Fragment-based development of HCV protease inhibitors for the treatment of hepatitis C.
A novel computational technology based on fragmentation of the chemical compounds has been used for the fast and efficient prediction of activities of prospective protease inhibitors of the hepatitis C virus. This study spans over a discovery cycle from the theoretical prediction of new HCV NS3 protease inhibitors to the first cytotoxicity experimental tests of the best candidates. The measured cytotoxicity of the compounds indicated that at least two candidates would be suitable further development of drugs.


Safety and efficacy of moxonidine in mild to moderate hypertension.
Moxonidine, a new imidazoline II receptor agonist was evaluated in patients with essential hypertension. After an informed consent, 16 patients with essential hypertension were recruited. After a wash out period, the patients received increasing doses of Moxonidine (maximum dose 0.6 mg) for four weeks to control the blood pressure. The mean reduction in blood pressure (systolic/diastolic) was 17.6/14.5 mm Hg in supine position and 18.2/12.6 mm Hg in standing position. The significant side effect was observed only in one patient. Moxonidine thus appears to be safe and effective antihypertensive drug in patients with essential hypertension.


Comparative evaluation of platelet counts in two hematology analyzers and potential effects on prophylactic platelet transfusion decisions.
Decisions on prophylactic platelet (PLT) transfusions are generally based on the recipient's PLT count, but few clinicians are aware of precision and accuracy of the PLT counting methods used by the clinical laboratory. Each PLT counting technology has its specific inaccuracy, especially in thrombocytopenic samples and therefore may impact decisions on PLT transfusions. Five routine PLT counting methods available in two hematology analyzers (Sysmex XN-2000 and Abbott CELL-DYN Sapphire) were investigated (impedance and optical on both analyzers and fluorescent on XN-2000), using the CD61 immunologic PLT method as a reference. The impact of counting inaccuracy on imaginary transfusion decisions was examined at various common PLT thresholds. In total 341 samples were analyzed, 178 of which had PLT counts of less than 35 × 109 /L. Despite excellent overall correlation with the reference method (r > 0.99), thrombocytopenic samples showed only modest correlation for impedance and XN-2000 optical methods. Sapphire optical and XN-2000 fluorescent methods correlated very well with the reference, albeit with bias in the very low range. We noticed potential risk of undertransfusion (ranging from 2% to 90%), depending on the threshold used. The risk of overtransfusion was small (<10%). The XN-2000 fluorescent PLT counting method showed excellent correlation with the CD61 reference count, closely followed by the CELL-DYN Sapphire optical method. XN-2000 impedance and optical and Sapphire impedance methods are not accurate enough for basing transfusion decisions on. Only XN-2000 fluorescent, Sapphire optical, and CD61 methods are sufficiently accurate for making appropriate clinical decisions in patients with severe thrombocytopenia.


Chromosomal aberrations induced in vitro by 3,7- and 3,9-dinitrofluoranthene.
The chromosomal aberration test using a Chinese hamster cell line (CHL) was carried out with 3,7- and 3,9-dinitrofluoranthene (DNF) with and without exogenous metabolic activation (rat liver S9 mix). The highest dose tested was limited to 20 micrograms/ml because of the compounds' insolubility in dimethyl sulfoxide. Both DNFs induced chromosomal aberrations in the absence of S9 mix; the frequency was not very high. Results were reproducible, but without clear dose-response relationships. Neither DNF induced chromosomal aberrations in the presence of S9 mix. Both DNFs did not induce polyploid cells under any conditions.


Apoptosis in meningoencephalitis of Angiostrongylus cantonensis-infected mice.
A hallmark of eosinophilic meningoencephalitis is infiltration of leukocytes into brain parenchyma and subarachnoid space infected by Angiostrongylus cantonensis. Apoptosis, a process that eliminates useless cells and counterbalances tissue homeostasis, is important for homeostasis of the immune system. In this study, we investigated the characteristics of cell death induced in BABL/c mice infected with A. cantonensis. We observed increased expression of the apoptotic proteins, caspase-3, caspase-8, caspase-9, and cytochrome c, and decreased expression of anti-apoptotic proteins, B-cell leukemia 2 and inhibitor of apoptosis protein 1. On immunohistochemistry, apoptotic proteins were localized within the leukocytes infiltrate. A terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling assay to detect DNA fragmentation confirmed these observations. The infiltration of leukocytes present in the brain parenchyma and subarachnoid space in vivo may also express these apoptotic regulatory molecules, which demonstrates the capacity of these cells to undergo apoptosis.


Doomed for carcinomatosis? An unusual presentation of abdominal tuberculosis.
A 35-year old male from Brazil presented with intermittent abdominal pain. Abdominal computed tomography revealed a nodule adjacent to splenic hilum and multiple abdominal nodules, suspicious of carcinomatosis. The patient underwent gastroscopy and endoscopic ultrasound (EUS), that revealed an ill-defined hypoechogenic lesion adjacent to the spleen and two hypoechogenic subepithelial lesions located in the 4th layer of the stomach and duodenal bulb. Biopsies revealed non-necrotizing granulomatous inflammation with multinucleated giant cells. Soon after, a 18cm palpable mass within the rectus abdominis muscle was identified, and the biopsy was positive for Mycobacterium tuberculosis DNA, confirming the diagnosis of disseminated abdominal tuberculosis.


Microwave-ultrasonic technique development coupled with natural deep eutectic solvents in anthocyanin extraction from perilla leaves (Perilla frutescens var. Acuta).
As potent antioxidants, anthocyanins can protect the body from free radicals. However, the traditional solvent extraction method has the disadvantages of requiring a high extraction temperature and long extraction time, so it is necessary to develop an efficient extraction method for anthocyanins. In this study, the technique of natural deep eutectic solvents (DESs) was applied to extract anthocyanins from purple perilla leaves with the aid of microwave-ultrasonic assisted extraction (MUAE). The response surface methodology (RSM), based on the Box-Behnken design (BBD), predicted the maximum extraction yield of anthocyanins to be 619.62 mg (100 g)-1 under the following conditions: x1 (ultrasonic extraction power) = 357.25 W, x2 (time) = 25.62 min, and x3 (temperature) = 57.80 °C. The biological activity of the extract obtained was evaluated by examining its radical-scavenging effect on 1,1-diphenyl-2-picrylhydrazyl, hydroxyl radical, and superoxide anion radicals. Its bacteriostatic impact was investigated on four typical bacteria: Shewanella putrefaciens (S. putrefaciens), Pseudomonas fluorescens (P. fluorescens), Escherichia coli (E. coli), and Staphylococcus aureus (S. aureus). The integrated extraction method of DESs with MUAE was efficient, energy-saving, green, and sustainable. © 2022 Society of Chemical Industry.


Tissue-restricted T cell alloresponses across HLA barriers: selection and identification of leukemia-restricted CTL in HLA-mismatched stimulator-responder pairs.
Exploiting the graft-versus-leukemia (GVL) effect in mismatched transplants requires its separation from graft-versus-host disease (GVHD). We generated leukemia-specific cytotoxic T lymphocytes (CTL) in three haplotype-mismatched, two class I-mismatched and two single HLA-A locus-matched stimulator-responder pairs. Six patients with chronic myelogenous leukemia and one patient with acute myeloid leukemia transformed from MDS were studied. CTL generated after 10 days stimulation with unselected leukemic peripheral blood mononuclear cells inhibited leukemic CFU-GM colony growth (>85% at 10:1 effector:target ratio) with no third-party colony inhibition. In five pairs, responders were cultured separately with leukemia cells, PHA-B or LCL from the stimulator. After 2-4 restimulations, the T cell repertoire was examined by flow analysis using Vbeta-specific antibodies. Test cultures (but not controls) showed preferential expansion of 1-4 Vbeta families either common to two or more stimulators or unique to a particular stimulator. Notably, we elicited leukemia-specific TCR Vbeta expansions on four out of five occasions. In two pairs, responder cells selected for the appropriate leukemia-specific Vbeta family were shown to have leukemia-specific cytotoxicity. These leukemia-restricted T-cells were CD8+ or CD4+ and CD25+ or CD57+. The results support the development of strategies to selectively deplete GVHD and conserve GVL reactivity in mismatched transplants.


Administering a Telemedicine Program.
The purpose of this review is to describe the process of administering a telemedicine program including reviewing telemedicine guidelines; discussing licensing, credentialing, and privileging of providers; outlining scheduling and recruitment of patients; and measuring outcomes of a telemedicine program. Recent literature findings suggest that telemedicine in specialty clinics continues to grow at a rapid pace. Medical specialty programs should prepare to adopt a practice that includes telemedicine to better serve their patients and families who have expressed significant satisfaction with the delivery of healthcare in this manner. With the appropriate support, any specialty clinic can provide their patients with a telemedicine option which has shown to be highly successful for Children's Mercy Allergy, Asthma, & Immunology Department.


Effects of smoking on the heart and peripheral circulation.
Cigarette smoking increases heart rate, arterial blood pressure, and plasma catecholamine levels. In healthy subjects the increase in heart rate occurs in the absence of peripheral vasoconstriction. In the studies reported here, short-term beta-blockade increased peripheral vascular resistance during smoking, more so for the nonselective beta-blocker propranolol than for the beta 1-selective blocker atenolol. However, after 3 months of continuous treatment of hypertensive patients with beta-blockers, smoking produced similar increases in blood pressure for atenolol and propranolol. Smoking attenuated the beneficial blood pressure-lowering effect of beta-blockers. Short-term clinical studies with the nonselective alpha-inhibitor phentolamine have suggested that blockade of arteriolar alpha-receptors may eliminate the increase in blood pressure response to smoking. Treatment with alpha-inhibitors also increased coronary sinus blood flow in patients with atherosclerotic artery disease. For patients who smoke, alpha-inhibiting drugs may provide beneficial therapy for hypertension and other cardiovascular disorders.


Visible-Light-Mediated Oxidative Coupling of Vinylarenes with Bromocarboxylates Leading to γ-Ketoesters.
A photocatalytic strategy for the synthesis of γ-ketoesters was reported. Using DMSO as both the solvent and terminal oxidant, oxidative coupling of vinylarenes with bromocarboxylates proceeded readily, giving a variety of γ-ketoesters in good isolated yields and with a broad functional-group tolerance.


Pharmaceutical and personal care products in tile drainage following land application of municipal biosolids.
Land application of municipal biosolids (sewage) is a common farming practice in many parts of the world. There is potential for transport of pharmaceuticals and personal care products (PPCPs) from agricultural fields to adjacent surface waters via tile drainage systems. In this study, liquid municipal biosolids (LMB) (total solids=11,933 mg L(-1)), supplemented with selected PPCPs and the fluorescent dye tracer rhodamine WT (RWT), were applied to tile drained fields using two land application approaches. Objectives included evaluating the relative benefits of land application practices with respect to reducing PPCP loadings to tile drains, evaluating PPCP persistence in tile water, and determining whether rhodamine WT can be used to estimate PPCP mass loads in tile. The PPCPs examined included an antibacterial agent used in personal care products (triclosan), a metabolite of nicotine (cotinine), and a variety of drugs including two sulfonamide antimicrobials (sulfapyridine, sulfamethoxazole), a beta-blocker (atenolol), an anti-epileptic (carbamazepine), an antidepressant (fluoxetine), analgesic/anti-inflammatories (acetaminophen, naproxen, ibuprofen), and a lipid-regulator (gemfibrozil). Maximum observed PPCP concentrations in the spiked LMB were about 10(3) ng g(-1) dry weight. PPCPs were shown to move rapidly via soil macropores to tile drains within minutes of the land application. Maximum observed PPCP concentrations in tile effluent associated with the LMB application-induced tile flow event were approximately 10(1) to 10(3) ng L(-1). PPCP mass loads, for the application-induced tile-hydrograph event, were significantly (p<0.1) higher for surface spreading over non-tilled soil (incorporation tillage occurring 20 h post-application), relative to aerating soil immediately prior to surface spreading using an AerWay slurry deposition system. PPCP concentrations that were detected above the limit of quantitation (LOQ) in tile water during several precipitation-induced tile flow events that occurred post-application, included: triclosan (max. approximately 1.5 x 10(2) ng L(-1)), carbamazepine (max. approximately 7 x 10(1) ng L(-1)), atenolol (max approximately 4 x 10(1) ng L(-1)), and cotinine (max approximately 2 x 10(1) ng L(-1)). In spite of their presence in biosolids, the other PPCPs were not observed above LOQ concentrations during these events. PPCP concentrations were predicted from RWT concentrations over a 40 day study period. Tile mass loads as a percent of PPCP mass applied to soil ranged from 4.2%+/-SD of 9.2% to 7.1%+/-10.9% for the AerWay system and surface spreading plus incorporation treatments, respectively.


rTMS treatment for abrogating intracerebral hemorrhage-induced brain parenchymal metabolite clearance dysfunction in male mice by regulating intracranial lymphatic drainage.
The discovery of the glymphatic system and meningeal lymphatic vessels challenged the traditional view regarding the lack of a lymphatic system in the central nervous system. It is now known that the intracranial lymphatic system plays an important role in fluid transport, macromolecule uptake, and immune cell trafficking. Studies have also shown that the function of the intracranial lymphatic system is significantly associated with neurological diseases; for example, an impaired intracranial lymphatic system can lead to Tau deposition and an increased lymphocyte count in the brain tissue of mice with subarachnoid hemorrhage. In this study, we assessed the changes in the intracranial lymphatic system after intracerebral hemorrhage and the regulatory effects of repeated transcranial magnetic stimulation on the glymphatic system and meningeal lymphatic vessels in an intracerebral hemorrhage (ICH) model of male mice. Experimental mice were divided into three groups: Sham, ICH, and ICH + repeated transcranial magnetic stimulation (rTMS). Three days after ICH, mice in the ICH+rTMS group were subjected to rTMS daily for 7 days. Thereafter, the function of the intracranial lymphatic system, clearance of RITC-dextran and FITC-dextran, and neurological functions were evaluated. Compared with the Sham group, the ICH group had an impaired glymphatic system. Importantly, rTMS treatment could improve intracranial lymphatic system function as well as behavioral functions and enhance the clearance of parenchymal RITC-dextran and FITC-dextran after ICH. Our results indicate that rTMS can abrogate ICH-induced brain parenchymal metabolite clearance dysfunction by regulating intracranial lymphatic drainage.


Cochleate drug delivery systems: An approach to their characterization.
Cochleate systems formed from phospholipids have very useful properties as drug delivery systems with sustained release capabilities, which are able to improve bioavailability and efficacy, reduce toxicity and increase the shelf-life of encapsulated molecules. These nanometric or micrometric structures are usually obtained after interaction of negatively charged liposomes with a positively charged bridging agent. Many different methods are now available to prepare cochleates and there are also numerous techniques that can be used to characterize them, some of which can be easily applied while others require more sophisticated equipment or analysis. The present review describes the important features of this drug delivery system; including their structural properties and potential applications, as well as a brief account of methods for their preparation and an extensive description of the techniques used for their characterization. This information could guide formulators in their choice of methods of characterization that would be best suited to their needs in terms of time, precision and technological difficulty.


Reconstituting National Institute for Biological Standards and Control (NIBSC) chemokines.
We observed significant discrepancies between immunoassay results when using different internally prepared reference preparations for interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) from the National Institute for Biological Standards and Control (NIBSC). To evaluate the reasons for this we prepared the chemokines using diluents that incorporated protein at different steps. This showed that even brief addition of water to these preparations, in the absence of additional protein, resulted in loss of immunoreactivity in assays. The data obtained emphasise the importance of adding protein at an early stage of preparation to avoid loss of material and potential loss of activity.


Insulin secretion in children with growth retardation.
The effect of tolbutamide administration on insulin secretion was studied in 69 children with growth retardation. Diminished insulin secretion was found in all the patients, compared to the control group. This insulin deficit was most evident in patients with isolated, total GH deficiency and least evident in children with idiopathic short stature. Intermediate values were found in dwarfism due to isolated, partial GH deficiency. These results favour the hypothesis that hypoinsulinism contributes to the somatotropin deficiency in causing growth retardation.


Post-burn squamous cell cancers in Nigerians.
Eighteen patients with post-burn skin cancers seen at Enugu over an 8-year period have been reviewed. All the tumours were well differentiated squamous cell carcinomas. The mean age of the patients was 36 years and the incubation period of 17.5 years was shorter than in previously reported studies. An aggressive policy in the treatment of these tumours by surgery alone has resulted in an 87% 5-year survival rate for lower limb tumours. When secondary tumours due to pre-existing dermatological diseases are excluded, post-burn skin epitheliomas constituted the largest group of squamous cell skin cancers seen in Enugu, Nigeria.


Evaluating the influence of different environmental water allocation schemes on the water level of a typical shallow lake in semiarid regions: From the perspective of an integrated modeling approach.
Many lakes in semiarid regions around the world rely on environmental water allocation to maintain the health of the lake ecosystem. However, under changing environments, the competition for water resources between human society and natural ecosystems has intensified. How to manage environmental water allocation more reasonably and precisely has become an important issue. The largest lake on the North China Plain, Baiyangdian Lake (BYDL), is a typical lake facing such challenges. To provide feasible strategies for sustainable water allocation to BYDL, with the proper parameterization of hydrological processes, this study developed a 10-day temporal scale lake water level prediction model to quantify how environmental water allocation regulates the BYDL water level under different hydroclimatic conditions. Evaluation of model performance revealed that environmental water allocation rather than natural climatic periodicity dominates the variation in the BYDL water level. The model structure could be further improved with consideration of more detailed observations of both the surface runoff entering BYDL and the water area beneath the canopy of the reeds in BYDL. Analysis of 72 model simulation scenarios indicated that water allocations from multiple sources are indispensable and that the water resources that guarantee maintaining the BYDL water level within the ecologically suitable range vary substantially under different hydroclimatic conditions. More elaborate allocation plans are required both to improve the water quality and health of the aquatic ecosystem of BYDL and to reduce the risk of flooding. The findings from this study are valuable for guiding the implementation of environmental water allocations to lakes in semiarid regions worldwide.


An accurate maxillary superior repositioning technique without intraoperative measurement in bimaxillary orthognathic surgery.
This article describes a simple and accurate technique for maxillary superior repositioning without any intraoperative measurement using reference points that can be the source of error in bimaxillary orthognathic surgery. A bilateral straight locking miniplates (SLMs)/screw system secured to the maxilla and mandible maintains the three-dimensional relationship between the mandible and the skull base precisely like the incisor pin of an articulator in model surgery. The maxilla can then be accurately moved into the planned position identical to that in model surgery by the SLMs technique.


The pathogenesis of amyotrophic lateral sclerosis: Mitochondrial dysfunction, protein misfolding and epigenetics.
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease with multiple complex mechanisms involved. Among them, mitochondrial dysfunction plays an important role in ALS. Multiple studies have shown that mitochondria are closely associated with reactive oxygen species production and oxidative stress and exhibit different functional states in different genetic backgrounds. In this review we explored the roles of Ca2+, autophagy, mitochondrial quality control in the regulation of mitochondrial homeostasis and their relationship with ALS. In addition, we also summarized and analyzed the roles of protein misfolding and abnormal aggregation in the pathogenesis of ALS. Moreover, we also discussed how epigenetic mechanisms such as DNA methylation and protein post-translational modification affect initiation and progression of ALS. Nevertheless, existing events still cannot fully explain the pathogenesis of ALS at present, more studies are required to explore pathological mechanisms of ALS.


Focal cooling for epilepsy: an alternative therapy that might actually work.
The therapy of focal epilepsy remains inadequate. Many patients who have localization-related seizures find themselves either overmedicated with anticonvulsants or suffering from frequent seizures. While surgical resection can lead to excellent outcomes in up to 60% of patients with neocortical epilepsy, there are obviously many who either fail surgery or are deemed inappropriate surgical candidates. We are currently determining the efficacy of local cooling for the therapy of certain focal epilepsies. We have attempted to adapt new technologies borrowed from electrical and mechanical engineering to develop cooling devices that will ultimately improve the diagnosis and therapy of these focal epilepsies. The present review describes the rationale for this research and our progress to date.


Sodium butyrate ameliorates deoxycorticosterone acetate/salt-induced hypertension and renal damage by inhibiting the MR/SGK1 pathway.
Our recent work demonstrates that infusion of sodium butyrate (NaBu) into the renal medulla blunts angiotensin II-induced hypertension and improves renal injury. The present study aimed to test whether oral administration of NaBu attenuates salt-sensitive hypertension in deoxycorticosterone acetate (DOCA)/salt-treated rats. Uninephrectomized male Sprague-Dawley (SD) rats were treated with DOCA pellets (150 mg/rat) plus 1% NaCl drinking water for 2 weeks. Animals received oral administration of NaBu (1 g/kg) or vehicle once per day. Our results showed that NaBu administration significantly attenuated DOCA/salt-increased mean arterial pressure from 156 ± 4 mmHg to 136 ± 1 mmHg. DOCA/salt treatment markedly enhanced renal damage as indicated by an increased ratio of kidney weight/body weight, elevated urinary albumin, extensive fibrosis, and inflammation, whereas kidneys from NaBu-treated rats exhibited a significant reduction in these renal damage responses. Compared to the DOCA/salt group, the DOCA/salt-NaBu group had ~30% less salt water intake and decreased Na+ and Cl- excretion in urine but no alteration in 24-h urine excretion. Mechanistically, NaBu inhibited the protein levels of several sodium transporters stimulated by DOCA/salt in vivo, such as βENaC, γENaC, NCC, and NKCC-2. Further examination showed that NaBu downregulated the expression of mineralocorticoid receptor (MR) and serum and glucocorticoid-dependent protein kinase 1 (SGK1) in DOCA/salt-treated rats or aldosterone-treated human renal tubular duct epithelial cells. These results provide evidence that NaBu may attenuate DOCA/salt-induced hypertension and renal damage by inhibiting the MR/SGK1 pathway.


The earliest history of eukaryotic life: uncovering an evolutionary story through the integration of biological and geological data.
While there is significant data on eukaryogenesis and the early development of the eukaryotic lineage, major uncertainties regarding their origins and evolution remain, including questions of taxonomy, timing, and paleoecology. Here we examine the origin and diversification of the eukaryotes in the Proterozoic Eon as viewed through fossils, organic biomarkers, molecular clocks, phylogenies, and redox proxies. Our interpretation of the integration of these data suggest that eukaryotes were likely aerobic and established in Proterozoic ecosystems. We argue that we must closely examine and integrate both biological and geological evidence and examine points of agreement and contention to gain new insights into the true origin and early evolutionary history of this vastly important group.


Prevalence of low alkaline phosphatase activity in laboratory assessment: Is hypophosphatasia an underdiagnosed disease?
Tissue-nonspecific alkaline phosphatase (TNSALP) encoded by the ALPL gene is of particular importance for bone mineralization. Mutation in the ALPL gene can lead to persistent low ALP activity resulting in the rare disease Hypophosphatasia (HPP) that is characterized by disturbed bone and dental mineralization. While severe forms are extremely rare with an estimated prevalence of 1/100.000, recent studies suggest that moderate form caused by heterozygous mutations are much more frequent with an estimated prevalence of 1/508. The purpose of this study was to estimate the prevalence of low AP levels in the population based on laboratory measurements. In this study, the prevalence of low AP activity and elevated pyridoxal-5-phosphate (PLP) levels was analyzed in 6.918.126 measurements from 2011 to 2016 at a single laboratory in northern Germany. Only laboratory values of subjects older than 18 years of age were included. Only the first measurement was included, all repeated values were excluded. In total, 8.46% of the measurements of a total of 6.918.126 values showed a value < 30 U/L. 0.59% of the subjects with an ALP activity below 30 U/L had an additional PLP measurement. Here, 6.09% showed elevated pyridoxal-5-phosphate (PLP) levels. This suggest that 0.52% (1:194) of subjects show laboratory signs of HPP. These data support the genetic estimation that the prevalence of moderate forms of HPP may be significantly higher than expected. Based on these data, we recommend automatically measurement of PLP in the case of low ALP activity and a notification to the ordering physician that HPP should be included in the differential diagnosis and further exploration is recommended.


Functional properties of proislet isografts in diabetic mice.
Fetal proislets (islet precursors) were transplanted to isogeneic diabetic CBA/H mice for analysis of their development and function post-transplant. Immunohistochemically, proislets contained scattered populations of insulin-, glucagon- and somatostatin-producing cells; following isotransplantation proislets developed into mature islet tissue with normal endocrine cell composition. Proislet isografts also developed normal endocrine function after transplantation to diabetic mice. Isografts of eight and four donor-equivalents of proislets reversed streptozotocin-induced diabetes by 30 +/- 7 and 74 +/- 26 days, respectively. Following stimulation with intragastric glucose (50 mg) mice carrying established isografts of four donor-equivalents of proislets exhibited normal blood glucose responses; those grafted with eight donor-equivalents of proislets showed decreased blood glucose levels. The beta cell mass of proislets continued to grow and differentiate post-transplant; established proislet grafts contained greater than 100-fold the total insulin content of the corresponding number of fetal pancreases from which the proislets were derived. The mean insulin content of eight and four donor-equivalent proislet grafts was 231% and 156%, respectively, that of the mean content of adult mouse pancreas. The data show that glucose homeostasis can be restored in diabetic mice following isotransplantation of fetal proislets and that the quantity of tissue transplanted is a critical parameter in achieving optimal control.


Two Dimensional Oblique Molecular Packing within a Model Peptide Ribbon Aggregate.
A10 K (A=alanine, K=lysine) model peptides self-assemble into ribbon-like β-sheet aggregates. Here, we report an X-ray diffraction investigation on a flow-aligned dispersion of these self-assembly structures. The two-dimensional wide-angle X-ray scattering pattern suggests that peptide pack in a two-dimensional oblique lattice, essentially identical to the crystalline packing of polyalanine, An (for n>4). One side of the oblique unit cell, corresponding to the anti-parallel β-sheet, is oriented along the ribbon's axis. Together with recently published small angle X-ray scattering data of the same system, this work thus yields a detailed description of the self-assembled ribbon aggregates, down to the molecular length scale. Notably, our results highlight the importance of the crystalline peptide packing within its self-assembly aggregates, which is often neglected.


Escaping an evolutionary trap: preference and performance of a native insect on an exotic invasive host.
Exotic plants may act as population sinks or evolutionary traps for native herbivores. The native butterfly Pieris oleracea lays eggs on garlic mustard, Alliaria petiolata, but larvae develop very poorly on this exotic invasive plant. We examined oviposition preference of individual females and larval performance of their offspring for individuals from one area where garlic mustard is well established and one where it is absent. These data were used to assess whether garlic mustard is being incorporated into or excluded from the diet. Females from the area without garlic mustard showed a wide range of preference, families had low larval survival on garlic mustard, and larval survivorship showed no correlation with mothers' preferences. Females from the area with garlic mustard preferred it to the native host, and larval survivorship on garlic mustard was positively correlated with the mother's preference. Individuals surviving on garlic mustard took longer to pupate and weighed >30% less compared to pupae reared on normal hosts. Our results suggest that where garlic mustard is well established P. oleracea may be adapting to this plant by both improved larval performance and increased adult female oviposition preference for it.


Engineering effect of oleogels with different structuring mechanisms on the crystallization behavior of cocoa butter.
As promising cocoa butter (CB) alternatives, oleogels have the potential to prevent fat blooms of chocolate. We aimed to explore possible reasons for the bloom resistance of oleogels by investigating the crystallization behavior of CB-oleogel blends, including crystallization kinetics, thermodynamic properties, crystal polymorphism, and oil distribution. Oleogels structured by monoglyceric stearate (MO), β-sitosterol/lecithin (SLO), and ethylcellulose (EO) were selected as representative oleogels with various structuring-mechanisms. Crystallization kinetic results showed that the crystallization dimension of CB-oleogel increased with the oleogel proportion (from one-dimensional to multi-dimensional), confirming that CB crystallization was inhibited. The presence of liquid oil and oleogelators in oleogels may increase the free energy barrier for CB crystallization. The proton mobility of liquid oil in CB-MO was lower because MO was more tightly bound to CB. The crystallization mechanism of the CB-oleogel suggested that the inhibitory effect of oleogels on CB crystallization delayed the polymorphic transition, thereby improving the bloom stability of chocolate.


The Change of Soluble Programmed Cell Death-Ligand 1 in Glioma Patients Receiving Radiotherapy and Its Impact on Clinical Outcomes.
The programmed cell death ligand 1 (PD-L1) plays a key role in glioma development. However, due to the specificity of glioma's anatomical position, the role of its expression as a tumor biomarker is limited. It has been proven that the levels of soluble programmed death-ligand 1 (sPD-L1) are associated with prognosis in many malignancies including glioma. However, the expression of sPD-L1 in glioma patients receiving radiotherapy (RT) remains unclear. The purpose of this study was to evaluate the concentration of sPD-L1 in the plasma of glioma patients before and after RT and to explore its relationship with clinical outcomes. Between October 2017 and September 2018, glioma patients treated with RT (30 ± 10 Gy, 2 Gy/f) were enrolled, and blood samples were collected before and after RT. We quantified the sPD-L1 levels by enzyme-linked immunosorbent assay (ELISA). The isocitrate dehydrogenase-1 (IDH-1) mutational status and Ki-67 expression of tumors were evaluated by immunohistochemistry. Glioma murine model were used to address whether circulating sPD-L1 molecules are directly targeted by an anti-PD-L1 antibody. The associations between sPD-L1 and clinical features were assessed with Pearson's or Spearman's correlation analysis. The progression-free survival (PFS) and overall survival (OS) were determined by the Kaplan-Meier method. Sixty glioma patients were included, with a median age of 52 years. The proportions of grade I, II, III, and IV gliomas were 6.7%, 23.3%, 28.4%, and 41.6%, respectively. The baseline sPD-L1 levels were significantly associated with tumor grade, IDH-1 mutation status and Ki-67 expression. Using 14.35 pg/ml as the cutoff, significantly worse PFS and OS were both observed in patients with higher baseline levels of sPD-L1 (P = 0.027 and 0.008, respectively). RT significantly increased the mean level of sPD-L1 (P < 0.001). Further analysis showed that the level of sPD-L1 in IDH-1 mutation patients was higher than that in wild-type patients. Furthermore, an analysis of glioma murine model indicated that anti-PD-L1 antibody combine with RT can be a potentially powerful cancer therapy. This study reported that sPD-L1 might be a potential biomarker to predict the outcome in glioma patients receiving RT. The elevated level of sPD-L1 after RT suggested that the strategy of a combination of immune checkpoint inhibitors and RT might be promising for glioma patients, especially for those with IDH-1 mutations.


Seasonal effects of Selank on the behavior of hibernating animals.
We studied the seasonal effects of peptide preparation Selank on orientation and exploratory activity in 36 arctic ground squirrels Citellus undulatus in the open-field and hole-board tests. Selank most significantly increased behavioral activity of hibernating animals in the spring and fall. The test peptide had no effect on locomotor activity of animals. The selective stimulatory effect of Selank on exploratory behavior of hibernating animals was season-dependent. The maximum effect was observed under conditions of seasonal depression-like state.


The functional requirements of mammalian hair: a compromise between crypsis and thermoregulation?
Mammalian fur often shows agouti banding with a proximal dark band near the skin and a lighter distal band. We examined the function of both bands in relation to camouflage, thermal properties of pelts, and thermal energetics of dunnarts (Sminthopsis crassicaudata), which are known to use torpor and basking. Although the distal band of dunnart fur darkened with increasing latitude, which is important for camouflage, it did not affect the thermal properties and the length of the dark band and total hair length were not correlated. In contrast, the length of the proximal dark band of preserved pelts exposed to sunlight was positively correlated (r (2) = 0.59) with the temperature underneath the pelt (T pelt). All dunnarts offered radiant heat basked by exposing the dark band of the hair during both rest and torpor. Basking dunnarts with longer dark bands had lower resting metabolism (r (2) = 0.69), warmed faster from torpor (r (2) = 0.77), required less energy to do so (r (2) = 0.32), and reached a higher subcutaneous temperature (T sub) at the end of rewarming (r (2) = 0.75). We provide the first experimental evidence on the possible dual function of the color banding of mammalian fur. The distal colored band appears to be important for camouflage, whereas the length of the dark proximal hair band facilitates heat gain for energy conservation and allows animals to rewarm quickly and economically from torpor.


Stratification of Patients with Chronic Obstructive Pulmonary Disease Using Volatile Organic Compounds.
Chronic obstructive pulmonary disease (COPD) is a common disease that causes long-term disability and death. Its natural history is punctuated by acute worsening of symptoms, called exacerbations, which are associated with increased mortality and hospitalization. In this work, we aim to stratify patients with COPD based on their risk for exacerbation; for this purpose, we employ non-invasive biomarkers, that is, volatile organic compounds (VOCs), acquired from the patients' exhaled breath coupled with their spirometry and age. We utilize a series of classification schemes with the best performing one achieving overall Accuracy = 93.5%. The yielded results are, therefore, encouraging and prompt for further investigation toward the utilization of VOCs in the management of COPD.


Pad test by mail for home evaluation of urinary incontinence.
To present a simple, cost-effective, and convenient method of home pad test using the mail system and evaluating change in pad weight over time. A series of nine kinds of commonly available commercial brands of urinary incontinence pads ranging from thin liners less than 10 g in dry weight to large diapers weighing over 100 g each were assembled. Two or three of each variety were individually weighed on an OHAUS LS2000 Portable Standard scale accurate to +/- 1 g. The pads were then wet uniformly with 20 cc of saline, placed individually in sealable plastic bags, sealed, and reweighed. Random groups of three pads were mailed by standard 1st class mail to the Urocenter of New York. The sealed pads were reweighed at 8 and 14 days from the original wetting. Concurrently, ten incontinence pads soiled with urine were similarly examined to confirm that there would be no detectable difference between urine and saline for the purpose of the study. In the second part of the study, 20 pads of the same type (13 inch-long pads with absorbent gel) were divided into 4 groups of 5 pads; each group was uniformly wet with 5, 10, 20, or 50 g of saline. These pads were mailed and reweighed at 7 and 14 days from the initial wetting. Twenty-four pads were used in the first part of the study. The dry weights of 22/24 (92%) of the pads were within 2 g of the average weight for their brand. At the first reweighing, 22 pads (92%) weighed within 2 g of their initial wet weights (Fig. 1). Only two pads on day 8 differed substantially from their initial weights: one pad appeared to have lost 4 g and another one 9 g. On day 14, 23/24 pads remained within 1 g of their day 8 weight and 1 differed by 2 g, and the total weight of the 24 pads on day 14 was only 4 g different from their initial weight (P = 0.71, Fig. 1). The soiled pads exhibited weight changes that were indistinguishable from the saline pads. The average cost of mailing the pads by 1st class mail was $4 and the average length of time in the mail was 5 days +/- 1 day. In the second part of the study, 18/20 pads had lost less than 1 g at 1 week, and at 2 weeks, 19/20 had lost less than 2 g when compared to their initial weights (Fig. 2). One pad had lost 3 g. Pads in the low volume groups (5 and 10 g) lost an average of 1.4 and 1.2 g, respectively, while pads in the high volume groups (20, 50 g) lost an average of 1.8 and 2 g at 2 weeks. Dry pads of any single brand have a relatively standard weight that varies insignificantly between pads. Up to 2 weeks of delay in the weighing of individually sealed pads does not significantly affect the clinical measurement of weight at a variety of low (5 g) or high (50 g) volumes of simulated incontinence. Home pad test using the mail system is a feasible, inexpensive, and clinically accurate method of evaluating incontinence. Patients can be instructed in techniques for home pad test, allowing for greater compliance, and convenience for both physicians and patients.


RNAi-mediated silencing of the Arabidopsis thaliana ULCS1 gene, encoding a WDR protein, results in cell wall modification impairment and plant infertility.
Ubiquitin mediated protein degradation constitutes one of the most complex post translational gene regulation mechanisms in eukaryotes. This fine-tuned proteolytic machinery is based on a vast number of E3 ubiquitin ligase complexes that mark target proteins with ubiquitin. The specificity is accomplished by a number of adaptor proteins that contain functional binding domains, including the WD40 repeat motif (WDRs). To date, only few of these proteins have been identified in plants. An RNAi mediated silencing approach was used here to functionally characterize the Arabidopsis thaliana ULCS1 gene, which encodes for a small molecular weight WDR protein. AtULCS1 interacts with the E3Cullin Ring Ligase subunit DDB1a, regulating most likely the degradation of specific proteins involved in the manifestation of diverse developmental events. Silencing of AtULCS1 results in sterile plants with pleiotropic phenotypes. Detailed analysis revealed that infertility is the outcome of anther indehiscence, which in turn is due to the impairment of the plants to accomplish secondary wall modifications. Furthermore, IREGULAR XYLEM gene expression and lignification is diminished in anther endothecium and the stem vascular tissue of the silenced plants. These data underline the importance of AtULCS1 in plant development and reproduction.


Nocturnal agitation in Huntington disease is caused by arousal-related abnormal movements rather than by rapid eye movement sleep behavior disorder.
Patients with Huntington disease (HD) and their spouses often complain of agitation during sleep, but the causes are mostly unknown. To evaluate sleep and nocturnal movements in patients with various HD stages and CAG repeats length. The clinical features and sleep studies of 29 patients with HD were retrospectively collected (11 referred for genotype-phenotype correlations and 18 for agitation during sleep) and compared with those of 29 age- and sex-matched healthy controls. All patients had videopolysomnography, but the movements during arousals were re-analyzed in six patients with HD with stored video. The patients had a longer total sleep period and REM sleep onset latency, but no other differences in sleep than controls. There was no correlation between CAG repeat length and sleep measures, but total sleep time and sleep efficiency were lower in the subgroup with moderate than milder form of HD. Periodic limb movements and REM sleep behavior disorders were excluded, although 2/29 patients had abnormal REM sleep without atonia. In contrast, they had clumsy and opisthotonos-like movements during arousals from non-REM or REM sleep. Some movements were violent and harmful. They might consist of voluntary movements inappropriately involving the proximal part of the limbs on a background of exaggerated hypotonia. Giant (>65 mcV) sleep spindles were observed in seven (24%) patients with HD and one control. The nocturnal agitation in patients with HD seems related to anosognostic voluntary movements on arousals, rather than to REM sleep behavior disorder and other sleep problems.


Structural biology of telomeres and telomerase.
Telomeres are protein-DNA complexes that protect chromosome ends from illicit ligation and resection. Telomerase is a ribonucleoprotein enzyme that synthesizes telomeric DNA to counter telomere shortening. Human telomeres are composed of complexes between telomeric DNA and a six-protein complex known as shelterin. The shelterin proteins TRF1 and TRF2 provide the binding affinity and specificity for double-stranded telomeric DNA, while the POT1-TPP1 shelterin subcomplex coats the single-stranded telomeric G-rich overhang that is characteristic of all our chromosome ends. By capping chromosome ends, shelterin protects telomeric DNA from unwanted degradation and end-to-end fusion events. Structures of the human shelterin proteins reveal a network of constitutive and context-specific interactions. The shelterin protein-DNA structures reveal the basis for both the high affinity and DNA sequence specificity of these interactions, and explain how shelterin efficiently protects chromosome ends from genome instability. Several protein-protein interactions, many provided by the shelterin component TIN2, are critical for upholding the end-protection function of shelterin. A survey of these protein-protein interfaces within shelterin reveals a series of "domain-peptide" interactions that allow for efficient binding and adaptability towards new functions. While the modular nature of shelterin has facilitated its part-by-part structural characterization, the interdependence of subunits within telomerase has made its structural solution more challenging. However, the exploitation of several homologs in combination with recent advancements in cryo-EM capabilities has led to an exponential increase in our knowledge of the structural biology underlying telomerase function. Telomerase homologs from a wide range of eukaryotes show a typical retroviral reverse transcriptase-like protein core reinforced with elements that deliver telomerase-specific functions including recruitment to telomeres and high telomere-repeat addition processivity. In addition to providing the template for reverse transcription, the RNA component of telomerase provides a scaffold for the catalytic and accessory protein subunits, defines the limits of the telomeric repeat sequence, and plays a critical role in RNP assembly, stability, and trafficking. While a high-resolution definition of the human telomerase structure is only beginning to emerge, the quick pace of technical progress forecasts imminent breakthroughs in this area. Here, we review the structural biology surrounding telomeres and telomerase to provide a molecular description of mammalian chromosome end protection and end replication.


Early chest tube removal after video-assisted thoracoscopic wedge resection of the lung.
Traditional management of chest tubes after a wedge resection of peripheral pulmonary tissue often lasts several days. We evaluated the safety and efficacy of early chest tube removal in the recovery room after uncomplicated video-assisted thoracoscopic surgical wedge resections of the lung. From December 1995 to July 1997, 59 patients underwent video-assisted thoracoscopic surgical wedge resection for indeterminate pulmonary nodules (n = 33) or interstitial lung disease (n = 26). We prospectively evaluated early chest tube removal in the last 33 patients; 18 patients with nodules and 15 with interstitial lung disease. Patients who were in the early removal group had chest tubes removed within 90 minutes of the surgical procedure. Criteria for early removal were established and met before chest tube removal. There was no difference between groups with respect to age, sex, comorbidities, or pathologic evaluation of resection specimens. Ninety-four percent (31 of 33) of patients considered for early chest tube removal met criteria for immediate tube removal. Air leak and excessive drainage prohibited early removal in 2 patients. Patients who were managed traditionally averaged 3.3 days with chest tubes--1.8 days on suction, 1.3 days on water seal. Patients who had early removal of their chest tubes had a shorter postoperative stay (2.0+/-1.0 versus 3.9+/-2.1 days, p = 0.001) and fewer chest roentgenograms (2.8+/-2.1 versus 5.1+/-2.0, p = 0.001). There were no differences in complications including small pneumothoraces (5 in the early removal group, 7 in the traditional management group), which were managed with observation alone. Total narcotic requirements were greater in the traditional management group (54+/-44.8 versus 24.6+/-22.9 morphine milligram equivalents, p = 0.005). Early chest tube removal after video-assisted thoracoscopic surgical wedge resection of peripheral pulmonary tissue appears to be a safe and cost-effective practice if strict criteria for removal are met.


Economics of using beef semen on dairy herds.
The economic value of using beef semen in dairy herds depends on the market value of calves (crossbred beef and dairy), market price of semen (beef, conventional, and sexed), herd reproductive performance, and semen combination strategies. Due to the complex interaction among all these factors and their inherent changing conditions, the quest for an optimal strategy is best served by the application of an integrated model and a decision support tool adaptable to ever-changing farm and market conditions. We have developed a model and a decision support tool to calculate the income from calves over semen costs (ICOSC) in response to user-defined beef semen crossbreeding strategies in combination with sexed and conventional semen utilization. The model follows a Markov-chain approach in which animal (heifer and cow) statuses (age, months after calving, lactation, pregnancy, calving) are simulated monthly. Replacement balance is calculated as the difference between demand and supply of calves in function of selected semen utilization protocols, which could include beef, sexed, or conventional semen. A case study was performed in a 1,000-cow virtual Holstein herd with 35% turnover rate and 7% stillbirth rate. Five strategies of beef semen utilization on adult cows (0 to 100% in 25-percentage-unit intervals) were combined with 6 strategies of sexed semen use [none (NS), first service in heifers (1H), first and second services in heifers (2H), 2H + 20% top genetic cows (TOP), 2H + first service in primiparous (1C), and 1C + first service in second-lactation cows (2C)]. All animals not bred to either sexed or beef semen were bred to conventional semen. Having a price of beef calves ~4 times greater than the price of a dairy calf and having the price of sexed semen ~2.3 times greater than the conventional or beef semen determined that the optimal breeding semen protocols that concurrently maximized the ICOSC and produced enough replacements were 100% beef semen use after 2C sexed semen protocol (ICOSC = $2,001) for medium reproductive performance (~20% 21-d pregnancy rate) and 100% beef semen after 1H sexed semen protocol (ICOSC = $6,215) for high reproductive performance (~30% 21-d pregnancy rate). These strategies were consistently the best options under several feasible market conditions for herds with medium and high reproductive performance. However, the optimal ICOSC was negative or marginally low for low-performance herds (~15% 21-d pregnancy rate), for which the opportunity to use beef semen is minimal or nonexistent.


Processing by inverted plasma membrane vesicles of in vitro synthesized major lipoprotein from Escherichia coli.
Synthesis, lipid modification and proteolytic processing of the major lipoprotein from Escherichia coli is shown to occur in a homologous in vitro transcription-translation system containing inverted plasma membrane vesicles. The primary translation product (cross-reacting with anti-lipoprotein antiserum) is a precursor which is converted into a lower molecular mass species of the size of mature lipoprotein by the addition of inverted membrane vesicles from E. coli. Conversion is prevented by globomycin, a specific inhibitor of the unique lipoprotein-signal peptidase II, which is active only on lipid-containing precursors. The inverted plasma membrane vesicles used here must therefore contain active lipid-modifying enzymes and signal peptidase II.


Retinol-binding protein-hijacking nanopolyplex delivering siRNA to cytoplasm of hepatic stellate cell for liver fibrosis alleviation.
Activated hepatic stellate cell (aHSC) is mainly responsible for deposition of extracellular collagen matrix that causes liver fibrosis. Although several siRNAs adequately inhibited HSC activation in vitro, they were demonstrated poor RNAi efficiency in vivo. Developing HSC-targeting and cytoplasmic delivery nanocarrier is highly essential to acquire a desirable siRNA therapeutic index for anti-liver fibrosis. Here, we developed a unique crosslinking nanopolyplex (called T-C-siRNA) modified by vitamin A (VA) with the well-designed natures, including the negative charge, retinol-binding protein (RBP) hijacking, and cytoplasmic siRNA release in response to ROS and cis diol molecules. The nanopolyplex was given a yolk-shell-like shape, camouflage ability in blood, and HSC-targeting capability by hijacking the endogenous ligand RBP via surface VA. PDGFR-β siRNA (siPDGFR-β) supplied via T-C-siPDGFR-β nanopolyplex dramatically reduced HSC activation and its production of pro-fibrogenic proteins in vitro and in vivo. Furthermore, T-C-siPDGFR-β nanopolyplex effectively alleviated CCl4-induced liver injury, decreased hepatic collagen sediment, and recovered liver function in mice. This study provides a sophisticated method for HSC-targeting cytoplasmic RNA delivery using endogenous ligand hijacking and dual sensitivity of ROS and cis diol compounds.


Cancer rates by county: how well does mortality relate to incidence?
The extent to which excess cancer mortality in a county is indicative of excess cancer incidence in the same county in the same time period was studied in white Iowa residents using routinely available 1973-1977 data. Evaluation consisted of population-weighted correlation analysis. The best correlations were obtained for cancers of the lung, pancreas, liver, stomach, brain (males), and kidney (males). These are the cancers for which excess mortality best indicates excess incidence. Correlations were low for prostate, female breast, and corpus uteri. The best indications of excess incidence are made for cancers with five year relative survival rates of 37.0 percent or less for males and 33.4 percent or less for females. Studies relating cancer mortality rates to occupational, environmental, or other demographic variables may be very misleading as to risk factors if the cancers selected have low correlations of incidence with mortality. Further development of the correlation model is recommended using data from all Surveillance, Epidemiology, and End Results Program counties for a ten-year period.


Serologic evidence for human ehrlichiosis in Africa.
Human ehrlichiosis is a recently recognized rickettsial disease. It is caused by Ehrlichia chaffeensis, an intraleucocytic Gram-negative, obligate intracellular bacterium, grouped within the genus Ehrlichiae. Most human cases of ehrlichiosis have been diagnosed in the USA. Two cases have been reported outside of the USA, one in Europe and one in Africa. From 1 January to 30 June 1992, 765 sera from blood donors or other asymptomatic subjects in 8 African countries, including Ivory Coast, Burkina Faso, Mali, Central African Republic, Angola, Zimbabwe, Mozambique and Commores Islands, were tested by indirect immunofluorescence for the presence of E. chaffeensis antibodies. Positive sera were confirmed by Western immunoblotting. Only two of 765 sera tested were positive. One serum obtained from Burkina Faso had an IgG titer of 1:200 and one from Mozambique had an IgG titer of 1:80. Human ehrlichiosis seems to occur infrequently in Africa, although many more sera from additional African countries need to be evaluated.


Priming and semantic memory loss in Alzheimer's disease.
Semantic memory (SM) was investigated in six patients with probable Alzheimer's disease (AD) by on-line measurement of semantic priming in a lexical decision task, and off-line tests of comprehension. Detailed assessment was carried out on naming, name comprehension, and probes of semantic knowledge with a battery of 150 items. The patients performed normally on perceptual tests and displayed an item-specific loss of knowledge on the semantic tests. In a primed lexical decision task, greater semantic priming was found relative to age-matched normals. The priming was substantially greater for items with "degraded" representations as determined by the off-line tests. Lexical decision was also performed more slowly on these items. These unexpected results demand a reevaluation of the concepts of the lexicon and semantic memory structure and their possible alteration in dementia.


Clinical characteristics and outcome of apical ballooning syndrome in Auckland, New Zealand.
Apical ballooning syndrome (ABS) mimics myocardial infarction but is characterised by transient left ventricular (LV) dysfunction without significant coronary artery obstruction. We prospectively identified 100 consecutive patients presenting in the Auckland region between March 2004 and July 2010 and obtained clinical, laboratory, electrocardiography, echocardiography, coronary angiography and long-term follow-up data. Chest pain or dyspnoea were the most common presenting symptom, 95% were women (mean age 65 ± 11 years). An associated stressor was identified in two-thirds of patients, troponin was elevated in all patients, and one-third had ECG ST-elevation. There was a similar range of initial LV ejection fraction (EF), myocardial damage, LV recovery and prognosis in those with and without ST-elevation, and with and without identifiable stressors. One-quarter had a complicated in-hospital course. Lower admission LVEF, but not peak troponin level or ECG ST-elevation, was associated with a complicated in-hospital course. The mean follow-up was 3.0 ± 1.7 years. One patient died in hospital. Four died late after discharge, all from non-cardiac causes. Seven had recurrent ABS. In this large, prospective, New Zealand ABS cohort a quarter of patients had a complicated in-hospital course, but almost all recovered, recurrence was infrequent and long-term prognosis dependent on associated non-cardiac disease.


Cerebral cavernous malformation: novel mutation in a Chinese family and evidence for heterogeneity.
Familial cerebral cavernous malformation (CCM) is an autosomal dominant disorder producing vascular anomalies throughout the central nervous system associated with seizures and hemorrhagic stroke. Linkage analysis has shown evidence for at least three genetic loci underlying this disorder with a founder mutation in the Mexican/Hispanic community. We report the first family of Chinese ethnic origin with CCM having a novel mutation in the CCM1 gene. The mutation in exon 19 causes a premature stop codon (Q698X) predicted to produce a truncated Krev1 interaction-trapped 1 (KRIT1) protein. Members of the family with this mutation have a wide range in age of onset with seizures, ataxia, spinal cord vascular malformation, headaches and skin lesions. An additional unrelated sporadic subject with brain lesions compatible with CCM as well as vascular skin findings suggesting the blue rubber bleb nevus (BRBN) syndrome has no mutation detected in the CCM1 gene. These findings expand the phenotype of and demonstrate further evidence for the heterogeneity in the CCM syndrome.


Behavioural characterization of interleukin-6 overexpressing or deficient mice during agonistic encounters.
Interleukin-6 (IL-6) is a cytokine released by activated immune cells which has been shown to affect brain function. In this experiment aggressive and affiliative behaviour exhibited during agonistic encounters by transgenic male mice either not expressing (IL-6 -/-) or overexpressing (NSE-hIL-6) IL-6 in the central nervous system was investigated. All subjects were isolated for 24 days before the aggressive encounter and were 52 days old at the time of testing. Subjects were placed for 5 consecutive days in a neutral cage for 15 min with an opponent of the Balb/c strain that had been previously isolated for the same amount of time. The first and the last test sessions were videotaped to evaluate the first approach and the establishment of the social role, respectively. A number of behavioural categories were later scored. When compared with wild-type controls, IL-6 -/- mice showed a higher degree of aggressive behaviour as indicated by a higher frequency of Offensive Upright Posture, an effect more pronounced on the fifth encounter. On the contrary, NSE-hIL-6 subjects showed a tendency to be more involved in affiliative-type social interactions, displaying a higher frequency and duration of behaviours such as Anogenital, Nose or Body Sniff. IL-6 -/- mice showed a clear tendency to exhibit less affiliative interactions compared with their controls while dopamine levels were found to be modified in a number of brain regions in these mice. Overall, these data suggest that IL-6 affects both aggressive and affiliative-type interactions, although the behaviour of the NSE-hIL-6 subjects was less affected than that of the IL-6 -/- group. The effects of the genetic background of the animal in screening the outcome of gene manipulations on behaviour are also discussed.


Successful transfer of ADA gene in vitro into human peripheral blood CD34+ cells by transfecting EBV-based episomal vectors.
We report a novel non-viral system for transfecting human immature hematopoietic cells in vitro. Epstein-Barr virus (EBV)-based episomal vectors carrying human adenosine deaminase (ADA) gene cDNA were transfected by electroporation into human peripheral blood (PB) CD34+ cells. The transgene-specific mRNA were detected from 37 to 100% of CFU-c (colony forming unit in culture) colonies derived from the transfected cells. A two-fold increase in enzyme activity was also found. These results indicate the successful transfer and expression of genes in human immature hematopoietic cells using the EBV-based episomal vector system.


Phylogenetic analysis of the bacterial community in a full scale autothermal thermophilic aerobic digester (ATAD) treating mixed domestic wastewater sludge for land spread.
The bacterial community associated with a full scale autothermal thermophilic aerobic digester (ATAD) treating sludge, originating from domestic wastewater and destined for land spread, was analysed using a number of molecular approaches optimised specifically for this high temperature environment. 16S rDNA genes were amplified directly from sludge with universally conserved and Bacteria-specific rDNA gene primers and a clone library constructed that corresponded to the late thermophilic stage (t = 23 h) of the ATAD process. Sequence analyses revealed various 16S rDNA gene sequence types reflective of high bacterial community diversity. Members of the bacterial community included α- and β-Proteobacteria, Actinobacteria with High G + C content and Gram-Positive bacteria with a prevalence of the Firmicutes (Low G + C) division (class Clostridia and Bacillus). Most of the ATAD clones showed affiliation with bacterial species previously isolated or detected in other elevated temperature environments, at alkaline pH, or in cellulose rich environments. Several phylotypes associated with Fe(III)- and Mn(IV)-reducing anaerobes were also detected. The presence of anaerobes was of interest in such large scale systems where sub-optimal aeration and mixing is often the norm while the presence of large amounts of capnophiles suggest the possibility of limited convection and entrapment of CO(2) within the sludge matrix during digestion. Comparative analysis with organism identified in other ATAD systems revealed significant differences based on optimised techniques. The abundance of thermophilic, alkalophilic and cellulose-degrading phylotypes suggests that these organisms are responsible for maintaining the elevated temperature at the later stages of the ATAD process.


Characterization of the BPI-like gene from a subtracted cDNA library of large yellow croaker (Pseudosciaena crocea) and induced expression by formalin-inactivated Vibrio alginolyticus and Nocardia seriolae vaccine challenges.
One expressed sequence tag (EST 64LF004 clone), which is from the subtracted cDNA library of the head kidney of large yellow croaker (Pseudosciaena crocea) stimulated with peptidoglycan (PG) by suppression subtractive hybridization (SSH) method, was cloned using RACE-PCR. The full length cDNA, which possesses typical structural features of a signal peptide, a conserved LPS binding domain and two bactericidal permeability-increasing (BPI) motifs as in higher vertebrates, was identified as a novel homologue, namely of the large yellow croaker BPI-like molecule (Pc-BPI-L). Phylogenetic analysis showed this Pc-BPI-L of large yellow croaker as the most ancestral branch in bony fish clade. The recombinant Pc-BPI-L protein expressed in the Tn-5B1-4 insect cells was successfully produced and confirmed to have the predicted size of 52 kDa by Western blot analysis. At the message level, Pc-BPI-L mRNA was ubiquitously expressed in all tissues examined. Following formalin-inactivated Vibrio alginolyticus and Nocardia seriolae treatment, Pc-BPI-L message was differentially up-regulated in primary immune organs. These results indicate that Pc-BPI-L might be involved in the immune response to bacterial infection.


Siblings reduce multiple sclerosis risk by preventing delayed primary Epstein-Barr virus infection.
Epstein-Barr virus infection, and perhaps almost exclusively delayed Epstein-Barr virus infection, seems to be a prerequisite for the development of multiple sclerosis. Siblings provide protection against infectious mononucleosis by occasionally preventing delayed primary Epstein-Barr virus infection, with its associated high risk of infectious mononucleosis. Each additional sibling provides further protection according to the age difference between the index child and the sibling. The closer the siblings are in age, the higher the protection, with younger siblings being more protective against infectious mononucleosis than older siblings. If the hypothesis that delayed Epstein-Barr virus infection is necessary for the development of multiple sclerosis is true, then the relative risk of multiple sclerosis as a function of sibship constellation should mirror the relative risk of infectious mononucleosis as a function of sibship constellation. Such an indirect hypothesis test is necessitated by the fact that age at primary Epstein-Barr virus infection is unknown for practically all people who have not experienced infectious mononucleosis. In this retrospective cohort study using nationwide registers, we followed all Danes born during the period 1971-2018 (n = 2 576 011) from 1977 to 2018 for hospital contacts with an infectious mononucleosis diagnosis (n = 23 905) or a multiple sclerosis diagnosis (n = 4442), defining two different end points. Relative risks (hazard ratios) of each end point as a function of sibship constellation were obtained from stratified Cox regression analyses. The hazard ratios of interest for infectious mononucleosis and multiple sclerosis could be assumed to be identical (test for homogeneity P = 0.19), implying that having siblings, especially of younger age, may protect a person against multiple sclerosis through early exposure to the Epstein-Barr virus. Maximum protection per sibling was obtained by having a 0-2 years younger sibling, corresponding to a hazard ratio of 0.80, with a 95% confidence interval of 0.76-0.85. The corresponding hazard ratio from having an (0-2 years) older sibling was 0.91 (0.86-0.96). Our results suggest that it may be possible essentially to eradicate multiple sclerosis using an Epstein-Barr virus vaccine administered before the teenage years. Getting there would require both successful replication of our study findings and, if so, elucidation of why early Epstein-Barr virus infection does not usually trigger the immune mechanisms responsible for the association between delayed Epstein-Barr virus infection and multiple sclerosis risk.


[Cross-Project Support of Registries in Development, Implementation and Operation].
The German Federal Ministry of Education and Research funded a project accompanying a funding initiative for registries in health services research. The aim was to provide cross-registry support initially for 16 and later 6 projects with regard to methodological, technical and structural standards. The 16 projects were initially guided in concept development, e. g., providing a template for a registry protocol. Furthermore, an expert consultation was organized and carried out. To assist in the selection of an IT solution, a challenge workshop was hosted where different vendors presented their software for registries. The catalogs of data elements of the projects were migrated into a metadata catalog and transferred to the standard model of ISO/IEC 11179. A set of quality indicators was defined for a cross-registry quality management approach to be implemented during the operational phase. To improve data quality, the indicators were to be transmitted and evaluated on a regular basis. The template for a registry protocol was used by the majority of projects when applying for funding of their operational phase. At the workshop on IT solutions, 12 products for registry software were presented; however, the projects opted for other solutions for different reasons. Transferring the catalogs of data elements into a standard model enabled a comparison of attributes and value sets, which in turn enabled formulation of recommendations for important elements. A set of five quality indicators was defined for quality management, for which an initial evaluation was carried out for 2020. The template of a registry protocol serves a systematic development of a concept. The use of a uniformly structured catalog of data elements supports compliance with the FAIR principles. Monitoring of data quality can be achieved by regularly identifying quality indicators across registries.


Outcomes of elective total laryngectomy for laryngopharyngeal dysfunction in disease-free head and neck cancer survivors.
Total laryngectomy (TL) can be offered for management of chronic aspiration, radionecrosis, and/or airway compromise after head and neck cancer (HNC) treatment. The objective of this study was to evaluate functional outcomes after TL in disease-free HNC survivors. Retrospective case series with chart review. The University of Texas MD Anderson Cancer Center. Twenty-three disease-free HNC survivors who underwent TL for laryngopharyngeal dysfunction. TL ± pharyngectomy. Post-TL swallowing-related (diet, gastrostomy dependence, and pneumonia rates) and communication outcomes. All patients who underwent TL for dysfunction were previously treated with radiotherapy (12/23, 52%) or chemoradiotherapy (11/23, 48%). Preoperative complications included aspiration (22/23, 96%), pneumonia (16/23, 70%), tracheostomy (9/23, 39%), and stricture (7/23, 30%); 17 patients (74%) required enteral/parenteral nutrition, and 13 of 23 (57%) were nothing per oral (NPO). Rates of pneumonia, NPO status, and feeding tube dependence significantly decreased after TL (P < .001). At last follow-up after TL, all patients tolerated oral intake, but 4 (17%) required supplemental enteral nutrition. Continued smoking after radiotherapy and a preoperative history of recurrent pneumonia were significantly (P < .05) associated with final tube dependence and/or diet level. Sixteen patients (70%) underwent tracheoesophageal (TE) puncture, and 57% (13 of 23) communicated using TE voice after TL. Salvage TL may improve health status by significantly decreasing the rate of pneumonia and improve quality of life by restoring oral intake in patients with refractory laryngopharyngeal dysfunction after HNC treatment. TE voice restoration may enhance functional outcomes in select patients treated with elective TL for dysfunction.


Egg age distribution, the direct way to cladoceran birth rates.
Cladoceran hatching frequency can easily be calculated from observed egg age distribution and estimated egg development time. The large timelag errors of models incorporating birth rates in the equation are avoided, and the population growth rate can be estimated directly.


Two- and three-dimensional distributions of opioid receptors on NG108-15 cells visualized with the aid of fluorescence confocal microscopy and anti-idiotypic antibodies.
Polyclonal anti-idiotypic antibodies previously shown to be specific for mu- and delta-opioid receptor subclasses on rat brain membrane preparations using radioreceptor assays have been labeled with fluorescent conjugates of avidin. The resulting complexes have been used to study the distribution and properties of opioid receptors on living and fixed NG108-15 cells using normal fluorescence, and confocal fluorescence, microscopy. From the confocal data three-dimensional views of immunofluorescent distributions have been obtained using a computer transformation of the data. From experiments involving co-incubation with media containing naloxone, selective opioid peptides and stereoisomeric synthetic opioid analogs as blocking agents, the specificity and stereospecificity of immunofluorescent binding is reported. The effects on immunofluorescent binding by co-incubation with media containing a sulfhydryl reagent, 100 mM-NaCl and sodium azide are also reported. The results have implications concerning the use of immunoprecipitation in the biochemical isolation of the receptors. Finally, the confocal two-dimensional sections and two-dimensional reconstructions obtained from fixed cells show internal immunofluorescence in the form of cytoplasmic clusters. The reagents and methods should be applicable to visualizing native opioid receptor distributions on neurons.


Enlarging the gas access channel to the active site renders the regulatory hydrogenase HupUV of Rhodobacter capsulatus O2 sensitive without affecting its transductory activity.
In the photosynthetic bacterium Rhodobacter capsulatus, the synthesis of the energy-producing hydrogenase, HupSL, is regulated by the substrate H2, which is detected by a regulatory hydrogenase, HupUV. The HupUV protein exhibits typical features of [NiFe] hydrogenases but, interestingly, is resistant to inactivation by O2. Understanding the O2 resistance of HupUV will help in the design of hydrogenases with high potential for biotechnological applications. To test whether this property results from O2 inaccessibility to the active site, we introduced two mutations in order to enlarge the gas access channel in the HupUV protein. We showed that such mutations (Ile65-->Val and Phe113-->Leu in HupV) rendered HupUV sensitive to O2 inactivation. Also, in contrast with the wild-type protein, the mutated protein exhibited an increase in hydrogenase activity after reductive activation in the presence of reduced methyl viologen (up to 30% of the activity of the wild-type). The H2-sensing HupUV protein is the first component of the H2-transduction cascade, which, together with the two-component system HupT/HupR, regulates HupSL synthesis in response to H2 availability. In vitro, the purified mutant HupUV protein was able to interact with the histidine kinase HupT. In vivo, the mutant protein exhibited the same hydrogenase activity as the wild-type enzyme and was equally able to repress HupSL synthesis in the absence of H2.


Incidence, determinants and the transient impact of cancer treatments on venous thromboembolism risk among lymphoma patients in Denmark.
Valid estimation of the incidence and risk factors for venous thromboembolism (VTE) among lymphoma patients has been limited by small studies focused on selected lymphoma subtypes and failure to account for death as a competing risk. Using a nationwide cohort of Danish lymphoma patients diagnosed from 2000 to 2010, we examined the incidence and risk factors for VTE and evaluated the transient impact of cancer treatments on VTE risk. Medical databases contained cancer, comorbidity, treatment, and VTE information. We computed VTE incidence rates (IRs) per 1000 person-years and 1- and 2-year incidence accounting for competing risks. Using Cox proportional hazards models, we identified factors associated with VTE risk. In a nested self-controlled design, we evaluated the transient effect of chemotherapy, radiation, central venous catheter use and rituximab on VTE risk using logistic regression models and adjusted odds ratios (aORs). VTE IRs were >40/1000 person-years within 180 days post-diagnosis, decreasing to 8/1000 person-years in year two. VTE risk was 2.9% and 3.5% at 1 and 2 years, respectively. Lymphoma subtype, central nervous system involvement, and elevated lactate dehydrogenase were associated with VTE risk. Central venous catheter use increased the transient odds of VTE (aOR=6.7 (1.2, 28.1)). We report a lower VTE incidence among lymphoma patients compared with prior studies. Lymphoma aggressiveness was the main driver of baseline VTE risk, whereas central venous catheter use increased transient risks. These accurate estimates may improve the identification of lymphoma subgroups at highest VTE risk, for whom future targeted prevention interventions may be beneficial.


Reduction in Nonabstinent WHO Drinking Risk Levels and Change in Risk for Liver Disease and Positive AUDIT-C Scores: Prospective 3-Year Follow-Up Results in the U.S. General Population.
Abstinence is often the treatment aim for alcohol use disorders (AUD), but this may deter individuals who prefer drinking reduction goals from entering treatment, and be an overly restrictive end point in alcohol clinical trials. Nonabstinent drinking reductions that predict improvement in how individuals feel or function may be useful clinical trial outcomes, for example, reductions in the 4-category World Health Organization (WHO) drinking risk levels. To investigate the clinical relevance of these reductions, we examined their relationship with 2 outcomes of interest to medical providers: liver disease, and positive scores on an alcohol screening measure. Current drinkers in a U.S. national survey (n = 21,925) were interviewed in 2001 to 2002 (Wave 1) and re-interviewed 3 years later (Wave 2). WHO drinking risk levels, liver disease, and the Alcohol Use Disorders Identification Test-Consumption (AUDIT-C) were assessed at both waves. Adjusted odds ratios (aORs) were used to indicate the association of change in WHO drinking risk levels with Wave 2 liver disease and AUDIT-C scores. Wave 1 very-high-risk drinkers who reduced 1, 2, or 3 WHO drinking risk levels had significantly lower odds of Wave 2 liver disease (aORs = 0.34, 0.23, 0.17) and positive AUDIT-C scores (aORs = 0.27, 0.09, 0.03). Wave 1 high-risk drinkers who reduced 1 or 2 WHO risk levels had significantly lower odds of positive AUDIT-C scores (aORs = 0.61, 0.25). Adjusting for alcohol dependence or AUDIT-C scoring variations did not affect results. In the highest-risk drinkers, reductions in WHO drinking risk levels predicted lower likelihood of liver disease and positive AUDIT-C scores. Results add to findings that reductions in the 4-category WHO drinking risk levels are a meaningful indicator of how individuals feel and function, and could serve as nonabstinent end points in clinical trials. Results also connect the WHO risk drinking levels to commonly used alcohol screening questions, which may be more familiar to healthcare providers.


Detrimental effects of cerium oxide nanoparticles on testis, sperm parameters quality, and in vitro fertilization in mice: An experimental study.
Cerium oxide nanoparticles (CeO 2 NPs) as an important nanomaterial have a wide range of applications in many fields and human beings' exposure to this nanomaterial is unavoidable. The effects of CeO 2 NPs on the male reproductive system are controversial. To determine the effects of the administration of CeO 2 NPs on the testis tissue, sperm parameters, and in vitro fertilization (IVF) in mice. Twenty-four male mice were divided into three groups (n = 8/each): one control and two experimental groups receiving CeO 2 NPs at doses of 50 and 100 mg/kg body weight, respectively, for 35 days. At the end of the experiment, the diameter of seminiferous tubules (SNTs), epithelial height of SNTs, spermiogenesis index in testes, sperm parameters (count, motility, viability, and morphology), sperm chromatin condensation, DNA integrity, and IVF assays were analyzed. Histological results showed that the tubular diameter, the epithelial height of the SNTs, and the spermiogenesis index were significantly decreased in the experimental groups receiving CeO 2 NPs. All sperm parameters in the experimental groups were significantly reduced and, additionally, the percentages of immature sperms and sperms with DNA damage were significantly increased in groups treated with CeO 2 NPs compared to the control. Furthermore, the rates of IVF and in vitro embryo development were decreased. Collectively, the current study showed that oral administration of CeO 2 NPs in mice had detrimental effects on the male reproductive system through inducing testicular tissue alterations, decreasing sperm parameters quality, and also diminishing the IVF rate and in vitro embryonic development.


Autophagy in sepsis-induced acute lung injury: Friend or foe?
Sepsis-induced acute lung injury (ALI) is a life-threatening syndrome with high mortality and morbidity, resulting in a heavy burden on family and society. As a key factor that maintains cellular homeostasis, autophagy is regarded as a self-digesting process by which damaged organelles and useless proteins are recycled for cell metabolism, and it thus plays a crucial role during physiological and pathological processes. Recent studies have indicated that autophagy is involved in the pathophysiological process of sepsis-induced ALI, including cell apoptosis, inflammation, and mitochondrial dysfunction, which indicates that regulating autophagy may be beneficial for this disease. However, the role of autophagy in the etiology and treatment of sepsis-induced ALI is not well characterized. This review summarizes the autophagy-related signaling pathways in sepsis-induced ALI, as well as focuses on the dual role of autophagy and its regulation by non-coding RNAs during disease progression, for the development of potential therapeutic strategies in this disease.


Rhabdomyosarcomas are oncogene addicted to the activation of AVIL.
Rhabdomyosarcoma (RMS) is one of the most common pediatric soft-tissue cancer. Previously, we discovered a gene fusion, MARS-AVIL formed by chromosomal inversion in RMS. Suspecting that forming a fusion with a housekeeping gene may be one of the mechanisms to dysregulate an oncogene, we investigated AVIL expression and its role in RMS. We first showed that MARS-AVIL translates into an in-frame fusion protein, which is critical for RMS cell tumorigenesis. Besides forming a gene fusion with the housekeeping gene, MARS, the AVIL locus is often amplified, and its RNA and protein expression are overexpressed in the majority of RMSs. Tumors with AVIL dysregulation exhibit evidence of oncogene addiction: Silencing MARS-AVIL in cells harboring the fusion, or silencing AVIL in cells with AVIL overexpression, nearly eradicated the cells in culture, as well as inhibited in vivo xenograft growth in mice. Conversely, gain-of-function manipulations of AVIL led to increased cell growth and migration, enhanced foci formation in mouse fibroblasts, and most importantly transformed mesenchymal stem cells in vitro and in vivo. Mechanistically, AVIL seems to serve as a converging node functioning upstream of two oncogenic pathways, PAX3-FOXO1 and RAS, thus connecting two types of RMS associated with these pathways. Interestingly, AVIL is overexpressed in other sarcoma cells as well, and its expression correlates with clinical outcomes, with higher levels of AVIL expression being associated with worse prognosis. AVIL is a bona fide oncogene in RMS, and RMS cells are addicted to its activity.


Amisulpride promotes cognitive flexibility in rats: the role of 5-HT7 receptors.
The antagonism of 5-HT7 receptors may contribute to the antidepressant and procognitive actions of the atypical antipsychotic drug, amisulpride. It has been previously demonstrated that the selective 5-HT7 receptor antagonist reversed restraint stress-induced cognitive impairments in a rat model of frontal-dependent attentional set-shifting task (ASST). Therefore, the first aim of the present study was to assess the effectiveness of amisulpride against stress-evoked cognitive inflexibility. The second goal was to elucidate whether the pro-cognitive effect of amisulpride could be due to the compound's action at 5-HT7 receptors. Rats repeatedly exposed (1 h daily for 7 days) to restraint stress demonstrated impaired performance on the extra-dimensional (ED) set-shifting stage of the ASST. Amisulpride (3 mg/kg) given to stressed rats 30 min before testing reversed this restraint-induced cognitive inflexibility and improved ED performance of the unstressed control group. The 5-HT7 receptor agonist, AS19 (10 mg/kg), abolished the pro-cognitive efficacy of amisulpride (3 mg/kg). The present study suggests that the antagonism of 5-HT7 receptors may contribute to the mechanisms underlining the pro-cognitive action of amisulpride. These results may have therapeutic implications in frontal-like deficits associated with stress-related disorders.


The murderer is the bed: an unusual case of death by traumatic asphyxia in a hotel folding bunk bed.
This paper presents the first referenced case on a death by traumatic asphyxia in a folding bunk bed. A middle-aged man was found dead in a hotel room trapped into a lower folding bunk bed where he had been sleeping after a party. The autopsy showed signs of asphyxia and excluded signs of struggle and sexual intercourse. Toxicological analyses revealed alcohol intoxication. A differential diagnosis of the manner of death including a technical study of the bed which contributed to understand the circumstances of death was made. The medico-legal investigation of the case strongly supported the hypothesis of an accidental death by traumatic asphyxia.


Differences in reendothelialization after balloon catheter removal of endothelial cells, superficial endarterectomy, and deep endarterectomy.
The process of reendothelialization was studied in a deendothelialized 3 cm segment of the canine descending thoracic aorta from which peripheral endothelial cell ingrowth had been prevented by impervious prosthetic graft sleeves. Three preparations were studied: (1) removal of only the endothelial cells, accomplished by flow surface drying and balloon catheter denudation, (2) removal of the superficial portion of the nonvasal media, accomplished by endarterectomy to a depth of 200 to 300 micron, and (3) removal of the entire inner (nonvasal) media, accomplished by endarterectomy to a depth of 500 to 600 micron to reach the outer (vasal) media. A total of 44 specimens were studied after implantation periods of 7, 14, 28, and 56 days. In all cases endothelial cell ingrowth from the host aorta into the test specimen was prevented by the impervious graft sleeves. In the deep endarterectomy group there were scattered areas of reendothelialization at 1 week, extensive reendothelialization at 2 weeks, almost complete reendothelialization at 4 weeks, and confluence by 8 weeks. However, in the superficial endarterectomy group scanning electron microscopy showed scattered areas of factor VIII/von Willebrand factor (FVIII/vWF)-negative, endothelial-like cells at 4 weeks, whereas at 8 weeks most of the surface was covered by endothelial cells identified by FVIII/vWF. In those specimens subjected to balloon catheter removal of endothelial cells only, reendothelialization was not seen, even at 8 weeks.


Faculty development initiatives for the integration of informatics competencies and point-of-care technologies in undergraduate nursing education.
Faculty members have a critical role in deciding the content that is taught to their nursing students. They must grasp the importance of using technology to facilitate learning and knowledge of informatics concepts and skills. This article describes a successful faculty development program that was aimed at upgrading the technology and informatics skills of the faculty while at the same time developing and threading informatics skills across the baccalaureate nursing curriculum.


Molecular dissection of mercury-responsive transcriptome and sense/antisense genes in Medicago truncatula.
We described a newly developed approach, namely next-generation tag sequencing, to identify global gene transcripts and complexity regulated by heavy metals in Medicago truncatula. Two cDNA libraries were generated from M. truncatula seedlings: treated and non-treated with the toxic heavy metal mercury Hg(II). With the large number of read-mapped genes generated, we observed that most of the genes were differentially expressed between the two libraries. In addition, several classes of new transcripts including transcription factors, antisense transcripts, and stress responsive genes were detected. The forty genes most altered in expression levels were associated with tolerance to environmental stress and secondary metabolism. Validation of genes by quantitative RT-PCR confirmed the results from deep-sequencing. Most of genes coding for metal transporters, sulfate metabolism, and cell wall solidification were significantly altered by Hg exposure. We also examined altered expression ratios of sense and antisense (S-AS) transcripts between the two libraries. By analyzing strand-specific information of read sequences, S-AS transcripts were found to be enriched with metal treatment. The transcriptome sequences were analyzed further with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) and showed diverse biological functions and metabolic pathways under the metal stress.


The gastric mucosal-associated microbiome in patients with gastric polyposis.
The characteristics of the gastric microbiota in patients with gastric polyposis (GP) remain unclear. Given this we collected gastric antrum and gastric body biopsies from healthy controls (HC.A and HC.B group) and gastric antrum, gastric body and polyp biopsies from patients with multiple gastric polyps (GP.A, GP.B and GP.P group) for 16S rDNA sequencing. The results showed that the diversity of the gastric flora in the GP group was significantly lower than that of the HC group. The gastric flora composition of the GP group was significantly different from the HC group. However, flora diversity and compositions in different parts of the stomach (gastric antrum, gastric body or polyp tissue) were not significantly different. H. pylori abundance could influence the composition of gastric microbiota. Meanwhile, patients with fundic gland polyps (FGPs) and those with hyperplastic polyps (HPs) had considerably similar gastric bacterial compositions. We constructed a microbial dysbiosis index (MDI) based on the gastric microbiota at the genus level as a predictive model, and it was able to distinguish between individuals in the GP and HC groups. These findings showed that intragastric flora dysbiosis may be closely related to the occurrence and development of gastric polyps.


Comparative bioavailability study with two amiodarone tablet formulations administered with and without food in healthy subjects.
The aim was to assess the comparative bioavailability of two formulations (200 mg tablet) of amiodarone (CAS 19774-82-4) in healthy volunteers of both sexes, with and without food. The study was conducted using an open, randomized, two-period crossover design with a 3-week washout interval, in two groups, with and without food. Plasma samples were obtained for up to 240 h post dose. Plasma amiodarone concentrations were analyzed by liquid chromatography coupled to tandem mass spectrometry (LC-MS-MS) with positive ion electrospray ionization using multiple reactions monitoring (MRM). From the amiodarone plasma concentration vs. time curves, the following pharmacokinetic parameters were obtained, with and without food: AUC(last), AUC(inf), AUC(0-240h), AUC(0-72h) and C(max). The limit of quantification was 1 ng/mL for plasma amiodarone analysis. The geometric mean and 90% confidence interval CI of Test/Reference percent ratios were, without and with food, respectively: 107.61 (92.73-124.89) and 100.6 (94.1-107.5) for C(max), 107.05 (95.88-119.51) and 100.2 (96.0-104.7) for AUC(last), 107.27 (95.78-120.15) and 100.8 (97.0-104.8) for AUC(0-72h), 106.76 (95.84-118.94) and 100.2 (96.0-104.7) for AUC(0-240h) and AUC(inf) 105.15 (94.18-117.41) and 100.7 (96.6-105.0). Since the 90% CI for AUC(0-72) and C(max) ratios were within the 80-125% interval proposed by the US FDA, it was concluded that the amiodarone 200 mg tablet (test formulation) with and without food was bioequivalent to the reference 200 mg tablet for both the rate and extent of absorption.


Self-reported assessment by medical students and interns of unprofessional practice.
Medical trainees should learn appropriate professional attitudes requisite for practicing medicine. Describe self-reported assessment of medical students and interns of unprofessional practice. A cross-sectional study was conducted on 88 participants (65 senior students and 23 interns) using self-administered questionnaire of 6 clinical and academic scenarios portraying a fictitious doctor engaged in unprofessional practice. Eighty-one (92.1 percent) participants believed professional misconduct is wrong, particularly failure to consult a specialist, disrespect to patients' autonomy, and research fraud. Seventy-three (83 percent) respondents would not perform these activities. There were no significant differences in responses by gender or year of study. Women and students, respectively, were significantly more lenient than men (p =.001) and interns (p =.002) regarding penalties appropriate for unprofessional behavior. Sixty-four (72.7 percent) participants would take action if colleagues failed to achieve professional standards. Perceptions of professional misconduct are not different in United Arab Emirates medical undergraduates from those observed elsewhere.


Low-Dose Endothelial Monocyte-Activating Polypeptide-II Induces Blood-Tumor Barrier Opening Via the cAMP/PKA/Rac1 Pathway.
Previous studies have demonstrated that low-dose endothelial monocyte-activating polypeptide-II (EMAP-II) induces blood-tumor barrier (BTB) hyperpermeability via both paracellular and transcellular pathways. In a recent study, we revealed that cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)-dependent signaling pathway is involved in EMAP-II-induced BTB hyperpermeability. This study further investigated the exact mechanisms through which the cAMP/PKA-dependent signaling pathway affects EMAP-II-induced BTB hyperpermeability. In an in vitro BTB model, low-dose EMAP-II (0.05 nM) induced a significant decrease in Rac1 activity in rat brain microvascular endothelial cells (RBMECs). Pretreatment with forskolin to elevate intracellular cAMP concentration completely blocked EMAP-II-induced inactivation of Rac1. Besides, pretreatment with 6Bnz-cAMP to activate PKA partially attenuated EMAP-II-induced Rac1 inactivation. Moreover, 6Bnz-cAMP pretreatment significantly diminished EMAP-II-induced changes in BTB permeability, myosin light chain (MLC) phosphorylation, expression and distribution of ZO-1, and actin cytoskeleton arrangement in RBMECs. These effects of 6Bnz-cAMP were completely blocked in the presence of NSC-23766 (the specific inhibitor of Rac1). In conclusion, this study demonstrates that low-dose EMAP-II induces BTB hyperpermeability via the cAMP/PKA/Rac1 signaling pathway.