Comparison of results of hormonal analysis of samples obtained from selected venous sites versus cervical ultrasonography for localizing parathyroid masses in dogs.
To compare a technique in which samples obtained from selected venous sites are analyzed for parathyroid hormone (PTH) concentration versus usefulness of cervical ultrasonography for localizing primary hyperparathyroidism (PHP) in dogs. Prospective study. 12 dogs with PHP. For each dog, blood samples were collected from the left and right jugular veins and 1 cephalic vein for determination of serum PTH concentration. Ultrasonography of the neck was performed in each dog. Each dog underwent exploratory surgery of the neck. Abnormal appearing parathyroid tissue was removed. Dogs were included in the study if serum calcium concentration decreased within 12 hours after surgery, hypercalcemia completely resolved within 96 hours after surgery, and serum calcium concentration was maintained within the reference range for at least 6 months after surgery. Serum PTH concentrations from the 3 veins were similar in 11 of 12 dogs with PHP. In 1 dog, the serum PTH concentration from the jugular vein ipsilateral to a parathyroid adenoma was greater than that from the contralateral jugular or cephalic vein. Ultrasonography correctly identified a parathyroid mass and its location in 10 of 11 dogs with a solitary abnormal parathyroid gland and in 1 dog in which both parathyroid glands were enlarged. Surgeons may benefit from knowing the location of abnormal parathyroid tissue in dogs with PHP before surgical exploration. Ultrasonography has potential value for identifying and localizing abnormal parathyroid tissue, whereas sample collection from selected sites for PTH analysis is not likely to be helpful.


First Report of Leaf Necrosis on Microstegium vimineum Caused by Bipolaris microstegii in Maryland.
Japanese stiltgrass (JSG, Microstegium vimineum) is an invasive weed causing significant ecological changes in the United States. Severely diseased plants in a shaded location 2 × 4 m in size were discovered in August 2012 at a residence on Indian Springs Rd., Frederick, MD (39.46747° N, 77.46106° W). JSG in larger monoculture stands at sunny locations within 6 to 10 m of diseased plants had a few, small, necrotic spots. Diseased plants had leaves with brown, often large, elliptical, necrotic spots up to 0.5 × 1.5 cm. Lesions were surrounded by a diffuse chlorotic margin, and larger lesions had tan centers. Diseased plants were smaller in stature than neighboring, non-symptomatic plants. Symptoms were similar to those on JSG reported by Kleczewski and Flory (2). Field samples of diseased leaves in moist chambers at room temperature and lighting produced dematiaceous conidiophores and conidia typical of Bipolaris within 2 days. Subcultures of the isolate (FDWSRU 12-049) were made from the conidia. Cultures growing on modified potato carrot agar (broth from 140 g each of potatoes and carrots and 20 g agar in 1 L water), were gray, velutinous or tomentose, and had clumps of short aerial hyphae on the upper surface. Healthy plants were grown in potting soil from seeds collected at the disease site. A minimum of five 4-week-old plants were spray-inoculated in each of three replications by conidia from detached leaves in a suspension of 105 spores/ml, given a 16-h dew period at 25°C, and placed in a 25°C greenhouse for observation. Non-inoculated plants were included for comparison. Brown or dark tan necrotic, irregular, often linear, spots with entire margins developed on all inoculated individual plants and not on control plants. The Bipolaris species was recovered from inoculated plant samples incubated in moist chambers, thus fulfilling Koch's postulates. Conidia were produced sympodially on dematiaceous conidiophores, often in clusters of two to three spores at the terminus, were medium to dark brown, straight or slightly curved, nearly fusiform with obtuse apices, and typically had 8 to 10 distoseptate cells that were 74.8 ± 2.3 × 16.4 ± 0.3 μm, and Q = 4.6 ± 0.1 (mean ± ci, P = 0.05; n = 100). A sequence of the ITS1-5.8S-ITS2 region of DNA, extracted using a DNeasy Plant Mini Kit (QIAGEN), was 100% identical to that of the type specimen of B. microstegii from M. vimineum (BPI 883727; GenBank Accession No. JX089579), using BLAST. On the basis of fungal morphology and molecular characteristics (1), along with symptomatology and published information (2), the causal agent of this disease has been determined to be B. microstegii. Dried specimens of both the isolate and diseased leaves were deposited in the U.S. National Fungus Collections (BPI 892680) and sequence information was submitted to GenBank (KF150215). Bipolaris leaf spot was also found on JSG in Howard and Prince Georges counties, MD, in 2009, but the causal agents were not formally characterized (K. Rane, Univ. Maryland; personal communication). This is the first confirmed identification of B. microstegii on JSG in Maryland, a plant that occurs as extensive monocultures in natural areas. These results provide a basis for characterization of this disease in the mid-Atlantic region. References: (1) P. W. Crous et al. Fungal Planet description sheets: 128-153. Persoonia 29:146, 2012. (2) N. M. Kleczewski and S. L. Flory. Plant Dis. 94:807, 2010.


Syntheses, structures, and magnetic properties of two unique Cu(ii)-based coordination polymers involving a crystal-to-crystal structural transformation from a 1D chain to a 3D network.
Two new coordination polymers, Cu(Hbsal)2(4,4'-bpy)2 (1) and Cu(bsal)(4,4'-bpy)·DMF (2), have been synthesized from 5-bromosalicylic acid (H2bsal) and the auxiliary ligand 4,4'-bipyridine (4,4'-bpy) through a slow diffusion method. Compound 1 is composed of parallel 1D linear chains, whereas compound 2 represents a non-interpenetrated 3D sod net which is constructed from left and right-handed helical chains alternately separated by 4,4'-bpy. The most interesting finding is that compound 1 can be irreversibly transformed into 2 in a crystal-to-crystal manner in the case that compound 1 is immersed in the DMF solvent. Particularly, the 1D-to-3D crystal-to-crystal structural transformation also incurs a drastic change in the magnetic properties of 1 and 2.


Confronting the Carbon-Footprint Challenge of Blockchain.
The distributed consensus mechanism is the backbone of the rapidly developing blockchain network. Blockchain platforms consume vast amounts of electricity based on the current consensus mechanism of Proof-of-Work (PoW). Here, we point out a different consensus mechanism named Proof-of-Stake (PoS) that can eliminate the extensive energy consumption of the current PoW-based blockchain. We comprehensively elucidate the current and projected energy consumption and carbon footprint of the PoW- and PoS-based Bitcoin and Ethereum blockchain platforms. The model of energy consumption of PoS-based Ethereum blockchain can lead the way toward the prediction of other PoS-based blockchain technologies in the future. With the widespread adoption of blockchain technology, if the current PoW mechanism continues to be employed, the carbon footprint of Bitcoin and Ethereum will push the global temperature above 1.5 °C in this century. However, a PoS-based blockchain can reduce the carbon footprint by 99% compared to the PoW mechanism. The small amount of carbon footprint from PoS-based blockchain could make blockchain an attractive technology in a carbon-constrained future. The study sheds light on the urgency of developing the PoS mechanism to solve the current sustainability problem of blockchain.


Preparation and in vitro/in vivo evaluation of vinpocetine elementary osmotic pump system.
Preparation and in vitro and in vivo evaluation of vinpocetine (VIN) elementary osmotic pump (EOP) formulations were investigated. A method for the preparation of VIN elementary osmotic pump tablet was obtained by adding organic acid additives to increase VIN solubility. VIN was used as the active pharmaceutical ingredient, lactose and mannitol as osmotic agent. Citric acid was used as increasing API solubility and without resulting in the API degradation. It is found that the VIN release rate was increasing with the citric acid amount at a constant range. Cellulose acetate 398-3 was employed as semipermeable membrane containing polyethylene glycol 6000 and diethyl-o-phthalate as pore-forming agent and plasticizer for controlling membrane permeability. In addition, a clear difference between the pharmacokinetic patterns of VIN immediate release and VIN elementary osmotic pump formulations was revealed. The area under the plasma concentration-time curve after oral administration of elementary osmotic pump formulations was equivalent to VIN immediate release formulation. Furthermore, significant differences found for mean residence time, elimination half-life, and elimination rate constant values corroborated prolonged release of VIN from elementary osmotic pump formulations. These results suggest that the VIN osmotic pump controlled release tablets have marked controlled release characters and the VIN osmotic pump controlled release tablets and the normal tablets were bioequivalent.


Phase-Field Model for the Simulation of Brittle-Anisotropic and Ductile Crack Propagation in Composite Materials.
In this work, a small-strain phase-field model is presented, which is able to predict crack propagation in systems with anisotropic brittle and ductile constituents. To model the anisotropic brittle crack propagation, an anisotropic critical energy release rate is used. The brittle constituents behave linear-elastically in a transversely isotropic manner. Ductile crack growth is realised by a special crack degradation function, depending on the accumulated plastic strain, which is calculated by following the J2-plasticity theory. The mechanical jump conditions are applied in solid-solid phase transition regions. The influence of the relevant model parameters on a crack propagating through a planar brittle-ductile interface, and furthermore a crack developing in a domain with a single anisotropic brittle ellipsoid, embedded in a ductile matrix, is investigated. We demonstrate that important properties concerning the mechanical behaviour of grey cast iron, such as the favoured growth of cracks along the graphite lamellae and the tension-compression load asymmetry of the stress-strain response, are covered by the model. The behaviour is analysed on the basis of a simulation domain consisting of three differently oriented elliptical inclusions, embedded in a ductile matrix, which is subjected to tensile and compressive load. The material parameters used correspond to graphite lamellae and pearlite.


Fundamental considerations of the design and function of intranasal antrostomies.
The natural history of intranasal antrostomy is poorly understood despite the popularity of the procedure. Researches have been conducted to examine this and in particular to establish factors which might be responsible for closure. A complete appraisal of the anatomy of the inferior meatus has been undertaken to determine factors which limit the dimensions of an antrostomy. Retrospective and prospective studies have been performed on patients undergoing the operation to assess patency and size. The results of these studies demonstrate that initial size is important in determining long-term patency in adults, and if an antrostomy is open at one year it usually remains open in the long-term unless infection supervenes; in children, however, antrostomies appear to close more rapidly.


Prolactin regulates maternal bile secretory function post partum.
The single-pass isolated perfused rat liver was used to assess bile secretory function in pregnancy and the post partum period and to evaluate the regulatory role of prolactin. Nonpregnant controls, post partum rats (48 hr post partum), pregnant rats (19-20 days of pregnancy) and pregnant rats treated with bromocriptine (3 mg/kg/day, s.c.) from day 20 of pregnancy until 48 hr post partum to block prolactin secretion were examined. In a separate set of experiments, ovariectomized rats were implanted with osmotic minipumps containing solvent alone or solvent plus varying concentrations of ovine prolactin (oPRL). Livers were isolated, [3H]taurocholate (TC) was infused at increasing concentrations and bile flow, bile acid secretory rate and hepatic clearance of TC from the perfusate were determined. Maximal bile flow (microliters/min/g liver) in response to infusion of TC was significantly higher in post partum rats (3.4 +/- 0.3) than in pregnant rats (1.4 +/- 0.3) and bromocriptine-treated post partum rats (1.7 +/- 0.3). The maximal bile acid secretory rate (SRm, nmol/min/g liver) in post partum rats was 276 +/- 19 and was significantly greater than SRm in pregnant (103 +/- 26) and bromocriptine-treated post partum rats (142 +/- 25). Hepatic clearance (ml/min) was significantly greater in post partum rats than in pregnant, control and bromocriptine-treated post partum rats at the highest TC concentration. Infusion of oPRL at doses of 100, 250 and 500 micrograms/day significantly increased maximal bile flow and SRm in a dose-dependent manner; these measures were increased to 4.38 +/- 0.21 microliters/min/g liver and 338.2 +/- 16.3 nmol/min/g liver, respectively, in rats treated with 500 micrograms/day oPRL.(ABSTRACT TRUNCATED AT 250 WORDS)


Molecular biology of KSHV lytic reactivation.
Kaposi's sarcoma-associated herpesvirus (KSHV) primarily persists as a latent episome in infected cells. During latent infection, only a limited number of viral genes are expressed that help to maintain the viral episome and prevent lytic reactivation. The latent KSHV genome persists as a highly ordered chromatin structure with bivalent chromatin marks at the promoter-regulatory region of the major immediate-early gene promoter. Various stimuli can induce chromatin modifications to an active euchromatic epigenetic mark, leading to the expression of genes required for the transition from the latent to the lytic phase of KSHV life cycle. Enhanced replication and transcription activator (RTA) gene expression triggers a cascade of events, resulting in the modulation of various cellular pathways to support viral DNA synthesis. RTA also binds to the origin of lytic DNA replication to recruit viral, as well as cellular, proteins for the initiation of the lytic DNA replication of KSHV. In this review we will discuss some of the pivotal genetic and epigenetic factors that control KSHV reactivation from the transcriptionally restricted latent program.


The Prevalence and Severity of Burnout among Physiotherapists in an Arabian Setting and the Influence of Organizational Factors: An Observational Study.
Burnout has been shown to be present in different health professions, but the prevalence among physiotherapists working in an Arabian setting has not been established. [Purpose] This study aimed to investigate the burnout levels of physiotherapists working in Saudi Arabia and the association of burnout with work and organization-related factors. [Subjects and Methods] A cross-sectional study was conducted at government hospitals in Saudi Arabia. One hundred and nineteen Saudi physiotherapists were included. They electronically completed a questionnaire that included the Maslach Burnout Inventory and the Areas of Worklife Survey. [Results] Participants showed a moderate degree of burnout as reflected by mean scores of the three subscales of the Maslach Burnout Inventory. The majority of participants demonstrated moderate to high burnout levels across the three subscales. A significant association was found between the exhaustion subscale and the subspecialty in which participants worked. A strong association was found between workload and exhaustion subscale scores. [Conclusion] This study was the first to explore burnout and related factors among physiotherapists in an Arabian setting. A moderate degree of burnout and associations of burnout with work and organizational factors were found. The findings may help human resource planning and managing the physiotherapy services.


C3a activates the respiratory burst in human polymorphonuclear neutrophilic leukocytes via pertussis toxin-sensitive G-proteins.
In contrast to C5a, which represents a well-established potent activator of the respiratory burst in polymorphonuclear neutrophilic granulocytes (PMN), the functional role of C3a in the activation of PMN is, so far, poorly understood. Herein, the potential role of human C3a in the activation of the respiratory burst in human PMN was investigated. The release of reactive oxygen species (ROS) of PMN from healthy donors was measured by lucigenin-dependent chemiluminescence. C3a dose-dependently induced the production of ROS in human PMN in the range between 10 ng/mL and 1,000 ng/mL, whereas C3a-desArg was inactive. Flow cytometric measurement of H2O2 by dihydrorhodamine-123 labeling of anti-CD16-stained PMN showed that predominantly neutrophilic PMN are responsible for the C3a-induced activation of the respiratory burst. To exclude that C3a stimulation was caused by contamination with C5a, the specificity of C3a-induced activation of PMN was shown using monoclonal antibodies (MoAbs). Accordingly, the effect of C3a was completely abolished in the presence of Fab fragments of a blocking anti-C3a MoAb. In addition, blockade of the C5a receptor by the anti-C5a receptor (anti-C5aR) MoAb, S5/1, totally inhibited the C5a-induced production of ROS, whereas the C3a response in the presence of the anti-C5aR MoAb was unaffected. The specificity of the response was further confirmed by homologous desensitization after restimulation with C3a. In contrast, no cross-desensitization was observed upon stimulation with C5a. The C3a-induced ROS production by PMN was inhibited by pertussis toxin, indicating the involvement of guanine nucleotide-binding proteins (Gi proteins) in the signal transduction process initiated by C3a. In addition, stimulation of PMN by C3a resulted in a transient increase in the cytosolic free calcium concentration ([Ca2+]i) in a dose-dependent manner. In contrast to C3a-induced ROS production, C3a did not induce a chemotactic response in PMN, indicating functional qualitative differences as compared with C5a. In summary, these results show that C3a is a potent activator of the respiratory burst in human PMN. Therefore, these findings point to a novel role of C3a in the pathogenesis of inflammatory diseases associated with increased C3a levels and PMN activation.


Comparison of real-time B-mode ultrasonography and bilateral ascending phlebography for detection of postoperative deep vein thrombosis following elective hip surgery. The Venous Thrombosis Group.
A prospective study compared real-time B-mode ultrasound examination with bilateral ascending phlebography in the diagnosis of postoperative deep vein thrombosis in 60 patients undergoing elective total hip replacement. The overall sensitivity and specificity of the ultrasound method were 54 and 91%, respectively, and the positive and negative predictive values were 83 and 69%, respectively. The rather low overall sensitivity of the ultrasound method in this study was due to difficulty in detecting thrombi smaller than 1 cm wherever they were located in the deep veins, and in diagnosing thrombi in the calf, regardless of their size. We conclude that real-time B-mode ultrasonography is a technique that can easily be used routinely for detection of postoperative DVT in hip surgery, but its sensitivity for proximal thrombosis (63%) is too low for it to be used alone.


Modulation of the activity of a human granulocyte-macrophage colony-stimulating factor/interleukin-3 fusion protein (pIXY 321) by the macrocyclic lactone protein kinase C activator bryostatin 1.
We have examined the effect of the macrocyclic lactone protein kinase C (PK-C) activator bryostatin 1 on the proliferative capacity and lineage commitment of CD34+ human bone marrow cells exposed to the granulocyte-macrophage colony-stimulating factor/interleukin-3 (GM-CSF/IL-3) fusion protein pIXY 321. pIXY 321 administered at a dose of 10 ng/mL was as effective as the combination of plateau concentrations of recombinant (r) IL-3 and rGM-CSF (e.g., 50 ng/mL) in stimulating the growth of day-14 committed myeloid progenitors (colony-forming units granulocyte/macrophage [CFU-GM]). In the large majority of samples tested, coadministration of 0.5 to 100 nM bryostatin 1 with either pIXY 321 or the combination of rIL-3 plus rGM-CSF led to modest but significant increases (e.g., 30 to 75%) in the number of CFU-GM, compared to administration of growth factors alone. The degree of bryostatin 1-induced potentiation, however, was considerably less than that previously observed in the case of cells exposed to either rIL-3 or rGM-CSF, where increases of 100 to 150% were regularly noted. While at least 50% of day-14 CFU-GM stimulated by either pIXY 321 or the combination of rIL-3 plus rGM-CSF were of the pure or mixed eosinophilic variety, coadministration of bryostatin 1 resulted in a dramatic inhibition of eosinophilic colonies and a corresponding increase in pure and mixed neutrophil and macrophage colonies. Although coadministration of recombinant granulocyte colony-stimulating factor (rG-CSF) or recombinant colony-stimulating factor-1 (rCSF-1) mimicked the capacity of bryostatin 1 to increase the total number of pIXY 321-induced day-14 CFU-GM, these growth factors, unlike bryostatin 1, were not capable of inhibiting eosinophilic colony formation. Furthermore, whereas addition of neutralizing antibodies to G-CSF or CSF-1 blocked the capacity of these growth factors to potentiate colony formation in the presence of pIXY 321, it did not abrogate the effect of bryostatin 1 on progenitor cell growth or lineage commitment. Finally, in contrast to its effects on committed myeloid progenitors, bryostatin 1 did not increase the growth of erythroid (burst-forming units-erythroid [BFU-E]) and multipotent (multipotent colony-forming units [CFU-GEMM]) progenitors stimulated by pIXY 321, but instead inhibited colony formation at higher concentrations (e.g., 10 to 100 nM).(ABSTRACT TRUNCATED AT 400 WORDS)


Numerical modelling of a three-phase internal air-lift circulating photocatalytic reactor.
The photocatalytic degradation process has been recognized as a low-cost, environmentally friendly and sustainable technology for water and wastewater treatment. As a key carrier of the photocatalytic process, the semiconductor TiO2 has been used in many studies. Analysis and modelling of hydrodynamics in the three-phase flow system can provide useful information for process design, operation and optimization of the three-phase flow photocatalytic reactor, which requires research on the mixing and flow characteristics of the interphase regions in the reactor. In this study, we modelled the hydrodynamics in an internal air-lift circulating photocatalytic reactor using an Eulerian multi-fluid approach. Localized information on phase holdup, fluid flow patterns and mixing characteristics was obtained. The simulation results revealed that the distribution of solid particle concentration depends on the flow field in the internal air-lift circulating photocatalytic reactor. The distance between the draft tube and wall of the reactor and changes in the superficial gas velocity (Ug) were found to be influential factors in reactor performance. The computational model developed could support optimizing reactor design to improve the hydrodynamics and provide guidance for scale-up.


Comparison of Free Radical Levels in the Aerosol from Conventional Cigarettes, Electronic Cigarettes, and Heat-Not-Burn Tobacco Products.
Aerosols from electronic cigarettes and heat-not-burn tobacco products have been found to contain lower levels of almost all compounds from the list of Harmful and Potentially Harmful Constituents known to be present in tobacco products and tobacco smoke than smoke from conventional cigarettes. Free radicals, which also pose potential health risks, are not considered in this list, and their levels in the different product types have not yet been compared under standardized conditions. We compared the type and quantity of free radicals in mainstream aerosol of 3R4F research cigarettes, two types of electronic cigarettes, and a heat-not-burn tobacco product. Free radicals and NO in the gas phases were separately spin trapped and quantified by electron paramagnetic resonance (EPR) spectroscopy by using a smoking machine for aerosol generation and a flow-through cell to enhance reproducibility of the quantification. Particulate matter was separated by a Cambridge filter and extracted, and persistent radicals were quantified by EPR spectroscopy. Levels of organic radicals for electronic cigarettes and the heat-not-burn product, as measured with the PBN spin trap, did not exceed 1% of the level observed for conventional cigarettes and were close to the radical level observed in air blanks. The radicals found in the smoke of conventional cigarettes were oxygen centered, most probably alkoxy radicals, whereas a signal for carbon-centered radicals near the detection limit was observed in aerosol from the heat-not-burn product and electronic cigarettes. The NO level in aerosol produced by electronic cigarettes was below our detection limit, whereas for the heat-not-burn product, it reached about 7% of the level observed for whole smoke from 3R4F cigarettes. Persistent radicals in particulate matter could be quantified only for 3R4F cigarettes. Aerosols from vaping and heat-not-burn tobacco products have much lower free radical levels than cigarette smoke, however, the toxicological implications of this finding are as yet unknown.


Neonatal Hypoxic-Ischemic Brain Injury Alters Brain Acylcarnitine Levels in a Mouse Model.
Hypoxic-ischemic brain injury (HIBI) leads to depletion of ATP, mitochondrial dysfunction, and enhanced oxidant formation. Measurement of acylcarnitines may provide insight into mitochondrial dysfunction. Plasma acylcarnitine levels are altered in neonates after an HIBI, but individual acylcarnitine levels in the brain have not been evaluated. Additionally, it is unknown if plasma acylcarnitines reflect brain acylcarnitine changes. In this study, postnatal day 9 CD1 mouse pups were randomized to HIBI induced by carotid artery ligation, followed by 30 min at 8% oxygen, or to sham surgery and normoxia, with subgroups for tissue collection at 30 min, 24 h, or 72 h after injury (12 animals/group). Plasma, liver, muscle, and brain (dissected into the cortex, cerebellum, and striatum/thalamus) tissues were collected for acylcarnitine analysis by LC-MS. At 30 min after HIBI, acylcarnitine levels were significantly increased, but the differences resolved by 24 h. Palmitoylcarnitine was increased in the cortex, muscle, and plasma, and stearoylcarnitine in the cortex, striatum/thalamus, and cerebellum. Other acylcarnitines were elevated only in the muscle and plasma. In conclusion, although plasma acylcarnitine results in this study mimic those seen previously in humans, our data suggest that the plasma acylcarnitine profile was more reflective of muscle changes than brain changes. Acylcarnitine metabolism may be a target for therapeutic intervention after neonatal HIBI, though the lack of change after 30 min suggests a limited therapeutic window.


Regulation and Role of Neuron-Derived Hemoglobin in the Mouse Hippocampus.
Hemoglobin (Hb) is the oxygen transport protein in erythrocytes. In blood, Hb is a tetramer consisting of two Hb-alpha (Hb-α) chains and two Hb-beta (Hb-β) chains. A number of studies have also shown that Hb-α is also expressed in neurons in both the rodent and human brain. In the current study, we examined for age-related regulation of neuronal Hb-α and hypoxia in the hippocampus and cerebral cortex of intact male and female mice. In addition, to confirm the role and functions of neuronal Hb-α, we also utilized lentivirus CRISPR interference-based Hb-α knockdown (Hb-α CRISPRi KD) in the non-ischemic and ischemic mouse hippocampus and examined the effect on neuronal oxygenation, as well as induction of hypoxia-inducible factor-1α (HIF-1α) and its downstream pro-apoptotic factors, PUMA and NOXA, and on neuronal survival and neurodegeneration. The results of the study revealed an age-related decrease in neuronal Hb-α levels and correlated increase in hypoxia in the hippocampus and cortex of intact male and female mice. Sex differences were observed with males having higher neuronal Hb-α levels than females in all brain regions at all ages. In vivo Hb-α CRISPRi KD in the mouse hippocampus resulted in increased hypoxia and elevated levels of HIF-1α, PUMA and NOXA in the non-ischemic and ischemic mouse hippocampus, effects that were correlated with a significant decrease in neuronal survival and increased neurodegeneration. As a whole, these findings indicate that neuronal Hb-α decreases with age in mice and has an important role in regulating neuronal oxygenation and neuroprotection.


Evidence for the Involvement of Pleckstrin Homology Domain-Containing Proteins in the Transport of Enterocin DD14 (EntDD14); a Leaderless Two-Peptide Bacteriocin.
Bacteriocins synthesis is initiated from an inactive precursor, which is composed of an N-terminal leader peptide attached to a C-terminal pro-peptide. However, leaderless bacteriocins (LLB) do not possess this N-terminal leader peptide nor undergo post-translational modifications. These atypical bacteriocins are observed to be immediately active after their translation in the cytoplasm. However, although considered to be simple, the biosynthetic pathway of LLB remains to be fully understood. Enterocin DD14 (EntDD14) is a two-peptide LLB produced by Enterococcus faecalis 14, which is a strain isolated from meconium. In silico analysis of DNA encoding EntDD14 located a cluster of 10 genes ddABCDEFGHIJ, where ddE and ddF encode the peculiar DdE and DdF proteins, carrying pleckstrin homology (PH) domains. These modules are quite common in Eucarya proteins and are known to be involved in intracellular signaling or cytoskeleton organization. To elucidate their role within the EntDD14 genetic determinants, we constructed deletion mutants of the ddE and ddF genes. As a result, the mutants were unable to export EntDD14 outside of the cytoplasm even though there was a clear expression of structural genes ddAB encoding EntDD14, and genes ddHIJ encoding an ABC transporter. Importantly, in these mutant strains (ΔddE and ΔddF), EntDD14 was detected by mass spectrometry in the intracellular soluble fraction exerting, upon its accumulation, a toxic effect on the producing strain as revealed by cell-counting and confocal microscopy analysis. Taken together, these results clearly indicate that PH domain-containing proteins, such as DdE and DdF, are involved in the transport of the leaderless two-peptide EntDD14.


Parental human leukocyte antigen-C allotypes are predictive of live birth rate and risk of poor placentation in assisted reproductive treatment.
To study the pregnancy, miscarriages, and live birth rates (LBRs) according to maternal killer cell immunoglobulin-like receptor (KIR) genes expressed by uterine natural killer cells and paternal or oocyte donor human leukocyte antigen-C (HLA-C) genes expressed by trophoblast cells in patients with recurrent reproductive failure. Prospective observational cohort study. Private infertility center. Participants included 204 women with recurrent miscarriage or recurrent implantation failure. The KIR and HLA-C genotypes of all women and HLA-C of their partners, gamete donors, miscarriage tissue, and babies were analyzed. All clinical variables (pregnancy, miscarriage, and LBRs) were analyzed and categorized based on KIR, oocyte origin, and single embryo transfer (SET)/double embryo transfer (DET). A higher miscarriage rate was observed after DETs in KIR AA mothers (47.8% egg donation and 37.5% in vitro fertilization [IVF]) compared with KIR AB (10.5% egg donation and 12.5% IVF) or KIR BB (6.7% egg donation and 0% IVF). A significantly decreased LBR was observed after DETs with oocyte donation in KIR AA patients (4.3%) compared with KIR AB (26.3%) or BB (46.7%). The LBR decreased significantly as the fetal HLA-C2 load increased in KIR AA women. Elective SET improves the reproductive outcomes compared with DET. An increased embryo HLA-C2 load has a negative impact on the LBR in KIR AA patients. The selection of HLA-C1 over HLA-C2 donors could have a positive impact on the LBR in KIR AA patients. NCT04052438.


Extremely rapid visual search: the maximum rate of scanning letters for the presence of a numeral.
Subjects searched a rapid sequence of computer-produced letter arrays for the presence of a numeral in one of the arrays. The subjects' scanning rates were computed from their precentage of correct detections of the location of the numeral. Scanning rates were very high and approximately the same for a wide variety of conditions; the highest scanning rates (125 and 75 letters per second for two subjects) occurred when there were 9 or 16 letters in each of the arrays and when new arrays were presented every 40 to 50 milliseconds. Giving the subject advance knowledge of the numeral to be presented made little difference in the scores.


Chronic hepatitis B in children. Natural history and treatment.
Hepatitis B virus (HBV) infection in children is worldwide in distribution, but the features of HBV-associated liver disease differ depending on the route of transmission and the time of acquisition of the infection. The degree of liver injury varies from a mild disease to the development of cirrhosis and hepatocellular carcinoma, and depends on the replicative status of the viral genome. It is believed that the immune function plays a key role in the severity of HBV disease, and the impact of HBV mutants needs to be assessed. The goals of antiviral therapy in children are therefore, the clearance of viremia and HBV sequences from infected tissues, together with an improvement in the liver disease. Administration of 10 MU/m2 b.s. 3 times weekly over 6 months resulted in a significantly higher clearance of viremia, with normalization of ALT values and greater improvement in liver histology in treated than in untreated patients. Long-term follow-up of these cases reveals the presence of the viral genome in serum and liver by PCR without clearance of HBsAg. Complete eradication of HBV might need more years of evolution as for adult patients. The combination of more than one antiviral agent, as well as the potentiation of the immune system, needs to be assessed to improve the actual response rate obtained with interferon-alpha.


Short limbs, cleft palate, and delayed formation of flat proliferative chondrocytes in mice with targeted disruption of a putative protein kinase gene, Pkdcc (AW548124).
During long bone development, round proliferative chondrocytes (RPCs) differentiate into flat proliferative chondrocytes (FPCs), and then into hypertrophic chondrocytes (HCs). FPCs and HCs support longitudinal bone growth. Here we show that a putative protein kinase gene, Pkdcc (AW548124), is required for longitudinal bone growth. We originally found Pkdcc expressed in the head organizer, but it is also expressed throughout embryogenesis and in various adult tissues. Pkdcc-/- embryos had no head organizer-related defects, but showed various morphological abnormalities at birth, including short limbs, cleft palate, sternal dysraphia, and shortened intestine. In the long bones of the limbs, only the mineralized regions were shortened, and the cartilage length was normal. In the humerus, Pkdcc was strongly expressed in the early FPCs, and FPC and HC formation was delayed in Pkdcc-/- mutants. Together, these data indicate that Pkdcc encodes a protein kinase that is required for the appropriate timing of FPC differentiation.


Response to corticosteroids in chronic airflow obstruction: relationship to emphysema and airways collapse.
We have studied the relationship between emphysema and airways collapse, and response to corticosteroids in patients with chronic airflow obstruction. One hundred and seven patients completed a placebo-controlled trial comparing 2 wks treatment with oral prednisolone 40 mg.day-1 to inhaled beclomethasone dipropionate 500 micrograms t.d.s. Response to corticosteroids was defined on the basis of changes in forced expiratory volume in one second (FEV1), and/or forced vital capacity (FVC), and/or mean peak expiratory flow (PEF) after treatment. Patients were categorized as those with physiologically defined emphysema (carbon monoxide transfer coefficient (KCO) less than 70% predicted and total lung capacity greater than 120% predicted), and those with pressure dependent airways collapse on the flow-volume loop (ratio of inspiratory to expiratory flow at 50% vital capacity [I:E50] greater than 10). The response to placebo showed a significant order effect, probably due to a carry-over effect of active treatment of at least 3 wks. Hence, the efficacy of active treatment over placebo in the subgroups defined was assessed by analysis of data generated from the first treatment phase of the trial. The presence or absence of physiologically defined emphysema did not affect the response to oral prednisolone. Inhaled beclomethasone dipropionate, however, was less effective in the emphysema group. Pressure dependent airways collapse did not affect the response to either prednisolone or beclomethasone. However, when data from all three treatment phases were analysed there was no significant difference in the response to either drug in any of the subgroups defined.(ABSTRACT TRUNCATED AT 250 WORDS)


Mitochondria Autoimmunity and MNRR1 in Breast Carcinogenesis: A Review.
We review here the evidence for participation of mitochondrial autoimmunity in BC inception and progression and propose a new paradigm that may challenge the prevailing thinking in oncogenesis by suggesting that mitochondrial autoimmunity is a major contributor to breast carcinogenesis and probably to the inception and progression of other solid tumors. It has been shown that MNRR1 mediated mitochondrial-nuclear function promotes BC cell growth and migration and the development of metastasis and constitutes a proof of concept supporting the participation of mitochondrial autoimmunity in breast carcinogenesis. The resemblance of the autoantibody profile in BC detected by IFA with that in the rheumatic autoimmune diseases suggested that studies on the autoantibody response to tumor associated antigens and the characterization of the mtDNA- and nDNA-encoded antigens may provide functional data on breast carcinogenesis. We also review the studies supporting the view that a panel of autoreactive nDNA-encoded mitochondrial antigens in addition to MNRR1 may be involved in breast carcinogenesis. These include GAPDH, PKM2, GSTP1, SPATA5, MFF, ncRNA PINK1-AS/DDOST as probably contributing to BC progression and metastases and the evidence suggesting that DDX21 orchestrates a complex signaling network with participation of JUND and ATF3 driving chronic inflammation and breast tumorigenesis. We suggest that the widespread autoreactivity of mtDNA- and nDNA-encoded mitochondrial proteins found in BC sera may be the reflection of autoimmunity triggered by mitochondrial and non-mitochondrial tumor associated antigens involved in multiple tumorigenic pathways. Furthermore, we suggest that mitochondrial proteins may contribute to mitochondrial dysfunction in BC even if mitochondrial respiration is found to be within normal limits. However, although the studies show that mitochondrial autoimmunity is a major factor in breast cancer inception and progression, it is not the only factor since there is a multiplex autoantibody profile targeting centrosome and stem cell antigens as well as anti-idiotypic antibodies, revealing the complex signaling network involved in breast carcinogenesis. In summary, the studies reviewed here open new, unexpected therapeutic avenues for cancer prevention and treatment of patients with cancer derived from an entirely new perspective of breast carcinogenesis.


Repair of critical size bone defects with porous poly(D,L-lactide)/nacre nanocomposite hollow scaffold.
To generate a novel porous poly(D,L-lactide)/nacre nanocomposite hollow scaffold. This study was performed in the Department of Spine Surgery, Southern Medical University, Guangzhou, China from September 2010 to September 2011. Nacre nanoparticles were prepared using a physical process and identified by x-ray diffraction and transmission electron microscopy, to generate a novel scaffold though the salt leaching processing technique. The morphology and structure properties of this scaffold were further investigated under scanning electron microscope and mechanical property testing. Additionally, the biological characteristics were evaluated by cell culture experiments in vitro. Thirty-six rabbits were randomly divided into 3 groups. The defects were implanted with/without poly(D,L-lactide)/nacre scaffold or poly(D,L-lactide) scaffold. The results were assessed by radiographs and bone mineral density to monitor bone repairing. The nacre nanoparticles were spherical in shape, with a diameter range from 45-95 nm. The scaffolds possessed an interconnected porous structure with an average pore size of 322.5+/-50.8 μm, and exhibited a high porosity (82.5 +/-0.8%), as well as good compressive strength of 4.5+/-0.25 Mpa. Primary biocompatibility experiments in vitro showed that cells adhered and proliferated well on the scaffolds. The animal study further demonstrated that the scaffolds could repair the critical size segmental bone defects in 12 weeks. Newly established scaffolds may serve as a promising biomaterial for bone tissue engineering.


Genome Wide Association Mapping of Seedling and Adult Plant Resistance to Barley Stripe Rust (Puccinia striiformis f. sp. hordei) in India.
Barley stripe rust is caused by Puccinia striiformis f.sp. hordei, (Psh), occurs worldwide, and is a major disease in South Asia. The aim of this work was to identify and estimate effects of loci underlying quantitative resistance to rust at seedling and adult plant stages. HI-AM panel of 261 barley genotypes consisting of released cultivars from North and South America, Europe, Australia, advanced breeding lines, and local landraces from ICARDA barley program were screened at seedling and adult plant stages for resistance to Psh. Seedling resistance was evaluated with the five prevalent Psh races in India. Screening for the adult plant stage resistance was also performed in two different locations by inoculating with a mixture of the five races used for seedling screeing. The panel was genotyped using DaRT-Seq high-throughput genotyping platform. The genome-wide association mapping (GWAM) showed a total of 45 QTL located across the seven barley chromosomes for seedling resistance to the five races and 18 QTL for adult plant stage resistance. Common QTL for different races at seedling stage were found on all chromosomes except on chromosome 1H. Four common QTL associated with seedling and adult plant stage resistance were found on chromosomes 2, 5, and 6H. Moreover, one of the QTL located on the long arm of chromosome 5H showed stable effects across environments for adult plant stage resistance. Several QTL identified in this study were also reported before in bi-parental and association mapping populations studies validating current GWAM. However 15 new QTL were found at adult plant stage on all chromosomes except the 4H, explaining up to 36.79% of the variance. The promising QTL detected at both stages, once validated, can be used for MAS in Psh resistance breeding program globally.


Prostate cancer screening: getting your patients to follow-up.
Nurses have the clinical knowledge and organizational ability to carry out a quality prostate cancer screening program. In addition, nurses possess the communication and patient education skills necessary for effective follow-up. With the dramatic increase in prostate screening programs, nurses are likely to be among the leaders in this effort. Motivating patients to follow up when they have an abnormal DRE or serum PSA can increase the early detection of prostate cancer and potentially cure the disease. After all, this is the primary purpose of the screening.


A TNFR2-Specific TNF Fusion Protein With Improved In Vivo Activity.
Tumor necrosis factor (TNF) receptor-2 (TNFR2) has attracted considerable interest as a target for immunotherapy. Indeed, using oligomeric fusion proteins of single chain-encoded TNFR2-specific TNF mutants (scTNF80), expansion of regulatory T cells and therapeutic activity could be demonstrated in various autoinflammatory diseases, including graft-versus-host disease (GvHD), experimental autoimmune encephalomyelitis (EAE) and collagen-induced arthritis (CIA). With the aim to improve the in vivo availability of TNFR2-specific TNF fusion proteins, we used here the neonatal Fc receptor (FcRn)-interacting IgG1 molecule as an oligomerizing building block and generated a new TNFR2 agonist with improved serum retention and superior in vivo activity. Single-chain encoded murine TNF80 trimers (sc(mu)TNF80) were fused to the C-terminus of an in mice irrelevant IgG1 molecule carrying the N297A mutation which avoids/minimizes interaction with Fcγ-receptors (FcγRs). The fusion protein obtained (irrIgG1(N297A)-sc(mu)TNF80), termed NewSTAR2 (New selective TNF-based agonist of TNF receptor 2), was analyzed with respect to activity, productivity, serum retention and in vitro and in vivo activity. STAR2 (TNC-sc(mu)TNF80 or selective TNF-based agonist of TNF receptor 2), a well-established highly active nonameric TNFR2-specific variant, served as benchmark. NewSTAR2 was assessed in various in vitro and in vivo systems. STAR2 (TNC-sc(mu)TNF80) and NewSTAR2 (irrIgG1(N297A)-sc(mu)TNF80) revealed comparable in vitro activity. The novel domain architecture of NewSTAR2 significantly improved serum retention compared to STAR2, which correlated with efficient binding to FcRn. A single injection of NewSTAR2 enhanced regulatory T cell (Treg) suppressive activity and increased Treg numbers by > 300% in vivo 5 days after treatment. Treg numbers remained as high as 200% for about 10 days. Furthermore, a single in vivo treatment with NewSTAR2 upregulated the adenosine-regulating ectoenzyme CD39 and other activation markers on Tregs. TNFR2-stimulated Tregs proved to be more suppressive than unstimulated Tregs, reducing conventional T cell (Tcon) proliferation and expression of activation markers in vitro. Finally, singular preemptive NewSTAR2 administration five days before allogeneic hematopoietic cell transplantation (allo-HCT) protected mice from acute GvHD. NewSTAR2 represents a next generation ligand-based TNFR2 agonist, which is efficiently produced, exhibits improved pharmacokinetic properties and high serum retention with superior in vivo activity exerting powerful protective effects against acute GvHD.


Episodic boundary cells in human medial temporal lobe during the free recall task.
A necessary condition for forming episodic memories is the construction of specific episodes demarcated from other episodes in space and time. Evidence from studies of episodic memory formation using rodent models suggest that the medial temporal lobe (MTL) supports the representation of boundary information. Building on recent work using human microelectrode recordings as well, we hypothesized of human MTL neurons with firing rates sensitive to episodic boundary information. We identified 27 episodic boundary neurons out of 736 single neurons recorded across 27 subjects. Firing of these neurons increased at the beginning and end of mnemonically relevant episodes in the free recall task. We distinguish episodic boundary neurons from a population of ramping neurons (n = 58), which are time-sensitive neurons whose activity provides complementary information during episodic representation. Episodic boundary neurons exhibited a U-shaped activity pattern demonstrating increased activity after both beginning and end boundaries of encoding and retrieval epochs. We also describe evidence that the firing of boundary neurons within episodic boundaries is organized by hippocampal theta oscillations, using spike-field coherence metrics.


ASC-J9 suppresses castration-resistant prostate cancer growth through degradation of full-length and splice variant androgen receptors.
Early studies suggested androgen receptor (AR) splice variants might contribute to the progression of prostate cancer (PCa) into castration resistance. However, the therapeutic strategy to target these AR splice variants still remains unresolved. Through tissue survey of tumors from the same patients before and after castration resistance, we found that the expression of AR3, a major AR splice variant that lacks the AR ligand-binding domain, was substantially increased after castration resistance development. The currently used antiandrogen, Casodex, showed little growth suppression in CWR22Rv1 cells. Importantly, we found that AR degradation enhancer ASC-J9 could degrade both full-length (fAR) and AR3 in CWR22Rv1 cells as well as in C4-2 and C81 cells with addition of AR3. The consequences of such degradation of both fAR and AR3 might then result in the inhibition of AR transcriptional activity and cell growth in vitro. More importantly, suppression of AR3 specifically by short-hairpin AR3 or degradation of AR3 by ASC-J9 resulted in suppression of AR transcriptional activity and cell growth in CWR22Rv1-fARKD (fAR knockdown) cells in which DHT failed to induce, suggesting the importance of targeting AR3. Finally, we demonstrated the in vivo therapeutic effects of ASC-J9 by showing the inhibition of PCa growth using the xenografted model of CWR22Rv1 cells orthotopically implanted into castrated nude mice with undetectable serum testosterone. These results suggested that targeting both fAR- and AR3-mediated PCa growth by ASC-J9 may represent the novel therapeutic approach to suppress castration-resistant PCa. Successful clinical trials targeting both fAR and AR3 may help us to battle castration-resistant PCa in the future.


Biosynthesis of Aurodox, a Type III Secretion System Inhibitor from Streptomyces goldiniensis.
The global increase in antimicrobial-resistant infections means that there is a need to develop new antimicrobial molecules and strategies to combat the issue. Aurodox is a linear polyketide natural product that is produced by Streptomyces goldiniensis, yet little is known about aurodox biosynthesis or the nature of the biosynthetic gene cluster (BGC) that encodes its production. To gain a deeper understanding of aurodox biosynthesis by S. goldiniensis, the whole genome of the organism was sequenced, revealing the presence of an 87 kb hybrid polyketide synthase/non-ribosomal peptide synthetase (PKS/NRPS) BGC. The aurodox BGC shares significant homology with the kirromycin BGC from S. collinus Tϋ 365. However, the genetic organization of the BGC differs significantly. The candidate aurodox gene cluster was cloned and expressed in a heterologous host to demonstrate that it was responsible for aurodox biosynthesis and disruption of the primary PKS gene (aurAI) abolished aurodox production. These data supported a model whereby the initial core biosynthetic reactions involved in aurodox biosynthesis followed that of kirromycin. Cloning aurM* from S. goldiniensis and expressing this in the kirromycin producer S. collinus Tϋ 365 enabled methylation of the pyridone group, suggesting this is the last step in biosynthesis. This methylation step is also sufficient to confer the unique type III secretion system inhibitory properties to aurodox. IMPORTANCE Enterohemorrhagic Escherichia coli (EHEC) is a significant global pathogen for which traditional antibiotic treatment is not recommended. Aurodox inhibits the ability of EHEC to establish infection in the host gut through the specific targeting of the type III secretion system while circumventing the induction of toxin production associated with traditional antibiotics. These properties suggest aurodox could be a promising anti-virulence compound for EHEC, which merits further investigation. Here, we characterized the aurodox biosynthetic gene cluster from Streptomyces goldiniensis and established the key enzymatic steps of aurodox biosynthesis that give rise to the unique anti-virulence activity. These data provide the basis for future chemical and genetic approaches to produce aurodox derivatives with increased efficacy and the potential to engineer novel elfamycins.


Ciprofloxacin is actively transported across bronchial lung epithelial cells using a Calu-3 air interface cell model.
Ciprofloxacin is a well-established broad-spectrum fluoroquinolone antibiotic that penetrates well into the lung tissues; still, the mechanisms of its transepithelial transport are unknown. The contributions of specific transporters, including multidrug efflux transporters, organic cation transporters, and organic anion-transporting polypeptide transporters, to the uptake of ciprofloxacin were investigated in vitro using an air interface bronchial epithelial model. Our results demonstrate that ciprofloxacin is subject to predominantly active influx and a slight efflux component.


Renal Cell Carcinoma with Venous Extension: Safety of Laparoscopic Surgery for Thrombus Levels I-IIIa.
Background: Novel studies are helping to consider minimally invasive surgery for treating patients with renal cell carcinoma and venous tumor thrombus. Evidence regarding its feasibility and safety is still sparse and does not include a subclassification for level III thrombi. We aim to compare the safety of laparoscopic vs open surgery in patients with levels I-IIIa thrombus. Materials and Methods: This is a cross-sectional comparative study using single-institutional data on adult patients treated surgically between June 2008 and June 2022. Participants were categorized into open and laparoscopic surgery groups. Primary outcome was difference in the incidence of 30-day major postoperative complications (Clavien-Dindo III-V) between groups. Secondary outcomes were differences in operative time, length of hospital stay, intraoperative blood transfusions, delta hemoglobin level, 30-day minor complications (Clavien-Dindo I-II), estimated overall survival, and progression-free survival between groups. A logistic regression model was performed including adjustment for confounding variables. Results: Overall, 15 patients in the laparoscopic group and 25 patients in the open group were included. Major complications occurred in 24.0% of patients within the open group and 6.7% of patients were treated laparoscopically (p = 0.120). Minor complications arose in 32.0% of patients treated with open surgery and in 13.3% of patients treated in the laparoscopic group (p = 0.162). Although not significant, there was a higher perioperative death rate within open surgery cases. The laparoscopic approach presented a crude odds ratio for major complications of 0.22 (95% confidence interval 0.02-2.1, p = 0.191) compared with open surgery. No differences were found between groups regarding oncologic outcomes. Conclusion: Laparoscopic approach for patients with venous thrombus levels I-IIIa seems to be as safe as open surgery.


Phytoextraction of Zn and Cu from sewage sludge and impact on agronomic characteristics.
The presence of elevated concentrations of heavy metals limits the usage of sewage sludge as a fertilizer and soil amendment. Experiments were carried out to examine the extent to which seven plant species phytoextracted Zn and Cu from dewatered sludge. The hyperaccumulators Thlaspi caerulescens and Sedum alfredii showed the greatest removal of Zn, while shoots and tubers of two species of Alocasia showed the greatest Cu removal. Cultivation of plants in the sludge resulted in significant decreases in total Zn and changes in the partitioning of Zn between soil pools. However, Cu levels were largely unchanged and remained associated predominantly with the organic matter pool. Agronomic characteristics of the sludge material, such as pH, organic matter content, and nitrogen, phosphorus, and potassium concentrations, did not change significantly during the four-month growth period, indicating that subsequent crops could be sustained by this material. These results suggest that Zn can be phytoextracted from sludge material, provided the rate of metal uptake exceeds the rate of mobilization to the exchangeable fraction. Since there was no appreciable accumulation of Zn and Cu in seeds of Zea mays in this study, some tissues from sludge-grown plants could potentially be used as animal fodder.


Cytochrome P450 CYP94B3 mediates catabolism and inactivation of the plant hormone jasmonoyl-L-isoleucine.
The phytohormone jasmonoyl-L-isoleucine (JA-Ile) signals through the COI1-JAZ coreceptor complex to control key aspects of plant growth, development, and immune function. Despite detailed knowledge of the JA-Ile biosynthetic pathway, little is known about the genetic basis of JA-Ile catabolism and inactivation. Here, we report the identification of a wound- and jasmonate-responsive gene from Arabidopsis that encodes a cytochrome P450 (CYP94B3) involved in JA-Ile turnover. Metabolite analysis of wounded leaves showed that loss of CYP94B3 function in cyp94b3 mutants causes hyperaccumulation of JA-Ile and concomitant reduction in 12-hydroxy-JA-Ile (12OH-JA-Ile) content, whereas overexpression of this enzyme results in severe depletion of JA-Ile and corresponding changes in 12OH-JA-Ile levels. In vitro studies showed that heterologously expressed CYP94B3 converts JA-Ile to 12OH-JA-Ile, and that 12OH-JA-Ile is less effective than JA-Ile in promoting the formation of COI1-JAZ receptor complexes. CYP94B3-overexpressing plants displayed phenotypes indicative of JA-Ile deficiency, including defects in male fertility, resistance to jasmonate-induced growth inhibition, and susceptibility to insect attack. Increased accumulation of JA-Ile in wounded cyp94b3 leaves was associated with enhanced expression of jasmonate-responsive genes. These results demonstrate that CYP94B3 exerts negative feedback control on JA-Ile levels and performs a key role in attenuation of jasmonate responses.


Characterization of Staphylococcus hyicus with the ATB 32 Staph system and with conventional tests.
The ATB 32 Staph system correctly identified 45 of 54 Staphylococcus hyicus cultures isolated from pigs and cattle. The biochemical profiles of the remaining nine cultures were not listed in the product data base. The 40 porcine and 14 bovine cultures resulted in three and seven different biochemical profiles, respectively. In parallel experiments, almost all S. hyicus cultures showed hemolytic reactions on chocolate agar, had CAMP-like reactivities in the zone of lysis of the staphylococcal beta-lysin, and had bacteriolytic properties on Micrococcus luteus cells. In addition, the S. hyicus cultures were unpigmented, were DNase positive, expressed an S. hyicus-specific teichoic acid, and were usually coagulase positive in porcine plasma. Most of the porcine strains were protein A positive, and two porcine cultures were clumping factor positive.


Adherence and HIV RNA Suppression in the Current Era of Highly Active Antiretroviral Therapy.
We examined trends in adherence to highly active antiretroviral therapy (HAART) and HIV RNA suppression and estimated the minimum cutoff of adherence to newer HAART formulations needed for HIV RNA suppression by regimen type. We used Veterans Affairs pharmacy dispensing data from the Veterans Aging Cohort Study Virtual Cohort between October 2000 and September 2010 and defined adherence as the duration of time the patient had the medications available, relative to the total number of days between refills for all antiretrovirals in a year. Temporal trends in adherence and viral load suppression were examined by the patient's most frequently used HAART regimen in the year. The minimum needed adherence was defined as the level at which the odds of suppression was not significantly different than that observed with ≥ 95% adherence using repeated-measures logistic regression. A total of 21,865 HAART users contributed 82,217 person-years of follow-up. There was a significant increase (P(trend) < 0.001) in the proportion virally suppressed even among those with <95% adherence (2001: 38% to 2010: 84%), and the trend was similar when restricting to their first HAART regimen. For nonnucleoside reverse transcriptase inhibitor multi-pill users, the odds of suppression did not differ for 85%-89% adherence compared to those with ≥ 95% adherence [odds ratios: 0.82 (0.64-1.04)], but for protease inhibitor users, the odds of suppression significantly differed if adherence levels were <95% compared to ≥ 95% adherence. Although all HIV-infected persons should be instructed to achieve perfect adherence, concerns of slightly lower adherence should not hinder prescribing new HAART regimens early in HIV infection.


[Alanine aminopeptidase from small intestinal mucosa of dogs, rabbits and trout. II. Behaviour of "detergent forms"].
Alanine aminopeptidases from the mucous membranes of the small intestines of dog, rabbit and trout were purified as detergent forms. The enzymes obtained were characterized with respect to substrate specificity, effector behaviour, Michaelis constants, molecular weight and temperature dependence. In comparison to data of the protease forms of the alanine aminopeptidases, differences could be demonstrated.


Nonintegrating foamy virus vectors.
Foamy viruses (FVs), or spumaviruses, are integrating retroviruses that have been developed as vectors. Here we generated nonintegrating foamy virus (NIFV) vectors by introducing point mutations into the highly conserved DD35E catalytic core motif of the foamy virus integrase sequence. NIFV vectors produced high-titer stocks, transduced dividing cells, and did not integrate. Cells infected with NIFV vectors contained episomal vector genomes that consisted of linear, 1-long-terminal-repeat (1-LTR), and 2-LTR circular DNAs. These episomes expressed transgenes, were stable, and became progressively diluted in the dividing cell population. 1-LTR circles but not 2-LTR circles were found in all vector stocks prior to infection. Residual integration of NIFV vectors occurred at a frequency 4 logs lower than that of integrase-proficient FV vectors. Cre recombinase expressed from a NIFV vector mediated excision of both an integrated, floxed FV vector and a gene-targeted neo expression cassette, demonstrating the utility of these episomal vectors. The broad host range and large packaging capacity of NIFV vectors should make them useful for a variety of applications requiring transient gene expression.


Effects of theophylline on erythropoietin production in normal subjects and in patients with erythrocytosis after renal transplantation.
Erythrocytosis occurs in 10 to 15 percent of renal-transplant recipients, and there is in vitro evidence that the production of erythropoietin is modulated by adenosine. We prospectively evaluated the effects of theophylline, a nonselective adenosine antagonist, in eight patients with erythrocytosis after renal transplantation and in five normal controls. After an eight-week course of theophylline treatment, the mean (+/- SEM) serum erythropoietin levels were significantly reduced in both the renal-transplant recipients (from 60 +/- 14 units per liter at base line to 9 +/- 7 units after treatment; P less than 0.05) and the normal subjects (from 6.9 +/- 0.8 units per liter at base line to 4.7 +/- 0.5 units per liter after treatment; P less than 0.05). Similarly, the hematocrits were reduced in both the transplant recipients (from 0.58 +/- 0.04 at base line to 0.46 +/- 0.03 after treatment; P less than 0.05) and the normal subjects (from 0.43 +/- 0.01 at base line to 0.39 +/- 0.01; P less than 0.05). In the renal-transplant recipients, red-cell mass was also reduced after eight weeks of theophylline (from 3197 +/- 82 ml at base line to 2273 +/- 69 ml after treatment; P less than 0.05). The previous requirement of weekly phlebotomy was eliminated in all recipients. Plasma and urinary cyclic AMP levels were not increased. These effects were reproducible when the subjects were rechallenged with theophylline after a recovery period. Theophylline attenuates the production of erythropoietin in both normal subjects and patients with erythrocytosis after renal transplantation and may be useful in the treatment of the latter condition.


Rapamycin and chloroquine: the in vitro and in vivo effects of autophagy-modifying drugs show promising results in valosin containing protein multisystem proteinopathy.
Mutations in the valosin containing protein (VCP) gene cause hereditary Inclusion body myopathy (hIBM) associated with Paget disease of bone (PDB), frontotemporal dementia (FTD), more recently termed multisystem proteinopathy (MSP). Affected individuals exhibit scapular winging and die from progressive muscle weakness, and cardiac and respiratory failure, typically in their 40s to 50s. Histologically, patients show the presence of rimmed vacuoles and TAR DNA-binding protein 43 (TDP-43)-positive large ubiquitinated inclusion bodies in the muscles. We have generated a VCPR155H/+ mouse model which recapitulates the disease phenotype and impaired autophagy typically observed in patients with VCP disease. Autophagy-modifying agents, such as rapamycin and chloroquine, at pharmacological doses have previously shown to alter the autophagic flux. Herein, we report results of administration of rapamycin, a specific inhibitor of the mechanistic target of rapamycin (mTOR) signaling pathway, and chloroquine, a lysosomal inhibitor which reverses autophagy by accumulating in lysosomes, responsible for blocking autophagy in 20-month old VCPR155H/+ mice. Rapamycin-treated mice demonstrated significant improvement in muscle performance, quadriceps histological analysis, and rescue of ubiquitin, and TDP-43 pathology and defective autophagy as indicated by decreased protein expression levels of LC3-I/II, p62/SQSTM1, optineurin and inhibiting the mTORC1 substrates. Conversely, chloroquine-treated VCPR155H/+ mice revealed progressive muscle weakness, cytoplasmic accumulation of TDP-43, ubiquitin-positive inclusion bodies and increased LC3-I/II, p62/SQSTM1, and optineurin expression levels. Our in vitro patient myoblasts studies treated with rapamycin demonstrated an overall improvement in the autophagy markers. Targeting the mTOR pathway ameliorates an increasing list of disorders, and these findings suggest that VCP disease and related neurodegenerative multisystem proteinopathies can now be included as disorders that can potentially be ameliorated by rapalogs.


Behavioral effects and receptor binding affinities of fentanyl derivatives in rhesus monkeys.
These studies examined the opioid receptor binding affinities and behavioral effects of several fentanyl derivatives in rhesus monkeys. OHM3295, OHM3296, OHM3326 and OHM3463 displayed high affinity for mu (IC50 = 7-66 nM) as compared to kappa (IC50 = 263-3255 nM) or delta (IC50 = 480-4500 nM) receptors as measured by their ability to displace [3H](D-Ala2-Me-Phe4,Glyol5)enkephalin, [3H](5,7,8[beta])-N-[2- (1-pyrrolidinyl)1-oxaspiro[4,5]dec-8-yl]benzeneacetamide and [3H](D-Pen2-D-Pen5)enkephalin, respectively. All four compounds maintained i.v. self-administration responding at rates above those maintained by the mu agonist alfentanil. In drug discrimination studies, OHM3463, OHM3326 and OHM3296 substituted completely for nalbuphine whereas OHM3295, and a related compound, mirfentanil, substituted partially for nalbuphine. In morphine-treated monkeys, OHM3295 substituted for naltrexone; in monkeys acutely deprived of morphine, only OHM3463 reversed naltrexone-lever responding. All four compounds had antinociceptive effects, although the extent to which these effects were accompanied by respiratory depression or modified by naltrexone, as well as the interactions between antinociceptive effects of fentanyl derivatives and alfentanil, varied markedly among compounds. Thus, OHM3463 shared effects with mu agonists (e.g., alfentanil) under all conditions; the other three compounds had opioid agonist effects under only a subset of conditions. Moreover, one of these compounds (OHM3295) antagonized the discriminative stimulus and antinociceptive effects of other mu agonists. Collectively, these compounds appear to vary on two dimensions: opioid efficacy and the contribution of nonopioid actions to their antinociceptive effects. Together with results obtained with other fentanyl derivatives (mirfentanil) under similar conditions, results of the current study suggest this chemical class might be especially fertile for the development of novel analgesics that might have reduced toxicity and abuse liability as compared to fentanyl and related compounds that are currently used in medicine.


Extracorporeal Membrane Oxygenation for Coronavirus Disease 2019: Crisis Standards of Care.
The coronavirus disease 2019 (COVID-19) pandemic has placed extraordinary strain on global healthcare systems. Use of extracorporeal membrane oxygenation (ECMO) for patients with severe respiratory or cardiac failure attributed to COVID-19 has been debated due to uncertain survival benefit and the resources required to safely deliver ECMO support. We retrospectively investigated adult patients supported with ECMO for COVID-19 at our institution during the first 80 days following New York City's declaration of a state of emergency. The primary objective was to evaluate survival outcomes in patients supported with ECMO for COVID-19 and describe the programmatic adaptations made in response to pandemic-related crisis conditions. Twenty-two patients with COVID-19 were placed on ECMO during the study period. Median age was 52 years and 18 (81.8%) were male. Twenty-one patients (95.4%) had severe ARDS and seven (31.8%) had cardiac failure. Fifteen patients (68.1%) were managed with venovenous ECMO while 7 (31.8%) required arterial support. Twelve patients (54.5%) were transported on ECMO from external institutions. Twelve patients were discharged alive from the hospital (54.5%). Extracorporeal membrane oxygenation was used successfully in patients with respiratory and cardiac failure due to COVID-19. The continued use of ECMO, including ECMO transport, during crisis conditions was possible even at the height of the COVID-19 pandemic.


Infant sleep and bedtime cereal.
We studied whether feeding infants rice cereal before bedtime promotes their sleeping through the night. One hundred six infants were randomly assigned to begin bedtime cereal feeding (1 tablespoon per ounce in a bottle) at 5 weeks or at 4 months of age. Caretakers recorded the infant's sleep from age 4 to 21 weeks for one 24-hour period per week. Sleeping through the night was defined as sleeping at least 8 consecutive hours, with the majority of time being between the hours of midnight and 6 AM. The results were also reviewed changing the requirement from 8 hours to 6 hours. There was no statistically significant trend or a consistent tendency of one group to have a higher proportion of sleepers than the other. Therefore, feeding infants rice cereal in the bottle before bedtime does not appear to make much difference in their sleeping through the night.


NK cell deficiency predisposes to viral-induced Th2-type allergic inflammation via epithelial-derived IL-25.
Severe respiratory syncytial virus (RSV) infection has long been associated with an increased risk for the development of childhood asthma and exacerbations of this disorder. Despite much research into the induction of Th2 responses by allergens and helminths, the factors associated with viral infection that predispose to Th2-regulated asthma remain unknown. Recently, clinical studies have shown reduced numbers of NK cells in infants suffering from a severe RSV infection. Here we demonstrate that NK cell deficiency during primary RSV infection of BALB/c mice results in the suppression of IFN-γ production and the development of an RSV-specific Th2 response and subsequent allergic lung disease. The outgrowth of the Th2 responses was dependent on airway epithelial cell-derived IL-25, which induced the upregulation of the notch ligand Jagged1 on dendritic cells. This study identifies a novel pathway underlying viral-driven Th2 responses that may have functional relevance to viral-associated asthma.


Ion Channel Dysfunction and Neuroinflammation in Migraine and Depression.
Migraine and major depression are debilitating disorders with high lifetime prevalence rates. Interestingly these disorders are highly comorbid and show significant heritability, suggesting shared pathophysiological mechanisms. Non-homeostatic function of ion channels and neuroinflammation may be common mechanisms underlying both disorders: The excitation-inhibition balance of microcircuits and their modulation by monoaminergic systems, which depend on the expression and function of membrane located K+, Na+, and Ca+2 channels, have been reported to be disturbed in both depression and migraine. Ion channels and energy supply to synapses not only change excitability of neurons but can also mediate the induction and maintenance of inflammatory signaling implicated in the pathophysiology of both disorders. In this respect, Pannexin-1 and P2X7 large-pore ion channel receptors can induce inflammasome formation that triggers release of pro-inflammatory mediators from the cell. Here, the role of ion channels involved in the regulation of excitation-inhibition balance, synaptic energy homeostasis as well as inflammatory signaling in migraine and depression will be reviewed.


Enigmas in sarcoidosis.
Sarcoidosis is a multisystem granulomatous disorder of unknown cause that presents most frequently in young adults with bilateral hilar adenopathy, pulmonary infiltrates, and skin or eye lesions. The multisystem clinical manifestations of this disease are a diagnostic challenge to all physicians. Although the clinical and pathologic characteristics of sarcoidosis are well described, the decision to treat and the optimal therapy are less well defined. This review focuses on the natural history, clinical manifestations, controversies in therapy, including steroid-sparing agents, and current concepts of how the disease's activity can be monitored.


A Review on Carotid Ultrasound Atherosclerotic Tissue Characterization and Stroke Risk Stratification in Machine Learning Framework.
Cardiovascular diseases (including stroke and heart attack) are identified as the leading cause of death in today's world. However, very little is understood about the arterial mechanics of plaque buildup, arterial fibrous cap rupture, and the role of abnormalities of the vasa vasorum. Recently, ultrasonic echogenicity characteristics and morphological characterization of carotid plaque types have been shown to have clinical utility in classification of stroke risks. Furthermore, this characterization supports aggressive and intensive medical therapy as well as procedures, including endarterectomy and stenting. This is the first state-of-the-art review to provide a comprehensive understanding of the field of ultrasonic vascular morphology tissue characterization. This paper presents fundamental and advanced ultrasonic tissue characterization and feature extraction methods for analyzing plaque. Additionally, the paper shows how the risk stratification is achieved using machine learning paradigms. More advanced methods need to be developed which can segment the carotid artery walls into multiple regions such as the bulb region and areas both proximal and distal to the bulb. Furthermore, multimodality imaging is needed for validation of such advanced methods for stroke and cardiovascular risk stratification.


Investigation of genetic susceptibility to non-small cell lung cancer by fragile site expression.
Fragile sites are non-staining gaps and breaks in specific points of chromosomes that are inducible by various culture conditions. Previous studies have shown that various clastogenic agents increase expression of fragile sites. In this study, the expression of common fragile sites induced by aphidicolin was evaluated on prometaphase chromosomes obtained from peripheral blood lymphocytes. Chromosomal aberrations and fragile site expression of 60 individuals, including 20 patients with non-small cell lung cancer (NSCLC), 20 of their clinically healthy family members, and 20 age-matched normal healthy controls without history of any cancer type were studied. Both the proportion of damaged cells (P < 0.001) and the mean number of gaps and breaks per cell (P < 0.001) were significantly higher in both the patients and relatives' groups when compared with the control group. However, they were insignificant when the patients were compared to their relatives (P > 0.05). We determined four aphidicolin type common fragile sites in our study. These sites in patients with NSCLC and relatives were the following: 1p21, 2q33, 3p14, and 16q23. In these fragile sites, 2q33, 3p14, and 16q23 sites were statistically significant when compared with control group (P < 0.001, P < 0.0005, and P < 0.05, respectively). Consequently, we believe that fragile site studies may be helpful to detection of cancer risk.


Seasonal variations in mortality caused by cardiovascular diseases in Norway and Ireland.
Seasonal variations in mortality resulting from cardiovascular diseases (CVD) have been demonstrated in many countries, with the highest levels observed during the coldest months of the year. We studied the seasonal changes in CVD mortality in Norway and the Republic of Ireland, two countries which are demographically quite similar, but climatically different; we also examined the relation between CVD mortality and air temperature. Registered monthly data for mortality from CVD for the period 1985-1995 were obtained from the Norwegian Central Bureau of Statistics and the Irish Central Statistics Office. Meteorological data were provided by the Norwegian Institute of Meteorology and Met Eireann, in Ireland. Monthly mortality ratio for both men and women aged 60 and older was calculated from the mortality date. Mean monthly air temperatures for the two countries were calculated from the meteorological data. For the 10-year period investigated, the lowest and highest monthly mortality ratios were on average found in August and January, respectively, and mean excess winter mortality, expressed as the difference between the August and January values for the entire 10-year period, was 22% (Norway) and 35% (Ireland). However, when the percentage difference in the months with the respective highest and lowest mortality ratios were calculated for each year, the average of these differences for each of the 10 individual years was 29% and 45%. Mortality ratio was found to increase much more steeply with decreasing air temperature in Ireland than in Norway. Although the seasonal variation between CVD mortality in both countries is similar, the different relation with climatic conditions may result from differences in housing standards, allowing outdoor temperatures to have a greater influence on indoor temperature in Ireland than in Norway.


Solitary Plasmacytoma of Tibia in a 29-Year-Old Woman.
Solitary plasmacytoma of the bone is a plasma cell tumor characterized by a single bone lesion with no evidence of myeloma elsewhere, including fewer than 5% plasma cells in the bone marrow. Although median age is about 14 years younger than that of patients with multiple myeloma, both diseases are extremely uncommon under 30 years of age. Solitary plasmacytoma of bone seem to have a prediction to occur in the axial skeleton, particularly in a vertebra and long bones are rarely affected. Herein, we report a case of solitary plasmacytoma of bone involving the tibia in a 29-year-old woman.


Maximizing capture efficiency and specificity of magnetic separation for Mycobacterium avium subsp. paratuberculosis cells.
In order to introduce specificity for Mycobacterium avium subsp. paratuberculosis prior to a phage amplification assay, various magnetic-separation approaches, involving either antibodies or peptides, were evaluated in terms of the efficiency of capture (expressed as a percentage) of M. avium subsp. paratuberculosis cells and the percentage of nonspecific binding by other Mycobacterium spp. A 50:50 mixture of MyOne Tosylactivated Dynabeads coated with the chemically synthesized M. avium subsp. paratuberculosis-specific peptides biotinylated aMp3 and biotinylated aMptD (i.e., peptide-mediated magnetic separation [PMS]) proved to be the best magnetic-separation approach for achieving 85 to 100% capture of M. avium subsp. paratuberculosis and minimal (<1%) nonspecific recovery of other Mycobacterium spp. (particularly if beads were blocked with 1% skim milk before use) from broth samples containing 10(3) to 10(4) CFU/ml. When PMS was coupled with a recently optimized phage amplification assay and used to detect M. avium subsp. paratuberculosis in 50-ml volumes of spiked milk, the mean 50% limit of detection (LOD(50)) was 14.4 PFU/50 ml of milk (equivalent to 0.3 PFU/ml). This PMS-phage assay represents a novel, rapid method for the detection and enumeration of viable M. avium subsp. paratuberculosis organisms in milk, and potentially other sample matrices, with results available within 48 h.


Cognitive Behavioral Group Therapy for Anger Management in Juvenile Delinquents: Effectiveness Study in a Turkey Child Prison.
The purpose of this study was to evaluate the effectiveness of cognitive-behavioral group therapy (CBGT) for anger management in Juvenile Delinquents. The therapy program consisted of eight sessions including relaxation, self-instruction, cognitive restructuring, and assertiveness training techniques. The therapy program has been conducted for juvenile delinquents in one of the juvenile prisons in Turkey. Sixty juvenile delinquents, aged between 14 and 18, who met the inclusion criteria (treatment = 30, control = 30), were included at the beginning of the study. To evaluate the effectiveness of the program, State Trait Anger and Expression Inventory (STAXI), and Brief Symptom Inventory (BSI) were administered as pre-test. At the end of the therapy program, final analysis was conducted on 35 participants (20 therapy group, 15 control group) due to participants dropout. Mixed ANOVA was conducted to examine the time (pre-test and post-test) and group effects (therapy and control group). Results showed that the participants in the therapy group showed a significant decrease in trait anger and anger expression scores, both subscales of STAXI and in the hostility subscale of BSI; a statistically significant increase in the anger control scores of the subscale of STAXI when compared to the control group. In conclusion, this culturally attuned CBGT for anger management may be applicable for Juvenile Delinquents in Turkey.


Co-designing a peer-led model of delivering behavioural activation for people living with depression or low mood in Australian farming communities.
Farmers face a range of factors that negatively influence their mental health and suicide risk, yet have limited access to appropriate support. Behavioural activation (BA) is an evidence-based therapy that can be effectively delivered by nonclinical workers. Working with members of farming communities to deliver BA to their peers has the potential to overcome many well-established barriers to mental health help-seeking and improve outcomes for this at-risk group. This paper describes the findings of a co-design phase informing the development of a peer (farmer)-led approach for delivering BA for farmers living with depression or low mood. This qualitative study used a co-design approach involving members of the target community. Focus groups were transcribed and analysed using Thematic Analysis and the Framework approach. Ten online focus groups with 22 participants were held over 3 months. Four overarching, interlinked themes were identified: (i) filling the gap in rural mental health support; (ii) alignment with the farming context-tailoring how, where and when we engage about mental health; (iii) the 'messenger' is as important as the message; and (iv) sustainability, governance and support. Findings suggest BA could be a contextually appropriate model of support for the farming community-given its practical and solution-focused approach-and could help improve access to support. Having peer workers deliver the intervention was viewed as appropriate. Ensuring governance structures are developed to support peers to deliver the intervention will be essential to facilitate effectiveness, safety and sustainability. Insights gained through co-design have been critical to the success of developing this new model of support for members of farming communities experiencing depression or low mood.


Tinkering with successful synapse regeneration in the leech: adding insult to injury.
In the leech, synapse regeneration in adults and synapse formation during embryonic development can be studied in single, identifiable cells that make precise connections with their targets. Certain cellular components, such as synaptic targets and glia, were selectively destroyed to study how the regenerating axons locate their targets, what triggers axons to start growing and what stops them. The results showed that glia and targets play only a limited role in synapse regeneration and in axon degeneration. For example, contact with the synaptic target may inhibit sprouting and availability of targets may promote it. Comparative studies on axon growth and synapse formation by interneurones in embryos showed that regeneration does not simply recapitulate embryonic development. There are clearly separate constraints on the two processes. Axon survival is a different problem. Although isolated axon segments can survive for up to a year in the leech, temperature is a major factor in survival. Axon segments in a tropical leech that regenerates synapses well at 31 degrees C degenerated within 2-3 weeks at this elevated temperature, even when regeneration was prevented. In similar leeches at room temperature (22 degrees C), segments survived for months. Overall, results in the leech support the idea that degeneration as well as regeneration share fundamental mechanisms with other invertebrates and the vertebrates, including mammals. Perhaps long-lived axon segments and other features of the leech that speed or encourage functional regeneration can now be made to operate in repair of the mammalian nervous system.


[Bioethics of Sigmund Freud´s death: euthanasia or appropriation?].
The death of Freud raises the ethical dilemma about euthanasia. It can be characterized as indirect active euthanasia according to the rule of double effect, or terminal sedation, or palliated death. The primacy of the principle of autonomy over non maleficence, conditioned the physician's attitude toward his patient Freud. The physician assisted death was and remains punishable in western medicine. Therefore, a fundamental tradition was infringed. In contrast, the present study attempts to characterize the final position of Freud himself to his death and called it appropriation of his finitude; he assumes his being-unto-death, that is, he now projects his being not as a being-at-his-end but as a being-unto-end, indicating thereby that he understood that the end always penetrated his whole existence.


Surgical treatment of craniopharyngiomas: experience with 168 patients.
The goal of this study was to assess the outcome of surgical management in 168 consecutive patients harboring craniopharyngiomas treated between January 1983 and April 1997. In 148 patients undergoing initial (primary) surgery, the pterional approach was most frequently used (39.2%), followed by the transsphenoidal approach (23.6%). For large retrochiasmatic craniopharyngiomas, the bifrontal interhemispheric approach was used increasingly over the pterional approach and led to improved surgical results. Total tumor removal was accomplished in 45.7% of transcranial and 85.7% of transsphenoidal procedures. The main reasons for incomplete removal were attachment to and/or infiltration of the hypothalamus, major calcifications, and attachment to vascular structures. The success rate in total tumor removal was inferior in the cases of tumor recurrence. The operative mortality rate in transcranial surgery was 1.1% in primary cases and 10.5% in cases of tumor recurrence. No patient died in the group that underwent transsphenoidal surgery. The rate of recurrence-free survival after total removal was 86.9% at 5 years and 81.3% at 10 years. In contrast, the 5-year recurrence-free survival rate was only 48.8% after subtotal removal and 41.5% after partial removal. Following primary surgery, the actuarial survival rate was 92.7% at 10 years, with the best results after complete tumor removal. At last follow up, 117 (79%) of 148 patients who underwent primary surgery were independent and without impairment. Total tumor removal while avoiding hazardous intraoperative manipulation provides favorable early results and a high rate of long-term control in craniopharyngiomas.


[In-house questioning on patient satisfaction in a maximum resource clinic--report from practice].
Since 1999, the statutory obligation of all German hospitals to establish an internal quality management system has resulted in an increase of patient surveys on inpatient care. Information on the subjective satisfaction of the patients is of particular interest because improvements may be realised. In the present paper, the objectives, the methodology and the choice of a tested survey instrument, the interpretation of the results and the practical problems in implementing a survey on patient satisfaction at the Leipzig University Hospital (a public institution comprising 1,464 beds in 2001) will be reported. This is intended to facilitate comparable projects in the future and to enable hospitals with fewer resources to organize and use such surveys. Altogether, 1,718 questionnaires (Hamburg questionnaires on patient stay in hospital, HFK) from 43 wards could be evaluated. The high satisfaction determined (mean values of the scores for 10 test parameters are between 1.72 and 2.02 on a 5-point scale) corresponds to the general tendency of patients to make socially desirable responses. Covert dissatisfaction was therefore analysed comparing the relative frequencies of complete and limited satisfaction at item level. Sociodemographic patient characteristics such as age and stay in hospital were of no substantial relevance. One important aim of the Leipzig survey project was to give detailed feedback within brief time periods. The project focussed on a differentiated and early monitoring of responses. If requested, an oral presentation of the results was made to each structural unit. This presentation included the spontaneous responses of the patient, which enhanced the acceptance of the investigation by the medical staff.


Use of a New Standardized Unfractionated Heparin Protocol for Left-Sided Ablation Procedures Improves Time in the Therapeutic ACT Range.
This study evaluated whether a novel standardized heparin dosing protocol used during atrial fibrillation catheter ablation resulted in a higher percentage of therapeutic activated clotting time (ACT) values compared to historic nonstandardized procedures. A retrospective cohort study SETTING: This study was conducted at Ochsner Medical Center, the largest tertiary-care teaching hospital in New Orleans, LA PARTICIPANTS: Patients undergoing catheter-based atrial fibrillation ablation INTERVENTIONS: The authors implemented a standardized heparin protocol, and enrolled 202 patients between November 2020 and March 2021. The historic controls consisted of 173 patients who underwent atrial fibrillation ablation between April 2020 and September 2020. Heparin administration in the control group was based on physician preference and was nonstandardized. The primary endpoint was the percentage of intraprocedural ACTs in therapeutic range (≥300 to <450 s). Secondary endpoints included first measured ACT at ≥300 s and percent of measured ACTs in the supratherapeutic range (>450 s). Comparisons were performed using chi-squared tests or Fisher exact tests. Patients in the intervention group had a higher mean percentage of ACTs in the therapeutic range compared to the control group (84.9% vs. 75.8%, p<0.001). More patients in the intervention group reached therapeutic ACT on the first measurement compared to the control group (70.3% vs. 31.2%, p<0.001). During catheter-based cardiac ablation procedures, a novel standardized unfractionated heparin dosing protocol resulted in a higher percentage of ACTs in the target range, and a higher proportion of initial ACTs in the therapeutic range compared with baseline nonstandardized heparin dosing.


Alpha-glucosidase inhibitors from clove (Syzgium aromaticum).
The ellagitannins, casuarictin and eugeniin, were isolated as rat intestinal maltase inhibitors from methanol extracts of clove (Syzgium aromaticum). Eugeniin showed inhibitory activity with an IC50 value of 10(-3) M. A structure-activity relationship study among the isolates and their related compound, penta-O-galloyl-beta-D-glucose, indicates that an increasing number of galloyl units in the molecule might lead to an increase in the inhibitory activity. Eugeniin also inhibited maltase activity toward the human intestinal epithelial cell line, Caco-2.


Partial agonist effects on the interaction between the atrial muscarinic receptor and the inhibitory guanine nucleotide-binding protein in a reconstituted system.
The mechanism of action of the partial muscarinic agonist pilocarpine was analyzed in a reconstituted system consisting of the purified porcine atrial muscarinic receptor and the purified porcine atrial inhibitory guanine nucleotide-binding protein, Gi. When GTPase activity was measured as a function of receptor.agonist complex concentration at saturating concentrations of either the full agonist carbachol or pilocarpine, both ligands gave similar values of kcat (4.3 +/- 0.2 min-1 for carbachol and 5.4 +/- 0.7 min-1 for pilocarpine); however, the observed dissociation constant for the ligand.receptor complex binding to Gi was about 4-fold lower for carbachol (0.81 +/- 0.19 nM) than for pilocarpine (3.02 +/- 0.83 nM). These results suggested that, in this system, the reduced activity of the partial agonist compared with the full agonist was the result of a decrease in affinity of the receptor.ligand complex for Gi, as opposed to differences in their relative abilities to activate the guanine nucleotide-binding protein. Several analogues of oxotremorine were also tested to determine their effects on the GTPase activity of Gi. Results from these studies indicate that the reconstituted system may be useful in determining structure-function relationships for muscarinic agonists with regard to receptor.Gi interactions.


Geochemical assessments and classification of coal mine spoils for better understanding of potential salinity issues at closure.
Coal mining wastes in the form of spoils, rejects and tailings deposited on a mine lease can cause various environmental issues including contamination by toxic metals, acid mine drainage and salinity. Dissolution of salt from saline mine spoil, in particular, during rainfall events may result in local or regional dispersion of salts through leaching or in the accumulation of dissolved salts in soil pore water and inhibition of plant growth. The salinity in coal mine environments is from the geogenic salt accumulations and weathering of spoils upon surface exposure. The salts are mainly sulfates and chlorides of calcium, magnesium and sodium. The objective of the research is to investigate and assess the source and mobility of salts and trace elements in various spoil types, thereby predicting the leaching behavior of the salts and trace elements from spoils which have similar geochemical properties. X-ray diffraction analysis, total digestion, sequential extraction and column experiments were conducted to achieve the objectives. Sodium and chloride concentrations best represented salinity of the spoils, which might originate from halite. Electrical conductivity, sodium and chloride concentrations in the leachate decreased sharply with increasing leaching cycles. Leaching of trace elements was not significant in the studied area. Geochemical classification of spoil/waste defined for rehabilitation purposes was useful to predict potential salinity, which corresponded with the classification from cluster analysis based on leaching data of major elements. Certain spoil groups showed high potential salinity by releasing high sodium and chloride concentrations. Therefore, the leaching characteristics of sites having saline susceptible spoils require monitoring, and suitable remediation technologies have to be applied.


A window to the world of global insect declines: Moth biodiversity trends are complex and heterogeneous.
Moths are the most taxonomically and ecologically diverse insect taxon for which there exist considerable time-series abundance data. There is an alarming record of decreases in moth abundance and diversity from across Europe, with rates varying markedly among and within regions. Recent reports from Costa Rica reveal steep cross-lineage declines of caterpillars, while other sites (Ecuador and Arizona, reported here) show no or only modest long-term decreases over the past two decades. Rates of decline for dietary and ecological specialists are steeper than those for ecologically generalized taxa. Additional traits commonly associated with elevated risks include large wingspans, small geographic ranges, low dispersal ability, and univoltinism; taxa associated with grasslands, aridlands, and nutrient-poor habitats also appear to be at higher risk. In temperate areas, many moth taxa limited historically by abiotic factors are increasing in abundance and range. We regard the most important continental-scale stressors to include reductions in habitat quality and quantity resulting from land-use change and climate change and, to a lesser extent, atmospheric nitrification and introduced species. Site-specific stressors include pesticide use and light pollution. Our assessment of global macrolepidopteran population trends includes numerous cases of both region-wide and local losses and studies that report no declines. Spatial variation of reported losses suggests that multiple stressors are in play. With the exception of recent reports from Costa Rica, the most severe examples of moth declines are from Northern Hemisphere regions of high human-population density and intensive agriculture.