Concentration dependence of alpha-synuclein fibril length assessed by quantitative atomic force microscopy and statistical-mechanical theory.
The initial concentration of monomeric amyloidogenic proteins is a crucial factor in the in vitro formation of amyloid fibrils. We use quantitative atomic force microscopy to study the effect of the initial concentration of human alpha-synuclein on the mean length of mature alpha-synuclein fibrils, which are associated with Parkinson's disease. We determine that the critical initial concentration, below which low-molecular-weight species dominate and above which fibrils are the dominant species, lies at approximately 15 muM, in good agreement with earlier measurements using biochemical methods. In the concentration regime where fibrils dominate, we find that their mean length increases with initial concentration. These results correspond well to the qualitative predictions of a recent statistical-mechanical model of amyloid fibril formation. In addition, good quantitative agreement of the statistical-mechanical model with the measured mean fibril length as a function of initial protein concentration, as well as with the fibril length distributions for several protein concentrations, is found for reasonable values of the relevant model parameters. The comparison between theory and experiment yields, for the first time to our knowledge, an estimate of the magnitude of the free energies associated with the intermolecular interactions that govern alpha-synuclein fibril formation.


Prediction of molecular properties including symmetry from quantum-based molecular structural formulas, VIF.
Structurally covariant valency interaction formulas, VIF, gain chemical significance by comparison with resonance structures and natural bond orbital, NBO, bonding schemes and at the same time allow for additional prediction such as symmetry of ring systems and destabilization of electron pairs with respect to reference energy of -1/2 Eh. Comparisons are based on three chemical interpretations of Sinanoğlu's theory of structural covariance: (1) sets of structurally covariant quantum structural formulas, VIF, are interpreted as the same quantum operator represented in linearly related basis frames; (2) structurally covariant VIF pictures are interpreted as sets of molecular species with similar energy; and (3) the same VIF picture can be interpreted as different quantum operators, one-electron density or Hamiltonian; for example. According to these three interpretations, bond pair, lone pair, and free radical electrons understood in terms of a localized orbital representation are recognized as having energies above, below, or equal to a predetermined reference, frequently-1/2 Eh. The probable position of electron pairs and radical electrons is predicted. The selectivity of concerted ring closures in allyl anion and cation is described. Symmetries of conjugated ring systems are predicted according to their numbers of pi-electrons and spin-multiplicity. The pi-distortivity of benzene is predicted.The 3c/2e- H-bridging bonds in diborane are derived in a natural way according to the notion that the bridging bonds will have delocalizing interactions between them consistent with results of the NBO method. Key chemical bonding motifs are described using VIF. These include 2c/1e-, 2c/2e-, 2c/3e-, 3c/2e-, 3c/3e-,3c/4e-, 4n antiaromatic, and 4n+2 aromatic bonding systems. Some common organic functional groups are represented as VIF pictures and because these pictures can be interpreted simultaneously as one-electron density and Hamiltonian operators, the valence shell electron pair repulsion method is applied toward understanding the energies of valence NBOs with respect to the reference energy of -1/2Eh.


Healthcare-seeking behaviour in case of influenza-like illness in the French general population and factors associated with a GP consultation: an observational prospective study.
GP consultation rates for influenza-like illness (ILI) are poorly known in France and there is a paucity of literature on this topic. In the few articles that have been published, the results are heterogeneous. The aim of the present study was to estimate the proportion of ILI inducing a GP consultation, and to assess its determinants. Participants of a French web-based cohort study who reported ≥1 ILI episode between 2012 and 2015 were included. Sociodemographic characteristics, access to health care, and health status variables were collected. Healthcare-seeking behaviour was analysed and factors associated with a GP consultation identified using a conditional logistic regression. Of the 6023 ILI episodes reported, 1961 (32.6%) led to a GP consultation, with no difference between those at risk of influenza complications and those not (P = 0.42). A GP consultation was more frequent for individuals living in a rural area (odds ratio [OR] = 1.21, 95% confidence interval [CI] = 1.02 to 1.43); those with a lower educational level (OR = 1.43, 95% CI = 1.18 to 1.74); those using the internet to find information about influenza (OR = 1.63, 95% CI = 1.30 to 2.03); patients presenting with worrying symptoms (fever, cough, dyspnoea, sputum, or asthenia); patients having a negative perception of their own health status (OR = 1.51, 95% CI = 1.07 to 2.13; and those having declared a personal doctor (OR = 2.86, 95% CI = 1.72 to 4.76). A GP consultation was less frequent for individuals using alternative medicine (OR = 0.68, 95% CI = 0.58 to 0.78). This study allows the identification of specific factors associated with GP consultation for an ILI episode. These findings may help to coordinate health information campaigns and to raise awareness, especially among individuals at risk of influenza complications.


Effectiveness and Efficiency of Non-drug Therapy Among Community-Dwelling Adults With Hypertension in China: A Protocol for Network Meta-Analysis and Cost-Effectiveness Analysis.
Introduction: The Chinese government has established a nationwide community-based chronic disease management program since 2009 with hypertension a vital part of it. Though drugs have been proven effective with hypertensive patients, they bring economic burden as well, especially for those who with elevated blood pressure and are potentially eligible for national programs. When the effectiveness of pharmacotherapy-only interventions remains uncertain on these patients, non-pharmacological interventions have demonstrated non-inferior effectiveness and may have economic advantages. To date, there rarely are evidences on the effectiveness and cost-effectiveness of non-pharmacological treatment in comparison with pharmacological interventions for patients with varying severity of blood pressure. This study aims to propose a study for a network meta-analysis and cost-effectiveness analysis to explore what kind of intervention is potentially effective and cost-effective to four specific patient groups, stage I-III hypertensive patients and patients with elevated blood pressure, and to provide recommendations for hypertensive management to Chinese decision makers. Methods: We will systematically search databases (MEDLINE, PubMed, Cochrane Library, etc.,) for randomized controlled trials and observational studies with qualified study design in recent decade that assess the effectiveness of non-pharmacological, pharmacological, or combined intervention aimed at adult populations who are diagnosed with the above four types of hypertension in China. The effectiveness outcomes will include changes in SBP/DBP, rate of comorbidities, mortality, and health related quality of life. We will use network meta-analysis to compare and rank effectiveness of different interventions. Subgroup analyses and meta-regression analyses will be performed to analyze and explain heterogeneity. The economic outcome will include cost-effectiveness based on simulation results from Markov models. Under study perspective of Chinese health system, life-time direct cost will be included. Discussion: This study aims to compare and rank the effectiveness and cost-effectiveness of pharmacological, non-pharmacological and combined interventions for stage I-III hypertensive patients and those who with elevated blood pressure. Compared to existing studies, this comprehensive synthesis of relevant evidences will influence future practice with better efficiency and generalizability for community-based hypertensive management programs in China. The study might also be valuable for other low- and middle-income countries to find their own solutions. PROSPERO registration number: CRD42020151518.


Microvascular abnormalities in capillaroscopy correlate with higher serum IL-18 and sE-selectin levels in patients with type 1 diabetes complicated by microangiopathy.
Microvascular abnormalities are one of the most important causes of persistent diabetic complications. The aim of this study was to compare microvascular changes examined by nailfold capillaroscopy with serum concentrations of soluble E-selectin (sE-selectin) and IL-18 in type 1 diabetic patients with and without microangiopathy. Serum levels of sE-selectin and IL-18 were determined by an enzyme-linked immunosorbent assay in 106 patients with type 1 diabetes and in 40 healthy controls. All diabetic patients were evaluated by extensive clinical, laboratory and capillaroscopic studies. Morphological changes were observed by nailfold capillaroscopy in 86 out of 106 (81%) diabetic patients. Severe capillaroscopic changes were seen in 32 out of 54 (59%) patients with microangiopathy, but in only seven out of 52 (13%) patients without microangiopathy. Higher serum levels of sE-selectin (p < 0.001) and IL-18 (p < 0.05) were demonstrated in diabetic patients compared to controls. Significant differences of sE-selectin (p , 0.001) and IL-18 (p < 0.01) serum concentrations were observed between diabetic patients with microangiopathy and controls. Moreover, comparison between patients with and without microangiopathic complications showed a significantly higher capillaroscopic score and sE-selectin serum concentration in the group with microangiopathy (p < 0.001). Furthermore, diabetic patients with severe microvascular changes in capillaroscopy showed significantly higher IL-18 (p < 0.001) and sE-selectin (p < 0.05) serum levels than subgroups without changes or with mild abnormalities. Our findings suggest that abnormalities in nailfold capillaroscopy may reflect the extent of microvascular involvement and are associated with higher sE-selectin and IL-18 serum levels, as well as with microangiopathic complications in diabetic patients.


Associations of Arterial Stiffness With Cognitive Performance, and the Role of Microvascular Dysfunction: The Maastricht Study.
The mechanisms underlying cognitive impairment are incompletely understood but may include arterial stiffness and microvascular dysfunction. In the population-based Maastricht Study, we investigated the association between arterial stiffness and cognitive performance, and whether any such association was mediated by microvascular dysfunction. We included cross-sectional data of 2544 participants (age, 59.7 years; 51.0% men; 26.0% type 2 diabetes mellitus). We used carotid-femoral pulse wave velocity and carotid distensibility coefficient as measures of aortic and carotid stiffness, respectively. We calculated a composite score of microvascular dysfunction based on magnetic resonance imaging features of cerebral small vessel disease, flicker light-induced retinal arteriolar and venular dilation response, albuminuria, and plasma biomarkers of microvascular dysfunction (sICAM-1 [soluble intercellular adhesion molecule-1], sVCAM-1 [soluble vascular adhesion molecule-1], sE-selectin [soluble E-selectin], and vWF [von Willebrand factor]). Cognitive domains assessed were memory, processing speed, and executive function. A cognitive function score was calculated as the average of these domains. Higher aortic stiffness (per m/s) was associated with lower cognitive function (β, -0.018 SD [95% CI, -0.036 to -0.000]) independent of age, sex, education, and cardiovascular risk factors, but higher carotid stiffness was not. Higher aortic stiffness (per m/s) was associated with a higher microvascular dysfunction score (β, 0.034 SD [95% CI, 0.014 to 0.053]), and a higher microvascular dysfunction score (per SD) was associated with lower cognitive function (β, -0.089 SD [95% CI, -0.124 to -0.053]). Microvascular dysfunction significantly explained 16.2% of the total effect of aortic stiffness on cognitive function. The present study showed that aortic stiffness, but not carotid stiffness, is independently associated with worse cognitive performance, and that this association is in part explained by microvascular dysfunction.


Association of Atrial Fibrillation With Cognitive Decline and Dementia Over 20 Years: The ARIC-NCS (Atherosclerosis Risk in Communities Neurocognitive Study).
Previous studies have reported that atrial fibrillation (AF) is associated with cognitive decline and dementia. These studies, however, had limited follow-up, were based mostly on white and highly selected populations, and did not account for attrition. We evaluated the association of incident AF with 20-year change in cognitive performance (accounting for attrition) and incident dementia in the ARIC (Atherosclerosis Risk in Communities) Study. We analyzed data from 12 515 participants (mean age, 56.9 [SD, 5.7] years in 1990-1992; 56% women and 24% black) from 1990 to 1992 through 2011 to 2013. Incident AF was ascertained from study ECGs and hospital discharge codes. Cognitive tests were performed in 1990 to 1992, 1996 to 1998, and 2011 to 2013. Incident dementia was clinician adjudicated. We used generalized estimating equations and Cox proportional hazards models to assess the association of time-dependent AF with change in Z scores of cognitive tests and incident dementia, respectively. During 20 years, 2106 participants developed AF and 1157 participants developed dementia. After accounting for cardiovascular risk factors, including ischemic stroke, the average decline over 20 years in global cognitive Z score was 0.115 (95% confidence interval, 0.014-0.215) greater in participants with AF than in those without AF. Further adjustment for attrition by multiple imputation by chained equations strengthened the association. In addition, incident AF was associated with an increased risk of dementia (hazard ratio, 1.23; 95% confidence interval, 1.04-1.45), after adjusting for cardiovascular risk factors, including ischemic stroke. AF is associated with greater cognitive decline and increased risk of dementia, independent of ischemic stroke. Because cognitive decline is a precursor to dementia, our findings prompt further investigation to identify specific treatments for AF that will delay the trajectory of cognitive decline and, thus, prevent dementia in patients with AF.


The combined and interactive effects of zinc, temperature, and phosphorus on the structure and functioning of a freshwater community.
Ecotoxicological studies mainly consist of single-species experiments evaluating the effects of a single stressor. However, under natural conditions aquatic communities are exposed to a mixture of stressors. The present study aimed to identify how the toxicity of zinc (Zn) is affected by increased temperature and increased phosphorus (P) supply and how these interactions vary among species, functional groups, and community structure and function. Aquatic microcosms were subjected to 3 Zn concentrations (background, no Zn added, and 75 and 300 μg Zn/L), 2 temperatures (16-19 and 21-24 °C), and 2 different P additions (low, 0.02, and high, 0.4 mg P L-1 wk-1 ) for 5 wk using a full factorial design. During the study, consistent interactions between Zn and temperature were only rarely found at the species level (4%), but were frequently found at the functional group level (36%), for community structure (100%) and for community function (100%; such as dissolved organic carbon concentrations and total chlorophyll). The majority of the Zn × temperature interactions were observed at 300 μg Zn/L and generally indicated a smaller effect of Zn at higher temperature. Furthermore, no clear indication was found that high P addition by itself significantly affected the overall effects of Zn on the community at any level of organization. Interestingly, though, 90% of all the Zn × temperature interactions observed at the species, group, and community composition level were found under high P addition. Collectively, the results of our study with the model chemical Zn suggest that temperature and phosphorus loading to freshwater systems should be accounted for in risk assessment, because these factors may modify the effects of chemicals on the structure and functioning of aquatic communities, especially at higher levels of biological organization. Environ Toxicol Chem 2018;37:2413-2427. © 2018 SETAC.


Analysis of the effects of Cu-MOFs on fungal cell inactivation.
Three dimensional (3D) copper metal organic frameworks (Cu-MOFs) containing glutarates and bipyridyl ligands (bpa = 1,2-bis(4-pyridyl)ethane, bpe = 1,2-bis(4-pyridyl)ethylene, or bpp = 1,3-bis(4-pyridyl)propane) were synthesized by using previously reported hydrothermal reactions or a layering method. All three Cu-MOFs contained well-defined one dimensional (1D) channels with very similar pore shapes and different pore dimensions. The bulk purities of the Cu-MOFs were confirmed using powder X-ray diffraction (PXRD) and infrared spectroscopy (IR) spectra. When the three types of Cu-MOFs were applied to Candida albicans cells and Aspergillus niger spores, an average of about 50-80% inactivation was observed at the highest concentration of Cu-MOFs (2 mg mL-1). The efficiency of the fungal inactivation was not significantly different among the three different types (bpa, bpe, bpp). Treatment of the fungi using Cu-MOFs induced an apoptosis-like death and this was more severe in A. niger than C. albicans. This may be due to elevation of the intracellular level of reactive oxygen species (ROS) in A. niger. Generation of the reactive species in solution by Cu-MOFs was observed. However, there was a dramatic variation in the levels observed among the three types. Our results suggest that Cu-MOFs can produce antifungal effects and induce apoptosis-like death of the fungi, which was probably caused by the elevated level of intracellular reactive species.


Muscle Synergies Control during Hand-Reaching Tasks in Multiple Directions Post-stroke.
Purpose: A muscle synergies model was suggested to represent a simplifying motor control mechanism by the brainstem and spinal cord. The aim of the study was to investigate the feasibility of such control mechanisms in the rehabilitation of post-stroke individuals during the execution of hand-reaching movements in multiple directions, compared to non-stroke individuals. Methods: Twelve non-stroke and 13 post-stroke individuals participated in the study. Muscle synergies were extracted from EMG data that was recorded during hand reaching tasks, using the NMF algorithm. The optimal number of synergies was evaluated in both groups using the Variance Accounted For (VAF) and the Mean Squared Error (MSE). A cross validation procedure was carried out to define a representative set of synergies. The similarity index and the K-means algorithm were applied to validate the existence of such a set of synergies, but also to compare the modulation properties of synergies for different movement directions between groups. The similarity index and hierarchical cluster analysis were also applied to compare between group synergies. Results: Four synergies were chosen to optimally capture the variances in the EMG data, with mean VAF of 0.917 ± 0.034 and 0.883 ± 0.046 of the data variances, with respective MSE of 0.007 and 0.016, in the control and study groups, respectively. The representative set of synergies was set to be extracted from movement to the center of the reaching space. Two synergies had different muscle activation balance between groups. Seven and 17 clusters partitioned the muscle synergies of the control and study groups. The control group exhibited a gradual change in the activation in the amplitude in the time domain (modulation) of synergies, as reflected by the similarity index, whereas the study group exhibited consistently significant differences between all movement directions and the representative set of synergies. The study findings support the existence of a representative set of synergies, which are modulated to execute movements in different directions. Conclusions: Post-stroke individuals differently modulate the activation of synergies to different movement directions than do non-stroke individuals. The conclusion was supported by different muscle activation balances, similarity values and different classifications of synergies among groups.


Effect of liposome composition and other factors on the targeting of liposomes to experimental tumors: biodistribution and imaging studies.
We have examined the distribution of radiolabeled liposomes in tumor-bearing mice after i.v. injection. Two mouse tumors (B16 melanoma, J6456 lymphoma) and a human tumor (LS174T colon carcinoma) inoculated i.m., s.c., or in the hind footpad were used in these studies. When various liposome compositions with a mean vesicle diameter of approximately 100 nm were compared using a radiolabel of gallium-67-deferoxamine, optimal tumor localization was obtained with liposomes containing a phosphatidylcholine of high phase-transition temperature and a small molar fraction of monosialoganglioside or hydrogenated phosphatidylinositol (HPI). At 24 h after injection, average values of tumor uptake higher than 10% of the injected dose per g and liver-to-tumor ratios close to 1 were reproducibly obtained. Increasing the molar fraction of HPI from 9% to 41% of the total phospholipid resulted in enhancement of liver uptake and decrease of tumor uptake. Methodological aspects that influence vesicle size appear to affect significantly liposome localization in the tumor. However, varying the phospholipid dose within a 10-fold range caused only minor changes in the percent of injected dose recovered in the tumor. A high uptake by tumors was also observed using other radiolabels [[3H]inulin and indium-111-labeled bleomycin (111In-Bleo)] in monosialoganglioside- and HPI-containing liposomes. In the case of 111In-Bleo, encapsulation in liposomes resulted in approximately 20- to 40-fold increase in tumor accumulation of the radiolabel at 24 h after injection. The marked localization of liposomes in the mouse footpad inoculated with tumor as opposed to the contralateral mock-injected footpad was also documented by imaging experiments with gallium-67-deferoxamine and 111In-Bleo-labeled liposomes. These results support the contention that some glycolipid-containing liposomes previously shown to have long circulating half-lives accumulate significantly in a variety of tumors and are promising tools for the delivery of anti-tumor agents.


Recent developments in manufacturing emulsions and particulate products using membranes.
Membrane emulsification (ME) is a relatively new technique for the highly controlled production of particulates. This review focuses on the recent developments in this area, ranging from the production of simple oil-in-water (O/W) or water-in-oil (W/O) emulsions to multiple emulsions of different types, solid-in-oil-in-water (S/O/W) dispersions, coherent solids (silica particles, solid lipid microspheres, solder metal powder) and structured solids (solid lipid microcarriers, gel microbeads, polymeric microspheres, core-shell microcapsules and hollow polymeric microparticles). Other emerging technologies that extend the capabilities into different membrane materials and operation methods (such as rotating membranes, repeated membrane extrusion of coarsely pre-emulsified feeds) are introduced. The results of experimental work carried out by cited researchers in the field together with those of the current authors are presented in a tabular form in a rigorous and systematic manner. These demonstrate a wide range of products that can be manufactured using different membrane approaches. Opportunities for creation of new and novel entities are highlighted for low throughput applications (medical diagnostics, healthcare) and for large-scale productions (consumer and personal products).


[Pelvic actinomycosis in Tunisia: five cases].
Actinomycosis is a rare suppurative disease due to Actinomyces species. These Gram-positive, non-acid fast anaerobic filamentous bacteria are normal inhabitants of the human body, tending to reside in the oropharynx and bowel but are occasionally found in the vagina. Pelvic actinomycosis is a rare bacterial disease in women. Clinical manifestations are various and non specific and may be acute or chronic. No consensus exists for treatment. We reviewed files and identified all five cases of pelvic actinomycosis managed at Obstetrics and Gynaecology department "A" at the Maternity Center of Tunis over an eight-year period (1998-2005). The women's average age was 39.2 years. One patient was menopausal and consulted for bleeding. The other four patients were younger and had all been using an intrauterine device (IUD) for contraception. They presented with acute clinical manifestations. Their main symptom was pelvic pain. Three women had fever, and two presented with urinary tract obstruction. All patients had surgery. A pelvic abscess was found in four cases. Laparoscopic management was possible in only one case. Laparotomy was necessary in the other four. Four women had adnexectomies, two with hysterectomy. Digestive complications occurred in three cases. Actinomycosis was diagnosed only after surgery, by the histological examination. This series confirms the difficulties encountered in the management of pelvic actinomycosis. We review the recent literature and describe the diagnostic and therapeutic procedures currently recommended. The relationship between pelvic actinomycosis and IUDs, the most common method of contraception in Tunisia, is clearly established. Clinical diagnosis of pelvic actinomycosis is difficult because the symptoms are non-specific. Laboratory tests can help by showing serious inflammation, however. Imaging findings are also non-specific and may suggest an abscess or an inflammatory or neoplastic process. Interventional radiology, specifically CT- or ultrasound-guided aspiration biopsy can facilitate diagnosis, which in any case requires a histological examination. Treatment of pelvic actinomycosis is not standardized and depends on clinical form. Medical treatment is based on long-term penicillin G. It is always necessary and may be used alone in cases of preoperative diagnosis. Surgery should be offered only in resistant cases and should always be followed by long-term antibiotic treatment. The prognosis of correctly treated pelvic actinomycosis is generally good.


Staff-student relationships and their impact on nursing students' belongingness and learning.
This paper is a report of selected findings from a study exploring the relationship between belongingness and placement experiences of preregistration nursing students. Staff-student relationships are an important influence on students' experiences of belongingness and their clinical learning. The need to belong is universal and pervasive, exerting a powerful influence on thought processes, emotions, behaviour, health and happiness. People deprived of belongingness are more likely to experience diminished self-esteem, increased stress and anxiety, depression and a decrease in general well-being. Nursing students' motivation and capacity to learn, self-concept, confidence, the extent to which they are willing to question or conform to poor practice and their future career decisions are influenced by the extent to which they experience belongingness. During 2006, 18 third year students from two Australian universities and one United Kingdom university participated in in-depth semi-structured interviews. Data were analysed thematically. Participants described placement experiences spanning a continuum from those promoting a high degree of belongingness to those provoking intense feelings of alienation. Staff-student relationships (including receptiveness, inclusion/exclusion, legitimization of the student role, recognition and appreciation, challenge and support) were the most important influence on students' sense of belonging and learning. Similarities between sites were remarkable, despite the differences in healthcare and higher education systems. Staff-student relationships are key to students' experience of belongingness. Understanding the types of interactions and behaviours that facilitate or impede students' belongingness and learning are essential to the creation of positive clinical experiences.


Internal carotid artery aneurysms occurring at the origin of fetal variant posterior cerebral arteries: surgical and endovascular experience.
A fetal variant posterior cerebral artery (fetal PCA) is an embryological remnant in which the PCA is primarily supplied via the anterior cerebral circulation. Internal carotid artery (ICA) aneurysms originating from the takeoff of fetal PCA vessels deserve special attention before surgical or endovascular obliteration because of a greater potential for ischemic injury. We present the first series of ICA-posterior communicating artery (PComA) aneurysms originating at the takeoff of fetal PCA vessels that were treated by surgical or endovascular intervention. A retrospective chart review was conducted for all patients who underwent surgical and endovascular treatment of an ICA-PComA aneurysm at Los Angeles County-University of Southern California Medical Center during a 15-year period (1991-2006) to identify cases with aneurysms originating from fetal variant PCAs. Data were retrospectively reviewed and analyzed. During a 15-year period, 271 patients were treated for 273 ICA-PComA aneurysms. Aneurysms occurring at the origin of fetal PCAs were identified in 30 patients (11%). There were 23 women (77%) and seven men (23%) (sex difference, P = 0.0035). Twenty-four patients underwent surgical clipping, whereas six patients underwent endovascular coiling. The mean aneurysm size was 7 mm. The mean ischemia time with temporary clipping (12 cases) was 4.5 minutes. Intraoperative rupture occurred in four surgical cases (17%). Postoperative angiography demonstrated occlusion of the fetal PCA in one case after clip ligation (3%), with an ensuing occipital infarct yet no clinical symptoms. ICA-PComA aneurysms originating from fetal PCA vessels may pose a more substantial risk for infarction and subsequent neurological sequelae with surgical or endovascular obliteration. Fetal variant circulations were identified at the PComA origin in 11% of ICA-PComA aneurysm patients and were more commonly encountered in women. The decision of surgical versus endovascular treatment of fetal PCA aneurysms must be carefully considered, given the greater potential for ischemic injury with parent vessel occlusion.


Identification of PCR-amplified genetically modified organisms (GMOs) DNA by peptide nucleic acid (PNA) probes in anion-exchange chromatographic analysis.
PCR products obtained by selective amplification of transgenic DNA derived from food samples containing Roundup Ready soybean or Bt-176 maize have been analyzed by anion-exchange HPLC. Peptide nucleic acids (PNAs), oligonucleotide analogues known to bind to complementary single-stranded DNA with high affinity and specificity, have been used as specific probes in order to assess the identity of the peaks observed. Two different protocols were adopted in order to obtain single-stranded DNA: amplification with an excess of one primer or digestion of one DNA strand. The single-stranded DNA was mixed with the PNA probe, and the presence of a specific sequence was revealed through detection of the corresponding PNA:DNA peak with significantly different retention time. Advantages and limits of this approach are discussed. The method was tested with reference materials and subsequently applied to commercial samples.


Behavioral Experimental Paradigms for the Evaluation of Drug's Influence on Cognitive Functions: Interpretation of Associative, Spatial/Nonspatial and Working Memory.
Currently, a large number of people throughout the world are affected by neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease and Huntington's disease which appear with a lapse in recall, attention and altered cognitive functions. Learning and memory, the fundamental indices defining cognitive functions, are the complex psychological processes governing acquisition, consolidation, and retrieval of stored information. These processes are synchronized by the coordination of various parts of the brain including hippocampus, striatum and amygdala. The present review is centered on different behavioral paradigms in rodents interpreting learning and memory both explicitly and implicitly. Furthermore, it is also emphasizing on the interaction of various brain structures during different stages of associative, spatial and non-spatial memory. We embarked on an objective review of literature relevant to screening methods for evaluation of drug's influence on a wide range of cognitive functions (learning and memory) as well as the underlying mechanism responsible for modulation of these functions. Our review highlighted the behavioral paradigms based on associative, spatial/nonspatial and working memory. The cited research acknowledged the hippocampal and striatal control on learning and memory. Since the neurodegenerative disorders and dementia have continuously been increasing, a wide range of therapeutic targets have been developed at the cellular and molecular level. This arises the necessity of screening of these targets in different cognitive behavioral paradigms which reflect their memory enhancing potential. The understanding of behavioral models and the involvement of brain structures in cognitive functions highlighted in the present review might be helpful to advance therapeutic interventions.


Improved Statistical Methods are Needed to Advance Personalized Medicine.
Common methods of statistical analysis, e.g. Analysis of Variance and Discriminant Analysis, are not necessarily optimal in selecting therapy for an individual patient. These methods rely on group differences to identify markers for disease or successful interventions and ignore sub-group differences when the number of sub-groups is large. In these circumstances, they provide the same advice to an individual as the average patient. Personalized medicine needs new statistical methods that allow treatment efficacy to be tailored to a specific patient, based on a large number of patient characteristics. One such approach is the sequential k-nearest neighbor analysis (patients-like-me algorithm). In this approach, the k most similar patients are examined sequentially until a statistically significant conclusion about the efficacy of treatment for the patient-at-hand can be arrived at. For some patients, the algorithm stops before the entire set of data is examined and provides beneficial advice that may contradict recommendations made to the average patient. Many problems remain in creating statistical tools that can help individual patients but this is an important area in which progress in statistical thinking is helpful.


Patterns of speciation in a parapatric pair of Saturnia moths as revealed by target capture.
The focus of this study has been to understand the evolutionary relationships and taxonomy of a widely distributed parapatric species pair of wild silk moths in Europe: Saturnia pavonia and Saturnia pavoniella (Lepidoptera: Saturniidae). To address species delimitation in these parapatric taxa, target enrichment and mtDNA sequencing was employed alongside phylogenetic, admixture, introgression, and species delimitation analyses. The dataset included individuals from both species close to and farther away from the contact zone as well as two hybrids generated in the lab. Nuclear markers strongly supported both S. pavonia and S. pavoniella as two distinct species, with hybrids forming a sister group to S. pavoniella. However, the Maximum Likelihood (ML) tree generated from mtDNA sequencing data presented a different picture, showing both taxa to be phylogenetically intermixed. This inconsistency is likely attributable to mitonuclear discordance, which can arise from biological factors (e.g., introgressive hybridization and/or incomplete lineage sorting). Our analyses indicate that past introgressions have taken place, but that there is no evidence to suggest an ongoing admixture between the two species, demonstrating that the taxa have reached full postzygotic reproductive isolation and hence represent two distinct biological species. Finally, we discuss our results from an evolutionary point of view taking into consideration the past climatic oscillations that have likely shaped the present dynamics between the two species. Overall, our study demonstrates the effectiveness of the target enrichment approach in resolving shallow phylogenetic relationships under complex evolutionary circumstances and that this approach is useful in establishing robust and well-informed taxonomic delimitations involving parapatric taxa.


Effects of pH and elevated glucose levels on the electrochemical behavior of dental implants.
Implant failure is more likely to occur in persons with medically compromising systemic conditions, such as diabetes related to high blood glucose levels and inflammatory diseases related to pH levels lower than those in healthy people. The aim of this study was to investigate the effects of lower pH level and simulated- hyperglycemia on implant corrosion as these effects are critical to biocompatibility and osseointegration. The electrochemical corrosion properties of titanium implants were studied in four different solutions: Ringer's physiological solution at pH = 7.0 and pH = 5.5 and Ringer's physiological solution containing 15 mM dextrose at pH = 7 and pH = 5.5. Corrosion behaviors of dental implants were determined by cyclic polarization test and electrochemical impedance spectroscopy. Surface alterations were studied using a scanning electron microscope. All test electrolytes led to apparent differences in corrosion behavior of the implants. The implants under conditions of test exhibited statistically significant increases in I(corr) from 0.2372 to 1.007 μAcm(-2), corrosion rates from 1.904 to 8.085 mpy, and a decrease in polarization resistances from 304 to 74 Ω. Implants in dextrose-containing solutions were more prone to corrosion than those in Ringer's solutions alone. Increasing the acidity also yielded greater corrosion rates for the dextrose-containing solutions and the solutions without dextrose.


Integrin α6β4 in colorectal cancer.
The ability of cells to interact with extracellular matrix macromolecules is at the forefront of the regulation of cell phenotype and organization. Indeed most if not all cells bear specific cell surface receptors for these molecules, namely the integrins, which are specific for the ligation of various macromolecules such as the laminins, fibronectins and tenascins. It is now well established that integrins can regulate a variety of biological activities, most notably cell cycle and tissue-specific gene expression. In the intestine, several observations suggest functional roles for cell-matrix interactions in the regulation of epithelial cell functions. This article focuses on integrin α6β4 as a paradigm to illustrate the importance as well as the complexity of integrins in the mediation of cell-matrix interactions. Indeed, α6β4 has been well-characterized for its involvement as a link between the cytoskeleton and extracellular matrix molecules as well as in the activation of a variety of intracellular signalization processes in cooperation with growth factor receptors. Furthermore, recent studies show that distinct forms of α6 and β4 subunits are expressed in the human intestine and, more importantly, recent work provides experimental evidence that various forms of α6β4 can differentially regulate intestinal epithelial cell functions under both normal and pathological conditions. For instance, it has been discovered that colorectal cancer cells express a hybrid form of α6β4 that is never seen in normal cells. Although further work is needed, integrin α6β4 is emerging as a key regulator of intestinal functions in both intestinal health and disease.


The application of waterworks sludge ash to stabilize the volume of cement paste.
In order to extend the recycling of waterworks sludge to engineering applications, this paper addresses the influence of nano-SiO2 on incinerated waterworks sludge ash (IWSA) cement paste attacked by sulfate. Tests were performed such as length measurement for volume change, compressive strength, weight loss, and micro-structural testing using scanning electron microscopy (SEM). The results indicate that when a portion of the cement in the paste was replaced by IWSA, the IWSA diluted the cementitious material C3A, and filled the capillary pores in the hardened paste. Moreover, since IWSA has potential pozzolanic activity, it can chemically react with Ca(OH)2 crystals in the paste and can consequently improve the resistance of the paste to sulfate attack. Test results also show that due to the fully developed pozzolanic effect of IWSA, the major reaction products of sulfate attack, gypsum and ettringite, were clearly reduced. Hence, the expansion rate in length decreased with the increase of IWSA replacement. Furthermore, the addition of nano-SiO2 to IWSA cement paste can also reduce the length expansion rate.


Cheek reconstruction: current concepts in managing facial soft tissue loss.
Significant defects of the cheek present a reconstructive challenge due to their extremely visible site, as well as limited local tissue supply. In addition, the cheek abuts several structures of expressive function, such as the eye, mouth, and local facial musculature. To achieve satisfactory functional and aesthetic results, reconstruction of such defects requires careful three-dimensional restoration of all missing components, adequate texture matching, as well as functional restoration. Aesthetic reconstruction of facial defects should adhere to the priority goals of first preserving function and second achieving cosmesis. According to the size of the defect, location on the cheek, relationship to adjacent structures, available donor tissue, and existing skin tension lines, a host of techniques is available for closure. As a well-established principle in facial reconstructive surgery, one should use local tissue whenever possible to provide the best tissue for color and contour restoration. However, thoughtful reliance upon the "reconstructive ladder," including direct closure, skin grafting, local flap creation, regional flap placement, and free-flap repair, will invariably guide the surgeon in an optimal approach to cheek reconstruction.


Valorisation of biodiesel production wastes: Anaerobic digestion of residual Tetraselmis suecica biomass and co-digestion with glycerol.
One of the principal opportunity areas in the development of the microalgal biodiesel industry is the energy recovery from the solid microalgal biomass residues to optimise the fuel production. This work reports the cumulative methane yields reached from the anaerobic digestion of the solid microalgal biomass residues using different types of inocula, reporting also the improvement of biogas production using the co-digestion of microalgal biomass with glycerol. Results demonstrate that the solid microalgal biomass residues showed better biogas production using a mesophilic inoculum, reaching almost two-fold higher methane production than under thermophilic conditions. Furthermore, the solid microalgal biomass residues methane production rate showed an increase from 173.78 ± 9.57 to 438.46 ± 40.50 mL of methane per gram of volatile solids, when the co-digestion with glycerol was performed. These results are crucial to improve the energy balance of the biodiesel production from Tetraselmis suecica, as well as proposing an alternative way to treat the wastes derived from the microalgae biodiesel production.


Tackling tuberculosis patients' internalized social stigma through patient centred care: an intervention study in rural Nicaragua.
We report a patient-centered intervention study in 9 municipalities of rural Nicaragua aiming at a reduction of internalized social stigma in new AFB positive tuberculosis (TB) patients diagnosed between March 2004 and July 2005. Five out of 9 municipal teams were coached to tailor and introduce patient-centered package. New TB patients were assigned to the intervention group when diagnosed in municipalities implementing effectively at least TB clubs and home visits. We compared the changes in internalized stigma and TB treatment outcome in intervention and control groups. The internalized stigma was measured through score computed at 15 days and at 2 months of treatment. The treatment results were evaluated through classical TB program indicators. In all municipalities, we emphasized process monitoring to capture contextual factors that could influence package implementation, including stakeholders. TB clubs and home visits were effectively implemented in 2 municipalities after June 2004 and in 3 municipalities after January 2005. Therefore, 122 patients were included in the intervention group and 146 in the control group. After 15 days, internalized stigma scores were equivalent in both groups. After 2 months, difference between scores was statistically significant, revealing a decreased internalized stigma in the intervention group and not in the control group. This study provides initial evidences that it is possible to act on TB patients' internalized stigma, in contexts where at least patient centered home visits and TB clubs are successfully implemented. This is important as, indeed, TB care should also focus on the TB patient's wellbeing and not solely on TB epidemics control.


"Remnants of feudalism"? Women's health and their utilization of health services in rural China.
Almost five decades ago, the Chinese Communist Party wished to abolish all "remnants of feudalism," including the patriarchal social order. Just one year after the revolution, the Marriage Law endorsed women's rights within the family, but no operative measures were taken to enforce it. Some of the economic reforms since independence even strengthened patrilocality and, possibly, patriarchal values. The purpose of this study was to explore the degree to which patrilocality served to maintain the traditional patriarchal stratification among women in the household by exploring women's health patterns and utilization of health services. Data were collected from 3859 women residing in rural Hebei, and variation in health and help seeking of six categories of relation to household head--mothers, wives, daughters, daughters-in-law, family heads, and other relatives--were explored. Utilization of health services is not dependent on women's position in the household, but primarily on per-capita income. Health patterns seem to indicate that mothers of the head of the household still have a considerable power to define their roles and share of household work. Women head of family, most of whom are married, appear to be under strain, which could be a result of their culturally "deviant" position. We conclude that old patriarchal values are intertwined with values of equality in current rural China.


Autonomic nervous system activation mediates the increase in whole-body glucose uptake in response to electroacupuncture.
A single bout of low-frequency electroacupuncture (EA) causing muscle contractions increases whole-body glucose uptake in insulin-resistant rats. We explored the underlying mechanism of this finding and whether it can be translated into clinical settings. Changes in glucose infusion rate (GIR) were measured by euglycemic-hyperinsulinemic clamp during and after 45 min of low-frequency EA in 21 overweight/obese women with polycystic ovary syndrome (PCOS) and 21 controls matched for age, weight, and body mass index (experiment 1) and in rats receiving autonomic receptor blockers (experiment 2). GIR was higher after EA in controls and women with PCOS. Plasma serotonin levels and homovanillic acid, markers of vagal activity, decreased in both controls and patients with PCOS. Adipose tissue expression of pro-nerve growth factor (proNGF) decreased, and the mature NGF/proNGF ratio increased after EA in PCOS, but not in controls, suggesting increased sympathetic-driven adipose tissue metabolism. Administration of α-/β-adrenergic receptor blockers in rats blocked the increase in GIR in response to EA. Muscarinic and dopamine receptor antagonist also blocked the response but with slower onset. In conclusion, a single bout of EA increases whole-body glucose uptake by activation of the sympathetic and partly the parasympathetic nervous systems, which could have important clinical implications for the treatment of insulin resistance.-Benrick, A., Kokosar, M., Hu, M., Larsson, M., Maliqueo, M., Marcondes, R. R., Soligo, M., Protto, V., Jerlhag, E., Sazonova, A., Behre, C. J., Højlund, K., Thorén, P., Stener-Victorin, E. Autonomic nervous system activation mediates the increase in whole-body glucose uptake in response to electroacupuncture.


[The choice of reinforcement quality depending on the delay time of the instrumental reaction in cats].
In experiments on seven cats instrumental delayed reflexes with different quality reinforcement were elaborated by method of "active choice", where the time of delay of the motor reaction served as a signal of quality of the reinforcement. Five cats used a strategy with long delay high-quality reinforcement (meat), and two cats preferred short delay and low-quality reinforcement (bread-meat mixture). In special experiments with the change of alimentary motivation (24-hour alimentary deprivation and saturation of animals) it was established that the level of alimentary motivation within the studied limits did not change the strategy of behaviour preferred by the given animal. Individually varying ability to choice of reaction determining more valuable reinforcement it is possible to consider as an experimental analogue of the mechanisms of will. The question is discussed about the role of typological properties of cats in choice of behaviour strategy.


FTIR Microspectroscopy Reveals Chemical Changes in Mice Fetus Following Phenobarbital Administration.
Phenobarbital is a phenobarbiturate used as a sedative, anticonvulsant or hypnotic with the doses prescribed and can cause teratogenic effects. The goal of this study was to examine an alternative method for the recognition of the mechanism or the bimolecular potential changes in mice fetus caused by Phenobarbital using FTIR micro spectroscopy. The mice were injected with Phenobarbital (120 mg/Kg) on gestation day 9. Fetuses were dissected on day 15 of gestation and morphological and histological studies on the fetus were carried out. Sections (10 µm) of normal and Phenobarbital treated fetus brains and livers were used for FTIR measurement in the wave number region of 400- 4000 cm(-1). The results were shown by 2(nd) derivatization of spectra and also subtracting from control spectra. In liver, the intensity at 1054 cm(-1), 1155 cm(-1), 1353 cm(-1), 1453cm(-1),1645 cm(-1), 1622 cm(-1), 2944 cm(-1), 2913 cm(-1) and 2845 cm(-1) were shifted and increased. In the brain, the intensity at 879 cm(-1), 911 cm(-1), 955 cm(-1), 1223 cm(-1), 1256 cm(-1), 1304 cm(-1), 1360 cm(-1), 1453 cm(-1), 1529 cm(-1), 1636 cm(-1), 2845 cm(-1), 2915 cm(-1) and 2950 cm(-1) were increased and shifted. The most important changes of the fetus brain tissue are on the β structure of proteins due to the amide I bands at 1636 cm(-1), while extensive effects on the DNA structure were obvious for the Phenobarbital treated liver tissues. As a conclusion, FTIR spectroscopy might well be assumed as a potentially powerful teratogenic measurement instrument with a unique ability to identify the modified bimolecular structures.


Association between antimicrobial usage, biosecurity measures as well as farm performance in German farrow-to-finish farms.
Antimicrobial usage in food producing animals is of major concern. A clear link between the extent of use and the development of antimicrobial resistance has already been demonstrated. To evaluate strategies that may reduce the antimicrobial usage while assuring pig health and welfare, it requires profound knowledge of factors that are associated with antimicrobial usage. Data on biosecurity and herd management practices are important parameters to identify risk factors which are related to a higher antimicrobial usage. To investigate between-farm variations of high and low usage the treatment incidence (TI) per age group in 60 German farrow-to-finish herds was qualitatively and quantitatively analysed and linked to biosecurity measures, and herd management characteristics. Weaned pigs received most of the treatments (median TI = 487.6), followed by suckling pigs (median TI = 138.9). Suckling pigs were treated with critically important antimicrobials (3rd and 4th generation cephalosporines) to a remarkable extent. The number of sows present at site (p < 0.01) and a low score for external biosecurity (p = 0.06) were associated with a higher antimicrobial usage in pigs from birth till slaughter. Herds with a higher treatment incidence in growing pigs (TI 200 days): i) were located in a region with a high pig density (p < 0.01), ii) had a less strict access check for visitors and personnel (p < 0.01) and iii) scored lower in the subcategory 'cleaning and disinfection' (internal biosecurity) (p < 0.01). Herds with a higher treatment incidence in breeding pigs weaned more piglets per sow and year and scored better in the internal biosecurity level (p = 0.02). With the main focus on the treatment incidence in pigs from birth till slaughter and in breeding pigs risk factors for a high usage in these age groups were identified. The level of biosecurity of a herd was associated with the amount of antimicrobials used. Therefore, the findings in this study indicate possible points of action in the reduction and prudent use of antimicrobials in Germany. The active improvement of biosecurity measures could be a promising alternative to reduce antimicrobial usage on herd level.


Substitution reactions in metal-organic frameworks and metal-organic polyhedra.
Substitution reaction, as one of the most powerful and efficient chemical reactions, has been widely used in various syntheses, including those for the design and preparation of functional molecules or materials. In the past decade, a class of newly developed inorganic-organic hybrid materials, namely metal-organic materials (MOMs), has experienced a rapid development. MOMs are composed of metal-containing nodes connected by organic linkers through strong chemical bonds, and can be divided into metal-organic frameworks (MOFs) and metal-organic polygons/polyhedra (MOPs) with infinite and discrete structural features, respectively. Recent research has shown that the substitution reaction can be used as a new strategy in the synthesis and modification of MOFs and MOPs, particularly for pre-designed ones with desired structures and functions, which are usually difficult to access by a direct one-pot self-assembly synthetic approach. This review highlights the implementation of the substitution reaction in MOFs and MOPs. Examples of substitution reactions at metal ions, organic ligands, and free guest molecules of MOFs and MOPs are listed and analyzed. The changes or modifications in the structures and/or properties of these materials induced by the substitutions, as well as the nature of the associated reaction, are discussed, with the conclusion that the substitution reaction is really feasible and powerful in synthesizing and tailoring MOMs.


Stereological investigation of the effect of Elaeagnus angustifolia fruit hydroalcoholic extract on osteoporosis in ovariectomized rats.
Postmenopausal osteoporosis is characterized by increased fracture risk. However, each approved treatment has specific side effects. Therefore, foods with plant origins have increasingly attracted attention as an alternative treatment. Studies have shown that Elaeagnus angustifolia (EA) has antioxidant properties. The present study aimed to investigate the effects of EA hydroalcoholic extract on ovariectomy-induced osteoporosis in rats using stereological methods. 55 female Sprague-Dawley rats were randomly assigned to control, sham operated (normal saline), ovariectomized (OVX), OVX + EA fruit extract (600 mg/kg BW/day), and OVX + estradiol benzoate (3 mg/kg BW) for 16 weeks. Blood samples were collected to measure calcium, phosphorus, and alkaline phosphatase (ALP) plasma levels. Then, specimens from tibia and fifth lumbar vertebra (L5) bones were prepared and stereological analysis was done. Ovariectomy significantly decreased the calcium level and increased the ALP level in the OVX group. In spite of improvement in calcium hemostasis in groups treated with estrogen and EA fruit extract (p<0.05), only treatment with estrogen was able to reduce ALP levels. Moreover, treatment with EA fruit extract and estrogen caused a significant increase in the number of osteoblasts in vertebra and tibia compared to the OVX group (p<0.05). Estrogen and EA fruit extract were also able to reduce the number of osteoclasts in tibia of the treated OVX rats (p<0.05). The results showed that EA extract exerted more effects, markedly, on osteoblastogenesis in the OVX rats. Thus, it could be considered as a potential agent to treat patients with osteoporosis.


Detection of circulating tumor cells in the cerebrospinal fluid of a patient with a solitary metastasis from breast cancer: A case report.
Brain lesions identified following the diagnosis and eradication of primary cancers are often ambiguous in origin, existing as a solitary metastasis or an independent primary brain tumor. The brain is a relatively common site of metastasis with breast cancer, although determining whether metastases have originated from the breast or brain is often not possible without invasive biopsies. In the current case report, a patient presented with a brain lesion identified by radiography and was without systemic disease. The patient had previously exhibited a complete response to chemotherapy and surgery for a poorly differentiated invasive ductal carcinoma. The origin of the brain lesion could not be determined by magnetic resonance imaging, giving rise to a diagnostic dilemma with diverging treatment options. We previously reported a method to isolate and enumerate tumor cells of epithelial origin in the cerebrospinal fluid (CSF). CSF tumor cell analysis of the patient revealed massive CSF tumor cell burden of epithelial origin, indicating that the brain lesion was likely of breast origin. The current case report highlights the use of CSF tumor cell detection as a differential diagnostic tool, in addition to its previously demonstrated use as a marker of disease burden and therapeutic response.


Development of Behaviorally Based Monitoring and Biosurveillance Tools for the Invasive Spotted Lanternfly (Hemiptera: Fulgoridae).
The spotted lanternfly, Lycorma delicatula White, is an invasive planthopper (Hemiptera: Fulgoridae) that was first detected in the United States in Berks County, PA, in 2014, and has since spread in the mid-Atlantic region. This phloem-feeding pest has a broad host range, including economically important crops such as grape where their feeding causes dieback of infested plants. Monitoring the presence and abundance of L. delicatula is of utmost importance to develop pest management approaches. Current monitoring practices include sticky bands deployed on tree trunks, sometimes paired with commercially available methyl salicylate lures. A drawback associated with sticky bands is the high numbers of nontarget captures. Here, we developed traps for L. delicatula based on a circle trap originally designed for weevils. These traps are comprised of a screen funnel that wraps around the trunk of a tree and guides individuals walking up the trunk into a collection device. In 2018 and 2019, we compared circle trap designs with sticky bands in Pennsylvania and Virginia. In both years, circle trap designs yielded captures that were equivalent to or exceeded captures of L. delicatula on sticky bands. Nontarget captures were significantly lower for circle traps compared with sticky bands. Presence of a methyl salicylate lure in association with traps deployed on host trees or vertical tree-mimicking posts did not increase L. delicatula captures compared with unbaited traps. Circle traps, modified using vinyl screen and a larger collection device, present an alternative to the current approach with reduced nontarget capture for monitoring L. delicatula.


Nerve regeneration through holey silicone tubes.
Recent studies focus on regeneration where nerve stumps are placed in a silicone tube. Since the tube is impermeable, the fluid and cells that collect from the stumps bath the axons. This is presumably beneficial. Making the tube permeable by making holes in its walls should change the patterns of regeneration. If this is done, the major cytologic change is an increase in the fascicular perineurium. There are more individual fascicles, more cells line each fascicle and the lining cells are coated by more prominent external laminae than after similar regeneration in a regular silicone tube or in the normal untransected nerve. For axonal numbers, there are more myelinated and unmyelinated axons in the gap and more unmyelinated axons in the distal stump than after regeneration in a regular silicone tube. The numbers in the holey tube regenerate are statistically different from normal but they are closer to normal than after similar regeneration in a regular silicone tube. There are significantly fewer myelinated and unmyelinated axons than in the normal sural nerve after regeneration through a holey tube, but there are more than after regeneration through a regular tube. The numbers of axons in the nerve to the medial gastrocnemius muscle are not significantly different from normal or from the other regeneration paradigms. These data allow the suggestion that regeneration through a silicone tube with macroscopic holes in its walls may be superior in certain respects to regeneration through a regular impermeable silicone tube.


Changing patterns in HIV-1 non-B clade prevalence and diversity in Italy over three decades.
HIV-1 non-B subtypes have recently entered Western Europe following immigration from other regions. The distribution of non-B clades and their association with demographic factors, over the entire course of the HIV-1 epidemic, have not been fully investigated in Italy. We carried out a phylogenetic analysis of HIV-1 pol sequences derived from 3670 patients followed at 50 Italian clinical centres over nearly three decades. Overall, 417 patients (11.4%) carried non-B subtypes. The prevalence of non-B strains increased from 2.6% in 1980-1992 to 18.9% in 1993-2008 (P<0.0001) in a subset of 2479 subjects with a known year of diagnosis. A multivariate analysis on a subset of 1364 patients for whom relevant demographic data were available indicated that African ethnicity, heterosexual route of infection and year of diagnosis were independently associated with non-B HIV-1 infection (P ≤ 0.0001). All pure subtypes, except for clade K, and seven circulating recombinant forms were detected, accounting for 56.6 and 34.1% of the non-B infections, respectively. The F1 subtype was the most prevalent non-B clade among Europeans and was acquired heterosexually in half of this patient population. Unique recombinant forms accounted for 9.4% of the non-B sequences and showed a B/F1 recombination pattern in one-third of cases. The circulation of non-B clades has significantly increased in Italy in association with demographic changes. Spread of the F1 subtype and B/F recombinants appears to predominate, which may result in a redistribution of the relative proportions of the different strains, and this could lead to overlapping epidemics. Thus, the HIV-1 landscape in Italy may in future be distinct from that of the rest of Europe.


Providing access to risk prediction tools via the HL7 XML-formatted risk web service.
Cancer risk prediction tools provide valuable information to clinicians but remain computationally challenging. Many clinics find that CaGene or HughesRiskApps fit their needs for easy- and ready-to-use software to obtain cancer risks; however, these resources may not fit all clinics' needs. The HughesRiskApps Group and BayesMendel Lab therefore developed a web service, called "Risk Service", which may be integrated into any client software to quickly obtain standardized and up-to-date risk predictions for BayesMendel tools (BRCAPRO, MMRpro, PancPRO, and MelaPRO), the Tyrer-Cuzick IBIS Breast Cancer Risk Evaluation Tool, and the Colorectal Cancer Risk Assessment Tool. Software clients that can convert their local structured data into the HL7 XML-formatted family and clinical patient history (Pedigree model) may integrate with the Risk Service. The Risk Service uses Apache Tomcat and Apache Axis2 technologies to provide an all Java web service. The software client sends HL7 XML information containing anonymized family and clinical history to a Dana-Farber Cancer Institute (DFCI) server, where it is parsed, interpreted, and processed by multiple risk tools. The Risk Service then formats the results into an HL7 style message and returns the risk predictions to the originating software client. Upon consent, users may allow DFCI to maintain the data for future research. The Risk Service implementation is exemplified through HughesRiskApps. The Risk Service broadens the availability of valuable, up-to-date cancer risk tools and allows clinics and researchers to integrate risk prediction tools into their own software interface designed for their needs. Each software package can collect risk data using its own interface, and display the results using its own interface, while using a central, up-to-date risk calculator. This allows users to choose from multiple interfaces while always getting the latest risk calculations. Consenting users contribute their data for future research, thus building a rich multicenter resource.


Epididymal scanning electron microscopy study in hypothyroid rats.
The purpose of this research was to investigate, using scanning electron microscopy, the effect of hypothyroidism on the ultrastructure of the rat cauda epididymis. Thyroidectomy was obtained by ip injection of 270 muCi of 131I per rat. One month later, several portions of cauda epididymis were examined. Morphological and physiological differences were detected in the cauda epididymis of the hypothyroid animals when compared to the control normal rats. The hypothyroid condition was associated with important changes in the luminal surface of the cauda epididymis epithelium. Broken, oblique, and loss of stereocilia, denuded epithelial cells, and flattening of the tubule were observed. The results confirm that hypothyroidism causes marked structural changes in the cauda ductus epididymis and could be adversely affect sperm maturation motility.


Validation of Assessing Arthroscopic Skill using the ASSET Evaluation.
The Accreditation Council for Graduate Medical Education and the American Board of Orthopaedic Surgery have implemented "milestones" to evaluate residents during their progression in medical education. The purpose of this study was to determine whether a validated evaluation tool correlates with surgical experience, year in training, and progression over time. This was a retrospective study of already collected curriculum assessment data where 2 unbiased, blinded orthopedic surgeons evaluated resident performance on basic diagnostic knee arthroscopy using the Arthroscopic Surgical Skills Evaluation Tool (ASSET) over 3 years. Residents also gained arthroscopy experience through a structured arthroscopy curriculum and clinical experience. The study was conducted at the TRIA Orthopaedic Center (Bloomington, Minnesota, USA), an institutional site for The University of Minnesota orthopedic surgery residency program. Eleven orthopedic surgery residents at postgraduate years 2 to 5 were evaluated using the ASSET. The Pearson's Correlation Coefficient was used to validate both the number of arthroscopic procedures performed by residents (r = 0.946) and their level in training (r = 0.89). Residents who were re-evaluated after undergoing the arthroscopy curriculum throughout the year displayed significant increases in total ASSET scores (p < 0.01). Resident performance on the ASSET correlated with arthroscopic experience based on year-in training. More importantly, performance improved with additional years of training, demonstrating validity over time. The data also demonstrates interobserver reliability. Due to these correlations between exposure to surgery and score on the ASSET, we believe the tool could serve as a suitable means for assessing residents' technical proficiency as required by The Accreditation Council for Graduate Medical Education program guidelines.


The evolution of male genitalia: patterns of genetic variation and covariation in the genital sclerites of the dung beetle Onthophagus taurus.
Three main hypotheses, have been invoked to explain divergent genital evolution, the lock and key, pleiotropy, and sexual selection hypotheses, each of which make different predictions about how genital traits are inherited. Here we used a half-sib breeding design to examine the patterns of genetic variation and covariation between male genital sclerites, and their covariance with general body morphology in the dung beetle Onthophagus taurus. We found CV(A)'s and CV(P)'s were similar for both genital and general morphological traits and that CV(R)'s were large for both trait types. We found that male genital sclerites were negatively genetically correlated with general morphological traits. Variation in male genital morphology has direct implications for a male's fertilization success and the resulting sexual selection acting on male genitalia is predicted to maintain high levels of additive genetic variance. Contrary to this prediction, we found that individual genital sclerites all had low levels of additive genetic variance and large maternal and environmental sources of variation. Our data suggest that the genital sclerites in O. taurus are not inherited independently but as a genetically integrated unit. More importantly, the way the different sclerites function to influence male fertilization success reflects this genetic integration. Even though levels of V(A) in individual genital sclerites may be low, there may still be sufficient V(A) in multivariate trait space for selection to generate evolutionary change in the overall morphology of male genitalia.


Succinate dehydrogenase inhibition with malonate during reperfusion reduces infarct size by preventing mitochondrial permeability transition.
Previous studies demonstrated that pre-treatment with malonate, a reversible inhibitor of succinate dehydrogenase, given before ischaemia, reduces infarct size. However, it is unknown whether administration of malonate may reduce reperfusion injury. Isolated mice hearts were treated, under normoxic conditions, with increasing concentrations of disodium malonate (0.03-30 mmol/L, n = 4). Malonate induced a concentration-dependent decrease in left ventricular developed pressure (LVdevP) (EC50 = 8.05 ± 2.11 mmol/L). In isolated hearts submitted to global ischaemia (35 min) followed by reperfusion (60 min), malonate 3 mmol/L given only during the first 15 min of reperfusion reduced lactate dehydrogenase release (125.41 ± 16.82 vs. 189.20 ± 13.74 U/g dry tissue/15 min in controls, P = 0.015) and infarct size (24.57 ± 2.32 vs. 39.84 ± 2.78%, P = 0.001, n = 7-8 per group) and improved recovery of LVdevP (20.06 ± 3.82 vs 7.76 ± 2.53% of baseline LVdevP, P = 0.017). (1)H NMR spectroscopy demonstrated marked changes in the metabolic profile of malonate-treated hearts, including increased accumulation of succinate. Furthermore, malonate reduced reactive oxygen species (ROS) production, as measured by MitoSOX staining in myocardial samples obtained after 5 min of reperfusion and in mitochondrial preparations from these samples, preserved mitochondrial respiration, and reduced mitochondrial permeabilization, assessed by calcein retention. Treatment with malonate did not result in activation of RISK or SAFE signalling pathways in tissue extracts obtained 5 min after reperfusion. Succinate dehydrogenase inhibition with malonate at the onset of reperfusion reduces infarct size in isolated mice hearts through reduction in ROS production and mitochondrial permeability transition pore opening.


[Experience with using atropine in comatose and non-comatose doses in psychiatric clinical practice].
Treatment with atropine in comatose doses was given to 11 patients suffering from schizophrenia with obsessive disturbances. A favourable effect was noted when there were no hypochondriacal symptoms in the disease clinical picture. Twenty-two patients with neurotic states received atropine in non-comatose doses. Positive results were noted in patients with obsessions in the disease clinical picture, as well as in patients with reactive depressions. In patients with hypochondriacal disturbances, this form of the therapy appeared to be of no effect.


Stimuli-responsive surface crystallization of phospholipids from bimodal colloidal particles.
These studies focus on the effect of phospholipids in the presence of ionic surfactants on the behavior of poly(methylmethactrylate/n-butyl acrylate) (p-MMA/nBA) colloidal particles during film formation. With the presence of two surfactants, it is possible to obtain particles that exhibit two distinct particle sizes. The presence of hydrogenated soybean phosphatidylcholine (HSPC) and sodium dioctyl sulfosuccinate (SDOSS), which stabilize these bimodal colloidal dispersions, has a significant effect on the mobility of individual components during coalescence. Specifically, the presence of HSPC inhibits migration of SDOSS to the film-air (F-A) interface. Furthermore, the presence of electrolyte species such as aqueous CaCl2 has a very pronounced effect on film formation. When the Ca2+/HSPC ratio is 0.1/1.0, SDOSS is released to the F-A interface during coalescence. At 2.0/1.0 Ca2+/HSPC, HSPC diffuses to the F-A interface and crystalline domains consisting of HSPC are formed. This stimuli-responsive behavior is confirmed using IRIR imaging that ultimately exhibits different surface morphologies. These studies illustrate for the first time that it is possible to control the release of two different surface-active species during coalescence that form crystalline domains.


The enteropathogenic Escherichia coli (EPEC) Map effector is imported into the mitochondrial matrix by the TOM/Hsp70 system and alters organelle morphology.
Enteropathogenic Escherichia coli (EPEC) is a human intestinal pathogen and a major cause of diarrhoea, particularly among infants in developing countries. EPEC target the Map and EspF multifunctional effector proteins to host mitochondria - organelles that play crucial roles in regulating cellular processes such as programmed cell death (apoptosis). While both molecules interfere with the organelles ability to maintain a membrane potential, EspF plays the predominant role and is responsible for triggering cell death. To learn more about the Map-mitochondria interaction, we studied Map localization to mitochondria with purified mitochondria (from mammalian and yeast cells) and within intact yeast. This revealed that (i) Map targeting is dependent on the predicted N-terminal mitochondrial targeting sequence, (ii) the N-terminal 44 residues are sufficient to target proteins to mitochondria and (iii) Map import involves the mitochondrial outer membrane translocase (Tom22 and Tom40), the mitochondrial membrane potential, and the matrix chaperone, mtHsp70. These results are consistent with Map import into the mitochondria matrix via the classical import mechanism. As all known, Map-associated phenotypes in mammalian cells are independent of mitochondrial targeting, this may indicate that import serves as a mechanism to remove Map from the cytoplasm thereby regulating cytoplasmic function. Intriguingly, Map, but not EspF, alters mitochondrial morphology with deletion analysis revealing important roles for residues 101-152. Changes in mitochondrial morphology have been linked to alterations in the ability of these organelles to regulate cellular processes providing a possible additional role for Map import into mitochondria.


Retrospective evaluation of the eye irritation potential of agrochemical formulations.
Multiple in vitro eye irritation methods have been developed and adopted as OECD health effects test guidelines. However, for predicting the ocular irritation/damage potential of agrochemical formulations there is an applicability domain knowledge gap for most of the methods. To overcome this gap, a retrospective evaluation of 192 agrochemical formulations with in vivo (OECD TG 405) and in vitro (OECD TG 437, 438, and/or 492) data was conducted to determine if the in vitro methods could accurately assign United Nations Globally Harmonized System for Classification and Labelling of Chemicals (GHS) eye irritation hazard classifications. In addition, for each formulation the eye irritation classification was derived from the classification of the contained hazardous ingredients and their respective concentration in the product using the GHS concentration threshold (CT) approach. The results herein suggest that the three in vitro methods and the GHS CT approach were highly predictive of formulations that would not require GHS classification for eye irritation. Given most agrochemical formulations fall into this category, methods that accurately identify non-classified agrochemical formulations could significantly reduce the use of animals for this endpoint.


The potential of mycelium and culture broth of Lignosus rhinocerotis as substitutes for the naturally occurring sclerotium with regard to antioxidant capacity, cytotoxic effect, and low-molecular-weight chemical constituents.
Previous studies on the nutritional and nutraceutical properties of Lignosus rhinocerotis focused mainly on the sclerotium; however, the supply of wild sclerotium is limited. In this investigation, the antioxidant capacity and cytotoxic effect of L. rhinocerotis cultured under different conditions of liquid fermentation (shaken and static) were compared to the sclerotium produced by solid-substrate fermentation. Aqueous methanol extracts of the mycelium (LR-MH, LR-MT) and culture broth (LR-BH, LR-BT) demonstrated either higher or comparable antioxidant capacities to the sclerotium extract (LR-SC) based on their radical scavenging abilities, reducing properties, metal chelating activities, and inhibitory effects on lipid peroxidation. All extracts exerted low cytotoxicity (IC50>200 µg/ml, 72 h) against selected mammalian cell lines. Several low-molecular-weight compounds, including sugars, fatty acids, methyl esters, sterols, amides, amino acids, phenolics, and triterpenoids, were identified using GC-MS and UHPLC-ESI-MS/MS. The presence of proteins (<40 kDa) in the extracts was confirmed by SDS-PAGE and SELDI-TOF-MS. Principal component analysis revealed that the chemical profiles of the mycelial extracts under shaken and static conditions were distinct from those of the sclerotium. Results from bioactivity evaluation and chemical profiling showed that L. rhinocerotis from liquid fermentation merits consideration as an alternative source of functional ingredients and potential substitute for the sclerotium.


The characteristics of alcoholics frequently lost to follow-up.
In a follow-up of 85 alcoholic men, 93% were interviewed or confirmed deceased 8 years after discharge from hospital. The sample was also followed up at 1 and 3 years postdischarge. Follow-up rates of other published studies are discussed. Scales to assess difficulty of interview and difficulty of location were developed. Factor analysis revealed the differential characteristics of subjects defined as difficult to locate, difficult to interview or missing. Subjects who are difficult to locate, or who in the extreme case will go missing, tended to have poorer social functioning prior to intake and to be residentially unstable during the follow-up period, characteristics that tend to correlate with worse drinking outcome. Subjects who are unwilling to be interviewed tended to be residentially stable and show better interpersonal adjustment at follow-up. Less intensive location procedures would have resulted in data loss from those classified as difficult to locate. Less persuasive interview techniques would have resulted in data loss from those classified as difficult to interview. Early termination of the follow-up would also have resulted in data loss from the latter group. The distinct types of data biases that would be introduced by loss of information from each of the above subgroups are examined.


Early lexical development in a self-organizing neural network.
In this paper we present a self-organizing neural network model of early lexical development called DevLex. The network consists of two self-organizing maps (a growing semantic map and a growing phonological map) that are connected via associative links trained by Hebbian learning. The model captures a number of important phenomena that occur in early lexical acquisition by children, as it allows for the representation of a dynamically changing linguistic environment in language learning. In our simulations, DevLex develops topographically organized representations for linguistic categories over time, models lexical confusion as a function of word density and semantic similarity, and shows age-of-acquisition effects in the course of learning a growing lexicon. These results match up with patterns from empirical research on lexical development, and have significant implications for models of language acquisition based on self-organizing neural networks.


Accretion and core formation: constraints from metal-silicate partitioning.
Experimental metal-silicate partitioning data for Ni, Co, V, Cr, Nb, Mn, Si and W were used to investigate the geochemical consequences of a range of models for accretion and core formation on Earth. The starting assumptions were chondritic ratios of refractory elements in the Earth and the segregation of metal at the bottom of a magma ocean, which deepened as the planet grew and which had, at its base, a temperature close to the liquidus of the silicate. The models examined were as follows. (i) Continuous segregation from a mantle which is chemically homogeneous and which has a fixed oxidation state, corresponding to 6.26 per cent oxidized Fe. Although Ni, Co and W partitioning is consistent with chondritic ratios, the current V content of the silicate Earth cannot be reconciled with core segregation under these conditions of fixed oxidation state. (ii) Continuous segregation from a mantle which is chemically homogeneous but in which the Earth became more oxidized as it grew. In this case, the Ni, Co, W, V, Cr and Nb contents of core and mantle are easily matched to those calculated from the chondritic ratios of refractory elements. The magma ocean is calculated to maintain a thickness approximately 35 per cent of the depth to the core-mantle boundary in the accreting Earth, yielding a maximum pressure of 44GPa. This model yields a Si content of the core of 5.7 per cent, in good agreement with cosmochemical estimates and with recent isotopic data. (iii) Continuous segregation from a mantle which is not homogeneous and in which the core equilibrates with a restricted volume of mantle at the base of the magma ocean. This is found to increase depth of the magma ocean by approximately 50 per cent. All of the other elements (except Mn) have partitioning consistent with chondritic abundances in the Earth, provided the Earth became, as before, progressively oxidized during accretion. (iv) Continuous segregation of metal from a crystal-melt mush. In this case, pressures decrease to a maximum of 31GPa and it is extremely difficult to match the calculated mantle contents of the highly incompatible elements Nb and W to those observed. Progressive oxidation is required to fit the observed mantle contents of vanadium. All of the scenarios discussed above point to progressive oxidation having occurred as the Earth grew. The Earth appears to be depleted in Mn relative to the chondritic reference.


Simultaneous quantification of four native estrogen hormones at trace levels in human cerebrospinal fluid using liquid chromatography-tandem mass spectrometry.
Estrogens are known to exhibit neuroprotective effects on the brain. Their importance in this regard and in others has been emphasized in many recent studies, which increases the need to develop reliable analytical methods for the measurement of estrogen hormones. A heart-cutting two-dimensional liquid chromatography separation method coupled with electrospray ionization-tandem mass spectrometry (ESI-MS/MS) has been developed for simultaneous measurement of four estrogens, including estriol (E3), estrone (E1), 17β-estradiol (17β-E2), and 17α-estradiol (17α-E2), in human cerebrospinal fluid (CSF). The method was based on liquid-liquid extraction and derivatization of estrogens with dansyl chloride to enhance the sensitivity of ESI-based detection in conjunction with tandem mass spectrometry. Dansylated estriol and estrone were separated in the first dimension by an amide-C18 column, while dansylated 17β- and 17α-estradiol were resolved on the second dimension by two C18 columns (175 mm total length) connected in series. This is the first report of a method for simultaneous quantification of all four endogenous estrogen compounds in their dansylated form. The detection limits for E1, 17α-E2, 17β-E2, and E3 were 19, 35, 26, and 61pg/mL, respectively. Due to matrix effects, validation and calibration was carried out in charcoal-stripped CSF. The precision and accuracy were more than 86% for the two E2 compounds and 79% for E1 and E3 while the extraction recovery ranged from 91% to 104%. The method was applied to measure estrogens obtained in a clinical setting, from the CSF of ischemic trauma patients. While 17β-estradiol was present at a significant level in the CSF of some samples, other estrogens were present at lower levels or were undetectable.


Liriodendrin exerts protective effects against chronic endometritis in rats by modulating gut microbiota composition and the arginine/nitric oxide metabolic pathway.
Chronic endometritis (CE), a gynecological disease, is characterized by inflammation. Liriodendrin is reported to exhibit anti-inflammatory properties. However, the therapeutic effects of liriodendrin on CE and the underlying molecular mechanisms have not been elucidated. This study aimed to investigate the therapeutic effects of liriodendrin on CE in rats and the underlying mechanisms. A CE rat model was established and administered with liriodendrin for 21 days. The serum levels of inflammatory cytokines were examined using enzyme-linked immunosorbent assay. The uterine mRNA levels of cytokines were examined using quantitative real-time polymerase chain reaction analysis. The activation of the Toll-like receptor 4 (TLR4)/NF-κB pathway was investigated using western blotting analysis. The effects of liriodendrin on intestinal flora and serum metabolites were examined using 16S rRNA sequencing and untargeted serum metabolomics, respectively. The protein and mRNA levels of arginase-2 (Arg-2) and the nitric oxide (NO) metabolic pathway-related factors were assessed. Molecular docking was performed to explore the interaction between liriodendrin and Arg-2. Liriodendrin alleviated the CE-induced pathological changes in the uterus, modulated the serum levels of inflammatory cytokines, and downregulated the mRNA and protein levels of TLR4/NF-κB pathway-related factors. Treatment with liriodendrin mitigated the CE-induced upregulation of Firmicutes/Bacteroidetes ratio and Lachnospiraceae abundance and downregulation of Ruminococcaceae abundance. Serum metabolomic analysis revealed that liriodendrin regulated the biosynthesis of choline metabolism pathway-related factors. Liriodendrin suppressed the CE-induced upregulation of Arg-2 and downregulation of inducible nitric oxide synthase (iNOS) expression, and NO levels by directly binding to the amino acid residues of Arg-2 through hydroxyl bonds. Liriodendrin exerted therapeutic effects on CE in rats through the alleviation of inflammation by modulating the gut microbiota structure, directly downregulating Arg-2, and regulating the arginine/NO metabolic pathway.


Effects of berberine, curcumin, resveratrol alone and in combination with chemotherapeutic drugs and signal transduction inhibitors on cancer cells-Power of nutraceuticals.
Over the past fifty years, society has become aware of the importance of a healthy diet in terms of human fitness and longevity. More recently, the concept of the beneficial effects of certain components of our diet and other compounds, that are consumed often by different cultures in various parts of the world, has become apparent. These "healthy" components of our diet are often referred to as nutraceuticals and they can prevent/suppress: aging, bacterial, fungal and viral infections, diabetes, inflammation, metabolic disorders and cardiovascular diseases and have other health-enhancing effects. Moreover, they are now often being investigated because of their anti-cancer properties/potentials. Understanding the effects of various natural products on cancer cells may enhance their usage as anti-proliferative agents which may be beneficial for many health problems. In this manuscript, we discuss and demonstrate how certain nutraceuticals may enhance other anti-cancer drugs to suppress proliferation of cancer cells.


Silicon Metal-oxide-semiconductor Quantum Dots for Single-electron Pumping.
As mass-produced silicon transistors have reached the nano-scale, their behavior and performances are increasingly affected, and often deteriorated, by quantum mechanical effects such as tunneling through single dopants, scattering via interface defects, and discrete trap charge states. However, progress in silicon technology has shown that these phenomena can be harnessed and exploited for a new class of quantum-based electronics. Among others, multi-layer-gated silicon metal-oxide-semiconductor (MOS) technology can be used to control single charge or spin confined in electrostatically-defined quantum dots (QD). These QD-based devices are an excellent platform for quantum computing applications and, recently, it has been demonstrated that they can also be used as single-electron pumps, which are accurate sources of quantized current for metrological purposes. Here, we discuss in detail the fabrication protocol for silicon MOS QDs which is relevant to both quantum computing and quantum metrology applications. Moreover, we describe characterization methods to test the integrity of the devices after fabrication. Finally, we give a brief description of the measurement set-up used for charge pumping experiments and show representative results of electric current quantization.


Detergent and antigen fragility affect the ELISA for measurement of anti-prothrombin autoantibodies.
Some investigators have reported that anti-prothrombin autoantibodies (aPT) in lupus anticoagulant positive sera were detectable by ELISA. Discrepancies in aPT ELISA were observed by some investigators. To clarify this situation, we tested the binding of aPT positive sera to purified prothrombin under various conditions. We performed aPT ELISA under different conditions. The variables we tested were: ELISA plate (untreated or gamma irradiated polystyrene plates), buffer (phosphate buffered saline or Tris buffered saline), detergent (presence or absence of Tween-20), and antigen condition (intact or fragmented prothrombin). Anti-PT from patients with lupus or antiphospholipid syndrome were similarly bound to prothrombin with both buffers. Addition of Tween-20 to the buffer increased reactivity in the irradiated plate assay, but decreased reactivity in the untreated plate assay. Reactivities in 90% of lupus sera were decreased by the use of fragmented prothrombin. In contrast, the reactivity of serum from a healthy subject was remarkably increased by antigen fragmentation. The discrepant ELISA results in measurement of aPT in the various reports may have been due to the use of detergent in the buffer and condition of the prothrombin used as antigen. In our experiments the best ELISA condition for measurement of aPT was achieved using buffer with Tween-20 detergent, with prothrombin directly coated onto irradiated polystyrene plates.


Cognitive Impairment Is Associated with Absolute Intraoperative Frontal α-Band Power but Not with Baseline α-Band Power: A Pilot Study.
Cognitive abilities decline with aging, leading to a higher risk for the development of postoperative delirium or postoperative neurocognitive disorders after general anesthesia. Since frontal α-band power is known to be highly correlated with cognitive function in general, we hypothesized that preoperative cognitive impairment is associated with lower baseline and intraoperative frontal α-band power in older adults. Patients aged ≥65 years undergoing elective surgery were included in this prospective observational study. Cognitive function was assessed on the day before surgery using six age-sensitive cognitive tests. Scores on those tests were entered into a principal component analysis to calculate a composite "g score" of global cognitive ability. Patient groups were dichotomized into a lower cognitive group (LC) reaching the lower 1/3 of "g scores" and a normal cognitive group (NC) consisting of the upper 2/3 of "g scores." Continuous pre- and intraoperative frontal electroencephalograms (EEGs) were recorded. EEG spectra were analyzed at baseline, before start of anesthesia medication, and during a stable intraoperative period. Significant differences in band power between the NC and LC groups were computed by using a frequency domain (δ 0.5-3 Hz, θ 4-7 Hz, α 8-12 Hz, β 13-30 Hz)-based bootstrapping algorithm. Of 38 included patients (mean age 72 years), 24 patients were in the NC group, and 14 patients had lower cognitive abilities (LC). Intraoperative α-band power was significantly reduced in the LC group compared to the NC group (NC -1.6 [-4.48/1.17] dB vs. LC -6.0 [-9.02/-2.64] dB), and intraoperative α-band power was positively correlated with "g score" (Spearman correlation: r = 0.381; p = 0.018). Baseline EEG power did not show any associations with "g." Preoperative cognitive impairment in older adults is associated with intraoperative absolute frontal α-band power, but not baseline α-band power.


Allergic eye disease.
Ocular allergy is one of the most common conditions encountered by pediatricians and ophthalmologists and is characterized by bilateral injection with itching as the predominant symptom. Risk factors include history of atopy (asthma, eczema, seasonal allergies). Basic and clinical research have provided insight into the immunologic mechanisms, clinical presentation, differential diagnosis, and pharmacologicmanagement of this condition. New pharmacologic agents have improved the efficacy and safety of ocularallergy treatment. This article discusses the classification of ocular allergy diagnosis and management, and addresses clinical symptoms and signs that indicate more severe allergic disease or alternative diagnosis that should prompt expeditious referral to an ophthalmologist.


Murine trophoblast organoids as a model for trophoblast development and CRISPR-Cas9 screening.
The placenta becomes one of the most diversified organs during placental mammal radiation. The main in vitro model for studying mouse trophoblast development is the 2D differentiation model of trophoblast stem cells, which is highly skewed to certain lineages and thus hampers systematic screens. Here, we established culture conditions for the establishment, maintenance, and differentiation of murine trophoblast organoids. Murine trophoblast organoids under the maintenance condition contain stem cell-like populations, whereas differentiated organoids possess various trophoblasts resembling placental ones in vivo. Ablation of Nubpl or Gcm1 in trophoblast organoids recapitulated their deficiency phenotypes in vivo, suggesting that those organoids are valid in vitro models for trophoblast development. Importantly, we performed an efficient CRISPR-Cas9 screening in mouse trophoblast organoids using a focused sgRNA (single guide RNA) library targeting G protein-coupled receptors. Together, our results establish an organoid model to investigate mouse trophoblast development and a practicable approach to performing forward screening in trophoblast lineages.