parkinson's disease peers (control, n = 40). Diffusion-weighted MRI data and deterministic tractography were used to generate connectomes. Connectome topological indices under study included weighted indices of node strength, path length, clustering coefficient, and small-worldness. Relative to controls, node strength was reduced 4.99% for PD-Well (p = 0.041) and 13.2% for PD-MI (p = 0.004). We found bilateral differences in the node strength between PD-MI and controls for inferior parietal, caudal middle frontal, posterior cingulate, precentral, and rostral middle frontal. Correlations between connectome and cognitive domains of interest showed that topological indices of global connectivity negatively associated with working memory and displayed more and larger negative correlations with neuropsychological indices of memory in PD-MI than in PD-Well and controls. These findings suggest that indices of network connectivity are reduced in PD-MI relative to PD-Well and control participants.


[Translation and cross-cultural adaptation of the Multiple Sclerosis Walking Scale (MSWS-12) into Brazilian Portuguese].
Poor walking performance is predictive of heart disease and osteoporosis and increases the risk of death in the elderly. Gait and vision have been identified as the most valuable physical functions according to multiple sclerosis patients' perceptions. The objective of this study was to perform a translation and cross-cultural adaptation of the Multiple Sclerosis Walking Scale (MSWS-12) into Brazilian Portuguese. A study of cross-cultural adaptation was conducted in ten steps. Participation in the study included four translators, two back-translators, twelve medical experts, twelve patients, twelve healthy subjects, and a Portuguese language expert. Only the question "Did standing make it more difficult to do things?" posed difficulty in the translation process. Maximum time for completion was less than three minutes (171 seconds). Internal consistency analyses showed high reliability (Cronbach's alpha = 0.94). The content validation and internal consistency stages were completed satisfactorily.


Two Pairs of New Bisabolane-Type Sesquiterpenoids from Aspergillus sydowii.
Two pairs of new bisabolane-type sesquiterpenoids, (+)-aspersydowin A (7S) [(+)-1], (-)-aspersydowin A (7R) [(-)-1], (+)-aspersydowin B (7S,11S) [(+)-2], (-)-aspersydowin B (7R,11R) [(-)-2], along with six known compounds (1-8) were isolated from the fungus Aspergillus sydowii. Compounds 1 and 2 are enantiomers resolved by the Chiralpak IC, using a hexane- propan-2-ol mobile phase. The structure of 1 and 2 with absolute configuration were assigned tentatively by 1D (1 H, 13 C, and DEPT) & 2D (HSQC, 1 H-1 H COSY, HMBC, and NOESY) NMR data analyses and ECD calculations. Compounds 1-8 were screened for the biological activities in vitro. The results showed that compounds 3, 4 and 8 exhibited immunosuppressive activities with IC50 values of 10.9, 17.6 and 13.4 μM, respectively.


Creativity in Narcolepsy Type 1: The Role of Dissociated REM Sleep Manifestations.
A higher creative potential has been reported in narcoleptic patients and linked to lucid dreaming. The aim of the present study was to explore the role of narcolepsy symptoms (presence and severity) in predicting creativity. Sixty-six consecutive type 1 narcolepsy patients (mean age 38.62 ± 17.05, 31 females) took part in this study. Creative achievement in different life domains and creative beliefs were assessed by a self-reported questionnaire and a scale measuring the creative self, respectively; creative performance was measured through a divergent thinking test (generation of alternative original solutions to an open problem). We found a key effect of hypnagogic hallucinations in modulating creative behaviour. We therefore tested at first whether hypnagogic hallucinations could interact with specific mental states associated with creativity and in particular mind wandering, a factor associated with both creative performance and achievement. Secondly, we verified if hypnagogic hallucinations could influence the definition of creative identity in type 1 narcolepsy patients, which in turn could predict their creative achievement and creative performance. Results showed that spontaneous mind wandering influenced creative achievement through a moderation effect of sleep paralysis and hypnagogic hallucinations. Moreover, sleep paralysis and hypnagogic hallucinations indirectly influenced, through creative identity, both creative achievement and performance (fluency score). Our results highlight the role of hypnagogic hallucinations in triggering the process of mind wandering which leads to greater creative success. In addition, this symptom affects creative identity in narcolepsy, leading in turn to higher creative success and creative potential of narcoleptic patients.


Novel protein kinase of Arabidopsis thaliana (APK1) that phosphorylates tyrosine, serine and threonine.
During the course of characterizing polymerase chain reaction products corresponding to protein kinases of a higher plant, Arabidopsis thaliana, we found a DNA fragment that potentially codes for a polypeptide with mosaic sequences of two classes of protein kinases, a tyrosine-specific and a serine/threonine-specific one. Overlapping complementary DNA (cDNA) clones coinciding with this fragment were isolated from an A. thaliana cDNA library. From their sequence analyses a protein kinase was predicted composed of 410 amino acid residues (APK1, Arabidopsis protein kinase 1), in which the kinase domain was flanked by short non-kinase domains. Upon expression of APK1 in Escherichia coli cells, several bacterial proteins became reactive with anti-phosphotyrosine antibody but not with the same antibody preincubated with phosphotyrosine, convincing us that APK1 phosphorylated tyrosine residues. APK1 purified from an over-producing E. coli strain showed serine/threonine kinase activity, and no tyrosine kinase activity, towards APK1 itself, casein, enolase, and myosin light chains. APK1 was thus concluded to be a novel type of protein kinase, which could phosphorylate tyrosine, serine, and threonine residues, though tyrosine phosphorylation seemed to occur only on limited substrates. Since the structure of the APK1 N-terminal portion was indicative of N-myristoylation, APK1 might associate with membranes and thereby contribute to signal transduction. The A. thaliana genome contained two APK1 genes close to each other (APK1a and APK1b).


A Single Amino Acid Residue at Transmembrane Domain 4 of the α Subunit Influences Carisoprodol Direct Gating Efficacy at GABAA Receptors.
The muscle relaxant carisoprodol has recently been controlled at the federal level as a Schedule IV drug due to its high abuse potential and consequences of misuse, such as withdrawal syndrome, delusions, seizures, and even death. Recent work has shown that carisoprodol can directly gate and allosterically modulate the type A GABA (GABAA) receptor. These actions are subunit-dependent; compared with other GABAA receptors, carisoprodol has nominal direct gating effects in α3β2γ2 receptors. Here, using site-directed mutagenesis and whole-cell patch-clamp electrophysiology in transiently transfected human embryonic kidney 293 cells, we examined the role of GABAA receptor α subunit transmembrane domain 4 (TM4) amino acids in direct gating and allosteric modulatory actions of carisoprodol. Mutation of α3 valine at position 440 to leucine (present in the equivalent position in the α1 subunit) significantly increased the direct gating effects of carisoprodol without affecting its allosteric modulatory effects. The corresponding reverse mutation, α1(L415V), decreased carisoprodol direct gating potency and efficacy. Analysis of a series of amino acid mutations at the 415 position demonstrated that amino acid volume correlated positively with carisoprodol efficacy, whereas polarity inversely correlated with carisoprodol efficacy. We conclude that α1(415) of TM4 is involved in the direct gating, but not allosteric modulatory, actions of carisoprodol. In addition, the orientation of alkyl or hydroxyl groups at this position influences direct gating effects. These findings support the likelihood that the direct gating and allosteric modulatory effects of carisoprodol are mediated via distinct binding sites.


Highly spectrally undersampled vowels can be classified by machines without supervision.
An unsupervised automatic clustering algorithm (k-means) classified 1282 Mel frequency cepstral coefficient (MFCC) representations of isolated steady-state vowel utterances from eight standard German vowel categories with fo between 196 and 698 Hz. Experiment I obtained the number of MFCCs (1-20) in connection with the spectral bandwidth (2-20 kHz) at which performance peaked (five MFCCs at 4 kHz). In experiment II, classification performance with different ranges of fo revealed that ranges with fo > 500 Hz reduced classification performance but it remained well above chance. This shows that isolated steady state vowels with strongly undersampled spectra contain sufficient acoustic information to be classified automatically.


Molecular optimization of rabies virus glycoprotein expression in Pichia pastoris.
In this work, different approaches were investigated to enhance the expression rabies virus glycoprotein (RABV-G) in the yeast Pichia pastoris; this membrane protein is responsible for the synthesis of rabies neutralizing antibodies. First, the impact of synonymous codon usage bias was examined and an optimized RABV-G gene was synthesized. Nevertheless, data showed that the secretion of the optimized RABV-G gene was not tremendously increased as compared with the non-optimized one. In addition, similar levels of RABV-G were obtained when α-factor mating factor from Saccharomyces cerevisiae or the acid phosphatase PHO1 was used as a secretion signal. Therefore, sequence optimization and secretion signal were not the major bottlenecks for high-level expression of RABV-G in P. pastoris. Unfolded protein response (UPR) was induced in clones containing high copy number of RABV-G expression cassette indicating that folding was the limiting step for RABV-G secretion. To circumvent this limitation, co-overexpression of five factors involved in oxidative protein folding was investigated. Among these factors only PDI1, ERO1 and GPX1 proved their benefit to enhance the expression. The highest expression level of RABV-G reached 1230 ng ml(-1). Competitive neutralizing assay confirmed that the recombinant protein was produced in the correct conformational form in this host.


Managing the U.S. health care workforce: creating policy amidst uncertainty.
Managing the health care workforce will have important implications for costs, quality, and access. Factors influencing supply include the production of new professionals, their relative effort, and rates of retirement. Demand will depend upon the development of new diseases, new drugs, and technologies, as well as the growth of managed care, which uses fewer physicians, fewer specialists, and more midlevel practitioners. There is a general consensus that there are too many physicians, especially specialists. Reducing the number of residency positions would reduce supply, predominantly by slowing importation of international physicians. Obstacles to workforce reform include a distrust of supply projections, skepticism about governmental planning, conservatism of established institutions, and the reality that some hospitals would lose positions and resources.


Photoexcitation dynamics in films of C60 and Zn phthalocyanine with a layered nanostructure.
We elucidate photoexcitation dynamics in C(60) and zinc phthalocyanine (ZnPc) from picoseconds to milliseconds by transient absorption and time-resolved terahertz spectroscopy. Autoionization of C(60) is a precursor to photocarrier generation. Decay of the terahertz signal is due to decreasing photocarrier mobility over the first 20 ps and thereafter reflects recombination dynamics. Singlet diffusion rates in C(60) are determined by modeling the rise of ground state bleaching of ZnPc absorption following C(60) excitation. Recombination dynamics transform from bimolecular to monomolecular as the layer thickness is reduced, revealing a metastable exciplex at the C(60)/ZnPc interface with a lifetime of 150 μs.


[Effect of commercial beta blockers and their combination with artificial tears on the duration of tear film stability].
The influences of 8 different beta-blockers on the tear film stability-measured on the breakup time (BUT) of normal and healthy volunteers-was compared. All beta-blockers led to BUT reduction, however, the lowest influence was observed with levobunolol. The duration of this effect was compared with that of levobunolol, timolol and metipranolol. While the minor BUT alterations induced by levobunolol recovered after 30 min and were completely restored after 1 h, those of the two other beta-blockers lasted for over 2 h. The combination of timolol 0.5% with different artificial tears containing mainly polyvinyl alcohol (PVA) 1.4%, methylcellulose and polyacrylic acid initially led also to BUT reduction. The best results were obtained with PVA 1.4%.


In situ cross-linked electrospun fiber scaffold of collagen for fabricating cell-dense muscle tissue.
Engineered muscle tissues used as transplant tissues in regenerative medicine should have a three-dimensional and cell-dense structure like native tissue. For fabricating a 3D cell-dense muscle tissue from myoblasts, we proposed the electrospun type I collagen microfiber scaffold of the string-shape like a harp. The microfibers were oriented in the same direction to allow the myoblasts to align, and were strung at low density with micrometer intervals to create space for the cells to occupy. To realize this shape of the scaffold, we employed in situ cross-linking during electrospinning process for the first time to collagen fibers. The collagen microfibers in situ cross-linked with glutaraldehyde stably existed in the aqueous media and completely retained the original shape to save the spaces between the fibers for over 14 days. On the contrary, the conventional cross-linking method by exposure to a glutaraldehyde aqueous solution vapor partially dissolved and damaged the fiber to lose a low-density shape of the scaffold. Myoblasts could penetrate into the interior of the in situ cross-linked string-shaped scaffold and form the cell-dense muscle tissues. Histochemical analysis showed the total area occupied by the cells in the cross section of the tissue was approximately 73 %. Furthermore, the resulting muscle tissue fabricated from primary myoblasts showed typical sarcomeric cross-striations and the entire tissue continuously pulsated by autonomous contraction. Together with the in situ cross-linking, the string-shaped scaffold provides an efficient methodology to fabricate a cell-dense 3D muscle tissue, which could be applied in regenerative medicine in future.


Hazard Exposure with Health and Safety Outcomes Hinder the Work Ability of Salt Farm Workers in Thailand.
Unsafe environments and occupational hazards have been documented and are contributing to health risks for salt farm workers in Thailand. Whether exposures with various health hazards and outcomes affect work ability among these workers is uncertain. This study aimed to examine the effects of hazard exposures with health risks and outcomes, including accidents, for work ability among salt farm workers. A cross-sectional study was completed with 120 salt farm workers recruited from the largest salt farm company in Samutsakorn Province during January and February 2014. Face to face interviews were conducted using a structured questionnaire comprised of worker characteristics, working conditions, safety and health problems related to work, and a work ability index (questionnaire reliability =0.84) through a worksite visit. The MIMIC model was used to determine direct and indirect causal effects of work-related health hazards and outcomes on work ability. Among 120 salt farmers, 61% were female and 77% had primary educations. Average age and work experience was 49.5 years (SD = 13.7) and 23.7 years (SD = 13.5), respectively. 66% worked more than 5 days per week and 21.3% had at least one accident in the past 3 months. The results of structure equation analysis revealed that healthy skin was related to heat exposure (β=.35, p=.01) and directly influenced work ability (β=.37, p=.01), Eye irritation was influenced by length of work time (β=.24, p=.05), and harmonized the predictive value of the model at a high level (χ2=94.51, df=73, p-value =0.05, χ2/df=1.29. RMSEA= 0.066). Results for the MIMIC model indicate that two factors, bright sunlight and strong wind exposure over an extended work period, could be having crucial impacts on skin and eye health of salt farm workers. These factors might be protected against using PPE and health examinations with positive health outcomes directly improving work ability. Substantial governmental action with community leadership is needed immediately so action plans are instituted to improve occupational safety and health.


Effects of Notch signalling on the expression of SEMA3C, HMGA2, CXCL14, CXCR7, and CCL20 in breast cancer.
Metastasis is the main reason for death in breast cancer. Understanding the molecular players in metastasis is crucial for diagnostic and therapeutic purposes. Notch signalling plays an oncogenic role in breast tumorigenesis and is involved in metastasis. Downstream mediators of Notch signalling in prometastatic processes are not yet fully discovered. Here we aimed to investigate whether Notch signalling regulates the expression of SEMA3C, HMGA2, CXCL14, CXCR7, and CCL20, which are involved in prometastatic processes, in breast cell lines. To this end, expression of the selected genes was analysed following Notch activation by overexpression of the Notch1 intracellular domain in the normal breast epithelial cell line MCF10A, and inhibition by silencing of the Notch transcriptional mediator RBPjκ in the breast cancer cell line MDA MB 231. SEMA3C and HMGA2 mRNA were decreased, while CXCL14 and CXCR7 mRNA were increased significantly in response to Notch activation in MCF10A cells. Notch inhibition in MDA MB 231 cells significantly decreased HMGA2 and CCL20 mRNA. Protein levels were not significantly altered by Notch modulation. In conclusion, we showed that Notch signalling regulates expression of SEMA3C, CXCL14, CCL20, CXCR7, and HMGA2, which are prominent candidate genes that might function downstream of Notch to induce prometastatic processes.


Mosquito species composition and phenology (Diptera, Culicidae) in two German zoological gardens imply different risks of mosquito-borne pathogen transmission.
Due to their large diversity of potential blood hosts, breeding habitats, and resting sites, zoological gardens represent highly interesting places to study mosquito ecology. In order to better assess the risk of mosquito-borne disease-agent transmission in zoos, potential vector species must be known, as well as the communities in which they occur. For this reason, species composition and dynamics were examined in 2016 in two zoological gardens in Germany. Using different methods for mosquito sampling, a total of 2,257 specimens belonging to 20 taxa were collected. Species spectra depended on the collection method but generally differed between the two zoos, while species compositions and relative abundances varied seasonally in both of them. As both sampled zoos were located in the same climatic region and potential breeding sites within the zoos were similar, the differences in mosquito compositions are attributed to immigration of specimens from surrounding landscapes, although the different sizes of the zoos and the different blood host populations available probably also have an impact. Based on the differences in species composition and the various biological characteristics of the species, the risk of certain pathogens to be transmitted must also be expected to differ between the zoos.


Inflammatory response modulated by pinealectomy: effect of light.
In view of the known interactions between pineal and adrenal glands and the importance of glucocorticoids on the acute phase of inflammatory process, we investigated the effect of the pineal gland on the paw edema response induced by carrageenin in female Wistar rats, weighing 170-220 g, measured by the pletysmographic method. Adrenalectomy increased the rate of the inflammatory response (149.02 +/- 2.93%, N = 7 vs 113.48 +/- 2.99%, N = 7) whereas pinealectomy and sham-pinealectomy under white light had an inhibitory effect (81.73 +/- 2.16%, N = 15 and 80.21 +/- 1.85%, N = 14 vs 113.48 +/- 2.99%, N = 7). Sham-pinealectomy under red light did not modify the rate of the inflammatory response. These results indicate that the pineal gland can be affected by light during sham-pinealectomy. In view of the unaltered adrenal gland weight and corticosterone levels among the different groups, we propose an antagonism between melatonin (pineal hormone) and glucocorticoids at the peripheral level. The predominant effect of glucocorticoids over melatonin may explain the inhibition of the inflammatory response caused by pinealectomy.


The role of health insurance on treatment for opioid use disorders: Evidence from the Affordable Care Act Medicaid expansion.
We estimate the effect of health insurance coverage on opioid use disorder treatment utilization and availability by exploiting cross-state variation in effective dates of Medicaid expansions under the Affordable Care Act. Using a difference-in-differences design, we find that aggregate opioid admissions to specialty treatment facilities increased 18% in expansion states, most of which involved outpatient medication-assisted treatment (MAT). Opioid admissions from Medicaid beneficiaries increased 113% without crowding out admissions from individuals with other health insurance. These effects appeared to be driven by market entry of select MAT providers and by greater acceptance of Medicaid payments among existing MAT providers. Moreover, effects were largest in expansion states with comprehensive MAT coverage. Our findings suggest that Medicaid expansions resulted in substantial utilization and availability gains to clinically efficacious and cost-effective pharmacological treatments, implying potential benefits of expanding Medicaid to non-expansion states and extending MAT coverage.


Cloning and characterization of overlapping DNA fragments of the toxin A gene of clostridium difficile.
Clostridium difficile, a human pathogen, produces two very large protein toxins, A and B (250-600 kDa), which resist dissociation into subunits. To clone the toxin A gene, a genomic library of 3-8 kb chromosomal DNA fragments of C. difficile strain VPI 10463 established in pUC12 was screened with a rabbit polyclonal toxin A antiserum. Thirty-five clones were isolated which carried 2.5-7.0 kb inserts representing a 10 kb region of the C. difficile genome. All the inserts were oriented in the same direction, suggesting that toxin A gene expression was under control of the lac promoter of the pUC12 vector. Western blot experiments revealed the presence of low amounts of fusion proteins of variable size (30-170 kDa) in Escherichia coli strains harbouring recombinant plasmids. As deduced from subcloning experiments, the DNA sequences encoding toxin A comprised about 4 kb, corresponding to about 140 kDa of the 300-600 kDa protein. This was either due to incomplete cloning of the gene or it might indicate a subunit composition of toxin A. No additional gene(s) with homology to the cloned toxin A gene was detected.


Mental Health Outcomes in Barcelona: The Interplay between Gentrification and Greenspace.
Greenspace is widely related to mental health benefits, but this relationship may vary by social group. Gentrification, as linked to processes of unequal urban development and conflict, potentially impacts health outcomes. This study explores the relationships between greenspace and mental health and between gentrification and mental health associations. It also further examines gentrification as an effect modifier in the greenspace-mental health association and SES as an effect modifier in the gentrification-mental health association. We used cross-sectional Barcelona (Spain) data from 2006, which included perceived mental health status and self-reported depression/anxiety from the Barcelona Health Survey. Greenspace exposure was measured as residential access to (1) all greenspace, (2) greenways and (3) parks in 2006. Census-tract level gentrification was measured using an index including changes in sociodemographic indicators between 1991 and 2006. Logistic regression models revealed that only greenways were associated with better mental health outcomes, with no significant relationship between mental health and parks or all greenspace. Living in gentrifying neighborhoods was protective for depression/anxiety compared to living in non-gentrifying neighborhoods. However, only residents of gentrifiable census tracts benefited from the exposure to greenways. SES was not found to be an effect modifier in the association between gentrification and mental health. Future research should tackle this study's limitations by incorporating a direct measure of displacement in the gentrification status indicator, accounting for qualitative aspects of greenspace and user's perceptions. Gentrification may undermine the health benefits provided by greenspace interventions.


Chromosome aberrations in pressure-induced triploid Atlantic salmon.
Triploid organisms have three sets of chromosomes. In Atlantic salmon, hydrostatic pressure treatment of newly fertilized eggs has been extensively used to produce triploids which are functionally sterile due to their unpaired chromosomes. These fish often perform poorly on commercial farms, sometimes without explanation. Inheritance patterns in individuals subjected to pressure treatment have not been investigated in Atlantic salmon thus far. However, work on other species suggests that this treatment can result in aberrant inheritance. We therefore studied this in Atlantic salmon by genotyping 16 polymorphic microsatellites in eyed eggs and juveniles which had been subjected to pressure-induction of triploidy. Communally reared juveniles including fish subjected to pressure-induction of triploidy and their diploid siblings were included as a control. No diploid offspring were detected in any of the eggs or juveniles which were subjected to hydrostatic pressure; therefore, the induction of triploidy was highly successful. Aberrant inheritance was nevertheless observed in 0.9% of the eggs and 0.9% of the juveniles that had been subjected to pressure treatment. In the communally reared fish, 0.3% of the fish subjected to pressure treatment displayed aberrant inheritance, while their diploid controls displayed 0% aberrant inheritance. Inheritance errors included two eyed eggs lacking maternal DNA across all microsatellites, and, examples in both eggs and juveniles of either the maternal or paternal allele lacking in one of the microsatellites. All individuals displaying chromosome aberrations were otherwise triploid. This is the first study to document aberrant inheritance in Atlantic salmon that have been subjected to pressure-induction of triploidy. Our experiments unequivocally demonstrate that even when induction of triploidy is highly successful, this treatment can cause chromosome aberrations in this species. Based upon our novel data, and earlier studies in other organisms, we hypothesize that in batches of Atlantic salmon where low to modest triploid induction rates have been reported, aberrant inheritance is likely to be higher than the rates observed here. Therefore, we tentatively suggest that this could contribute to the unexplained poor performance of triploid salmon that is occasionally reported in commercial aquaculture. These hypotheses require further investigation.


The Chinese version of the self-efficacy scale for daily life activities among older adults: translation, validity and reliability.
To translate and cross-culturally adapt the Self-efficacy Scale for Daily Life Activities among Older Adults (SSDLAOA) into Chinese, as well as to examine the psychometric properties of the Chinese version of the SSDLAOA (C-SSDLAOA) in a Chinese community-based older adult population. This study used a cross-sectional survey. The psychometric properties of the C-SSDLAOA were evaluated among 355 older adults recruited from three communities in Huzhou, including internal reliability, test-retest reliability, face validity, content validity, construct validity, and concurrent validity. The C-SSDLAOA was consistent with the original version, which consisted of five dimensions and 23 items. The confirmatory factor analysis indicated a good fit for each indicator (χ2 /df=1.387, RMSEA = 0.043, CFI = 0.967, TLI = 0.962, and SRMR = 0.042). The Cronbach's alpha was 0.909, and the test-retest reliability was 0.966, with good internal consistency and stability. The content validity index of the items ranged from 0.83 to 1.00, while the content validity index of the scale was 0.96. The correlation between the C-SSDLAOA and GSES was 0.886, with good concurrent validity. The C-SSDLAOA is a reliable and valid instrument for assessing self-efficacy for daily life activities in Chinese community-dwelling older adults.


The dopaminergic system in peripheral blood lymphocytes: from physiology to pharmacology and potential applications to neuropsychiatric disorders.
Besides its action on the nervous system, dopamine (DA) plays a role on neural-immune interactions. Here we review the current evidence on the dopaminergic system in human peripheral blood lymphocytes (PBL). PBL synthesize DA through the tyrosine-hydroxylase/DOPA-decarboxylase pathway, and express DA receptors and DA transporter (DAT) on their plasma membrane. Stimulation of DA receptors on PBL membrane contributes to modulate the development and initiation of immune responses under physiological conditions and in immune system pathologies such as autoimmunity or immunodeficiency.The characterization of DA system in PBL gave rise to a further line of research investigating the feasibility of PBL as a cellular model for studying DA derangement in neuropsychiatric disorders. Several reports showed changes of the expression of DAT and/or DA receptors in PBL from patients suffering from several neuropsychiatric disorders, in particular parkinsonian syndromes, schizophrenia and drug- or alcohol-abuse. Despite some methodological and theoretical limitations, these findings suggest that PBL may prove a cellular tool with which to identify the derangement of DA transmission in neuropsychiatric diseases, as well as to monitor the effects of pharmacological treatments.


HLA class I and II antigens are partially co-clustered in the plasma membrane of human lymphoblastoid cells.
Major histocompatibility complex (MHC) class II molecules displayed clustered patterns at the surfaces of T (HUT-102B2) and B (JY) lymphoma cells characterized by interreceptor distances in the micrometer range as detected by scanning force microscopy of immunogold-labeled antigens. Electron microscopy revealed that a fraction of the MHC class II molecules was also heteroclustered with MHC class I antigens at the same hierarchical level as described by the scanning force microscopy data, after specifically and sequentially labeling the antigens with 30- and 15-nm immunogold beads. On JY cells the estimated fraction of co-clustered HLA II was 0.61, whereas that of the HLA I was 0.24. Clusterization of the antigens was detected by the deviation of their spatial distribution from the Poissonian distribution representing the random case. Fluorescence resonance energy transfer measurements also confirmed partial co-clustering of the HLA class I and II molecules at another hierarchical level characterized by the 2- to 10-nm Förster distance range and providing fine details of the molecular organization of receptors. The larger-scale topological organization of the MHC class I and II antigens may reflect underlying membrane lipid domains and may fulfill significant functions in cell-to-cell contacts and signal transduction.


Hepatitis B screening and vaccination strategies for newly arrived adult Canadian immigrants and refugees: a cost-effectiveness analysis.
Immigrants have increased mortality from hepatocellular carcinoma as compared to the host populations, primarily due to undetected chronic hepatitis B virus (HBV) infection. Despite this, there are no systematic programs in most immigrant-receiving countries to screen for chronic HBV infection and immigrants are not routinely offered HBV vaccination outside of the universal childhood vaccination program. A cost-effective analysis was performed to compare four HBV screening and vaccination strategies with no intervention in a hypothetical cohort of newly-arriving adult Canadian immigrants. The strategies considered were a) universal vaccination, b) screening for prior immunity and vaccination, c) chronic HBV screening and treatment, and d) combined screening for chronic HBV and prior immunity, treatment and vaccination. The analysis was performed from a societal perspective, using a Markov model. Seroprevalence estimates, annual transition probabilities, health-care costs (in Canadian dollars), and utilities were obtained from the published literature. Acute HBV infection, mortality from chronic HBV, quality-adjusted life years (QALYs), and costs were modeled over the lifetime of the cohort of immigrants. Costs and QALYs were discounted at a rate of 3% per year. Screening for chronic HBV infection, and offering treatment if indicated, was found to be the most cost-effective intervention and was estimated to cost $40,880 per additional QALY gained, relative to no intervention. This strategy was most cost-effective for immigrants < 55 years of age and would cost < $50,000 per additional QALY gained for immigrants from areas where HBV seroprevalence is ≥ 3%. Strategies that included HBV vaccination were either prohibitively expensive or dominated by the chronic HBV screening strategy. Screening for chronic HBV infection from regions where most Canadian immigrants originate, except for Latin America and the Middle East, was found to be reasonably cost-effective and has the potential to reduce HBV-associated morbidity and mortality.


[Transport of low molecular weight substances in protein crystals and films].
Diffusion of synthetic dyes in lysozyme crystals and bovine serum albumin (BSA) amorphous films is shown to depend on the size of diffusate molecule far more strongly than in solutions. The diffusion coefficient increases from 10(-8) to 10(-13) cm2 sec-1 on about 1.5 divided by 2--fold increase in diffusate dimensions, the diffusion being completely arrested on reaching the limiting dimensions of about 14 A and 12 A for BSA films and lysozyme crystals, respectively. The diffusion in lysozyme crystals is anisotropic, with a diffusion maximum directed along their screw axes. The diffusion coefficient dependence on the charge of diffusate and its binding to proteins is discussed.


Review of the Application of PAT in the Pharmaceutical Continuous Crystallization Process.
As an important pharmaceutical process, crystallization greatly impacts the final product. In recent years, the continuous crystallization process has attracted more attention from researchers, with the promotion of continuous manufacturing (CM) by the Food and Drug Administration (FDA). The continuous crystallization process has the advantages of high economic benefit, stable and uniform quality, a short production cycle, and personalization. To carry out continuous crystallization, some related process analytical technology (PAT) tools have become the focus of breakthroughs. Infrared (IR) spectroscopy, Raman spectroscopy, and focused beam reflection measurement (FBRM) tools have gradually become research hotspots due to their fast, non-destructive, and real-time monitoring characteristics. This review compared the advantages and disadvantages of the three technologies. Their applications in the upstream mixed continuous crystallization process, the middle reaches of crystal nucleation and growth, and the process of the downstream refining were discussed to provide corresponding guidance for the practice and further development of these three technologies in the continuous crystallization process and promote the development of CM in the pharmaceutical industry.


Real-time monitoring of PCR amplification of proto-oncogene c-MYC using a Ta₂O₅ electrolyte-insulator-semiconductor sensor.
We present a new approach for real-time monitoring of PCR amplification of a specific sequence from the human c-MYC proto-oncogene using a Ta(2)O(5) electrolyte-insulator-semiconductor (EIS) sensor. The response of the fabricated EIS sensor to cycle DNA amplification was evaluated and compared to standard SYBR-green fluorescence incorporation, showing it was possible to detect DNA concentration variations with 30 mV/μM sensitivity. The sensor's response was then optimized to follow in real-time the PCR amplification of c-MYC sequence from a genomic DNA sample attaining an amplification profile comparable to that of a standard real-time PCR. Owing to the small size, ease of fabrication and low-cost, the developed Ta(2)O(5) sensor may be incorporated onto a microfluidic device and then used for real-time PCR. Our approach may circumvent the practical and economical obstacles posed by current platforms that require an external fluorescence detector difficult to miniaturize and incorporate into a lab-on-chip system.


Guanidinium/ammonium competition and proton transfer in the interaction of the amino acid arginine with the tetracarboxylic 18-crown-6 ionophore.
The recognition of arginine plays a central role in modern proteomics and genomics. Arginine is unique among natural amino acids due to the high basicity of its guanidinium side chain, which sustains specific interactions and proton exchange biochemical processes. The search for suitable macrocyclic ionophores constitutes a promising route towards the development of arginine receptors. This study evaluates the conformational features involved in the binding of free arginine by the polyether macrocycle (18-crown-6)-tetracarboxylic acid. Infrared action vibrational spectroscopy and quantum-chemical computations are combined to characterize the complexes with net charges +1 and +2. The spectrum of the +1 complex can be explained in terms of a configuration predominantly stabilized by a robust bidentate coordination of guanidinium with a carboxylate group formed from the deprotonation of one side group of the crown ether. The released proton is transferred to the amino terminus of arginine, which then coordinates with the crown ether ring. In an alternative type of conformation, partly consistent with experiment, the amino terminus is neutral and the guanidinium group inserts into the crown ether cavity. In the +2 complexes, arginine is always doubly protonated and the most stable conformations are characterized by a tripodal coordination of the ammonium -NH3+ group of arginine with the oxygen atoms of the macrocycle ring, while the interactions of the amino acid with the side carboxylic acid groups of the crown ether acquire a remarkable lesser role.


The beta-lipoprotein doublet in type 3 hyperlipoproteinemia.
We used a previously reported agarose-gel electrophoresis system to study the lipoprotein patterns in patients with type 3 hyperlipoproteinemia. All samples from 20 subjects tested revealed the presence of two bands with beta-lipoprotein mobility, irrespective of quantitative changes in plasma lipids induced by diet or by the administration of drugs. If these observations are confirmed, the method will permit the diagnosis of type 3 hyperlipoproteinemia without resort to preparative ultracentrifugation and chemical measurement of the very-low-density lipoproteins.


Evaluation of CYP450 inhibitory effects and steady-state pharmacokinetics of genistein in combination with cholecalciferol and citrated zinc bisglycinate in postmenopausal women.
The combination of genistein 27 mg, cholecalciferol 200 IU, citrated zinc bisglycinate (4 mg elemental zinc) 20 mg per capsule in Fosteum(®), a prescription medical food regulated by the FDA and indicated for the dietary management of osteopenia and osteoporosis, was tested for drug interactions and to determine the pharmacokinetic profile for genistein, the principal bone-modulating ingredient in the product. In vitro human liver microsome cytochrome P450 (CYP450) assays were used to test the product for potential drug interactions with the isoforms 1A2, 2C8, 2C9, 2C19, 2D6, and 3A4. Due to specific 2C8 and 2C9 inhibition, a steady-state pharmacokinetic study was performed to assess serum genistein concentrations by high-pressure liquid chromatography-coupled mass spectroscopy in healthy fasting (n = 10) and fed (n = 10) postmenopausal women. The product showed minimal inhibition of 1A2, 2C19, 2D6, and 3A4, exhibiting IC(50) > 10 μM, but 2C8 and 2C9 yielded IC(50) of 2.5 μM and 2.8 μM, respectively, concentrations which are theroretically achievable when dosing the product twice daily. After seven days of administration in a steady-state pharmacokinetic study, significant differences were found for unconjugated genistein (including free and protein-bound), regarding time to peak concentration (1.88 ± 1.36 hours), maximum concentration reached (0.052 ± 0.055 μM), elimination half-life (2.3 ± 1.6 hours), and area under the concentration-time curve (53.75 ± 17.59 ng · hour/mL) compared with results for total genistein (including glucuronidated and sulfonated conjugates) time to peak concentration (2.22 ± 1.09 hours), maximum concentration reached (2.95 ± 1.64 μM), elimination half-life (10.4 ± 4.1 hours), and area under the concentration-time curve (10424 ± 6290 ng · hour/mL) in fasting subjects. Coadministration of food tended to extend the time and extent of absorption as well as slow elimination of genistein, but not in a statistically significant manner. Because the serum genistein concentrations achieved during pharmacokinetic testing at therapeutic doses were well below those required for enzyme inhibition in the in vitro liver microsome assays, these results indicate a low potential for drug interactions.


Clinical Outcomes Comparing Two Prosthetic Knee Designs in Individuals with Unilateral Transfemoral Amputation in Turkey.
Clinical outcome assessments provide important input for the rehabilitation of individuals with transfemoral amputation. Differences in prosthetic knee designs may influence clinical outcomes. The aim of this study was to compare functional mobility, balance, prosthetic satisfaction and quality of life in individuals with unilateral transfemoral amputation with microprocessor-controlled (MPK) and non-microprocessor knee designs (Non-MPK). The study included ten experienced MPK (Rheo Knee) users (Group 1) and ten experienced Non-MPK (Total Knee® 2000) users (Group 2). For mobility; the 6 Minute Walk Test (6MWT), for balance; the Berg Balance Scale (BBS), Single Leg Stand Test (SLST) and Four Square Step Test (FSST), for quality of life; the Nottingham Health Profile (NHP) and for prosthetic satisfaction; the Satisfaction with Prosthesis Questionnaire (SATPRO) were administered. 6MWT results of the MPK group were significantly higher than Non-MPK group (p<0.05). In the MPK group a strong negative correlation was found between the FSST and the 6MWT (r=-0.661, p=0.038). No statistically significant differences were found between the groups (p>0.05) comparing balance, prosthesis satisfaction and quality of life values. The findings will inform about the patient's prognosis and the expected clinical outcomes when prescribing an MPK or an Non-MPK. Individuals with unilateral transfemoral amputation covered longer distances using an MPK compared to Non-MPK.


Counseling the international traveler. Update '93.
The itinerary of international travelers will largely determine the amount of pretravel counseling, number of immunizations and type of malaria prophylaxis they will need. The countries visited are also the best predictor of traveler's diarrhea. Only yellow fever and cholera vaccines are required for entry into certain countries; the latter generally given only to satisfy entry requirements. Polio vaccine is important for some areas and is frequently neglected. For most malarious areas, chloroquine once per week is recommended. Mefloquine should be prescribed weekly for travelers going to chloroquine resistant malarious areas. Traveler's diarrhea is best prevented by avoiding high risk foods and beverages. Antibiotics, generally not recommended for prophylaxis, are very effective in treatment. Travelers should be reminded to advise physicians of their travel history during future medical encounters so that otherwise exotic tropical diseases, possibly contracted during travel, may be considered in diagnoses.


Effect of dynamical instability on timing jitter in passively mode-locked quantum-dot lasers.
We study the effect of noise on the dynamics of passively mode-locked semiconductor lasers both experimentally and theoretically. A method combining analytical and numerical approaches for estimation of pulse timing jitter is proposed. We investigate how the presence of dynamical features such as wavelength bistability in a quantum-dot laser affects timing jitter.


Method for designing an arbitrary continuous refracting surface with specified property of wavefronts conversion.
A method is proposed for designing an arbitrary continuous refracting surface that can realize conversion between two specified distributions of wavefront. Optimization technique is avoided by solving a partial differential equation governing the surface-profile function. This equation is obtained using the property of congruence and the law of refraction, and its solution in this special case is found to be the same as that obtained in wavefront conjugation. A method based on finding the local coefficients of the surface is developed to solve the equation numerically, and an example is given to illustrate its feasibility.


Azathioprine, mercaptopurine and birth outcome: a population-based cohort study.
Data on the safety of azathioprine and mercaptopurine during pregnancy are very sparse. To examine the risk of adverse birth outcomes in women who took up prescriptions for azathioprine or mercaptopurine during pregnancy. This is a Danish cohort study based on data from a population-based prescription registry, the Danish Birth Registry and the Hospital Discharge Registry. To examine the risk of congenital malformations, we included nine pregnancies exposed 30 days before conception or during the first trimester. To examine perinatal mortality, pre-term birth and low birth weight, we included 10 pregnancies exposed during the entire pregnancy. Eleven different exposed women were included in the study. Outcomes were compared with those of 19 418 pregnancies in which no drugs were prescribed to the mothers. Fifty-five per cent of the exposed women had inflammatory bowel disease and 45% other diseases. Adjusted odds ratios for congenital malformations, perinatal mortality, pre-term birth and low birth weight were 6.7 (95% confidence interval, 1.4-32.4), 20.0 (2.5-161.4), 6.6 (1.7-25.9) and 3.8 (0.4-33.3), respectively. Our results suggest that there is an increased risk of congenital malformations, perinatal mortality and pre-term birth in children born to women treated with azathioprine or mercaptopurine during pregnancy. More data are needed to determine whether the associations are causal or occur through confounding.


Accurate determination of isthmus of lumbar pedicle: a morphometric study using reformatted computed tomographic images.
Lumbar pedicle isthmus, the narrowest section of pedicle, was investigated. To determine the lumbar pedicle isthmus accurately and measure related parameters. Accurate anatomic knowledge of pedicle parameters is critical for a successful transpedicular procedure. Many studies on lumbar pedicle dimensions have been conducted, yet less is known about how to obtain these data and their veracity. In fact, the narrowest section is present in the pedicle, i.e., the isthmus of the pedicle. This is the bottleneck of the pedicle; to determine and measure it could illustrate the true morphologic characteristics of the lumbar pedicle. Ultra high-speed spiral CT scan of lumbar spine was performed in 41 patients of Chinese origin. After reformation of the original images, isthmus and transverse plane of pedicle axis were determined. Eleven dimensions of isthmus plane and three dimensions of transverse plane were calculated using software. Pedicle endosteal width and height in female patients who were over the age of 50 were greater than those of female patients younger than 50 years old. The isthmus endosteal width from L1 to L5 was 5.2, 6.0, 7.5, 7.5, and 8.7 mm and 4.0, 4.1, 5.4, 5.7, and 7.1 mm, respectively, in men and women. Isthmus inclination inclined to midline with less than 10 degrees above L3 but increased in L4 and 30 degrees in L5. An objective narrowest section, the pedicle isthmus, exists in lumbar pedicle axis. The parameters of isthmus, especially isthmus endosteal width, show the morphologic characteristics of the lumbar pedicle and are the most important data provided for transpedicular procedures. The small pedicles of female patients over the age of 50 also show certain primary osteoporosis. Isthmus inclination puts the pedicle in a more complicated space position. A better understanding of the complicated structure of the pedicle isthmus guarantees success in transpedicular procedures.


Multichromatic Control of Signaling Pathways in Mammalian Cells.
The precise control of signaling proteins is a prerequisite to decipher the complexity of the signaling network and to reveal and to study pathways involved in regulating cellular metabolism and gene expression. Optogenetic approaches play an emerging role as they enable the spatiotemporal control of signaling processes. Herein, a multichromatic system is developed by combining the blue light cryptochrome 2 system and the red/far-red light phytochrome B system. The use of three wavelengths allows the orthogonal control of the RAF/ERK and the AKT signaling pathway. Continuous exposure of cells to blue light leads to activation of AKT while simultaneous pulses of red and far-red light enable the modulation of ERK signaling in cells with constantly active AKT signaling. The optimized, orthogonal multichromatic system presented here is a valuable tool to better understand the fine grained and intricate processes involved in cell fate decisions.


Isolinderanolide B, a butanolide extracted from the stems of Cinnamomum subavenium, inhibits proliferation of T24 human bladder cancer cells by blocking cell cycle progression and inducing apoptosis.
Isolinderanolide B (IOB), a butanolide extracted from the stems of Cinnamomum subavenium, was investigated for its antiproliferative activity in T24 human bladder cancer cells. To identity the anticancer mechanism of IOB, its effect on apoptosis, cell cycle distribution, and levels of p53, p21 Waf1/Cip1, Fas/APO-1 receptor, and Fas ligand was assayed. Enzyme-linked immunosorbent assay showed that the G0/G1 phase arrest is because of increase in the expression of p21 Waf1/Cip1. An enhancement in Fas/APO-1 and membrane-bound Fas ligand (mFasL) might be responsible for the apoptotic effect induced by IOB. This study reports the novel finding that the induction of p21 Waf1/Cip1 and activity of the Fas/mFas ligand apoptotic system may participate in the antiproliferative activity of IOB in T24 cells.


Comparison of in vitro toxicity of aerosolized engineered nanomaterials using air-liquid interface mono-culture and co-culture models.
Airborne engineered nanomaterials (ENMs) can readily enter the human body through inhalation potentially leading to adverse health effects such as cardiovascular and pulmonary diseases. Our group has previously utilized and validated an integrated low flow system capable of generating and depositing airborne ENMs directly onto cells at an air-liquid interface (ALI). To further improve this ALI method for an even closer representation of the in vivo system, a co-culture model containing epithelial, endothelial and macrophage cell lines (A549, EA.hy 926, and THP-1 differentiated macrophages) was established and validated for testing ENMs toxicity. In the co-culture model, cells were exposed to citrate-capped gold (Au), 15% silver on silica (Ag-SiO2) and copper oxide (CuO) ENMs under the same protocol (4 h ALI exposure with a target concentration of 3.5 mg/m3) and compared to responses with A549 cells only or THP-1 differentiated cells only. The toxicological profile was assessed by measuring cell viability, reactive oxygen species (ROS) production, lactate dehydrogenase (LDH) release, and interleukin (IL)-8 concentration. Results showed that 15% Ag-SiO2 induced more oxidative stress-related toxicity in the co-culture than in A549 cells alone. Both 15% Ag-SiO2 and CuO exposure produced significantly higher levels of IL-8 in the co-culture compared with A549 cells alone. Citrate-capped Au was largely inert. Further exposures of CuO on macrophages alone provided evidence of cell-cell interaction in the co-culture model. In addition, the co-culture model exhibited a similar response to primary human bronchial epithelial cells in terms of ROS and IL-8 responses after CuO exposure, suggesting a more advanced refinement of the conventional model for in vitro inhalation study.


Isokinetic aerobic power output testing of the quadriceps muscle.
A stepwise increasing exercise protocol to measure the maximal aerobic power output of the quadriceps muscle is proposed for use in rehabilitation medicine. Sixteen healthy volunteers performed isokinetic knee extension/flexion exercises at an angular velocity of 180 degrees x s(-1). The protocol consisted of stages of 200 knee movements from 100 flexion to full extension, starting at 10% peak torque (PT) and increasing by 10% PT each following stage until exhaustion. Quadriceps work, oxygen uptake, heart rate, plasma lactate concentration and surface electromyography were monitored. Quadriceps power output (Pext) was highly related (r = 0.95) to the extension torque at which the subjects were instructed to exercise. The test-retest (r = 0.82) and left to right (r = 0.94) correlations of maximum quadriceps power output (Pmax) were high. Both sexes (males 43+/-9W, females 36+/-8W) achieved their Pmax at 47% PT. At submaximal power output stages oxygen uptake (r = 0.85) and EMG-amplitude (r = 0.88) were linearly related to Pext. Mechanical efficiency (optimum at 60% Pmax) showed a large variation between power output stages and between subjects. The relationship between relative oxygen uptake and heart rate or plasma lactate concentration was similar to that of whole body exercise, e.g. running. We conclude that the proposed test is reliable for assessing the maximal aerobic power output of the quadriceps muscle. This parameter of the muscle function may add useful information for assessing the rehabilitation process after knee immobilization.


Updates on Wound Infiltration Use for Postoperative Pain Management: A Narrative Review.
Local anesthetic wound infiltration (WI) provides anesthesia for minor surgical procedures and improves postoperative analgesia as part of multimodal analgesia after general or regional anesthesia. Although pre-incisional block is preferable, in practice WI is usually done at the end of surgery. WI performed as a continuous modality reduces analgesics, prolongs the duration of analgesia, and enhances the patient's mobilization in some cases. WI benefits are documented in open abdominal surgeries (Caesarean section, colorectal surgery, abdominal hysterectomy, herniorrhaphy), laparoscopic cholecystectomy, oncological breast surgeries, laminectomy, hallux valgus surgery, and radical prostatectomy. Surgical site infiltration requires knowledge of anatomy and the pain origin for a procedure, systematic extensive infiltration of local anesthetic in various tissue planes under direct visualization before wound closure or subcutaneously along the incision. Because the incidence of local anesthetic systemic toxicity is 11% after subcutaneous WI, appropriate local anesthetic dosing is crucial. The risk of wound infection is related to the infection incidence after each particular surgery. For WI to fully meet patient and physician expectations, mastery of the technique, patient education, appropriate local anesthetic dosing and management of the surgical wound with "aseptic, non-touch" technique are needed.


The ultrastructural features of plasmablastic lymphoma.
Plasmablastic lymphoma was originally described in 1997 by Delecluse et al. and is an aggressive variant of diffuse large B-cell non-Hodgkin lymphoma seen predominantly in a setting of acquired immunodeficiency syndrome and nearly always in extranodal sites. The authors have seen 10 cases in their department between 2001 and 2005. The patients' ages ranged from 24 to 39 years and there were 7 females and 3 males. In 7 cases where human immunodeficiency virus had been tested it was positive. Eight cases were extranodal and 2 cases involved lymph nodes. Five cases were followed up and 4 were confirmed dead within 5 months of diagnosis, verifying the aggressive nature of this condition. Histology showed large, polygonal tumor cells some of which had a slightly plasmacytic appearance. Six cases had a "starry sky" background. Immunohistochemical stains were negative in 1 case, while the other 9 cases were positive for CD138. Electron microscopy showed concentrically arranged rough endoplasmic reticulum in the cytoplasm in 9 cases. In 1 case the cells were too degenerate for evaluation. This study shows that the ultrastructural features are well developed and can help in distinguishing plasmablastic lymphoma from other light microscopically undifferentiated tumors.


Current state and prospects of artificial intelligence in allergy.
The field of medicine is witnessing an exponential growth of interest in artificial intelligence (AI), which enables new research questions and the analysis of larger and new types of data. Nevertheless, applications that go beyond proof of concepts and deliver clinical value remain rare, especially in the field of allergy. This narrative review provides a fundamental understanding of the core concepts of AI and critically discusses its limitations and open challenges, such as data availability and bias, along with potential directions to surmount them. We provide a conceptual framework to structure AI applications within this field and discuss forefront case examples. Most of these applications of AI and machine learning in allergy concern supervised learning and unsupervised clustering, with a strong emphasis on diagnosis and subtyping. A perspective is shared on guidelines for good AI practice to guide readers in applying it effectively and safely, along with prospects of field advancement and initiatives to increase clinical impact. We anticipate that AI can further deepen our knowledge of disease mechanisms and contribute to precision medicine in allergy.


[Changes in prescription patterns to patients with bipolar syndromes. Increased use of lamotrigine and decreased use of lithium].
Lithium is a first line option in the maintenance treatment of bipolar disorder, but several alternative treatment regimens have been introduced in recent years, among them treatment with antiepileptic compounds and atypical antipsychotic drugs. Little is known about if and how this has changed the prescription patterns of mood stabilizers. We analysed trends in prescription of mood stabilisers in Sweden using the national quality register for bipolar disorder (BipoläR), the Prescribed Drug Register, and the Patient Register during the years 2007-2011. We found that lithium use decreased while lamotrigine use increased in bipolar patients. These changes could not be ex-plained by differences in bipolar subtypes; lithium use decreased in both bipolar type I and type II, and the use of lamotrigine increased in bipolar type II. Lithium use was more common in men, whereas lamotrigine use was more common in women. The prescription of other mood stabilisers did not change during these years.