Diagnostic imaging of the diabetic foot.
Diabetic foot syndrome is a significant complication of diabetes. Diagnostic imaging is a crucial factor determining surgical decision and extent of surgical intervention. At present the gold standard is MRI scanning, whilst the role of bone scanning is decreasing, although in some cases it brings valuable information. In particular, in early stages of osteitis and Charcot neuro-osteoarthropathy, radionuclide imaging may be superior to MRI. Additionally, a significant contribution of inflammation-targeted scintigraphy should be noted. Probably the role of PET scanning will grow, although its high cost and low availability may be a limiting factor. In every case, vascular status should be determined, at least with Doppler ultrasound, with following conventional angiography or MR angiography.


[Convulsive disorder in celiac disease].
Several recent reports have described convulsions in patients with celiac disease, and in some, folic acid deficiency and brain calcifications. A 40-year-old woman with celiac disease, hypocalcemia and generalized tonic-clonic seizures is reported. Hypocalcemia was corrected and convulsions disappeared, but the EEG showed persistent occipital epileptiform activity. Patients with celiac disease and hypocalcemia due to malabsorption are particularly at increased risk for convulsions. Therefore, even a mild degree of hypocalcemia in these patients should be corrected as soon as possible.


Delta ribozyme benefits from a good stability in vitro that becomes outstanding in vivo.
The stability of a trans-acting delta ribozyme was studied under various conditions. Although in vitro (i.e., in the presence of protein extracts) this delta ribozyme appears to be only slightly more stable than a hammerhead ribozyme, in vivo (i.e., after cell transfection) it exhibits an outstanding stability that manifests itself in the calculated half-life of over 100 h regardless of the means of transfection. The P2 stem, which includes both the 5' and 3' ends, is shown to play a critical role in this stability. Direct mutagenesis of the most nuclease susceptible nucleotides failed to generate a more stable ribozyme that retained the same catalytic potential. Clearly, delta ribozyme appears to be well adapted to the human cell environment, and is therefore ideal for the development of a gene-inactivation system.


Central opioid control of feeding behavior in the white-crowned sparrow, Zonotrichia leucophrys gambelii.
Many behavioral responses to stress do not appear to be mediated by glucocorticoids, suggesting another mechanism. We tested the effects of intracerebroventricular administration of beta-endorphin, a neuropeptide implicated in the stress response, on feeding behavior in captive, wild-caught white-crowned sparrows (Zonotrichia leucophrys gambelii). The amount of time spent feeding and the number of feeding bouts were higher after infusion with beta-endorphin than after saline infusion. Beta-endorphin decreased the latency to feed compared with saline. Naloxone, an opioid antagonist, suppressed feeding behavior and increased latency to feed. These results support our hypothesis that neuropeptides associated with stress may initiate adaptive responses to natural stressors in wild species.


Computed tomographic assessment of vascular invasion and resectability of mediastinal masses in dogs and a cat.
To assess the sensitivity of non-angiographic contrast-enhanced computed tomography (CT) to determine the presence of vascular invasion of cranial mediastinal masses in dogs and a cat, and to evaluate the association between vascular invasion and peri-operative mortality. A retrospective study was conducted on 25 dogs and one cat. CT scans were completed with slices ranging from 2 to 10 mm. CT images were evaluated by a board-certified radiologist blinded to previous diagnoses and surgical findings. Each CT study was evaluated for vascular invasion, defined as disruption of the vessel wall and extension of the mass into the vessel lumen. Data retrieved from the surgery reports included surgical approach, whether vascular invasion was present, the surgeon's decision on operability, and post-operative complications. Computed tomographic evaluation revealed 25/26 masses had no evidence of vascular invasion. During surgical exploration, 10/26 masses were found to invade major regional vasculature; the cranial vena cava (CVC) was the vessel most commonly invaded (7/10 animals), and 4/7 (57%) patients with invasion of the CVC were euthanised or died in the peri-operative period, from surgical or disease-related problems, which was significantly higher than patients without vascular invasion (p=0.045). Non-angiographic contrast-enhanced CT was significantly less sensitive for detecting vascular invasion of cranial mediastinal masses when compared with surgical evaluation. If the CVC was invaded by a tumour there was a significant risk of death peri-operatively when compared with non-invasive cases. Due to the significantly higher mortality risk associated with invasion of the CVC, a more sensitive method than CT should be investigated to determine vascular invasion of mediastinal masses pre-operatively.


Spectral embedding based active contour (SEAC) for lesion segmentation on breast dynamic contrast enhanced magnetic resonance imaging.
Segmentation of breast lesions on dynamic contrast enhanced (DCE) magnetic resonance imaging (MRI) is the first step in lesion diagnosis in a computer-aided diagnosis framework. Because manual segmentation of such lesions is both time consuming and highly susceptible to human error and issues of reproducibility, an automated lesion segmentation method is highly desirable. Traditional automated image segmentation methods such as boundary-based active contour (AC) models require a strong gradient at the lesion boundary. Even when region-based terms are introduced to an AC model, grayscale image intensities often do not allow for clear definition of foreground and background region statistics. Thus, there is a need to find alternative image representations that might provide (1) strong gradients at the margin of the object of interest (OOI); and (2) larger separation between intensity distributions and region statistics for the foreground and background, which are necessary to halt evolution of the AC model upon reaching the border of the OOI. In this paper, the authors introduce a spectral embedding (SE) based AC (SEAC) for lesion segmentation on breast DCE-MRI. SE, a nonlinear dimensionality reduction scheme, is applied to the DCE time series in a voxelwise fashion to reduce several time point images to a single parametric image where every voxel is characterized by the three dominant eigenvectors. This parametric eigenvector image (PrEIm) representation allows for better capture of image region statistics and stronger gradients for use with a hybrid AC model, which is driven by both boundary and region information. They compare SEAC to ACs that employ fuzzy c-means (FCM) and principal component analysis (PCA) as alternative image representations. Segmentation performance was evaluated by boundary and region metrics as well as comparing lesion classification using morphological features from SEAC, PCA+AC, and FCM+AC. On a cohort of 50 breast DCE-MRI studies, PrEIm yielded overall better region and boundary-based statistics compared to the original DCE-MR image, FCM, and PCA based image representations. Additionally, SEAC outperformed a hybrid AC applied to both PCA and FCM image representations. Mean dice similarity coefficient (DSC) for SEAC was significantly better (DSC = 0.74 ± 0.21) than FCM+AC (DSC = 0.50 ± 0.32) and similar to PCA+AC (DSC = 0.73 ± 0.22). Boundary-based metrics of mean absolute difference and Hausdorff distance followed the same trends. Of the automated segmentation methods, breast lesion classification based on morphologic features derived from SEAC segmentation using a support vector machine classifier also performed better (AUC = 0.67 ± 0.05; p < 0.05) than FCM+AC (AUC = 0.50 ± 0.07), and PCA+AC (AUC = 0.49 ± 0.07). In this work, we presented SEAC, an accurate, general purpose AC segmentation tool that could be applied to any imaging domain that employs time series data. SE allows for projection of time series data into a PrEIm representation so that every voxel is characterized by the dominant eigenvectors, capturing the global and local time-intensity curve similarities in the data. This PrEIm allows for the calculation of strong tensor gradients and better region statistics than the original image intensities or alternative image representations such as PCA and FCM. The PrEIm also allows for building a more accurate hybrid AC scheme.


Rapid oxidation of iodide and hypoiodous acid with ferrate and no formation of iodoform and monoiodoacetic acid in the ferrate/I-/HA system.
Toxic and odorous iodinated disinfection byproducts (I-DBPs) could form in the chemical oxidation of iodine-containing water. A critical step for controlling the hazardous I-DBPs is to convert the iodine species into stable and harmless iodate (IO3-) while inhibiting the accumulation of highly reactive hypoiodous acid (HOI). Herein, the oxidation of I- and HOI with ferrate was investigated, and the formation profile of HOI was determined based on 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) coloring method through a stopped-flow spectrophotometer. The second-order rate constants (kapp) of ferrate with HOI decreased from 1.6 × 105 M-1s-1 to 8.3 × 102 M-1s-1 as the solution pH varied from 5.3 to 10.3, which were 7.5, 7.2 and 13.8 times higher than that of ferrate with I- at pH 6.0, 7.0 and 8.0, respectively. Compared with other oxidants such as ozone, hypochlorous acid, chloramine and potassium permanganate, ferrate would swiftly oxidize HOI formed in the I- oxidation process. For the ferrate oxidation of I-containing water, HOI was swiftly oxidized to IO3- from pH 5.0 to 9.0. Phosphate buffer promoted the oxidation of I- while inhibited the oxidation of HOI with ferrate. When 5 mgC/L of humic acids (HA) existed in the solution, no formation of iodoform and monoiodoacetic acid (MIAA) was observed in the oxidation of iodide (20 μM) with ferrate (from 10 μM to 80 μM). These results suggested that ferrate oxidation could be an effective method for the control of I-DBPs in iodine-containing water treatment.


Functional neutralization of HIV-1 Vif protein by intracellular immunization inhibits reverse transcription and viral replication.
Human immunodeficiency virus type 1 (HIV-1)-encoded Vif protein is important for viral replication and infectivity. Vif is a cytoplasmic protein that acts during virus assembly by an unknown mechanism, enhancing viral infectivity. The action of Vif in producer cells is essential for the completion of proviral DNA synthesis following virus entry. Therefore, Vif is considered to be an important alternative therapeutic target for inhibition of viral infectivity at the level of viral assembly and reverse transcription. To gain insight into this process, we developed a Vif-specific single-chain antibody and expressed it intracellularly in the cytoplasm. This intrabody efficiently bound Vif protein and neutralized its infectivity-enhancing function. Intrabody-expressing cells were shown to be highly refractory to challenge with different strains of HIV-1 and HIV-1-infected cells. Inhibition of Vif by intrabody expression in the donor cell produced viral particles that do not complete reverse transcription in the recipient cell. The anti-Vif scFv was shown to be specific for Vif protein because its function was observed only in nonpermissive cells (H9, CEM, and U38). Moreover, transduction of peripheral blood mononuclear cells with an HIV-derived retroviral vector expressing Vif intrabody was shown to confer resistance to laboratory-adapted and primary HIV strains. This study provides biochemical evidence for the role of Vif in the HIV-1 lifecycle and validates Vif as a target for the control of HIV-1 infection.


The interaction of healthcare service quality and community-based health insurance in Ethiopia.
Community-based health insurance (CBHI) as a demand-side intervention is presumed to drive improvements in health services quality, and the quality of health services is an important supple-side factor in motivating CBHI enrollment and retention. There is, however, limited evidence on this interaction. This study examined the interaction between quality of health services and CBHI enrollment and renewal. A mixed-method comparative study was conducted in four agrarian regions of Ethiopia. The study followed the Donabedian model to compare quality of health services in health centers located in woredas/districts that implemented CBHI with those that did not. Data was collected through facility assessments, client-exit interviews, and key informant interviews. In addition to manual thematic analysis of qualitative data, quantitative descriptive and inferential analyses were done using SPSS vs 25. The process related (composite index including provider-client interpersonal communication) and outcome related (client satisfaction) measures of service quality in CBHI woreda/districts differed significantly from non-CBHI woredas/districts, but there were no significant differences in overall measures of structural quality between the two. The study found better diagnostic test capacity, availability of tracer drugs, provider interpersonal communication, and service quality standards in CBHI woredas. A higher proportion of clients at CBHI health centers gave high ratings of overall satisfaction with services. Individual and household factors including family size, age, household health care-related expenditures, and educational status, played a more significant role in CBHI enrollment and renewal decisions than health service quality. Key-informants reported in interviews that participation in the scheme increased accountability of health facilities in CBHI woredas/districts, because they promised to provide quality services using the CBHI premium collected at the beginning of the year from all enrolled households. This study indicates a need for follow-up research to understand the nuanced linkages between quality of care and CBHI enrollment.


Stable isotope techniques in early drug development: an economic evaluation.
Stable isotope labeled (SIL) drug methods are compared with standard methods for performing early (phases I and IIa) drug development studies (mass balance, bioavailability, single-dose volunteer and patient, multiple-dose volunteer and patient). SIL methods offer considerable reduction in the cost (> 50%) and number of subjects (67%) required for bioavailability and multiple-dose patient studies. Moreover, a complete early drug development program is described for optimally combining SIL and standard studies, which can reduce cost by 23% and number of subjects by 36% compared with a program using standard methods. These reductions should result in development time savings of at least one year.


Chemoselective Asymmetric Intramolecular Dearomatization of Phenols with α-Diazoacetamides Catalyzed by Silver Phosphate.
We report asymmetric dearomatization of phenols using Ag carbenoids from α-diazoacetamides. The Ag catalyst promoted intramolecular dearomatization of phenols, whereas a Rh or Cu catalyst caused C-H insertion and a Büchner reaction. Studies indicated Ag carbenoids have a carbocation-like character, making their behavior and properties unique. Highly enantioselective transformations using Ag carbenoids have not been reported. We achieved a Ag carbenoid-mediated chemo- and highly enantioselective phenol dearomatization with substrate generality for the first time.


Children's hospital nurses 'outstanding'.
Nurses at Birmingham Children's Hospital NHS Foundation Trust won praise after it became the first children's hospital to be rated outstanding overall by the Care Quality Commission.


Two new protoberberine quaternary alkaloids from Corydalis yanhusuo.
Two new protoberberine quaternary alkaloids, 13-methyl-palmatrubine (1) and 13-methyl-dehydrocorydalmine (2), were isolated from the tubers of Corydalis yanhusuo W.T. Wang. 1 was isolated as a natural product for the first time, and 2 a new compound. Their structures were elucidated on the basis of spectroscopic evidence, especially 1D- and 2D-NMR experiments.


Sauchinone, a lignan from Saururus chinensis, inhibits staurosporine-induced apoptosis in C6 rat glioma cells.
Neuronal apoptosis may contribute to pathologic neuronal loss in certain disease states such as neurodegenerative diseases. Staurosporine (ST), a nonselective protein kinase inhibitor, has been shown to induce apoptosis in a variety of cells including nerve cell lines. In this study, we investigated the neuroprotective effect of sauchinone, which is a unique lignan from Saururus chinensis, on ST-induced apoptosis in C6 rat glioma cells. Sauchinone attenuated ST-induced apoptosis of C6 glioma cells as evidenced by DNA fragmentation. We also provide evidence that the inhibitory effect of sauchinone on ST-induced apoptosis involves a dose-dependent upregulation of an antiapoptotic protein, Bcl-2. Mounting evidence shows that the activation of caspases, especially caspase-3, triggers the apoptotic process. The activity of caspase-3 of ST-pretreated cells was significantly decreased upon sauchinone treatment in a dose-dependent manner. Taken together, the data demonstrate that sauchinone protects C6 glioma cells from ST-induced apoptosis in a caspase-3 dependent manner. Our findings may be critical for developing a strategy to protect nerve cells from apoptosis, suggesting the potential development of sauchinone as a neuroprotective agent.


Putative candidate genes responsible for leaf rolling in rye (Secale cereale L.).
Rolling of leaves (RL) is a phenomenon commonly found in grasses. Morphology of the leaf is an important agronomic trait in field crops especially in rice; therefore, majority of the rice breeders are interested in RL. There are only few studies with respect to RL of wheat and barley; however, the information regarding the genetic base of RL with respect to the shape of leaf in rye is lacking. To the best of our knowledge, this is the first study on the localization of loci controlling RL on high density consensus genetic map of rye. Genotypic analysis led to the identification of 43 quantitative trait loci (QTLs) for RL, grouped into 28 intervals, which confirms the multigenic base of the trait stated for wheat and rice. Four stable QTLs were located on chromosomes 3R, 5R, and 7R. Co-localization of QTL for RL and for different morphological, biochemical and physiological traits may suggests pleiotropic effects of some QTLs. QTLs for RL were associated with QTLs for such morphological traits as: grain number and weight, spike number per plant, compactness of spike, and plant height. Two QTLs for RL were found to coincide with QTLs for drought tolerance (4R, 7R), two with QTLs for heading earliness (2R, 7R), one with α-amylase activity QTL (7R) and three for pre-harvest sprouting QTL (1R, 4R, 7R). The set of molecular markers strongly linked to RL was selected, and the putative candidate genes controlling the process of RL were identified. Twelve QTLs are considered as linked to candidate genes on the base of DArT sequences alignment, which is a new information for rye. Our results expand the knowledge about the network of QTLs for different morphological, biochemical and physiological traits and can be a starting point to studies on particular genes controlling RL and other important agronomic traits (yield, earliness, pre-harvest sprouting, reaction to water deficit) and to appoint markers useful in marker assisted selection (MAS). A better knowledge of the rye genome and genes could both facilitate rye improvement itself and increase the efficiency of utilizing rye genes in wheat breeding.


Dietary and hormonal stimulation of rat exocrine pancreatic function regulates CRHSP-28 phosphorylation in vivo.
Dietary regulation of digestive enzyme secretion from the pancreas is essential for the breakdown of macronutrients in the gastrointestinal tract. Ca(2+)-responsive heat stable protein (CRHSP)-28 is a regulatory protein that modulates the exocytosis of digestive enzymes from pancreatic acinar cells. In the present study, isoelectric focusing and immunoblotting were used to characterize CRHSP-28 phosphorylation in isolated rat acinar cells and also after hormonal and dietary stimulation of rat pancreas in vivo. CRHSP-28 was highly phosphorylated in isolated acini after stimulation with a physiologic range of concentrations of cholecystokinin-octapeptide (CCK-8). Activation of the high affinity state of the CCK-A receptor with the synthetic peptide JMV-180 confirmed the physiologic relevance of the response. CRHSP-28 phosphorylation was contingent on elevated cellular Ca2+ because it was maximally stimulated by Ca2+ ionophore, but unchanged after protein kinase C, cAMP or cyclic guanosine monophosphate activation. Intravenous infusion of rats with a secretory concentration of the CCK analog, caerulein, stimulated CRHSP-28 phosphorylation by 100% over control (P < 0.01) within 15 min of dosing. Moreover, CRHSP-28 phosphorylation was stimulated by 150% over control (P < 0.05) immediately after consumption of a semipurified AIN-93 diet. These data demonstrate that CRHSP-28 phosphorylation occurs in vivo and can be used as a functional indicator of nutrient-driven acinar cell activation.


Functional lipid characteristics of Turkish Tombul hazelnut (Corylus avellana L.).
The quality of crude oil extracted from Tombul (Round) hazelnut, grown in the Giresun province of Turkey, was evaluated for its fatty acid, triacylglycerol (TAG), tocol, and phytosterol compositions. Oleic acid contributed 82.78% to the total fatty acids, followed by linoleic, palmitic, and stearic acids. Among 12 TAGs separated, 11 were identified (including one unknown): LLL, OLL, PLL, OOL, POL, PPL, OOO, POO, PPO, SOO, and PSO (where P, palmitoyl; S, stearoyl; O, oleoyl; and L, linoleoyl). The main components were OOO (71.31%), OOL (12.26%), and POO (9.45%), reflecting the high content of oleic acid present in hazelnut oil. Seven tocol isoforms (four tocopherols and three tocotrienols) and eight phytosterols as well as cholesterol were positively identified and quantified; among these, alpha-tocopherol (40.40 mg/100 g) and beta-sitosterol (134.05 mg/100 g) were predominant in hazelnut oil and contributed 78.74 and 81.28% to the total tocols and phytosterols present, respectively. Tocotrienols were detected in small amounts (1.02% to the total tocols). The crude hazelnut oil extracted from Turkish Tombul hazelnut, thus, serves as a good source of nutrients, bioactives, and health-promoting components.


Analysis of Closed Claims in the Clinical Management of Rheumatoid Arthritis in Japan.
Despite an increasing awareness of the risk of medical errors, few data sources are available to highlight the characteristics and patterns of medical errors in the clinical management of rheumatoid arthritis (RA). The present study aimed to evaluate medical malpractice claims associated with the management of RA and other autoimmune connective tissue diseases (ACTDs). We analyzed 38 ACTD-associated closed claims extracted from a total of 8530 claims processed between July 2004 and June 2014 by the Tokyo headquarters office of Sompo Japan Nipponkoa Incorporated, a leading malpractice insurer in Japan. RA was the most common ACTD assessed in this study, accounting for 20 cases. Although the male-to-female ratio among these cases was 5:15, in accordance with the general demographic distribution of RA, the proportion of patients older than 60 years (77.8%) was relatively high as the general range of RA susceptibility is 30-50 years. The analysis of allegation types among RA cases revealed statistically significant differences from non-RA cases (Fisher's exact test) as well as the following key findings: diagnosis-related allegations were absent (P < 0.01), whereas medication-related allegations were distinctively common (P = 0.02). Clinical processes related to the assessment process were most vulnerable to breakdown and leading to negligence identified with subsequent medication-related allegations, particularly among RA cases. The characteristics of malpractice claims associated with RA management, including the high frequency of medication-related allegations, breakdowns in the assessment process, and high claim numbers among patients older than 60 years, suggest the importance of caution exercised by physicians when administering immunosuppressants for the clinical treatment of RA.


Mouse mast cell protease-1 is required for the enteropathy induced by gastrointestinal helminth infection in the mouse.
The relationship between intestinal pathology and immune expulsion of gastrointestinal nematodes remains controversial. Immune expulsion of gastrointestinal helminth parasites is usually associated with Th2 responses, but the effector mechanisms directly responsible for parasite loss have not been elucidated. Mast cell hyperplasia is a hallmark of infection with gastrointestinal nematodes, in particular Trichinella spiralis. Although the precise mechanism by which mast cells induce expulsion of these parasites has not been elucidated, it has been proposed that mast cell mediators, including cytokines and granule chymases, act to create an environment inhospitable to the parasite, part of this being the induction of intestinal inflammation. Therefore, the aims of this study were to dissect the role of mast cells and mast cell proteases in the induction of parasite-induced enteropathy. Mast cell-deficient W/Wv and mouse mast cell protease-1 (mMCP-1)-deficient mice were infected with T. spiralis, and parasite expulsion, enteropathy, and Th2 responses were determined. Expulsion of the parasite was delayed in both strains of mice compared with wild-type controls; additionally, in both cases, the enteropathy was significantly ameliorated. Although Th2 responses were significantly reduced in mast cell-deficient W/Wv mice, those from mMCP-1-deficient mice were similar to wild-type mice. Additionally, levels of TNF-alpha and nitric oxide were significantly reduced in both W/Wv and mMCP-1 deficient mice. These results imply that mast cells may contribute to the induction of protective Th2 responses and, importantly, that the intestinal inflammation associated with gastrointestinal helminths is partly mediated by mMCP-1.


Role of the maternal environment in determining susceptibility to transplacentally induced chemical carcinogenesis in mouse fetuses.
Treatment of pregnant mice with 3-methylcholanthrene (MC) causes lung and liver tumors in the offspring, the incidences of which are greatly influenced by the Ah locus regulated induction phenotype for aryl hydrocarbon hydroxylase activity (AHH) in both the mother and fetuses. In order to examine the biochemical and molecular mechanisms responsible for the modulating effect of maternal environment on tumor susceptibility, reciprocal crosses between responsive C57BL/6 and non-responsive DBA/2 mice were made and the pregnant mothers were treated i.p. on the 17th day of gestation with either olive oil alone, 30 mg/kg of MC, or 30 mg/kg of beta-naphthoflavone (beta NF). At various times after injection, the mothers were killed and the fetuses removed for enzymatic and molecular blot analysis. In fetal lung tissues, the absolute levels and relative induction ratios of AHH activity from D2B6F1 fetuses were very similar to those obtained in B6D2F1 fetuses during the first 24 h following a transplacental exposure to either inducing agent. This was also the case 48 h after an injection of beta NF. However, 48 h after exposure to MC, the AHH activity in fetal lungs from B6 mothers had declined to practically control values, whereas fetal lungs from D2 mothers still exhibited a high level of AHH activity. Similar induction kinetics for the CYPIA1 gene were obtained in fetal livers. These results were confirmed at the RNA level by quantitative slot-blot analysis of fetal RNA preparations. In both organs, treatment with inducing agents for the P450IA1 gene resulted in a rapid and early induction of CYPIA1 RNA by 4 h. Fetuses from D2 mothers, however, showed a more sustained induction of CYPIA1 RNA following exposure to MC than did fetuses from B6 mothers. These results suggest that the observed increase in tumor susceptibility observed in the offspring of D2 mothers compared to the offspring of B6 mothers was due, at least in part, to the differences in the persistence of induction of the CYPIA1 gene locus, and may be the result of differences in the clearance rates of MC from the fetal and maternal compartments or its pharmacokinetic distribution in the two types of maternal environments.


[Molecular diagnosis of mitochondrial disorders].
Mitochondrial disorders are caused by either nuclear or mitochondrial DNA abnormalities. The latter but not the former can be examined as a means of diagnosis at present. The peculiarities and problems of the mtDNA test depend strongly on the characteristics of mtDNA itself, such as the multiplicity in number, vulnerability to mutation, and maternal transmission. The mtDNA abnormalities observed in patients with mitochondrial disorders indicate heteroplasmy, tissue/cell specificity, and threshold effects. It should be stressed that the presence of mutant mtDNA in an individual does not mean that he or she is ill. The problems of the mtDNA test could be solved if the genotype-phenotype relationship is farther elucidated in the future study.


Bending loss and thermo-optic effect of a hybrid PDMS/silica photonic crystal fiber.
In this paper, we demonstrate and report a photonic crystal fiber (PCF) infiltrated with PDMS elastomer which is sensitive to external bending and temperature perturbations. Numerical simulations and experimental measurements were carried out to investigate the fundamental TIR-based guiding mechanism of the hybrid PDMS/silica PCF in terms of effective index, effective modal area and loss. Wavelength dependence of bending losses was also measured for different bend diameters as well as the temperature dependence of the fundamental guiding mode for a range of temperatures from 20°C to 75°C. Experimental measurements have shown a ~6% power recovery of the bend-induced loss for a 6-cm long PDMS-filled PCF at 4 cm bend diameter.


Associations between loneliness, depressive symptoms and perceived togetherness in older people.
This study explores the associations of loneliness with depressive symptoms in a five-year follow-up and describes how the six dimensions of perceived togetherness explain loneliness and depressive symptoms at baseline. The data were collected on 207 residents of Jyväskylä, central Finland, who at baseline in 1990 were aged 80; and 133 residents who at follow-up in 1995 were aged 85. Loneliness was assessed using a questionnaire item with four preset response options, perceived togetherness using the Social Provisions Scale, and depressive symptoms using the CES-D scale. A recursive structural equation model showed that in women but not in men, depressive symptoms predicted more experiences of loneliness. Those who were lonely were more depressed (CES-D score 16 or over) and experienced less togetherness than those who were not. Loneliness was explained by reliable alliance, social integration and attachment; and depressive symptoms were explained by guidance, reassurance of worth, reliable alliance and attachment. A common feature in both loneliness and depressive symptoms was a lower level of perceived emotional togetherness in social interaction.


Ichthyotherapy as alternative treatment for patients with psoriasis: a pilot study.
Ichthyotherapy (therapy with the so-called 'Doctorfish of Kangal', Garra rufa) has been shown to be effective in patients with psoriasis in the Kangal hot springs in Turkey. This study evaluates the efficacy and safety of ichthyotherapy in combination with short-term ultraviolet A sunbed radiation in the treatment of psoriasis under controlled conditions. We retrospectively analyzed 67 patients diagnosed with psoriasis who underwent 3 weeks of ichthyotherapy at an outpatient treatment facility in Lower Austria between 2002 and 2004. Main outcome measures are as follows: overall relative reduction in Psoriasis Area Severity Index (PASI) score; proportion of patients with an improvement in their PASI score of >/=75% (PASI-75) and >/=50% (PASI-50); patient-reported outcomes assessed with a custom questionnaire; and patient follow-up with a questionnaire sent out in March 2005. Safety was evaluated by reviewing adverse events and vital signs. Overall there was a 71.7% reduction in PASI score compared to baseline (P < 0.0001). Of the 67 patients studied, 31 (46.3%) achieved PASI-75 and 61 patients (91%) achieved at least PASI-50. Patients reported substantial satisfaction with the treatment. The reported mean remission period was 8.58 months [95% confidence interval (CI) 6.05-11.11]. A total of 87.5% of patients reported a more favorable outcome with ichthyotherapy, when asked to compare ichthyotherapy to other previously tried therapies. Sixty-five percent stated that after the relapse their symptoms were less severe than before treatment. There were no significant adverse events. The benefit demonstrated in this study along with the favorable safety profile suggests that ichthyotherapy could provide a viable treatment option for patients with psoriasis.


Morphological analysis of the tongue and the digestive tube of saffron finch (Sicalis flaveola brasiliensis, LINNAUES 1766) apprehended by CETAS/IBAMA-PB.
The saffron finch (Sicalis flaveola brasiliensis) is a Passeriforme of the Brazilian wildlife. There are scarcely any morphological studies on it, although it is frequently trafficked for its birdsong abilities. Its peculiarities, such as territorialism and developed syrinx that provides outstanding song, draw attention towards its domestication. Thus, this study aimed to morphologically describe the tongue and digestive tube organs of this species to furnish subsidies for nutritional, clinical and conservation studies. Forty-one birds from the Wild Animal Screening Center (CETAS)/Brazilian Institute of Environment (IBAMA)/city of Cabedelo, state of Paraíba (PB) were used. Samples were collected, identified and sent to standard light microscopy; samples of proventriculus and gizzard were sent to Scanning Electron Microscopy (SEM). The samples showed stratification similar to that of other domestic and wild birds, confirmed in the scanning electron microscopy; however, they differed in the absence of dermal papillae in the tongue, lack of ingluvial glands and lack of muscular mucosa and sub-mucosa in the large intestine.


Effect of Cell Inner Pressure on Deposition Volume in Microinjection.
Microinjection is a widely used technique for introducing exogenous materials into cells. Many applications of microinjection, such as gene editing and drug testing, rely on the accurate control of the deposition volume. However, the deposition volume in microinjection is presently calibrated in an open medium without considering the cell inner pressure effect, which we experimentally show in this paper that it can induce an error as large as 30% between the actual deposition volume and the set volume. In this work, the relationship between the cell inner pressure and the deposition volume was analytically modeled and experimentally validated. On the basis of the developed model, the cell inner pressure of a given cell type can be well estimated from the injection pressure and the resulting deposition volume. The quantitated cell inner pressure is then used to reduce the error between the set volume and the actual deposition volume. Experiments conducted on human bladder cancer cells (T24 and RT4) showed that T24 cells have a higher inner pressure than RT4 cells (405 ± 45 Pa vs 341 ± 34 Pa), and after compensating for the cell inner pressure, the error between the intended set volume and the actual deposition volume into a cell became less than 3%.


Angiotensin-(1-7) prevents systemic hypertension, attenuates oxidative stress and tubulointerstitial fibrosis, and normalizes renal angiotensin-converting enzyme 2 and Mas receptor expression in diabetic mice.
We investigated the relationship between Ang-(1-7) [angiotensin-(1-7)] action, sHTN (systolic hypertension), oxidative stress, kidney injury, ACE2 (angiotensin-converting enzyme-2) and MasR [Ang-(1-7) receptor] expression in Type 1 diabetic Akita mice. Ang-(1-7) was administered daily [500 μg/kg of BW (body weight) per day, subcutaneously] to male Akita mice from 14 weeks of age with or without co-administration of an antagonist of the MasR, A779 (10 mg/kg of BW per day). The animals were killed at 20 weeks of age. Age-matched WT (wild-type) mice served as controls. Ang-(1-7) administration prevented sHTN and attenuated kidney injury (reduced urinary albumin/creatinine ratio, glomerular hyperfiltration, renal hypertrophy and fibrosis, and tubular apoptosis) without affecting blood glucose levels in Akita mice. Ang-(1-7) also attenuated renal oxidative stress and the expression of oxidative stress-inducible proteins (NADPH oxidase 4, nuclear factor erythroid 2-related factor 2, haem oxygenase 1), pro-hypertensive proteins (angiotensinogen, angiotensin-converting enzyme, sodium/hydrogen exchanger 3) and profibrotic proteins (transforming growth factor-β1 and collagen IV), and increased the expression of anti-hypertensive proteins (ACE2 and MasR) in Akita mouse kidneys. These effects were reversed by A779. Our data suggest that Ang-(1-7) plays a protective role in sHTN and RPTC (renal proximal tubular cell) injury in diabetes, at least in part, through decreasing renal oxidative stress-mediated signalling and normalizing ACE2 and MasR expression.


Staged Hard and Soft Tissue Reconstruction Followed by Implant Supported Restoration in the Aesthetic Zone: A Case Report.
Alveolar ridge deficiency is a common clinical consequence following tooth loss due to chronic periodontitis complicating ideal implant placement. Advanced hard and soft tissue augmentation procedures have been developed in the recent past with predictable clinical outcomes. A male patient presented with a Grade III mobile upper right central incisor associated with advanced bone loss and soft tissue deficit. Following extraction of tooth #11, socket augmentation was done using an autogenous cortico-cancellous block graft and subsequent soft tissue augmentation was done with palatal connective tissue graft. At the end of six months, a tapered self tapping implant fixture was placed with adequate primary stability and after eight weeks, second stage implant surgery was done with the Misch technique in order to recreate papillae and the implant was prosthetically restored. The alveolar ridge was adequately recontoured following the staged surgical protocol. The implant was well integrated at the end of 15 months. Execution of sequential surgical procedures in a highly deficient edentulous site made it possible to achieve of optimal pink and white aesthetics with stable implant supported fixed prosthesis.


Preparation of 3-Iodoquinolines from N-Tosyl-2-propynylamines with Diaryliodonium Triflate and N-Iodosuccinimide.
4-Aryl and 4-alkyl substituted 3-iodoquinolines could be smoothly obtained in one pot by treating N-tosyl-2-propynylamines with diaryliodonium triflate in the presence of K3PO4 and a catalytic amount of CuCl at room temperature, followed by treatment with N-iodosuccinimide and BF3·OEt2 at 0 °C, and then NaOH in methanol solution. The product, 3-iodo-4-phenylquinoline was smoothly transformed into 4-phenylquinoline with zinc; 4-phenyl-3-toluenesulfenylquinoline with toluenethiol, K2CO3, and CuI; 4-phenyl-3-phenylethynylquinoline with the Sonogashira coupling reaction; 4-phenyl-3-styrylquinoline with the Heck coupling reaction; 3,4-diphenylquinoline with the Suzuki-Miyaura coupling reaction; 2-cyclohexyl-3-iodo-4-phenylquinoline with cyclohexanecarboxylic acid, Ag2CO3, and K2S2O8; and 3-iodo-2-(2',5'-dioxan-1'-yl)-4-phenylquinoline with benzoyl peroxide in dioxane.


A comparative study of the opinions, experiences and individual innovativeness characteristics of operating room nurses on robotic surgery.
To compare and evaluate operating room nurses' opinions about robotic surgery with their individual innovativeness metric scores. The aim was also to identify the experiences, adaptation processes and influencing factors of operating room nurses working in robotic surgery rooms in Turkey. This was a comparative descriptive study. The sample included 114 operating room nurses working in 12 institutions that performed robotic surgery in Turkey. Data were collected between January 2018 and September 2019 using the Demographic Characteristics of Nurses and Robotic Surgery Evaluation Form and the Individual Innovativeness Scale, and were analysed using a quantitative statistical approach (independent samples t-test, one-way analysis of variance and Kruskal-Wallis H test). The data obtained from the Robotic Surgery Evaluation section, which consisted of open-ended and closed-ended questions, were analysed with appropriate steps. In particular, answers to open-ended questions were grouped and coded according to their content. Operating room nurses held positive opinions about robotic surgery. Only 35.8% of the nurses had received training before joining the robotic team, while 55.2% had individually searched for information. Operating room nurses with robotic surgery experience had significantly higher (p < .001) individual innovativeness scores. Over 85% of nurses who received training adapted to robotic surgery in 3 months or less, while nurses with higher individual innovativeness scores adapted in a statistically significantly (p < .05) shorter period. Training, teamwork, and practical experience were mentioned as facilitating factors; inadequate training and technical problems were reported as obstructing factors. Operating room nurses hold positive opinions about robotic surgery; nurses with robotic surgery experience have higher levels of individual innovativeness; and nurses who have received training are significantly better adapted. What problem did the study address? This study addresses the need for a better understanding of operating room nurses' opinions and experiences about robotic surgery and the influencing factors of adapting to it. This study also offers an evaluation and comparison of the nurses' individual innovativeness characteristics and the correlation with their adaptation processes to the new role. What were the main findings? The main findings show a correlation between specific education/training and nurses' adaptation to the new role of robotic surgery, the individual innovativeness characteristics metric of nurses with or without robotic surgery experience and the time frame of their adaptation. Where and on whom will the research have an impact? This research traces the profile of current operating room nurses working in the robotic surgery field and the factors influencing their experience. These findings and conclusions have a much broader impact than in Turkey alone. The findings raise awareness of the importance of educating and preparing operating room nurses before introducing them to the new roles and responsibilities inherent to robotic surgery.


Structural origin of chirality and properties of a remarkable helically pillared solid.
A new helically pillared and chiral solid, Cu(pzc)2AgReO4 (I, pzc = pyrazinecarboxylate), was synthesized from hydrothermal reactions at 95-125 degrees C. The structural origin of its chirality, relative to the achiral M(pzc)2(H2O)2AgReO4 (II, M = Co; III, M = Ni) analogues, arises from significantly tilted pillars and hydrogen bonds to the AgReO4 layers. The new pillared structure exhibits second harmonic generation activity, CO2 absorption, thermal stability to approximately 250 degrees C, and Curie-Weiss magnetism expected for isolated Cu2+.


Kidney microRNA Expression Pattern in Type 2 Diabetic Nephropathy in BTBR Ob/Ob Mice.
Diabetic nephropathy (DN) is the main leading cause of chronic kidney disease worldwide. Although remarkable therapeutic advances have been made during the last few years, there still exists a high residual risk of disease progression to end-stage renal failure. To further understand the pathogenesis of tissue injury in this disease, by means of the Next-Generation Sequencing, we have studied the microRNA (miRNA) differential expression pattern in kidneys of Black and Tan Brachyury (BTBR) ob/ob (leptin deficiency mutation) mouse. This experimental model of type 2 diabetes and obesity recapitulates the key histopathological features described in advanced human DN and therefore can provide potential useful translational information. The miRNA-seq analysis, performed in the renal cortex of 22-week-old BTBR ob/ob mice, pointed out a set of 99 miRNAs significantly increased compared to non-diabetic, non-obese control mice of the same age, whereas no miRNAs were significantly decreased. Among them, miR-802, miR-34a, miR-132, miR-101a, and mir-379 were the most upregulated ones in diabetic kidneys. The in silico prediction of potential targets for the 99 miRNAs highlighted inflammatory and immune processes, as the most relevant pathways, emphasizing the importance of inflammation in the pathogenesis of kidney damage associated to diabetes. Other identified top canonical pathways were adipogenesis (related with ectopic fatty accumulation), necroptosis (an inflammatory and regulated form of cell death), and epithelial-to-mesenchymal transition, the latter supporting the importance of tubular cell phenotype changes in the pathogenesis of DN. These findings could facilitate a better understanding of this complex disease and potentially open new avenues for the design of novel therapeutic approaches to DN.


A mutation in the TMPRSS6 gene, encoding a transmembrane serine protease that suppresses hepcidin production, in familial iron deficiency anemia refractory to oral iron.
Hepcidin plays a key role in body iron metabolism by preventing the release of iron from macrophages and intestinal cells. Defective hepcidin synthesis causes iron loading, while overproduction results in defective reticuloendothelial iron release and iron absorption. We studied a Sardinian family in which microcytic anemia due to defective iron absorption and utilization is inherited as a recessive character. Five members showed iron deficiency anemia that was not responsive to oral iron and only partially responsive to parenteral iron administration. To investigate the involvement of known genes implicated in iron metabolism we carried out linkage analysis with microsatellite markers mapping close to these genes. Afterwards, a genome-wide search was performed. No linkage was found between the phenotype of the patients and several known human genes involved in iron metabolism (DMT1, TF, TFRC, ZIRTL, HAMP, HJV). Genome-wide scanning by microsatellites and single nucleotide polymorphisms showed a multipoint LOD score of 5.6 on chromosome 22q12.3-13.1, where the matriptase-2 (also known as transmembrane protease, serine 6 or TMPRSS6) gene is located. Its murine counterpart (Tmprss6) has recently been found to be an essential component of a pathway that detects iron deficiency and suppresses hepcidin production. Sequencing analysis of TMPRSS6 revealed a homozygous causal mutation, predicting a splicing error and a truncated TMPRSS6 protein in affected members. Homozygous subjects had inappropriately elevated levels of serum and urinary hepcidin. The findings of this study suggest that the observed TMPRSS6 mutation leads to overproduction of hepcidin and, in turn, to defective iron absorption and utilization. More generally, they confirm in humans the inhibitory effect of matriptase-2 on hepcidin synthesis already demonstrated in mice.


Quantification of allele E198A in beta-tubulin conferring benzimidazole resistance in Monilinia fructicola using real-time PCR.
Thiophanate-methyl, a member of the benzimidazole class of fungicides, is used in California to control brown rot of stone fruit caused by Monilinia fructicola (G. Wint.) Honey. The goal of this study was to develop a real-time polymerase chain reaction (PCR) assay as an efficient method to quantify the E198A allele of beta-tubulin that confers benzimidazole resistance. Using the real-time PCR assay, the frequency of allele E198A (FEA) in a population was determined from the quantities of DNA amplified with the E198A allele-specific primer pair HRF/HRR and the M. fructicola-specific primer pair MfF6/MfR6. The average proportions of highly resistant isolates determined with the conventional fungicide sensitivity method were within the range of average FEA values determined with the real-time PCR assay. We also determined the FEAs of M. fructicola populations sampled from 21 stone fruit orchards in California. Only one orchard showed a high FEA over 0.20, seven orchards had values between 0.01 and 0.1, and 13 orchards had values less than 0.01. The real-time PCR assay developed in this study provides a potentially useful tool to efficiently quantify benzimidazole resistance for large M. fructicola populations.


[Modern methods for cancer external radiation therapies].
Radiotherapy has been in constant progress for the past century. New technologies are based on modern imaging modalities, efficient 3D treatment planning systems, sophisticated immobilization systems and rigorous quality assurance and treatment verification. The central objective of conformal radiotherapy is to ensure a high dose distribution tailored to the limits of the target volume while reducing exposure of healthy tissues. These techniques would then allow a further tumor dose escalation. New systems like CyberKnife, tomotherapy and hadrontherapy represent major potential progress for the treatment of complex tumours that are very difficult to treat with conventional radiotherapy techniques.


Activity Landscapes, Information Theory, and Structure - Activity Relationships.
Activity landscapes provide a comprehensive description of structure-activity relationships (SARs). An information theoretic assessment of their features, namely, activity cliffs, similarity cliffs, smooth-SAR, and featureless regions, is presented based on the probability of occurrence of these features. It is shown that activity cliffs provide highly informative SARs compared to smooth-SAR regions, although the latter are the basis for most QSAR studies. This follows since small structural changes in the former are coupled with relatively large changes in activity, thus pinpointing specific structural features associated with the changes in activity. In contrast, Smooth-SAR regions are typically associated with relatively small changes in both structure and activity. Surprisingly, similarity cliffs, which occur when both compounds in a compound-pair have approximately equal activities but significantly different structures, are the most prevalent feature of activity landscapes. Hence, from an information theoretic point of view, they are the least informative landscape feature. Nevertheless, similarity cliffs do provide SAR information on potentially new active compound classes, and in that sense they are quite useful in drug discovery programs since they provide alternative possibilities should ADMET or other issues arise during the discovery and earlier preclinical development phases of drug research.


Congenital upper lid colobomas: management and visual outcome.
Congenital upper lid colobomas may be associated with ocular and systemic anomalies. The ophthalmologist's role involves both the diagnosis and management of their various ocular structural defects and management of the visual development. All cases of congenital eyelid colobomas referred to the Singapore National Eye Centre between July 1992 and July 1995 were assessed for the extent of the eyelid defect, associated ocular anomalies, status of the cornea, and any systemic anomalies. Four infants were tested during this period. The mean follow-up was 25 months (range, 16 to 30 months). All required surgical correction of their eyelid defects before 2 years of age. The patients also had refractive errors requiring amblyopia management. Three of the babies also needed other surgical procedures. Congenital upper eyelid colobomas are a potential threat to vision at an early age and a significant cosmetic blemish later in life. Early surgical intervention is usually required when the defect is larger than one third of the eyelid margin. Close monitoring of the visual development of patients with congenital upper eyelid colobomas is also essential since the risk of amblyopia in these patients is high.


Gas sorption in solid surfaces: a computational study using rigid and Einstein-solid models.
The reactive Monte Carlo (RxMC) method was proposed to describe the sorption of gases in solid materials due to the chemical reaction A + B ⇌ C. Two models were used to simulate the solid; the first model considered simulations with rigid particles in the solid whereas in the second model the particles were allowed to vibrate inside the solid with a given spring constant, i.e. an Einstein solid was used to simulate the substrate. In both models not only physisorption but also chemisorption of the fluid was observed. Sorption curves, at different spring constants, were simulated and it was noted that sorption was always enhanced with the Einstein solid model. Moreover, an inverse dependent function of the spring constant with the temperature was found. Finally, the second model might be used to explain the unusual sorption behavior observed in actual experimental reactions such as CO2 + Li2O ⇌ Li2CO3.


Comparing and improving chronic illness primary care in Sweden and the USA.
Purpose - The purpose of this paper is to identify opportunities for improving primary care services for people with chronic illnesses by comparing how Sweden and US services use evidence-based practices (EBPs), including digital health technologies (DHTs). Design/methodology/approach - A national primary healthcare center (PHCC) heads surveys in 2012-2013 carried out in both countries in 2006. Findings - There are large variations between the two countries. The largest, regarding effective DHT use in primary care centers, were that few Swedish primary healthcare compared to US heads reported having reminders or prompts at the point of care (38 percent Sweden vs 84 percent USA), despite Sweden's established electronic medical records (EMR). Swedish heads also reported 30 percent fewer centers receiving laboratory results (67 percent Sweden vs 97 percent USA). Regarding following other EBPs, 70 percent of Swedish center heads reported their physicians had easy access to diabetic patient lists compared to 14 percent in the USA. Most Swedish PHCC heads (96 percent) said they offered same day appointment compared to 36 percent in equivalent US practices. Practical implications - There are opportunities for improvement based on significant differences in effective practices between the countries, which demonstrates to primary care leaders that their peers elsewhere potentially provide better care for people with chronic illnesses. Some improvements are under primary care center control and can be made quickly. There is evidence that people with chronic illnesses in these two countries are suffering unnecessarily owing to primary care staff failing to provide proven EBP, which would better meet patient needs. Public finance has been invested in DHT, which are not being used to their full potential. Originality/value - The study shows the gaps between current and potential proven effective EBPs for services to patients with chronic conditions. Findings suggest possible explanations for differences and practical improvements by comparing the two countries. Many enhancements are low cost and the proportionate reduction in suffering and costs they bring is high.


ImmCluster: an ensemble resource for immunology cell type clustering and annotations in normal and cancerous tissues.
Single-cell transcriptome has enabled the transcriptional profiling of thousands of immune cells in complex tissues and cancers. However, subtle transcriptomic differences in immune cell subpopulations and the high dimensionality of transcriptomic data make the clustering and annotation of immune cells challenging. Herein, we introduce ImmCluster (http://bio-bigdata.hrbmu.edu.cn/ImmCluster) for immunology cell type clustering and annotation. We manually curated 346 well-known marker genes from 1163 studies. ImmCluster integrates over 420 000 immune cells from nine healthy tissues and over 648 000 cells from different tumour samples of 17 cancer types to generate stable marker-gene sets and develop context-specific immunology references. In addition, ImmCluster provides cell clustering using seven reference-based and four marker gene-based computational methods, and the ensemble method was developed to provide consistent cell clustering than individual methods. Five major analytic modules were provided for interactively exploring the annotations of immune cells, including clustering and annotating immune cell clusters, gene expression of markers, functional assignment in cancer hallmarks, cell states and immune pathways, cell-cell communications and the corresponding ligand-receptor interactions, as well as online tools. ImmCluster generates diverse plots and tables, enabling users to identify significant associations in immune cell clusters simultaneously. ImmCluster is a valuable resource for analysing cellular heterogeneity in cancer microenvironments.


Magnetic resonance image-guided hypofractionated ablative radiation therapy for extrahepatic cholangiocarcinoma: Plan adaptation in changing anatomy.
Extrahepatic cholangiocarcinoma is an aggressive malignancy of biliary duct epithelium which typically has a poor prognosis. Although surgical resection improves overall survival, many patients are deemed medically inoperable or have unresectable tumors. Herein, we report a case of an 84-year-old Caucasian male who was diagnosed with medically inoperable extrahepatic cholangiocarcinoma. Magnetic resonance image-guided hypofractionated ablative radiotherapy was administered which has the advantages of superior soft tissue resolution, better visualization of the target and organ at risk, daily online adaptive planning and continuous cine MR tracking of the target during irradiation. Concomitant chemotherapy was used. On the first fraction the patient presented with a broken arm that forced the patient take a treatment position much different than the one used for simulation CT and planning. The patient was able to finish the treatment without the need of another simulation by adapting the plan according to the new anatomy. The patient is being followed up until today and is alive with no evidence of disease.


Structural studies of Fc receptors. IV. Structure required for phospholipids for reconstitution of the delipidated Fc receptor of macrophages.
To analyze the interaction of the macrophage Fc receptor with phospholipids, we established an experimental system for delipidation of Fc receptor fraction and reconstitution of the Fc receptor activity in phospholipid vesicles. The separation of FcR from membrane phospholipids was achieved by ion exchange chromatography on DEAE-cellulose of the anionic detergent-lysate of the crude membrane fraction of guinea pig macrophages in the presence of detergent. The separation was based on the difference in charge between the complex of FcR and the anionic detergent and that of phospholipids and the detergent. The FcR fraction free of phospholipids showed no FcR activity as assessed in terms of its ability to inhibit the binding of labeled soluble immune complex of IgG2 antibody to macrophages, but the same fraction showed a definite activity when associated with phospholipids. This fraction was shown to contain a component of 44,000 daltons that is susceptible to surface-labeling and binds to IgG2-Sepharose in the affinity chromatography, indicating this component to be the Fc receptor. Reconstitution experiments with this fraction showed that phosphatidylcholine is the most effective phospholipid to reconstitute the FcR activity among those tested. Phosphatidylserine, phosphatidylinositol, and sphingomyelin were ineffective, while phosphatidylethanolamine showed a moderate effect. The inactivating effect of phospholipase C treatment on the Fc receptor activity of the membrane was shown to be due to the cleavage of phospholipids in the membrane but not due to modification of the Fc receptor molecule itself.


ATP binding and hydrolysis disrupt the high-affinity interaction between the heme ABC transporter HmuUV and its cognate substrate-binding protein.
Using the energy of ATP hydrolysis, ABC transporters catalyze the trans-membrane transport of molecules. In bacteria, these transporters partner with a high-affinity substrate-binding protein (SBP) to import essential micronutrients. ATP binding by Type I ABC transporters (importers of amino acids, sugars, peptides, and small ions) stabilizes the interaction between the transporter and the SBP, thus allowing transfer of the substrate from the latter to the former. In Type II ABC transporters (importers of trace elements, e.g. vitamin B12, heme, and iron-siderophores) the role of ATP remains debatable. Here we studied the interaction between the Yersinia pestis ABC heme importer (HmuUV) and its partner substrate-binding protein (HmuT). Using real-time surface plasmon resonance experiments and interaction studies in membrane vesicles, we find that in the absence of ATP the transporter and the SBP tightly bind. Substrate in excess inhibits this interaction, and ATP binding by the transporter completely abolishes it. To release the stable docked SBP from the transporter hydrolysis of ATP is required. Based on these results we propose a mechanism for heme acquisition by HmuUV-T where the substrate-loaded SBP docks to the nucleotide-free outward-facing conformation of the transporter. ATP binding leads to formation of an occluded state with the substrate trapped in the trans-membrane translocation cavity. Subsequent ATP hydrolysis leads to substrate delivery to the cytoplasm, release of the SBP, and resetting of the system. We propose that other Type II ABC transporters likely share the fundamentals of this mechanism.


Chromosomal translocations and non-B DNA structures in the human genome.
The mechanisms of chromosomal translocations in mammalian cells have been largely undefined. Recent progress on the most common translocation in human cancer, t(14;18), highlights interesting issues in DNA structure and in the enzymes involved in the cutting and joining phases of the process.


A Van Gogh/Vangl tyrosine phosphorylation switch regulates its interaction with core Planar Cell Polarity factors Prickle and Dishevelled.
Epithelial tissues can be polarized along two axes: in addition to apical-basal polarity they are often also polarized within the plane of the epithelium, known as planar cell polarity (PCP). PCP depends upon the conserved Wnt/Frizzled (Fz) signaling factors, including Fz itself and Van Gogh (Vang/Vangl in mammals). Here, taking advantage of the complementary features of Drosophila wing and mouse skin PCP establishment, we dissect how Vang/Vangl phosphorylation on a specific conserved tyrosine residue affects its interaction with two cytoplasmic core PCP factors, Dishevelled (Dsh/Dvl1-3 in mammals) and Prickle (Pk/Pk1-3). We demonstrate that Pk and Dsh/Dvl bind to Vang/Vangl in an overlapping region centered around this tyrosine. Strikingly, Vang/Vangl phosphorylation promotes its binding to Prickle, a key effector of the Vang/Vangl complex, and inhibits its interaction with Dishevelled. Thus phosphorylation of this tyrosine appears to promote the formation of the mature Vang/Vangl-Pk complex during PCP establishment and conversely it inhibits the Vang interaction with the antagonistic effector Dishevelled. Intriguingly, the phosphorylation state of this tyrosine might thus serve as a switch between transient interactions with Dishevelled and stable formation of Vang-Pk complexes during PCP establishment.


Chemoinformatic modelling of the antioxidant activity of phenolic compounds.
Antioxidants are chemicals used to protect foods from deterioration by neutralizing free radicals and inhibiting the oxidative process. One approach to investigate the antioxidant activity is to develop quantitative structure-activity relationships (QSARs). A curated database of 165 structurally heterogeneous phenolic compounds with the Trolox equivalent antioxidant capacity (TEAC) was developed. Molecular geometries were optimized by means of the GFN2-xTB semiempirical method and diverse molecular descriptors were obtained afterwards. For model development, V-WSP unsupervised variable reduction was used before performing the genetic algorithms-variable subset selection (GAs-VSS) to construct the best five-descriptor multiple linear regression model. The coefficient of determination and the root mean square error were used to measure the performance in calibration (R2 = 0.789 and RMSEC = 0.381), and test set prediction (Q2 = 0.748 and RMSEP = 0.416), along several cross-validation criteria. To thoroughly understand the TEAC prediction, a fully explained mechanism of action of the descriptors is provided. In addition, the applicability domain of the model defined a theoretical chemical space for reliable predictions of new phenolic compounds. This in silico model conforms to the five principles stated by the Organisation for Economic Co-operation and Development. The model might be useful for virtual screening of the antioxidant chemical space and for identifying the most potent molecules related to an experimental measurement of TEAC activity. In addition, the model could assist chemists working on computer-aided drug design for the synthesis of new targets with improved activity and potential uses in food science. © 2023 Society of Chemical Industry.


In vivo enhancement of 5-fluorouracil cytotoxicity to AKR leukemia cells by thymidine in mice.
The spleen colony assay was used to examine the effect of thymidine (dThd) on 5-fluorouracil (FUra) cytotoxicity in two transplantable leukemias, AKR (in AKR mice) and L1210 [in (BALB/c x DBA/2)F1 mice], in vivo. A large dose of dThd (10 mg/mouse) could not rescue these cell lines from FUra toxicity. Instead, when dThd was given within 1 hour before FUra, it enhanced FUra cytotoxicity by a factor between 100 and 1,000 in AKR leukemia. That dThd increased the cytotoxicity of FUra only by a factor of 3 in L1210 leukemia suggested a different mechanism of interaction of the two drugs in the two cell lines. Examination in hybrid mice capable of supporting the growth of both leukemias showed the enhancement to be tumor related rather than host related. We also demonstrated a dose-dependent effect of dThd injection 15 minutes before FUra in AKR leukemia. Concerning the kinetics of killing of AKR leukemia colony-forming units (LCFU) following the administration of dThd 15 minutes before FUra, LCFU survival continued to decrease for 24--36 hours following drug administration.


Sperm viability, serological, molecular, and modified seminal plasma agglutination tests in the diagnosis of Leptospira in the semen and serum of bovine bulls.
The present study investigated the serum microscopic agglutination test (MAT) among 203 bovine bulls with reproduction by natural means, without apparent signs of orchitis or inflammation of accessory reproductive glands. Simultaneously, the semen of all bulls was subjected to sperm viability analysis and PCR based on the 16S rRNA gene. PCR-positive results of semen samples were confirmed by sequencing. A modified seminal plasma agglutination (MSPA) test, replacing the blood serum of all bulls in the MAT with seminal plasma was performed as well. Eight (8/203 = 3.9%) semen samples from bulls were considered nonviable (necrospermia and azoospermia) without relation to the PCR diagnosis. No agglutinin titers were identified in MSPA test. A high frequency (132/203 = 65%) of leptospiral agglutinin titers was identified in the MAT, particularly for the Sejroe serogroup (Hardjo CTG, 100/203 = 49.3%; Wolffi 74/203 = 36.4%; Guaricura 72/203 = 35.5%; and Hardjoprajitno 56/203 = 27.6%). Three (3/203 = 1.5%) semen samples of bulls were positive in the PCR, but these results were not confirmed by sequencing. The high frequency of serovars from the Sejroe serogroup typically adapted to bovines indicates the need for measures for the prophylaxis/control of the pathogen on the sampled farms. Discrepancies among the MAT, sperm viability, and molecular detection of leptospires in semen highlight the need for a combination of methods to diagnose leptospirosis in bovine bulls. To our knowledge, modified seminal plasma agglutination is described for the first time here to investigate anti-Leptospira antibodies produced locally in the genital tract in the diagnosis of bovine leptospirosis among bulls that reproduce by natural means.


Hexaazatriphenylene-Based Two-Dimensional Conductive Covalent Organic Framework with Anisotropic Charge Transfer.
The development of covalent organic frameworks (COFs) with efficient charge transport is of immense interest for applications in optoelectronic devices. To enhance COF charge transport properties, electroactive building blocks and dopants can be used to induce extended conduction channels. However, understanding their intricate interplay remains challenging. We designed and synthesized a tailor-made COF structure with electroactive hexaazatriphenylene (HAT) core units and planar dioxin (D) linkages, denoted as HD-COF. With the support of theoretical calculations, we found that the HAT units in the HD-COF induce strong, eclipsed π-π stacking. The unique stacking of HAT units and the weak in-plane conjugation of dioxin linkages leads to efficient anisotropic charge transport. We fabricated HD-COF films to minimize the grain boundary effect of bulk COFs, which resulted in enhanced conductivity. As a result, the HD-COF films showed an electrical conductivity as high as 1.25 S cm-1 after doping with tris(4-bromophenyl)ammoniumyl hexachloroantimonate.


Construction of a bioreporter by heterogeneously expressing a Vibrio natriegens recA::luxCDABE fusion in Escherichia coli, and genotoxicity assessments of petrochemical-contaminated groundwater in northern China.
Here, we constructed an Escherichia coli recA::luxCDABE bioreporter for genotoxicity assessments. The recA promoter was cloned from the marine bacterium Vibrio natriegens. This bioreporter showed a dose-response relationship following induction by mitomycin C, and other pollutants or environmental samples could be calculated as mitomycin C equivalents, which provided a way to quantitatively compare the genotoxicities of different environmental samples. This bioreporter was used to evaluate the genotoxicity under a wide range of external environmental conditions, like temperatures ranging from 15 °C to 42 °C, pH between 4.0 and 9.0, and salinity ranging from 0% to 3%. This successfully extended its application from the laboratory to the field, and allowed the bioreporter to assess the genotoxicity and bioavailability of genotoxins in various environmental media, including surface water, groundwater, seawater, and soil matrix. Expression of V. natriegens recA in E. coli indicated a LexA-like regulator in V. natriegens, and the putative SOS box of V. natriegens recA was similar to that of E. coli. The genotoxicities of groundwater samples from a petrochemical-contaminated site in northern China were evaluated by this bioreporter assay, and the genotoxic levels were in accordance with contamination levels obtained by chemical analyses.


Application of magnetic nanoparticles to gene delivery.
Nanoparticle technology is being incorporated into many areas of molecular science and biomedicine. Because nanoparticles are small enough to enter almost all areas of the body, including the circulatory system and cells, they have been and continue to be exploited for basic biomedical research as well as clinical diagnostic and therapeutic applications. For example, nanoparticles hold great promise for enabling gene therapy to reach its full potential by facilitating targeted delivery of DNA into tissues and cells. Substantial progress has been made in binding DNA to nanoparticles and controlling the behavior of these complexes. In this article, we review research on binding DNAs to nanoparticles as well as our latest study on non-viral gene delivery using polyethylenimine-coated magnetic nanoparticles.


Black Sea hydrate production value and options for clean energy production.
Natural gas hydrates of Bulgaria and Romania in the Black Sea have been subject to studies by several European research projects. The current understanding of the hydrate distribution, and the total amounts of hydrate in the region, makes it interesting to evaluate in terms of commercial potential. In this study, we have evaluated some well-known hydrate production methods. Thermal stimulation and adding chemicals are considered as not economically feasible. Pressure reduction may not be efficient due to the endothermic dissociation of hydrates and long-term cooling of the sediments. Chemical work due to pressure reduction is an additional mechanism but is too slow to be commercially feasible. Adding CO2/N2, however, has a dual value. In the future, CO2 can be stored at a price proportional to a CO2 tax. This is deducted from the value of the released natural gas. The maximum addition of N2 is around 30 mol% of the CO2/N2 mixture. A minor addition (in the order of 1 mol%) of CH4 increases the stability of the hydrate created from the injection gas. The maximum N2 amount is dictated by the demand for the creation of a new hydrate from injection gas but also the need for sufficient heat release from this hydrate formation to dissociate the in situ CH4 hydrates. An additional additive is needed to accelerate the formation of hydrate from injection gas while at the same time reducing the creation of blocking hydrate films. Based on reasonable assumptions and approximations as used in a verified kinetic model it is found that CH4/CO2 swapping is a feasible method for Black Sea hydrates. It is also argued that the technology is essentially conventional petroleum technology combined with learning from projects on aquifer storage of CO2, and a thermodynamic approach for design of appropriate injection gas. It is also argued that the CH4/CO2 swap can be combined with well-known technology for steam cracking of produced hydrocarbons to H2 and CO2 (for re-injection).


Application of computer texture analysis to the Singh Index.
The Singh Index, which describes trabecular patterns in the proximal femur, has been used as a predictor for hip fractures and as an indicator of osteopenia. Evidence suggests that its contribution to the assessment of hip fracture risk is in its description of the structural properties of the femur. However, subjectivity in its determination may limit its practical value. In this study, we have used computer texture analysis to characterize the textural changes in trabecular bone which occur in osteoporosis. Texture analysis of the proximal femur was used to rank radiographs of 25 cadaver femora by Singh grading. These rankings were compared with those performed by two experienced radiologists. The computer method performed better than the least consistent individual radiologist, but not as well as the two radiologists in combination. The method required no radiologist's time after the system had been 'trained' from a set of example radiographs. The method has potential for use in large studies in which speed and consistency of measurement are important.


BitQT: a graph-based approach to the quality threshold clustering of molecular dynamics.
Classical Molecular Dynamics (MD) is a standard computational approach to model time-dependent processes at the atomic level. The inherent sparsity of increasingly huge generated trajectories demands clustering algorithms to reduce other post-simulation analysis complexity. The Quality Threshold (QT) variant is an appealing one from the vast number of available clustering methods. It guarantees that all members of a particular cluster will maintain a collective similarity established by a user-defined threshold. Unfortunately, its high computational cost for processing big data limits its application in the molecular simulation field. In this work, we propose a methodological parallel between QT clustering and another well-known algorithm in the field of Graph Theory, the Maximum Clique Problem. Molecular trajectories are represented as graphs whose nodes designate conformations, while unweighted edges indicate mutual similarity between nodes. The use of a binary-encoded RMSD matrix coupled to the exploitation of bitwise operations to extract clusters significantly contributes to reaching a very affordable algorithm compared to the few implementations of QT for MD available in the literature. Our alternative provides results in good agreement with the exact one while strictly preserving the collective similarity of clusters. The source code and documentation of BitQT are free and publicly available on GitHub (https://github.com/LQCT/BitQT.git) and ReadTheDocs (https://bitqt.readthedocs.io/en/latest/), respectively. Supplementary data are available at Bioinformatics online.


A miR-29b Byproduct Sequence Exhibits Potent Tumor-Suppressive Activities via Inhibition of NF-κB Signaling in KRAS-Mutant Colon Cancer Cells.
We previously demonstrated that miR-29b-3p is a hopeful miRNA-based therapy against colorectal cancer. In this study, we aimed to clarify a value of miR-29b-1-5p as a next-generation treatment, especially for KRAS-mutant colorectal cancer. RT-PCR assay showed that the expression of miR-29b-3p was high, and its partner strand, miR-29b-1-5p, level was only negligible in clinical colorectal cancer samples. Mimic-miR-29b-1-5p significantly inhibited proliferation of KRAS-mutant colorectal cancer cell lines DLD1 and SW480 and KRAS wild-type HT29 cells. Proliferative activity was further examined by either miR-29b-1-5p strand or its opposite complementary sequence because miR-29b-1-5p is a passenger miRNA and may have no physiologic function. We found that completely opposite complementary strand to miR-29b-1-5p, but not miR-29b-1-5p, possessed a potent antitumor effect and named this byproduct miRNA sequence "MIRTX." MIRTX directly targeted the 3'-UTR of CXCR2 and PIK3R1 mRNA and suppressed the NF-κB signaling pathway in KRAS-mutated colorectal cancer cells. MIRTX induced apoptosis in DLD1 with downregulation of antiapoptotic BCL2, BCL-xL, and MCL1 and upregulation of cleaved caspase-3 and cleaved PARP. In mouse xenograft models, systemic administration of MIRTX using a super carbonate apatite as a delivery vehicle significantly inhibited tumor growth of DLD1 and HT29 cells without any particular toxicities. In conclusion, these findings indicate that inhibition of NF-κB signaling by this novel miRNA-based therapeutic could be a promising treatment against refractory KRAS-mutant colorectal cancer and KRAS wild-type colorectal cancer. Mol Cancer Ther; 17(5); 977-87. ©2018 AACR.


Improved NMR experiments with ¹³C-isotropic mixing for assignment of aromatic and aliphatic side chains in labeled proteins.
Three improved ¹³C-spinlock experiments for side chain assignments of isotope labelled proteins in liquid state are presented. These are based on wide bandwidth spinlock techniques that have become possible with contemporary cryogenic probes. The first application, the H(C(ali)C(aro))H-TOCSY, is an HCCH-TOCSY in which all CHn moieties of a protein are detected in a single experiment, including the aromatic ones. This enables unambiguous assignment of aromatic and aliphatic amino acids in a single, highly sensitive experiment. In the second application, the ¹³C-detected C(all)-TOCSY, magnetization transfer comprises all carbons--aliphatic, aromatic as well as the carbonyl carbons--making the complete carbon assignment possible using one spectrum only. Thirdly, the frequently used HC(CCO)NH experiment was redesigned by replacing the long C-carbonyl refocused INEPT transfer step by direct ¹³C-¹³C-TOCSY magnetization transfer from side chain carbons to the backbone carbonyls. The resulting HC(CCO)NH experiment minimizes relaxation losses because it is shorter and represents a more sensitive alternative particularly for larger proteins. The performance of the experiments is demonstrated on isotope labeled proteins up to the size of 43 kDa.


Neuropsychological test performance of successful brain injury simulators.
This study provided an examination of the performance characteristics of successful brain injury simulators (SBIS). Coached (n = 56) and uncoached (n = 35) brain injury simulators received instructions to fake cognitive impairment; controls were asked to do their best. The Test of Memory Malingering (TOMM) was administered along with standard neuropsychological measures (e.g., Wisconsin Card Sorting Test). The TOMM identified 80% of uncoached and 60% of coached brain injury simulators. SBIS were participants from the brain injury simulation groups whose TOMM performance indicated adequate effort. A total of 32% of all brain injury simulators scored above the TOMM cutoff scores for adequate effort (the SBIS group). Significantly more coached than uncoached participants composed the SBIS group (76% vs. 24%, respectively). SBIS performed significantly worse than controls and significantly better than unsuccessful brain injury simulators on select standard neuropsychological measures. The SBIS scores were lowered compared to controls; in some instances this lowered performance was at a clinically relevant level.


Quasi-Racemic X-ray Structures of K27-Linked Ubiquitin Chains Prepared by Total Chemical Synthesis.
Quasi-racemic crystallography has been used to determine the X-ray structures of K27-linked ubiquitin (Ub) chains prepared through total chemical synthesis. Crystal structures of K27-linked di- and tri-ubiquitins reveal that the isopeptide linkages are confined in a unique buried conformation, which provides the molecular basis for the distinctive function of K27 linkage compared to the other seven Ub chains. K27-linked di- and triUb were found to adopt different structural conformations in the crystals, one being symmetric whereas the other triangular. Furthermore, bioactivity experiments showed that the ovarian tumor family de-ubiquitinase 2 significantly favors K27-linked triUb than K27-linked diUb. K27-linked triUb represents the so-far largest chemically synthesized protein (228 amino acids) that has been crystallized to afford a high-resolution X-ray structure.


The Spatial Diffusion of Racial and Ethnic Diversity Across U.S. Counties.
Although increasing racial and ethnic diversity is a demographic trend with society-wide implications, it has advanced farther in some parts of the United States than others. Our research seeks to understand this unevenness at the local level. Drawing on 1980-2010 census data, we use an innovative spatial analytic approach to examine the spread or diffusion of diversity across counties in the 48 contiguous states. Three perspectives-locational persistence, spatial assimilation, and institutional hub-offer different expectations about the nature of the diffusion process. The perspectives are evaluated by mapping changes in the magnitude and structure of diversity and by tracing county transitions between types of diversity clusters. We document considerable stability in diversity patterns over a 30-year period, consistent with the logic of locational persistence. But support is also found for the spatial assimilation and institutional hub models in the form of cluster-type transitions that reflect contagious diffusion and hierarchical diffusion, respectively.


Development of A Liquid Chromatography-Mass Spectrometry Technique for Evaluation of Multi-class Pesticides in Rice Samples.
In the current study, a liquid chromatography coupled mass detector was set up to detect and quantify 108 pesticide residues in rice samples. QuEChERS method was applied for sample preparation and different validation parameters were determined to ensure the suitability of the method. The calibration curves were linear in the concentration 0.01-1.00 mg/kg with a coefficient of determination (R2) of more than 0.990 for all compounds. Based on signal to noise studies, the calculated LODs and LOQs were 0.005-0.060 mg/kg and 0.018-0.199 mg/kg, respectively; and acquired mean recoveries at three spiked levels (0.025, 0.200 and 0.800 mg/kg) were 72% - 117% with RSD < 20%. The developed method was used to investigate the occurrence of the studied pesticides in 65 internal and 65 foreign rice samples. The results showed that 14 internal and 15 imported samples were found to be contaminated 12 pesticides in the amounts between 0.027 mg/kg to 0.078 mg/kg and 0.031 mg/kg to 0.081 mg/kg, respectively. According to the Iranian regulations, with the exception of nine prohibited pesticides for rice production in Iran, bioallethrin, cypermethrin, deltamethrin, flutriafol, foramsulfuron, imazalil, phosphamidon, TCMTB, and triasulfuron, three permitted pesticides, cinosulfuron, triadimenol, and tricyclazole, found in positive rice samples were below MRLs established by Iranian National Standard Organization (INSO).


Effect of Cannabidiolic Acid, N-Trans-Caffeoyltyramine and Cannabisin B from Hemp Seeds on microRNA Expression in Human Neural Cells.
Given the increasing interest in bioactive dietary components that can modulate gene expression enhancing human health, three metabolites isolated from hemp seeds-cannabidiolic acid, N-trans-caffeoyltyramine, and cannabisin B-were examined for their ability to change the expression levels of microRNAs in human neural cells. To this end, cultured SH-SY5Y cells were treated with the three compounds and their microRNA content was characterized by next-generation small RNA sequencing. As a result, 31 microRNAs underwent major expression changes, being at least doubled or halved by the treatments. A computational analysis of the biological pathways affected by these microRNAs then showed that some are implicated in neural functions, such as axon guidance, hippocampal signaling, and neurotrophin signaling. Of these, miR-708-5p, miR-181a-5p, miR-190a-5p, miR-199a-5p, and miR-143-3p are known to be involved in Alzheimer's disease and their expression changes are expected to ameliorate neural function. Overall, these results provide new insights into the mechanism of action of hemp seed metabolites and encourage further studies to gain a better understanding of their biological effects on the central nervous system.


Bulk fluid phase behaviour of colloidal platelet-sphere and platelet-polymer mixtures.
Using a geometry-based fundamental measure density functional theory, we calculate bulk fluid phase diagrams of colloidal mixtures of vanishingly thin hard circular platelets and hard spheres. We find isotropic-nematic phase separation, with strong broadening of the biphasic region, upon increasing the pressure. In mixtures with large size ratio of platelet and sphere diameters, there is also demixing between two nematic phases with differing platelet concentrations. We formulate a fundamental measure density functional for mixtures of colloidal platelets and freely overlapping spheres, which represent ideal polymers, and use it to obtain phase diagrams. We find that, for low platelet-polymer size ratio, in addition to isotropic-nematic and nematic-nematic phase coexistence, platelet-polymer mixtures also display isotropic-isotropic demixing. By contrast, we do not find isotropic-isotropic demixing in hard-core platelet-sphere mixtures for the size ratios considered.


Clinical experience with enlargement of the aortic anulus by extension of the aortic incision into the anterior mitral leaflet.
Implantation of a small-sized biological prosthesis into a hypoplastic aortic annulus causes significant resting gradients. To make insertion of a suitable biological valve possible, we recommend enlargement of the aortic annulus by extension of the aortic incision into the anterior mitral leaflet for approximately 2 cm. A woven Dacron patch is sutured into the V-shaped defect of the anterior mitral leaflet and the aortic ring. This procedure results in enlargement of the aortic circumference of approximately 20 mm. We have applied this technique in 13 patients with severe aortic valvular disease and a hypoplastic aortic annulus. In all patients a prosthesis with a diameter of at least 25 mm could be implanted. The maximal postoperative pressure gradient measured 18 mmHg. There was no early death related to this operative method. All survivors have excellent clinical results with no evidence of mitral regurgitation. This operation represents a practicable alternative method to left ventricular apico-aortic conduit implantation.


Efferent Vestibular Neurons Show Homogenous Discharge Output But Heterogeneous Synaptic Input Profile In Vitro.
Despite the importance of our sense of balance we still know remarkably little about the central control of the peripheral balance system. While previous work has shown that activation of the efferent vestibular system results in modulation of afferent vestibular neuron discharge, the intrinsic and synaptic properties of efferent neurons themselves are largely unknown. Here we substantiate the location of the efferent vestibular nucleus (EVN) in the mouse, before characterizing the input and output properties of EVN neurons in vitro. We made transverse serial sections through the brainstem of 4-week-old mice, and performed immunohistochemistry for calcitonin gene-related peptide (CGRP) and choline acetyltransferase (ChAT), both expressed in the EVN of other species. We also injected fluorogold into the posterior canal and retrogradely labelled neurons in the EVN of ChAT:: tdTomato mice expressing tdTomato in all cholinergic neurons. As expected the EVN lies dorsolateral to the genu of the facial nerve (CNVII). We then made whole-cell current-, and voltage-clamp recordings from visually identified EVN neurons. In current-clamp, EVN neurons display a homogeneous discharge pattern. This is characterized by a high frequency burst of action potentials at the onset of a depolarizing stimulus and the offset of a hyperpolarizing stimulus that is mediated by T-type calcium channels. In voltage-clamp, EVN neurons receive either exclusively excitatory or inhibitory inputs, or a combination of both. Despite this heterogeneous mixture of inputs, we show that synaptic inputs onto EVN neurons are predominantly excitatory. Together these findings suggest that the inputs onto EVN neurons, and more specifically the origin of these inputs may underlie EVN neuron function.


Temporal relation of ATP-sensitive potassium-channel activation and contractility before cardioplegia.
Pharmacologic treatment using potassium-channel openers (PCOs) before cardioplegic arrest has been demonstrated to provide beneficial effects on left ventricular performance with subsequent reperfusion and rewarming. However, the PCO treatment interval necessary to provide protective effects during cardioplegic arrest remains to be defined. The present study was designed to determine the optimum period of PCO treatment that would impart beneficial effects on left ventricular myocyte contractility after simulated cardioplegic arrest. Left ventricular porcine myocytes were assigned randomly to three groups: (1) normothermic control = 37 degrees C for 2 hours; (2) cardioplegia = K+ (24 mEq/L) at 4 degrees C for 2 hours followed by reperfusion and rewarming; and (3) PCO and cardioplegia = 1 to 15 minutes of treatment with the PCO aprikalim (100 micromol/L) at 37 degrees C followed by hypothermic (4 degrees C) cardioplegic arrest and subsequent rewarming. Myocyte contractility was measured after rewarming by videomicroscopy. A minimum of 50 myocytes were examined at each treatment and time point. Myocyte velocity of shortening was reduced after cardioplegic arrest and rewarming compared with normothermic controls (63+/-3 microm/s versus 32+/-2 microm/s, respectively; p < 0.05). With 3 minutes of PCO treatment, myocyte velocity of shortening was improved after cardioplegic arrest to values similar to those of normothermic controls (56+/-3 microm/s). Potassium channel opener treatment for less than 3 minutes did not impart a protective effect, and the protective effect was not improved further with more prolonged periods of PCO treatment. A brief interval of PCO treatment produced beneficial effects on left ventricular myocyte contractile function in a simulated model of cardioplegic arrest and rewarming. These results suggest that a brief period of PCO treatment may provide a strategy for myocardial protection during prolonged cardioplegic arrest in the setting of cardiac operation.


[123I-alpha-methyltyrosine scintigraphy in malignant melanoma].
The aim of the study was to evaluate the ranking of the scintigraphy with L-3-123I-alpha-methyltyrosine (123I-AMT) in metastasized melanoma. 26 metastases and one primary tumor of a malignant melanoma in six patients were examined with 123I-AMT whole-body scintigraphy and SPECT. Positron Emission Tomography with 2-18F-fluoro-2-desoxy-D-glucose (18F-FDG) was used as the golden standard. With 123I-AMT-SPECT 8/10 metastases in the thorax > 1.6 cm were detected (ratio T/NT 1.2-1.8), metastases < 1.6 cm were not detectable with SPECT. In 123I-AMT whole-body scintigraphy not one lesion showed a positive tumor uptake. In single cases 123I-AMT scintigraphy can be helpful in staging of malignant melanoma.


Pentraxin 3 (PTX-3) Levels in Bronchoalveolar Lavage Fluid as a Lung Cancer Biomarker.
In this study, we investigated the utility of pentraxin 3 (PTX-3) in bronchoalveolar lavage fluid (BALF) as lung cancer (LCa) diagnostic. A total of 89 LCa patients and 84 non-LCa patients who received bronchoscopy in the First Affiliated Hospital of Xi'an Jiaotong University from December 2014 to February 2015 were enrolled. LCa was subdivided according to pathological type (scale, gland, and small cell lung cancer). BALF samples were obtained during bronchoscopy and PTX-3 levels assayed by ELISA. t-tests, Mann-Whitney, and Kruskal-Wallis tests were performed for the comparison of PTX-3 levels between the different groups. Correlation analysis and receiver operating characteristic (ROC) analysis were used to analyze clinical data. The levels of PTX-3 increased in the LCa groups. PTX-3 levels were higher in the small cell lung cancer (SCLC) compared to non-small-cell lung cancer (NSCLC) groups. In LCa patients, obstructive pneumonia could upregulate the expression of PTX-3 in BALF. The area under the ROC curve of PTX-3 in BALF during LCa diagnosis, SCLC, and LCa with obstructive pneumonia was 0.949 (p ≤ 0.001), 0.672 (p < 0.05), and 0.838 (p < 0.01), respectively. In conclusion, PTX-3 in BALF has a potential value as an LCa biomarker, particularly in cases of SCLC and LCa with obstructive pneumonia.


[Influence of the ovarian cycle on the central nervous system].
Estradiol, progesterone and some of their metabolites modulate the activity of neurotransmitters and neuropeptides in the CNS. The distribution and concentrations of sex steroids in the various CNS regions is partly dependent on the serum levels, but also on the local synthesis of the steroids. In general, estradiol and testosterone exert a stimulatory, progesterone an inhibitory effect on neuronal activities which are mediated by excitatory (e.g. glutamate, aspartate), and inhibitory amino acids (e.g. GABA) and neuropeptides (e.g. beta-endorphin), respectively. Gonadotropin release is primarily governed by the rhythm of pulsatile secretion of GnRH in the hypothalamus which is controlled by estradiol and progesterone by means of inhibitory or stimulatory modulation of the amplitude and frequency of GnRH pulses. The discharges of GnRH neurons triggered by excitatory amino acids are modulated by estradiol, while the inhibitory effect of progesterone is mediated by GABA and beta-endorphin which cause hyperpolarization of the GnRH neurons and consequently a reduced pulse frequency. The pulse amplitudes are primarily influenced by estradiol, but neuropeptide Y, neurotensin and noradrenaline contribute to their preovulatory enhancement. The postovulatory rise in core temperature is caused by the increasing level of progesterone and its metabolite 3 alpha-pregnanolone, respectively. Despite of this, up to 20% of ovulatory cycles do not show any rise in body temperature. Although 3 alpha-pregnanolone has sedative activities, there is no change in sleep quality during the luteal phase due to their low serum levels. It could be demonstrated that performance on tests of articulatory and fine motor skills are enhanced in the late follicular phase as compared to the menstruation phase, while spatial ability was better during menses. Estrogens may influence mood and well-being in a favorable manner, while in predisposed women progesterone may cause symptoms of premenstrual syndrome. In most women there are, however, no cycle-dependent mood changes. An increase in appetite can be observed during the periovulatory phase and before menses, while sexual interest increases in the follicular phase. Somatic complaints (back pain, abdominal pain, breast tenderness) which are highest before and during menstruation, are probably associated with a lowered pain threshold due to a fall in the beta-endorphin levels in the CNS.