Characterization of Americium and Curium Complexes with the Protein Lanmodulin: A Potential Macromolecular Mechanism for Actinide Mobility in the Environment.
Anthropogenic radionuclides, including long-lived heavy actinides such as americium and curium, represent the primary long-term challenge for management of nuclear waste. The potential release of these wastes into the environment necessitates understanding their interactions with biogeochemical compounds present in nature. Here, we characterize the interactions between the heavy actinides, Am3+ and Cm3+, and the natural lanthanide-binding protein, lanmodulin (LanM). LanM is produced abundantly by methylotrophic bacteria, including Methylorubrum extorquens, that are widespread in the environment. We determine the first stability constant for an Am3+-protein complex (Am3LanM) and confirm the results with Cm3LanM, indicating a ∼5-fold higher affinity than that for lanthanides with most similar ionic radius, Nd3+ and Sm3+, and making LanM the strongest known heavy actinide-binding protein. The protein's high selectivity over 243Am's daughter nuclide 239Np enables lab-scale actinide-actinide separations as well as provides insight into potential protein-driven mobilization for these actinides in the environment. The luminescence properties of the Cm3+-LanM complex, and NMR studies of Gd3+-LanM, reveal that lanmodulin-bound f-elements possess two coordinated solvent molecules across a range of metal ionic radii. Finally, we show under a wide range of environmentally relevant conditions that lanmodulin effectively outcompetes desferrioxamine B, a hydroxamate siderophore previously proposed to be important in trivalent actinide mobility. These results suggest that natural lanthanide-binding proteins such as lanmodulin may play important roles in speciation and mobility of actinides in the environment; it also suggests that protein-based biotechnologies may provide a new frontier in actinide remediation, detection, and separations.


The diverse biological properties of the chemically inert noble gases.
The noble gases represent an intriguing scientific paradox. They are extremely inert chemically but display a remarkable spectrum of clinically useful biological properties. Despite a relative paucity of knowledge of their mechanisms of action, some of the noble gases have been used successfully in the clinic. Studies with xenon have suggested that the noble gases as a class may exhibit valuable biological properties such as anaesthesia; amelioration of ischemic damage; tissue protection prior to transplantation; analgesic properties; and a potentially wide range of other clinically useful effects. Xenon has been shown to be safe in humans, and has useful pharmacokinetic properties such as rapid onset, fast wash out etc. The main limitations in wider use are that: many of the fundamental biochemical studies are still lacking; the lighter noble gases are likely to manifest their properties only under hyperbaric conditions, impractical in surgery; and administration of xenon using convectional gaseous anaesthesia equipment is inefficient, making its use very expensive. There is nonetheless a significant body of published literature on the biochemical, pharmacological, and clinical properties of noble gases but no comprehensive reviews exist that summarize their properties and the existing knowledge of their models of action at the molecular (atomic) level. This review provides such an up-to-date summary of the extensive, useful biological properties of noble gases as drugs and prospects for wider application of these atoms.


6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in rat brain.
The concentration of fructose 2,6-bisphosphate in the brain remained stable during starvation and early stages of ischaemia, but decreased in diabetes or after lengthened ischaemia. 6-Phosphofructo-1-kinase activity was also decreased in diabetic and ischaemic animals, whereas 6-phosphofructo-2-kinase was not modified. The concentration of the bisphosphorylated metabolite seems to be remarkably constant under a wide variety of experimental conditions, suggesting that it plays an essential role in the basal activation of 6-phosphofructo-1-kinase. Purified 6-phosphofructo-2-kinase also showed fructose-2,6-bisphosphatase activity with an activity ratio similar to that of the purified heart isoenzyme. The brain enzyme also has a net charge similar to that of the heart isoenzyme. Its activity is not modified by sn-glycerol 3-phosphate, and it is more sensitive to citrate than the liver or muscle isoenzyme. Moreover, the enzyme from brain, similarly to that from heart and muscle, is not modified by the cyclic AMP-dependent protein kinase or protein kinase C. A near-full-length cDNA probe from liver hybridized with RNA from brain and heart. In both cases, a major band of 6.8 kb of RNA and a minor one of 4 kb of RNA were detected. All these properties support the hypothesis that brain contains a different isoenzymic form from that of liver and muscle, and it is probably related to the heart isoform.


Biological reactivity of nanoparticles: mosaics from optical microscopy videos of giant lipid vesicles.
Emerging fields such as nanomedicine and nanotoxicology, demand new information on the effects of nanoparticles on biological membranes and lipid vesicles are suitable as an experimental model for bio-nano interaction studies. This paper describes image processing algorithms which stitch video sequences into mosaics and recording the shapes of thousands of lipid vesicles, which were used to assess the effect of CoFe(2)O(4) nanoparticles on the population of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine lipid vesicles. The applicability of this methodology for assessing the potential of engineered nanoparticles to affect morphological properties of lipid membranes is discussed.


Impact of Social Vulnerability on Diabetes-Related Cardiovascular Mortality in the United States.
Background Social vulnerability impacts the natural history of diabetes as well as cardiovascular disease (CVD). However, there are little data regarding the social vulnerability association with diabetes-related CVD mortality. Methods and Results County-level mortality data (where CVD was the underlying cause of death with diabetes among the multiple causes) extracted from the Centers for Disease Control multiple cause of death (2015-2019) and the 2018 Social Vulnerability Index databases were aggregated into quartiles based on their Social Vulnerability Index ranking from the least (first quartile) to the most vulnerable (fourth quartile). Stratified by demographic groups, the data were analyzed for overall CVD, as well as for ischemic heart disease, hypertensive disease, heart failure, and cerebrovascular disease. In the 5-year study period, 387 139 crude diabetes-related cardiovascular mortality records were identified. The age-adjusted mortality rate for CVD was higher in the fourth quartile compared with the first quartile (relative risk [RR], 1.66 [95% CI, 1.64-1.67]) with an estimated 39 328 excess deaths. Among the youngest age group (<55 years), those with the highest social vulnerability had 2 to 4 times the rate of cardiovascular mortality compared with the first quartile: ischemic heart disease (RR, 2.07 [95% CI, 1.97-2.17]; heart failure (RR, 3.03 [95% CI, 2.62-3.52]); hypertensive disease (RR, 3.79 [95% CI, 3.45-4.17]; and cerebrovascular disease (RR, 4.39 [95% CI, 3.75-5.13]). Conclusions Counties with greater social vulnerability had higher diabetes-related CVD mortality, especially among younger adults. Targeted health policies that are designed to reduce these disparities are warranted.


Accumulation of trehalose by Escherichia coli K-12 at high osmotic pressure depends on the presence of amber suppressors.
When grown at high osmotic pressure, some strains of Escherichia coli K-12 synthesized substantial levels of free sugar and accumulated proline if it was present in the growth medium. The sugar was identified as trehalose by chemical reactivity, gas-liquid chromatography, and nuclear magnetic resonance spectroscopy. Strains of E. coli K-12 could be divided into two major classes with respect to osmoregulation. Those of class A showed a large increase in trehalose levels with increasing medium osmolarity and also accumulated proline from the medium, whereas those in class B showed no accumulation of trehalose or proline. Most class A strains carried suppressor mutations which arose during their derivation from the wild type, whereas the osmodefective strains of class B were suppressor free. When amber suppressor mutations at the supD, supE, or supF loci were introduced into such sup0 osmodefective strains, they became osmotolerant and gained the ability to accumulate trehalose in response to elevated medium osmolarity. It appears that the original K-12 strain of E. coli carries an amber mutation in a gene affecting osmoregulation. Mutants lacking ADP-glucose synthetase (glgC) accumulated trehalose normally, whereas mutants lacking UDP-glucose synthetase (galU) did not make trehalose and grew poorly in medium of high osmolarity. Trehalose synthesis was repressed by exogenous glycine betaine but not by proline.


[The effect of the type of agar on the growth of the fungus Blakeslea trispora].
The relationship between the type of agar used to prepare solid nutrient media and the growth of Bl. trispora strains (+)8A and (-)8A was under study. Bl. trispora culture virtually failed to grow in the media prepared with powder agar, GOST 16280-70: it died after 2-month storage with 2 reinoculations. Bl. trispora grew fairly well in culture media with white frozen lamellar agar (GOST 16280-70) or with white frozen microbiologic agar, GOST 17206-71; the characteristic features of the culture were preserved after 10 monthly reinoculations.


Biochemical characterization of autologous fibrinogen adhesive.
Fibrinogen-based adhesive, derived from pooled human plasma, has been used in Europe with great success in otologic surgery, but has not been approved for use in the U.S. because of the risk of transmitting hepatitis. Autologous fibrinogen, derived by polyethylene glycol precipitation from the blood of an individual patient would avoid this risk, and has been shown to be relatively safe to the ear in animal studies. A study of the biochemical composition of this autologous fibrinogen concentrate derived from 15 human volunteer donors was performed. The mean starting plasma fibrinogen was 2.12 mg/ml (range 1.59-3.22 mg/ml). When 10% polyethylene glycol was used to precipitate the fibrinogen, the concentrate contained, on the average, 31.8 mg of fibrinogen/ml. The percent yield averaged 54.9%, and the protein in the final product was 91.9% fibrinogen. Increasing the polyethylene glycol concentration in the precipitation process to as high as 15% resulted in an increased yield as high as 91%, but the protein in the final product was only 42.5% fibrinogen. Polyacrylamide gel electrophoresis confirmed that the predominant protein in the 10% polyethylene glycol precipitate was fibrinogen. These data suggest that highly concentrated fibrinogen can be derived with relative ease from single donor human plasma, and that the product is relatively pure. When combined with thrombin and calcium chloride, this concentrate should provide an adhesive that avoids the risks associated with fibrinogen adhesive derived from pooled blood.


The role of breathing training in asthma management.
There is considerable public interest in the use of breathing modification techniques in the treatment of asthma. Surveys suggest many people with asthma use them, often without the knowledge of their medical attendants. Extravagant claims have been made about the effectiveness of some techniques, resulting in scepticism from orthodox clinicians. The evidence supporting breathing training for asthma was previously weak, and limited by the small size and methodological limitations of published research. The evidence base for the effectiveness of breathing training has recently improved, with reports from several larger and more methodologically robust controlled trials. These trials are reviewed in this study, and the findings placed in context. Trials have investigated a variety of breathing training programmes delivered by different therapists in different ways. All incorporate some instruction in breathing pattern, usually focusing on slow, regular, nasal, abdominal breathing and reduced ventilation, with patients instructed to practise exercises at home and when symptomatic. Current evidence suggests that breathing training programmes can be effective in improving patient-reported outcomes such as symptoms, quality of life and psychological impact; and may reduce the use of rescue bronchodilator medication. There is little evidence that airways physiology, hyper-responsiveness or inflammation is affected by such training. The optimal way of providing breathing training within the context of routine asthma care is still uncertain.


Planar-chiral (2E,7Z)- and (2Z,7E)-cyclonona-2,7-dien-1-yl carbamates by asymmetric, bis-allyic, alpha, alpha'-cycloalkylation--studies on their conformational stability.
Enantiomerically enriched (>80 % ee) (M,1R,2Z,7E)- and (M,1R,2E,7Z)-cyclonona-2,7-dienyl carbamates have been synthesized by an intramolecular cycloalkylation of the corresponding 1-lithio-9-chloronona-2,7-dienyl carbamates. The enantioenriched precursors were generated by asymmetric deprotonation of the dienyl carbamates by means of n-BuLi/(-)-sparteine. The primarily obtained cyclononadienes, each bearing one element of planar and centre chirality, were formed by an , coupling of both allylic moieties. These are the thermodynamically less favoured epimers, which arise from those allyllithiums bearing the carbamoyloxy residue in a 1-endo position. Both (M,R)-cyclononadienes epimerize slowly (t(1/2) = 209 min and 328 min at 308 K) with inversion of the planar chirality to the corresponding, more stable (P)-epimers (ratios 3:97 and 30:70). The kinetics were measured by 1H NMR spectroscopy, the activation energies EA were found to be 26.4 and 27.5 kcal mol(-1). From quantum-chemical calculations with the B3LYP density functional, the epimerization process was shown to consist of two coupled major conformational changes with high energy barriers. The calculated values match well with the observed ones.


The Theory of Ultra-Coarse-Graining. 3. Coarse-Grained Sites with Rapid Local Equilibrium of Internal States.
When viewed through a coarse-grained lens, important molecular and biophysical systems can appear to undergo discrete, switch-like state changes in addition to more continuous configurational motions. One of our recent papers described a theory for bottom-up coarse-graining of the equilibrium statistics of models with such behavior, called ultra-coarse-grained (UCG) models, and a follow up paper described an implementation when the states of the coarse-grained sites or "beads" change rarely. However, not all systems with this discrete behavior fall under that special limit. This article develops the general UCG theory for the opposite limit, that is, where the internal states of the CG particles or beads adjust rapidly so as to always remain effectively at quasi-equilibrium no matter what the positions of the coarse-grained particles. This rapid local equilibrium allows ultra-coarse-graining to mix standard coarse-grained force fields by using local order parameters to control the degree of mixing, which adds an environmental dependence and many-body effects to the coarse-grained model while requiring minimal new coding. This article first presents the definition of such UCG force fields as well as their fitting procedures from atomistic-scale data, and then it presents three examples of UCG simulations with an approach that we call UCG with rapid local equilibrium (UCG-RLE). We then present an application of UCG-RLE using the full bottom-up methodology to coarse-grain and simulate cooperative hydrophobic association of neopentane in methanol solvent. UCG-RLE force matching does a superior job of matching solute-solute correlation functions and solute cluster size distributions compared to the more standard force-matched models not having coarse-grained sites with discrete internal states.


The impact of simultaneous pancreas-kidney transplantation on long-term patient survival.
Simultaneous pancreas-kidney transplantation (SPK) ameliorates the progression of microvascular diabetic complications but the procedure is associated with excess initial morbidity and an uncertain effect on patient survival when compared with solitary cadaveric or living donor renal transplantation. We evaluated mortality risks associated with SPK, solitary renal transplantation, and dialysis treatment in a national cohort of type 1 diabetics with end-stage nephropathy. A total of 13,467 adult-type 1 diabetics enrolled on the renal and renal-pancreas transplant waiting list between 10/01/88 and 06/30/97 were followed until 06/30/98. Time-dependent mortality risks and life expectancy were calculated according to the treatment received subsequent to wait-list registration: SPK; cadaveric kidney only (CAD); living donor kidney only (LKD) transplantation; and dialysis [wait-listed, maintenance dialysis treatment (WLD)]. Adjusted 10-year patient survival was 67% for SPK vs. 65% for LKD recipients (P=0.19) and 46% for CAD recipients (P<0.001). The excess initial mortality normally associated with renal transplantation and the risk of early infectious death was 2-fold higher in SPK recipients. The time to achieve equal proportion of survivors as the WLD patients was 170, 95, and 72 days for SPK, CAD, and LKD recipients, respectively (P<0.001). However, the adjusted 5-year morality risk (RR) using WLD as the reference and the expected remaining life years were 0.40, 0.45, and 0.75 and 23.4, 20.9, and 12.6 years for SPK, LKD, and CAD, respectively. There was no survival benefit in SPK recipients > or =50 years old (RR=1.38, P=0.81). Among patients with type 1 DM with end-stage nephropathy, SPK transplantation before the age of 50 years was associated with long-term improvement in survival compared to solitary cadaveric renal transplantation or dialysis.


The Timing of Raf/ERK and AKT Activation in Protecting PC12 Cells against Oxidative Stress.
Acute brain injuries such as ischemic stroke or traumatic brain injury often cause massive neural death and irreversible brain damage with grave consequences. Previous studies have established that a key participant in the events leading to neural death is the excessive production of reactive oxygen species. Protecting neuronal cells by activating their endogenous defense mechanisms is an attractive treatment strategy for acute brain injuries. In this work, we investigate how the precise timing of the Raf/ERK and the AKT pathway activation affects their protective effects against oxidative stress. For this purpose, we employed optogenetic systems that use light to precisely and reversibly activate either the Raf/ERK or the AKT pathway. We find that preconditioning activation of the Raf/ERK or the AKT pathway immediately before oxidant exposure provides significant protection to cells. Notably, a 15-minute transient activation of the Raf/ERK pathway is able to protect PC12 cells against oxidant strike that is applied 12 hours later, while the transient activation of the AKT pathway fails to protect PC12 cells in such a scenario. On the other hand, if the pathways are activated after the oxidative insult, i.e. postconditioning, the AKT pathway conveys greater protective effect than the Raf/ERK pathway. We find that postconditioning AKT activation has an optimal delay period of 2 hours. When the AKT pathway is activated 30min after the oxidative insult, it exhibits very little protective effect. Therefore, the precise timing of the pathway activation is crucial in determining its protective effect against oxidative injury. The optogenetic platform, with its precise temporal control and its ability to activate specific pathways, is ideal for the mechanistic dissection of intracellular pathways in protection against oxidative stress.


Pandinus imperator scorpion venom blocks voltage-gated potassium channels in nerve fibers.
We have examined the effects of venom from the scorpion Pandinus imperator on the membrane currents of voltage-clamped frog myelinated nerve fibers using the Vaseline-gap method. Crude venom, applied externally in concentrations from 50 to 500 micrograms/ml, selectively blocked the voltage-gated potassium currents without affecting nodal sodium currents or resting conductances. Block of potassium channels by Pandinus venom was highly dependent on the membrane voltage, being greater at negative potentials than at positive potentials. The blocking effects of Pandinus venom were irreversible on the time scale of our experiments; however, even high concentrations of venom failed to block potassium currents completely at positive potentials. These results suggest that Pandinus venom contains a component(s) that interacts specifically and strongly with a subpopulation of axonal potassium channels.


Intravascular hemolysis and ultrastructural changes of erythrocytes in lethal canine endotoxin shock.
Intravascular hemolysis and ultrastructure of erythrocytes from liver sinusoids were studied during and after infusion of Escherichia coli endotoxin in Labrador retriever dogs. Endotoxin infusion caused hemoconcentration, and induced disseminated intravascular coagulation (DIC), characterized by a rapid drop of platelet numbers, a gradual consumption of coagulation factors and activation of fibrinolysis. Advanced DIC and circulatory shock gradually developed, and the animals died after 7-15 h. Plasma hemoglobin concentrations did not rise for several hours, but late in the experimental period a significant intravascular hemolysis was constantly found. Circulating adenosine diphosphate (ADP) did not appear. During shock, liver biopsies revealed accumulation of erythrocytes often disintegrated within distended sinusoidal lumina. In advanced shock the fragmented erythrocyte seemed to form occlusive masses within the vessels. A fibrin-like material frequently appeared adjacent to the red cells. However, it did not have the periodicity characteristic for fibrin, and the ultrastructure of the material was very similar to that inside the erythrocytes. None of these changes were induced by saline infusion in control animals. The lack of fibrin formation and the late development of intravascular hemolysis indicate that red cell breakdown was of little importance for the initiation and progress of DIC.


Encouraging appropriate, evidence-based use of oral nutritional supplements.
With the considerable cost of disease-related malnutrition to individuals and to society (estimated to be >£13×109 for the UK, 2007 prices), there is a need for effective and evidence-based ways of preventing and treating this condition. The wide range of oral nutritional supplements that may be prescribed for the dietary management of malnutrition and other conditions account for only about 1% (about £99×106, 2007 data) of the prescribing budget in England. Systematic reviews and meta-analyses consistently suggest that ready-made, multi-nutrient liquids which may be prescribed can improve energy and nutritional intake, body weight and have a variety of clinical and functional benefits in a number of patient groups. Meta-analyses have repeatedly shown that oral nutritional supplements produce significant reductions in complications (e.g. infections) and mortality, and a recent meta-analysis shows a reduction in hospital admissions (OR 0·56 (95% CI 0·41, 0·77), six randomised controlled trials). Such benefits suggest that the appropriate use of oral nutritional supplements should form an integral part of the management of malnutrition, particularly as there is currently a lack of evidence for alternative oral nutrition strategies (e.g. food fortification and counselling). As with all therapies, compliance to oral nutritional supplements needs to be maximised and the use monitored. To make sure that those at risk of malnutrition are identified and treated appropriately, there is a need to embed national and local policies into routine clinical practice. In doing so, the economic burden of this costly condition can be curtailed. As recently suggested by the National Institute for Health and Clinical Excellence, substantial cost savings could be made if screening and treatment of malnourished patients was undertaken.


Lack of Effectiveness of Antiretroviral Therapy in Preventing HIV Infection in Serodiscordant Couples in Uganda: An Observational Study.
We examined the real-world effectiveness of ART as an HIV prevention tool among HIV serodiscordant couples in a programmatic setting in a low-income country. We enrolled individuals from HIV serodiscordant couples aged ≥18 years of age in Jinja, Uganda from June 2009 - June 2011. In one group of couples the HIV positive partner was receiving ART as they met clinical eligibility criteria (a CD4 cell count ≤250 cells/ μL or WHO Stage III/IV disease). In the second group the infected partner was not yet ART-eligible. We measured HIV incidence by testing the uninfected partner every three months. We conducted genetic linkage studies to determine the source of new infections in seroconverting participants. A total of 586 couples were enrolled of which 249 (42%) of the HIV positive participants were receiving ART at enrollment, and an additional 99 (17%) initiated ART during the study. The median duration of follow-up was 1.5 years. We found 9 new infections among partners of participants who had been receiving ART for at least three months and 8 new infections in partners of participants who had not received ART or received it for less than three months, for incidence rates of 2.09 per 100 person-years (PYRs) and 2.30 per 100 PYRs, respectively. The incidence rate ratio for ART-use was 0.91 (95% confidence interval 0.31-2.70; p=0.999). The hazard ratio for HIV seroconversion associated with ART-use by the positive partner was 1.07 (95% CI 0.41-2.80). A total of 5/7 (71%) of the transmissions on ART and 6/7 (86%) of those not on ART were genetically linked. Overall HIV incidence was low in comparison to previous studies of serodiscordant couples. However, ART-use was not associated with a reduced risk of HIV transmission in this study.


Drinking and DSM-IV alcohol and drug dependence among white and Mexican-American DUI offenders.
To report data on drinking patterns and DSM-IV alcohol and drug dependence among whites and Mexican Americans in treatment for driving under the influence of alcohol (DUI). Interviews were conducted with a sample of 250 whites and 250 Mexican Americans admitted to one of five DUI treatment programs in a Northern California county. Interviews averaged 1 hour in length and were conducted in the programs by trained interviewers. The response rate was 72% for whites and 83% for Mexican Americans. Whites drink most frequently, followed by U.S.-born Mexican Americans and, finally, by Mexican Americans born in Mexico. However, rates of the amount of alcohol usually consumed are higher for Mexican Americans born in Mexico than for the other two groups. Prevalence rates of DSM-IV alcohol dependence are: 29% for whites, 27% for U.S.-born Mexican Americans and 9% for those born in Mexico. Prevalence rates of DSM-IV drug dependence are: 23% for whites, 24% for U.S.-born Mexican Americans and 6% for those born in Mexico. Rates of alcohol and drug dependence in these DUI treatment programs are much higher than rates of dependence in the general population, independent of ethnicity. Patterns of alcohol consumption vary dramatically among whites, U.S.-born Mexican Americans and Mexican Americans born in Mexico. The large amounts of alcohol ingested per occasion by Mexican-born Mexican Americans is particularly noteworthy and may put them at more risk for DUI. However, drug use and drug dependence are higher among whites and U.S.-born Mexican Americans than among Mexican Americans born in Mexico.


Phylogeography of an Island endemic, the Puerto Rican Freshwater Crab (Epilobocera sinuatifrons).
The endemic Puerto Rican crab, Epilobocera sinuatifrons (Pseudothelphusidae), has a freshwater-dependant life-history strategy, although the species has some capabilities for terrestrial movement as adults. In contrast to all other freshwater decapods on the island (e.g., caridean shrimp), E. sinuatifrons does not undertake amphidromous migration, and is restricted to purely freshwater habitats and adjacent riparian zones. As Puerto Rico has a dynamic geologic history, we predicted that both the life history of E. sinuatifrons and the geological history of the island would be important determinants of phylogeographic structuring in the species. Using a fragment of the cytochrome c oxidase subunit 1 mtDNA (mitochondrial DNA) gene, we tested for deviations from panmixia among and within rivers draining Puerto Rico and used statistical phylogeography to explore processes that may explain extant patterns of genetic variation in the species. While populations of E. sinuatifrons were significantly differentiated among rivers, they were likely to be recently derived because nested clade analysis (NCA) indicated evolutionarily recent restricted gene flow with isolation by distance (IBD) and contiguous range expansion at various spatial scales. Ongoing drainage rearrangements associated with faulting and land slippage were invoked as processes involved in sporadic gene flow among rivers throughout the Pleistocene. Patterns of genetic differentiation conformed to IBD and population demographic statistics were nonsignificant, indicating that although recently derived, populations from different rivers were in drift-mutation equilibrium. A shallow (0.6 million years ago), paraphyletic split was observed in the haplotype network, which NCA indicated arose via allopatric fragmentation. This split coincides with an area of high relief in central Puerto Rico that may have experienced relatively little drainage rearrangements. Shallow but significant genetic isolation of populations of E. sinuatifrons among Puerto Rican rivers suggests phylogeographic patterns that are intermediate to terrestrial habitat specialists (highly divergent populations) and other freshwater biota, such as amphidromous species and insects with aerial adult dispersal (highly connected populations).


Changes in nitric oxide, superoxide, and blood circulation in muscles over time after warm ischaemic reperfusion in rabbit rectus femoris muscle.
We present the sequence of changes in nitric oxide (NO) and superoxide (O2-) over time in reperfusion injuries. We examined both the changes in NO and O2- over time and the blood flow in an isolated ischaemia-reperfusion muscle model in rabbits. The ischaemic group comprised 8 animals which had had vascular pedicles clamped on the their rectus femoris muscles for 4 hours. The control group (n = 6) had a sham operation. Blood samples from the femoral vein proximal to the clamping point were collected before the operation, before clamping, before reperfusion, immediately after reperfusion, and 5, 15, 30, 60, and 120 minutes after reperfusion. NO was measured by Griess' method, and O2- by chemiluminescence. Blood flow was measured with a laser Doppler flowmeter. The amount of NO increased significantly immediately after reperfusion, and 15 and 30 minutes after reperfusion in the ischaemic group, compared with the control group (p < 0.05). O2- increased significantly at 5, 15, 60, 90 and 120 minutes after reperfusion, compared with the control group (p < 0.05). The blood flow volume curve increased by 1.4 times about four minutes after reperfusion compared with previously. After this it gradually decreased. The adverse effects of O2- became apparent when NO was extinguished by O2-.


Epidemiology and factors associated with all-terrain vehicle accidents in children: A retrospective cross-sectional study of a trauma registry in Saudi Arabia.
All-terrain vehicle (ATV) accidents have a substantial impact on the pediatric population in Saudi Arabia; however, few local studies are available. An in-depth study of this issue and adequate implementation of regulations are required to prevent additional casualties. The aim of this study was to describe the epidemiology of ATV injuries in the pediatric population and the outcomes associated with the injuries. We conducted a retrospective chart review at a Level 1 trauma center in Riyadh. All patients, aged ≤14 years and admitted due to an ATV accident, from 2004 to 2018 were included in this study. Demographic information, hospital course, and injury outcomes were extracted from the King Abdulaziz Medical City trauma registry. Each medical record was reviewed for short-term complications and the mechanism of injury. The primary outcome measure was the type of injury associated with ATV accidents; the secondary outcomes were injury site and mechanism of injury, and the association between the impact of injury and the clinical and demographic variable. In total, 165 patients were involved in ATV accidents and met our inclusion criteria. The mean age was 8 ± 4 years, and 79% (131/165) were boys. Over 50% (84/165) of the sample had lower limb injuries. The majority of patients had fracture injuries (37%, 61/165), followed by amputations (30%, 50/165). Of the amputation group, the majority (86%, 43/50) was from 1 to 5 years compared to the no amputation group (P < 0.001). For the amputation group, 67% (33/50) had a limb trapped in the chain of the vehicle as to the mechanism of injury. The majority of patients had lower-extremity injuries, specifically fractured (37%) or amputated (30%) with children from age 1-5 years having a significantly higher proportion of hospital admission compared to the rest of the study population. Despite the existing legislation for ATV use in children, they are not enforced. The finding of this study recommends urgent implementation of these regulations for both ATV retailers and users and promotes public awareness about the severity of such injuries.


Overlapping territories between the thalamostriatal and nigrothalamic projections in cats.
Fluorescent tracers were injected into different regions of the caudate nucleus and HRP-WGA in the substantia nigra of the cat in order to analyse the thalamic distribution of retrogradely labelled thalamostriatal neurones and anterogradely labelled nigrothalamic terminals within the thalamus. Overlapping thalamic territories between the thalamostriatal neurones projecting to areas of the caudate nucleus and the nigrothalamic connections were observed in the rostral nuclei of the central thalamic group (ventral anterior nucleus, ventral anterior-ventral lateral complex and ventral medial nucleus) and, more restricted, in the rostral (rhomboid, paracentral, ventral lateral, dorsal mediodorsal nuclei) and caudal intralaminar nuclei (centromedian-parafascicular complex). This study provides evidence of the existence of thalamic areas in which the input and output of the basal ganglia converge.


Age-related hearing loss and the factors determining continued usage of hearing aids among elderly community-dwelling residents.
While hearing aids are recommended for people with age-related hearing loss, many with impaired hearing do not use them. In this study, we investigated how many elderly people in the study area needed hearing aids, and the factors that determined continued wearing of the devices. The study area was Kurabuchi Town, Japan, where 1,437 residents (those aged 65 years or over) were eligible for participation in the study; 1,414 participated, of whom, 103 (7.3%) were already using hearing aids at the start of the study. After the primary screening, hearing aids were lent to 68 participants (4.8%) who did not already have one, 38 of whom (60.3% of the borrowers, representing 2.7% of the total aged population) went on to wear the hearing aid continuously. The Hearing Handicap Inventory for the Elderly (HHIE) score was significantly elevated among these 38 participants. This study indicated that hearing aids are of potential benefit to many local residents. Multivariate logistic regression revealed that HHIE scores were associated with the extent of HA usage. The adjusted odds ratio for a 1-unit increase in HHIE score was 1.08 (95% confidence interval: 1.02-1.14). Programs like this, in which people with impaired hearing are identified at the local level and given appropriate assistance, are useful models for future use in societies with aging populations.


Pervasive brain monitoring and data sharing based on multi-tier distributed computing and linked data technology.
EEG-based Brain-computer interfaces (BCI) are facing basic challenges in real-world applications. The technical difficulties in developing truly wearable BCI systems that are capable of making reliable real-time prediction of users' cognitive states in dynamic real-life situations may seem almost insurmountable at times. Fortunately, recent advances in miniature sensors, wireless communication and distributed computing technologies offered promising ways to bridge these chasms. In this paper, we report an attempt to develop a pervasive on-line EEG-BCI system using state-of-art technologies including multi-tier Fog and Cloud Computing, semantic Linked Data search, and adaptive prediction/classification models. To verify our approach, we implement a pilot system by employing wireless dry-electrode EEG headsets and MEMS motion sensors as the front-end devices, Android mobile phones as the personal user interfaces, compact personal computers as the near-end Fog Servers and the computer clusters hosted by the Taiwan National Center for High-performance Computing (NCHC) as the far-end Cloud Servers. We succeeded in conducting synchronous multi-modal global data streaming in March and then running a multi-player on-line EEG-BCI game in September, 2013. We are currently working with the ARL Translational Neuroscience Branch to use our system in real-life personal stress monitoring and the UCSD Movement Disorder Center to conduct in-home Parkinson's disease patient monitoring experiments. We shall proceed to develop the necessary BCI ontology and introduce automatic semantic annotation and progressive model refinement capability to our system.


Tumor Budding Is Independently Predictive for Lymph Node Involvement in Early Gastric Cancer.
The most important prognostic factor for early gastric cancer (EGC) is the lymph node status. It is important to predict early lesions without lymph node metastasis (LNM) before proceeding to radical surgery in locally excised lesions. Tumor budding is a feature known to be related to aggressive tumor behavior in several solid tumors. We aimed to assess the predictive value of tumor budding for LNM in pT1a and pT1b gastric cancer. We retrospectively investigated radical gastrectomy specimens for of 126 EGC patients and assess the possible relation between the clinicopathologic features, including age, gender, tumor location, tumor size, macroscopic tumor type, histologic differentiation, depth and width of submucosal invasion, lymphovascular invasion, and tumor budding with lymph node involvement. Among the 126 EGCs, 38 were stages as pT1a and 88 as pT1b. LNM rate in pT1a tumors was 13% whereas it was 33% in pT1b tumors. Tumor budding was the only factor significantly and independently related to LNM in pT1a patients. Female gender and tumor budding were found to be independent risk factors in pT1b group. Other clinicopathologic features were not related to LNM. Based on these results, we suggest that budding is a promising parameter to assess for prediction of LNM in EGC removed by endoscopic surgery, and to decide on the appropriate surgical approach.


Acute liver failure associated with prolonged use of bromfenac leading to liver transplantation. The Acute Liver Failure Study Group.
Bromfenac, a nonnarcotic analgesic nonsteroidal anti-inflammatory drug, was associated with reversible, minor elevations in serum aminotransferase levels during clinical trials. The aim of this study is to describe the clinical, laboratory, and histological features of 4 patients with severe bromfenac hepatotoxicity identified at 3 tertiary care centers participating in the US Acute Liver Failure Study Group. Bromfenac was administered for chronic musculoskeletal disorders to 4 women in therapeutic doses of 25 to 100 mg/d for a minimum of 90 days. All patients reported a prodrome of malaise and fatigue and presented with severe, symptomatic hepatocellular injury with associated hypoprothrombinemia. None of the subjects had underlying liver or kidney disease, and there was no evidence of a hypersensitivity reaction. Other identifiable causes of acute liver failure were uniformly excluded. Despite supportive measures, all the subjects developed progressive liver failure over 5 to 37 days, leading to emergency liver transplantation in 3 patients and death in 1 patient while awaiting transplantation. Extensive confluent parenchymal necrosis that appeared to begin in the central zones and was accompanied by a predominantly lymphocytic infiltrate was noted in all the livers examined. Nodular regeneration was seen in the 2 patients with a more protracted clinical course. Administration of therapeutic doses of bromfenac for greater than 90 days was associated with the development of acute liver failure leading to liver transplantation or death in 4 adult women. The poor outcomes observed in this series, coupled with the inability to identify individuals at risk for severe, idiosyncratic bromfenac hepatotoxicity, preclude further use of bromfenac in the medical community.


Epithelial apical glycosylation changes associated with thin endometrium in women with infertility - a pilot observational study.
Low endometrial receptivity is one of the major factors affecting successful implantation in assisted reproductive technologies (ART). Infertile patients with thin endometrium have a significantly lower cumulative clinical pregnancy rate than patients with normal endometrium. Molecular pathophysiology of low receptivity of thin endometrium remains understudied. We have investigated composition of glycocalyx of the apical surface of luminal and glandular epithelial cells in thin endometrium of infertile women. Thirty-two patients with tubal-peritoneal infertility undergoing in vitro fertilization (IVF) were included in the study. Endometrial samples were obtained in a natural menstrual cycle. Patients were divided into two groups: patients with normal endometrium (≥8 mm) and with thin endometrium (< 8 mm). Histochemical and immunohistochemical analysis of paraffin-embedded endometrial samples was performed using six biotinylated lectins (UEA-I, MAL-II, SNA, VVL, ECL, Con A) and anti-LeY and MECA-79 monoclonal antibodies (MAbs). Complex glycans analysis taking into account the adjusted specificity of glycan-binding MAbs revealed 1.3 times less expression of MECA-79 glycans on the apical surface of the luminal epithelial cells of thin endometrium compared to normal endometrium; this deficiency may adversely affect implantation, since MECA-79 glycans are a ligand of L-selectin and mediate intercellular interactions. The glycans containing a type-2 unit Galβ1-4GlcNAcβ (LacNAc) but lacking sulfo-residues at 6-OH of GlcNAcβ, and binding to MECA-79 MAbs were found; they can be considered as potential markers of endometrium receptivity. Expression of the lectins-stained glycans on the apical surfaces of the luminal and glandular epithelial cells did not differ significantly. Correlation between the expression of difucosylated oligosaccharide LeY on the apical surfaces of the luminal and glandular epithelial cells was found in patients with thin endometrium and recurrent implantation failure. A similar relationship was shown for mannose-rich glycans. Specific features of key glycans expression in epithelial compartments of thin endometrium may be essential for morphogenesis of the endometrial functional layer and explain its low receptivity.


Visual Perception of British Women's Skin Color Distribution in Two Nonindustrialized Societies, the Maasai and the Tsimane'.
In women with lightly pigmented skin in particular, facial skin color homogeneity decreases with age, primarily due to chronic exposure to solar ultraviolet radiation (UVR), leading to a decrease in perceived health and attractiveness. Perception of female skin may be influenced by continuous exposure to, and thus familiarity with, age-related changes in visible skin condition in a given society. Men and women of two traditional societies, the Maasai (Tanzania) and the Tsimane' (Bolivia), unfamiliar with lighter colored skin, judged images of British women's facial skin for age, health, and attractiveness. In both samples, images with homogeneous skin color (from the cheeks of younger women) were judged to be younger and healthier and received a stronger preference than corresponding images with heterogeneous skin color (from older women). We suggest that (i) human sensitivity for quality-related information from facial skin color distribution is universal and independent of any known age-dependent variation in skin in a given population and (ii) skin discoloration is universally associated with less positive judgment.


Statin Therapy and the Incidence of Thromboembolism and Vascular Events Following Liver Transplantation.
Statin therapy may reduce the risk of venous thromboembolism (VTE), which may impact solid organ transplant outcomes. We evaluated the incidence of VTE and other complications after liver transplantation stratified by hyperlipidemia status and statin use using a retrospective cohort study approach. We reviewed all primary orthotopic liver transplantation (OLT) records from January 2014 to December 2019 from our center. Intraoperative deaths were excluded. Recipient, donor clinical and demographic data were collected. We developed risk-adjusted models to assess the effect of statin use on the occurrence of VTE, hepatic artery complications (HACs), graft failure, and death, accounting for clinical covariates and competing risks. A total of 672 OLT recipients were included in the analysis. Of this cohort, 11.9% (n = 80) received statin therapy. A total of 47 patients (7.0%) had VTE events. HACs occurred in 40 patients (6.0%). A total of 42 (6.1%) patients experienced graft loss, whereas 9.1% (n = 61) of the cohort died during the study interval. Eighty OLT recipients (29.8%) were treated with statins. In the statin treated group, 0% of patients had VTE versus 7.9% of those not on statins (P = 0.02). HACs were identified in 1.2% of the statin group and 6.8% of the nonstatin group. Untreated hyperlipidemia was associated with a 2.1-fold higher risk of HACs versus patients with no hyperlipidemia status (P = 0.05). Statin therapy was associated with significantly better risk-adjusted thromboembolic event-free survival (absence of VTE, cerebrovascular accident, myocardial infarction, HACs, and death); hazard ratio, 2.7; P = 0.01. These data indicate that statin therapy is correlated with a lower rate of VTE and HACs after liver transplantation.


[Epidemiologic study of infection by hepatitis C virus in a hemodialysis unit].
To analyze the prevalence of infection, the frequency of HCV genotypes and the epidemiology characteristics among the patients in hemodialysis treatment in one 25 years old hospital hemodialysis center and one 15 years old secondary unit by a transversal cross-section study in 1998. 171 hemodialyzed-patients were studied. Patients sera were analyzed by the presence of HCV antibodies anti-VHC by a enzymoimmunoassay (Abbott Cientifica) and the presence of antibodies was confirmed by a line immunoassay (Inno-LIA HCV AbIII) and by the presence of VHC-RNA by reverse transcriptase PCR (Cobas Amplicor HCV). Genotypes were determinate by reverse hybridization (Inno-LIA HCV III). Fifty (29.2%) of the patients were HCV antibody positive. Forty-five (26.3%) were HCV-RNA positive, all of them with antibodies positive. The distribution of genotypes was: 1b, 34 (75.5%); 4f, 4 (8.9%); 1a, 3 (6.7%); 1, 3 (6.7%) and 1 case could not be typed (2.2%). In 14 patients (28.0%), seroconversions were documented Twenty-one patients (42.0%) were diagnosed when the routine tests were available and 15 patients (30.0%) were diagnosed pre-dialysis. The multivariate analysis showed that the risk of HCV infection was greater for patients who had been more 8 years on dialysis (OR: 6.22; 95% CI: 1.24-31.07). Data presented indicate that the prevalence of HCV infections in our hemodialysis units and the number of seroconversions were high and the HCV subtype 1b was more frequent; because of this, the screening by both serological and molecular methods is necessary, at least twice a year, to identify all the infected patients. Besides, we think that is necessary to increase the control of the completion of the Universal Precautions.


Health Visitors' experiences of Family Group Conferences in relation to child protection planning: a phenomenological study.
The purpose of this study was to explore Health Visitors' experiences of Family Group Conferences as part of Child Protection Planning in Hampshire, England. The aim was to identify good practice, recognizing the challenges of the approach and enabling recommendations for improved collaboration to be framed. The Family Group Conferences model is based on partnership, decision-making and family involvement and presents an alternative to case conferences. A Husserlian phenomenological approach was adopted, using taped semi-structured interviews with four health visitors who had experience of Family Group Conferences. Colaizzi's seven stages of phenomenological analysis were used. The four key categories related to the ability of the Family Group Conference model to empower families; the need for health visitors to receive appropriate education and training; organizational; and professional issues. Health visitors believed that Family Group Conferences could empower families, but they felt unprepared to attend. Concerns were identified regarding confidentiality and responsibility. Although the health visitors supported the principles underpinning Family Group Conferences, they were unsure about how to put theory into practice. The need for more education and training was strongly supported to enable the model to move from marginal to mainstream use. They also considered that Family Group Conferences could threaten interagency working, associated with issues relating to professional responsibility. The results identified training and procedural issues that need to be addressed if Family Group Conferences are to be introduced successfully within mainstream child protection practice. Insights from this study have led to inclusion of Family Group Conferences in the local child protection guidelines, with emphasis applied to interdisciplinary working, empowerment of families and professional staff, and education and training.


Immobilization-dependent bone collagen breakdown appears to increase with time: evidence for a lack of new bone equilibrium in response to reduced load during prolonged bed rest.
The purpose of this study was to evaluate the effect of prolonged immobilization on bone, in order to investigate how skeletal turnover adapts to bed rest. We examined indices of bone formation and bone resorption in the serum and urine of fifty-four patients (26 males and 28 females) immobilized after an episode of paralytic stroke. The length of immobilization ranged from 30 to 180 days. A significant, time-dependent increase in markers of resorption - urinary pyridinoline (Pyr) and deoxypyridinoline (D-Pyr), serum Type I collagen cross-linked C-telopeptide (ICTP) - was observed in immobilized patients, as compared to free-living healthy subjects. The positive correlation between resorption markers increase and the length of immobilization suggests that the rate of bone resorption did not decrease with time. On the other hand, the levels of markers of bone formation - bone-specific alkaline phosphatase (B-ALP), and the carboxyl-terminal propeptide of Type I procollagen (PICP) - remained within the normal range in all patients, regardless the length of immobilization. Our results would indicate an uncoupling between bone formation and bone resorption during bed rest, and suggest that the bone collagen break-down was not a self-limiting process in immobilized patients, and that a new equilibrium or "steady state" in response to the reduced load was not reached in the skeleton.


Approach to an experimental model of Mal de Debarquement Syndrome.
Mal de Debarquement Syndrome (MdDS) is the rare condition of enduring rocking sensations and subjective unsteadiness following a lengthy exposure to passive motion. The pathogenesis of MdDS is unknown and the available treatment is limited. Here, we developed an experimental model of MdDS that may facilitate systematic inquiry of MdDS pathophysiology and the development of prevention or treatment strategies for this condition. In an initial series of pilot experiments, suitable stimulation devices and conditions were evaluated. The final paradigm consisted of a low-frequency oscillatory motion stimulation, simultaneously deployed as roll and pitch rotation as well as heave on a six-degrees-of-freedom motion platform. Twelve healthy participants were stimulated under this condition for 30 min during free stance. Aftereffects with respect to rocking sensations and posturographic sway were monitored up to 60 min post-stimulation and compared to an initial pre-stimulation assessment as well as to posturographic recordings of spontaneous sway in ten patients with MdDS. Motion stimulation consistently evoked MdDS-like rocking sensations and postural alterations that lasted up to 45 min after cessation of passive motion exposure. Body sway alterations were most pronounced in anterior-posterior dimension during standing with eyes closed and primarily characterized by a distinct peak in the low-frequency sway spectrum close to stimulation frequency. These postural aftereffects further closely resembled spontaneous oscillatory low-frequency sway observed in patients with MdDS. Subsequent neurophysiological and imaging examinations are required to investigate whether the model of transient, experimental MdDS actually shares a common substrate with the enduring pathological condition of MdDS.


HRS/SRp40-mediated inclusion of the fibronectin EIIIB exon, a possible cause of increased EIIIB expression in proliferating liver.
Serine-arginine (SR)-rich proteins are believed to be important in mediating alternative pre-mRNA splicing. HRS/SRp40 expression is elevated in liver cell proliferation during development, regeneration, and oncogenesis. We tested whether HRS expression correlates with the appearance of alternatively spliced fibronectin transcripts during liver growth. HRS was highly expressed during the proliferative phase of liver development, correlating with expression of the fibronectin EIIIB alternative exon. In regenerating liver, HRS protein was induced in a time course consistent with the observed increase in fibronectin transcripts containing the EIIIB exon, particularly in nonparenchymal liver cells. Furthermore, in an in vivo assay, HRS, and not other SR proteins, directly mediated EIIIB exon inclusion in the fibronectin transcript. This alternative splicing was dependent on a purine-rich region within the EIIIB exon to which HRS specifically bound. We have established that HRS has the potential to contribute to the regulation of fibronectin pre-mRNA splicing during liver growth. Changes in fibronectin forms may be important in modifying liver architecture during the proliferative response, thus providing a potential mechanism by which SR proteins may participate in cellular growth control.


Food production and the energy crisis.
The principal raw material of modern U.S. agriculture is fossil fuel, whereas the labor input is relatively small (about 9 hours per crop acre). As agriculture is dependent upon fossil energy, crop production costs will also soar when fuel costs increase two- to fivefold. A return of 2.8 kcal of corn per 1 kcal of fuel input may then be uneconomical. Green revolution agriculture also uses high energy crop production technology, especially with respect to fertilizers and pesticides. While one may not doubt the sincerity of the U.S. effort to share its agricultural technology so that the rest of the world can live and eat as it does, one must be realistic about the resources available to accomplish this mission. In the United States we are currently using an equivalent of 80 gallons of gasoline to produce an acre of corn. With fuel shortages and high prices to come, we wonder if many developing nations will be able to afford the technology of U.S. agriculture. Problems have already occurred with green revolution crops, particularly problems related to pests (57). More critical problems are expected when there is a world energy crisis. A careful assessment should be made of the benefits, costs, and risks of high energy-demand green revolution agriculture in order to be certain that this program will not aggravate the already serious world food situation (58). To reduce energy inputs, green revolution and U.S. agriculture might employ such alternatives as rotations and green manures to reduce the high energy demand of chemical fertilizers and pesticides. U.S. agriculture might also reduce energy expenditures by substituting some manpower currently displaced by mechanization. While no one knows for certain what changes will have to be made, we can be sure that when conventional energy resources become scarce and expensive, the impact on agriculture as an industry and a way of life will be significant. This analysis is but a preliminary investigation of a significant agricultural problem that deserves careful attention and greater study before the energy situation becomes more critical.


South African serial rapists: the offenders, their victims, and their offenses.
Serial rapists have the propensity to cause harm to a significant number of victims, meaning that they are of concern to the police as well as to treatment providers. Despite the serious nature of their offending, there are surprisingly few studies that provide information regarding their characteristics, the types of victim they target, or the nature of the sexual offenses they commit, and those studies that do exist are varied in their findings. This study provides a descriptive analysis of serial rape in South Africa. One hundred and nineteen sexual offenses committed by 22 serial rapists were sampled. Information regarding the victims, the offenders, and the crimes they had committed were extracted from police files. The characteristics of victims and offenders are reported as well as the frequencies for 114 different crime scene behaviors. When compared with samples of serial sex offenders from other countries, differences emerged in victim characteristics and crime scene behaviors, including how the victims were targeted, the sexual behaviors engaged in, and the incidence of physical violence. The implications of these observed differences for practice are discussed.


Dissociable effects of lesions to orbitofrontal cortex subregions on impulsive choice in the rat.
The orbitofrontal cortex (OFC) is implicated in a variety of adaptive decision-making processes. Human studies suggest that there is a functional dissociation between medial and lateral OFC (mOFC and lOFC, respectively) subregions when performing certain choice procedures. However, little work has examined the functional consequences of manipulations of OFC subregions on decision making in rodents. In the present experiments, impulsive choice was assessed by evaluating intolerance to delayed, but economically optimal, reward options using a delay-discounting paradigm. Following initial delay-discounting training, rats received bilateral neurotoxic or sham lesions targeting whole OFC (wOFC) or restricted to either mOFC or lOFC subregions. A transient flattening of delay-discounting curves was observed in wOFC-lesioned animals relative to shams--differences that disappeared with further training. Stable, dissociable effects were found when lesions were restricted to OFC subregions; mOFC-lesioned rats showed increased, whereas lOFC-lesioned rats showed decreased, preference for the larger-delayed reward relative to sham-controls--a pattern that remained significant during retraining after all delays were removed. When locations of levers leading to small-immediate versus large-delayed rewards were reversed, wOFC- and lOFC-lesioned rats showed retarded, whereas mOFC-lesioned rats showed accelerated, trajectories for reversal of lever preference. These results provide the first direct evidence for dissociable functional roles of the mOFC and lOFC for impulsive choice in rodents. The findings are consistent with recent human functional imaging studies and suggest that functions of mOFC and lOFC subregions may be evolutionarily conserved and contribute differentially to decision-making processes.


Optical guiding by photoreceptor cells.
The potential for any morphological part of a photoreceptor cell to function as an optical waveguide is determined 1) by the dimensionless parameter V as shown by Snyder (IEEE Trans., Microwave Theory Tech. 17: 1133, 1969) and by Snyder, Pask and Mitchell (J. Opt. Soc. Am. 63: 59, 1973), and 2) by the nature of the optical excitation for that part of the cell. The effects of these two factors are considered for the myoids and outer segments of vertebrate photoreceptor cells and for the crystalline tracts and rhabdomers of invertebrate photoreceptor cells. Examples of both guiding and nonguiding myoids, outer segments, rhabdomers and tracts are cited and some consequences for vision are analyzed.


Structure of the siphophage neck-Tail complex suggests that conserved tail tip proteins facilitate receptor binding and tail assembly.
Siphophages have a long, flexible, and noncontractile tail that connects to the capsid through a neck. The phage tail is essential for host cell recognition and virus-host cell interactions; moreover, it serves as a channel for genome delivery during infection. However, the in situ high-resolution structure of the neck-tail complex of siphophages remains unknown. Here, we present the structure of the siphophage lambda "wild type," the most widely used, laboratory-adapted fiberless mutant. The neck-tail complex comprises a channel formed by stacked 12-fold and hexameric rings and a 3-fold symmetrical tip. The interactions among DNA and a total of 246 tail protein molecules forming the tail and neck have been characterized. Structural comparisons of the tail tips, the most diversified region across the lambda and other long-tailed phages or tail-like machines, suggest that their tail tip contains conserved domains, which facilitate tail assembly, receptor binding, cell adsorption, and DNA retaining/releasing. These domains are distributed in different tail tip proteins in different phages or tail-like machines. The side tail fibers are not required for the phage particle to orient itself vertically to the surface of the host cell during attachment.


Amyotrophic Lateral Sclerosis and soccer: an internet survey of 29 Italian players.
Previous epidemiological studies reported a significantly higher risk of Amyotrophic Lateral Sclerosis (ALS) in Italian male soccer players. As a consequence, sports newspapers and news agencies focused on this issue and spread the news of 51 male soccer players with a reported diagnosis of ALS. We searched news on male Italian national soccer players with a reported diagnosis of ALS quoted from January 1, 1950 to July 31, 2016 in at least two Internet web sites or in books by journalists. A total of 39 male soccer players with a reported diagnosis of ALS were identified. Subjects were born from 1905 to 1973, 32 were currently deceased, 6 were still living, while the status of 1 player was unknown. All gathered information was available for 29 soccer players. The group had a mean age at diagnosis of 45.3 ± 12.2 years, a mean age at onset of symptoms of 46.4 ± 12.1 years, and a mean age at death of 50.9 ± 12.3 years. A significant inverse correlation between year of birth and age at onset of symptoms was observed, with a younger age at onset of symptoms in soccer players born in more recent years (r = -0.65, p < 0.01). Italian male soccer players with a reported diagnosis of ALS have a significantly younger age at diagnosis when compared to other European patients with ALS. Results support a possible relationship between soccer and the risk of ALS. We believe that further research is urgently needed in this field.


Induction of nerve growth factor expression and release by mechanical and inflammatory stimuli in chondrocytes: possible involvement in osteoarthritis pain.
Nerve growth factor (NGF) level is increased in osteoarthritis (OA) joints and is involved in pain associated with OA. Stimuli responsible for NGF stimulation in chondrocytes are unknown. We investigated whether mechanical stress and proinflammatory cytokines may influence NGF synthesis by chondrocytes. Primary cultures of human OA chondrocytes, newborn mouse articular chondrocytes or cartilage explants were stimulated by increasing amounts of IL-1β, prostaglandin E₂ (PGE₂), visfatin/nicotinamide phosphoribosyltransferase (NAMPT) or by cyclic mechanical compression (0.5 Hz, 1 MPa). Before stimulation, chondrocytes were pretreated with indomethacin, Apo866, a specific inhibitor of NAMPT enzymatic activity, or transfected by siRNA targeting visfatin/NAMPT. mRNA NGF levels were assessed by real-time quantitative PCR and NGF released into media was determined by ELISA. Unstimulated human and mouse articular chondrocytes expressed low levels of NGF (19.2 ± 8.7 pg/mL, 13.5 ± 1.0 pg/mL and 4.4 ± 0.8 pg/mL/mg tissue for human and mouse articular chondrocytes and costal explants, respectively). Mechanical stress induced NGF release in conditioned media. When stimulated by IL-1β or visfatin/NAMPT, a proinflammatory adipokine produced by chondocytes in response to IL-1β, a dose-dependent increase in NGF mRNA expression and NGF release in both human and mouse chondrocyte conditioned media was observed. Visfatin/NAMPT is also an intracellular enzyme acting as the rate-limiting enzyme of the generation of NAD. The expression of NGF induced by visfatin/NAMPT was inhibited by Apo866, whereas IL-1β-mediated NGF expression was not modified by siRNA targeting visfatin/NAMPT. Interestingly, PGE₂, which is produced by chondrocytes in response to IL-1β and visfatin/NAMPT, did not stimulate NGF production. Consistently, indomethacin, a cyclooxygenase inhibitor, did not counteract IL-1β-induced NGF production. These results show that mechanical stress, IL-1β and extracellular visfatin/NAMPT, all stimulated the expression and release of NGF by chondrocytes and thus suggest that the overexpression of visfatin/NAMPT and IL-1β in the OA joint and the increased mechanical loading of cartilage may mediate OA pain via the stimulation of NGF expression and release by chondrocytes.


Mortality in People with Type 2 Diabetes Following SARS-CoV-2 Infection: A Population Level Analysis of Potential Risk Factors.
Research is ongoing to increase our understanding of how much a previous diagnosis of type 2 diabetes mellitus (T2DM) affects someone's risk of becoming seriously unwell following a COVID-19 infection. In this study we set out to determine the relative likelihood of death following COVID-19 infection in people with T2DM when compared to those without T2DM. This was conducted as an urban population study and based in the UK. Analysis of electronic health record data was performed relating to people living in the Greater Manchester conurbation (population 2.82 million) who had a recorded diagnosis of T2DM and subsequent COVID-19 confirmed infection. Each individual with T2DM (n = 13,807) was matched with three COVID-19-infected non-diabetes controls (n = 39,583). Data were extracted from the Greater Manchester Care Record (GMCR) database for the period 1 January 2020 to 30 June 2021. Social disadvantage was assessed through Townsend scores. Death rates were compared in people with T2DM to their respective non-diabetes controls; potential predictive factors influencing the relative likelihood of admission were ascertained using univariable and multivariable logistic regression. For individuals with T2DM, their mortality rate after a COVID-19 positive test was 7.7% vs 6.0% in matched controls; the relative risk (RR) of death was 1.28. From univariate analysis performed within the group of individuals with T2DM, the likelihood of death following a COVID-19 recorded infection was lower in people taking metformin, a sodium-glucose cotransporter 2 inhibitor (SGLT2i) or a glucagon-like peptide 1 (GLP-1) agonist. Estimated glomerular filtration rate (eGFR) and hypertension were associated with increased mortality and had odds ratios of 0.96 (95% confidence interval 0.96-0.97) and 1.92 (95% confidence interval 1.68-2.20), respectively. Likelihood of death following a COVID-19 infection was also higher in those people with a diagnosis of chronic obstructive pulmonary disease (COPD) or severe enduring mental illness but not with asthma, and in people taking aspirin/clopidogrel/insulin. Smoking in people with T2DM significantly increased mortality rate (odds ratio of 1.46; 95% confidence interval 1.29-1.65). In a combined analysis of patients with T2DM and controls, multiple regression modelling indicated that the factors independently relating to a higher likelihood of death (accounting for 26% of variance) were T2DM, age, male gender and social deprivation (higher Townsend score). Following confirmed infection with COVID-19 a number of factors are associated with mortality in individuals with T2DM. Prescription of metformin, SGLT2is or GLP-1 agonists and non-smoking status appeared to be associated with a reduced the risk of death for people with T2DM. Age, male sex and social disadvantage are associated with an increased risk of death.


Modeling the spatiotemporal spread of beneficial alleles using ancient genomes.
Ancient genome sequencing technologies now provide the opportunity to study natural selection in unprecedented detail. Rather than making inferences from indirect footprints left by selection in present-day genomes, we can directly observe whether a given allele was present or absent in a particular region of the world at almost any period of human history within the last 10,000 years. Methods for studying selection using ancient genomes often rely on partitioning individuals into discrete time periods or regions of the world. However, a complete understanding of natural selection requires more nuanced statistical methods which can explicitly model allele frequency changes in a continuum across space and time. Here we introduce a method for inferring the spread of a beneficial allele across a landscape using two-dimensional partial differential equations. Unlike previous approaches, our framework can handle time-stamped ancient samples, as well as genotype likelihoods and pseudohaploid sequences from low-coverage genomes. We apply the method to a panel of published ancient West Eurasian genomes to produce dynamic maps showcasing the inferred spread of candidate beneficial alleles over time and space. We also provide estimates for the strength of selection and diffusion rate for each of these alleles. Finally, we highlight possible avenues of improvement for accurately tracing the spread of beneficial alleles in more complex scenarios.


Anterior retropharyngeal plate screw fixation with bilateral anterior transarticular screws for odontoid fractures ... a new comprehensive technique.
A certain group of odontoid fractures (Anderson and D' Alonzo Type-2) are usually offered surgical treatment. Common surgical option is an anterior odontoid screw. Some of the fractures are not suitable for anterior odontoid screw (anterior oblique, displaced distal fragments and those with atlantoaxial instability) and these are usually offered posterior transarticular screws (Magerl's) or posterior atlantoaxial screw rod/plate fixation (Goel-Harms technique). Posterior surgery involves atlantoaxial fixation with an indirect attempt to reduce and fuse the fracture . Posterior surgery has a risk of injury to the vertebral arteries, hemorrhage from the paravertebral venous plexus and the C2 root ganglion. A direct anterior submandibular retropharyangeal approach with open reduction and fixation (ORIF) using a customized variable screw placement (VSP) plate was used to realign and fix the fracture fragments in compression mode under direct vision. Twenty patients of type-II odontoid fractures (unsuitable for anterior odontoid screw) underwent an anterior retropharyngeal approach with anterior variable screw position (VSP) plate and screw fixation and eight amongst them, who had associated atlantoaxial instability underwent additional bilateral anterior transarticular screws. All patients treated by this technique had 100% fracture site bone union without any implant failure. Longest follow-up has been for 3 years. Anterior retropharyangeal approach allows direct fracture fragment realignment under vision with an opportunity to fix in compression mode using the VSP plate, which ensures early fusion across the type-II odontoid fracture. Any associated instability can be treated by additional bilateral anterior transarticular screws. The approach is simple and safe without any risk to the vertebral arteries and biomechanically appealing.


Blocking Fcα receptor I on granulocytes prevents tissue damage induced by IgA autoantibodies.
IgA represents the most prominent Ab class at mucosal surfaces and the second most prevalent Ab in human blood after IgG. We recently demonstrated that cross-linking of the granulocyte IgA FcR (FcαRI) by IgA induces a chemotactic-driven positive-feedback migration loop, hereby amplifying recruitment of granulocytes to IgA deposits. Therefore, we postulated that aberrant IgA-Ag complexes, which can be found in tissues in IgA-mediated diseases, are responsible for tissue damage by inducing continuous granulocyte migration and activation. Using an IgA-dependent skin-blistering disease as a model system, we demonstrated colocalization of FcαRI-positive granulocyte infiltrates with IgA in cryosections of lesional skin of patients suffering from this disease. Furthermore, we showed granulocyte migration to IgA deposits injected in human skin explants and in murine skin of FcαRI transgenic mice in vivo. Importantly, ex vivo migration and tissue damage were inhibited by blocking FcαRI, indicating that these events are dependent on the interaction of IgA autoantibodies with FcαRI. Thus, interrupting the granulocyte migration loop by blocking FcαRI reduces tissue damage in diseases with aberrant IgA-immune complexes. As such, our results may lead to development of new therapies for IgA-mediated chronic inflammatory diseases, hereby decreasing severe morbidity and improving quality of life for these patients.


The lasso segment is required for functional dimerization of the Plasmodium formin 1 FH2 domain.
Apicomplexan parasites, such as the malaria-causing Plasmodium species, utilize a unique way of locomotion and host cell invasion. This substrate-dependent gliding motility requires rapid cycling of actin between the monomeric state and very short, unbranched filaments. Despite the crucial role of actin polymerization for the survival of the malaria parasite, the majority of Plasmodium cellular actin is present in the monomeric form. Plasmodium lacks most of the canonical actin nucleators, and formins are essentially the only candidates for this function in all Apicomplexa. The malaria parasite has two formins, containing conserved formin homology (FH) 2 and rudimentary FH1 domains. Here, we show that Plasmodium falciparum formin 1 associates with and nucleates both mammalian and Plasmodium actin filaments. Although Plasmodium profilin alone sequesters actin monomers, thus inhibiting polymerization, its monomer-sequestering activity does not compete with the nucleating activity of formin 1 at an equimolar profilin-actin ratio. We have determined solution structures of P. falciparum formin 1 FH2 domain both in the presence and absence of the lasso segment and the FH1 domain, and show that the lasso is required for the assembly of functional dimers.


Estimation of genetic parameters for milk yield in Murrah buffaloes by Bayesian inference.
Random regression models were used to estimate genetic parameters for test-day milk yield in Murrah buffaloes using Bayesian inference. Data comprised 17,935 test-day milk records from 1,433 buffaloes. Twelve models were tested using different combinations of third-, fourth-, fifth-, sixth-, and seventh-order orthogonal polynomials of weeks of lactation for additive genetic and permanent environmental effects. All models included the fixed effects of contemporary group, number of daily milkings and age of cow at calving as covariate (linear and quadratic effect). In addition, residual variances were considered to be heterogeneous with 6 classes of variance. Models were selected based on the residual mean square error, weighted average of residual variance estimates, and estimates of variance components, heritabilities, correlations, eigenvalues, and eigenfunctions. Results indicated that changes in the order of fit for additive genetic and permanent environmental random effects influenced the estimation of genetic parameters. Heritability estimates ranged from 0.19 to 0.31. Genetic correlation estimates were close to unity between adjacent test-day records, but decreased gradually as the interval between test-days increased. Results from mean squared error and weighted averages of residual variance estimates suggested that a model considering sixth- and seventh-order Legendre polynomials for additive and permanent environmental effects, respectively, and 6 classes for residual variances, provided the best fit. Nevertheless, this model presented the largest degree of complexity. A more parsimonious model, with fourth- and sixth-order polynomials, respectively, for these same effects, yielded very similar genetic parameter estimates. Therefore, this last model is recommended for routine applications.


Juvenile hormone (JH) esterase: why are you so JH specific?
Juvenile hormone esterases (JHEs) from six insects belonging to three orders (Lepidoptera, Coleoptera, and Diptera) were compared in terms of their deduced amino acid sequence and biochemical properties. The four lepidopteran JHEs showed from 52% to 59% identity to each other and about 30% identity to the coleopteran and dipteran JHEs. The JHE of Manduca sexta was remarkably resistant to the addition of organic co-solvents and detergent; in some cases, it demonstrated significant activation of activity. Trifluoromethylketone (TFK) inhibitors with chain lengths of 8, 10 or 12 carbons were highly effective against both lepidopteran and coleopteran JHEs. The coleopteran JHE remained sensitive to TFK inhibitors with a chain length of 6 carbons, whereas the lepidopteran JHEs were significantly less sensitive. When the chain was altered to a phenethyl moiety, the coleopteran JHE remained moderately sensitive, while the lepidopteran JHEs were much less sensitive. The lepidopteran and coleopteran JHEs did not show dramatic differences in specificity to alpha-naphthyl and rho-nitrophenyl substrates. However, as the chain length of the alpha-naphthyl substrates increased from propionate to caprylate, there was a trend towards reduced activity. The JHE of M. sexta was crystallized and the properties of the crystal suggest a high-resolution structure will follow.


Tendon Collagen Crosslinking Offers Potential to Improve Suture Pullout in Rotator Cuff Repair: An Ex Vivo Sheep Study.
The suture-tendon interface is often the weakest link in tendon to bone repair of massive rotator cuff tears. Genipin is a low-toxicity collagen crosslinker derived from the gardenia fruit that has been shown to augment collagen tissue strength and mechanically arrest tendon-tear progression. The purpose of the current study was to evaluate whether genipin crosslinking can sufficiently augment the suture-tendon interface to improve suture pullout strength using simple single-loop sutures and the modified Mason-Allen technique. The study also aimed to assess whether time of genipin treatment is a relevant factor in efficacy. In an ex vivo (cadaveric) sheep rotator cuff tendon model, a total of 142 suture pullout tests were performed on 32 infraspinatus tendons. Each tendon was prepared with three single-loop stitches. Two groups were pretreated by incubation in genipin solution for either 4 hours or 24 hours. Two corresponding control groups were incubated in phosphate buffered saline for the same periods. The same test protocol was applied to tendons using modified Mason-Allen technique stitch patterns. Each suture was loaded to failure on a universal materials testing machine. Suture pullout force, stiffness, and work to failure were calculated from force-displacement data, and then compared among the groups. Median single-loop pullout force on tendons incubated for 24 hours in genipin yielded an approximately 30% increase in maximum pullout force for single-loop stitches with a median of 73 N (range, 56-114 N) compared with 56 N (range, 40-69 N; difference of medians = 17 N; p = 0.028), with corresponding increases in the required work to failure but not stiffness. Genipin treatment for 4 hours showed no added benefit for suture-pullout behavior (46 N, [range, 35-95 N] versus 45 N, [range, 28-63 N]; difference of medians, 1 N; p = 1). No tested genipin crosslinking conditions indicated benefit for tendons grasped using the modified Mason-Allen technique after 4 hours (162 N, [range, 143-193 N] versus 140 N, [range, 129-151 N]; difference of medians, 22 N; p = 0.114) or after 24 hours of crosslinking (172 N, [range, 42-183 N] versus 164 N [range, 151-180 N]; difference of medians, 8 N; p = 0.886). Exogenous collagen crosslinking in genipin can markedly improve resistance to pullout at the tendon-suture interface for simple stitch patterns while the modified Mason-Allen stitch showed no benefit in an ex vivo animal model. Tendon strength augmentation by genipin pretreatment offers the potential to improve suture retention properties. Future studies are warranted for the development of clinically viable intraoperative delivery strategies and in vivo testing for safety and efficacy.


Benzodiazepine Use and Cognitive Decline in Elderly With Normal Cognition.
Benzodiazepine (BZD) use may be associated with dementia. However, differing opinions exist regarding the effect of BZDs on long-term changes in cognition. We evaluated the association between BZD use and cognitive decline in the elderly with normal cognition from the National Alzheimer's Disease Coordinating Center's Uniform Data Set. The study exposure, BZD use, was classified 2 ways: any-use [reported BZD use at a minimum of 1 Alzheimer's disease center (ADC) visit] and always-use (reported BZD use at all ADC visits). The reference group included participants without any declared BZD use at any Alzheimer's Disease Center (ADC) visit. The main outcome measures were Clinical Dementia Rating Sum of Boxes score and Mini-Mental State Examination score. We observed a decline in cognitive status over time in the 2 comparison groups. All participants who reported taking BZDs had poorer cognitive performance at all visits than nonusers. However, cognitive decline was statistically similar among all participants. We found no evidence of an association between BZD use and cognitive decline. The poor cognitive performance in BZD users may be due to prodromal symptoms caused by preclinical dementia processes.


Identification of a novel conserved HLA-A*0201-restricted epitope from the spike protein of SARS-CoV.
The spike (S) protein is a major structural glycoprotein of coronavirus (CoV), the causal agent of severe acute respiratory syndrome (SARS). The S protein is a potent target for SARS-specific cell-mediated immune responses. However, the mechanism CoV pathogenesis in SARS and the role of special CTLs in virus clearance are still largely uncharacterized. Here, we describe a study that leads to the identification of a novel HLA-A*0201-restricted epitope from conserved regions of S protein. First, different SARS-CoV sequences were analyzed to predict eight candidate peptides from conserved regions of the S protein based upon HLA-A*0201 binding and proteosomal cleavage. Four of eight candidate peptides were tested by HLA-A*0201 binding assays. Among the four candidate peptides, Sp8 (S958-966, VLNDILSRL) induced specific CTLs both ex vivo in PBLs of healthy HLA-A2+ donors and in HLA-A2.1/Kb transgenic mice immunized with a plasmid encoding full-length S protein. The immunized mice released IFN-gamma and lysed target cells upon stimulation with Sp8 peptide-pulsed autologous dendritic cells in comparison to other candidates. These results suggest that Sp8 is a naturally processed epitope. We propose that Sp8 epitope should help in the characterization of mechanisms of virus control and immunopathology in SARS-CoV infection.


The role of intracellular pathways in the proliferation of human K562 cells mediated by muscarinic receptors.
Muscarinic acetylcholine receptors (mAChRs) are members of the superfamily of G protein coupled receptors (GPCRs). Muscarinic receptors are relatively abundant in the central nervous system and in the peripheral parasympathetic nervous system. Several studies have suggested that muscarinic receptors also mediate some cellular events in hematopoietic cells. K562 erythroleukemia cells contain muscarinic receptors M2, M3 and M4, and activation of muscarinic receptors changes cell proliferation. We examined the effects of several compounds on cell proliferation in K562 erythroleukemia cells. These included a muscarinic receptor agonist carbachol (CCh), a protein kinase inhibitor staurosporine; the phospholipase C inhibitor U73122, the MEK 1-2 inhibitor UO126, the PI3-kinase inhibitor wortmannin, the Ca(2+) chelators BAPTA/AM and 2-aminoethoxy-diphenylborate (2APB). In addition, we also investigated muscarinic receptor mediated protein kinase C (PKC) expression in K562 cells. CCh caused a decrease in DNA synthesis in K562 cells supplemented with 1% fetal bovine serum after starvation. Pre-treatment of K562 cells with U73122 and BAPTA/AM antagonized the inhibitory effect of CCh, suggesting that phospholipase C and intracellular calcium are involved in CCh-mediated inhibition of proliferation in K562 cells. Our data also suggest that the regulatory roles of protein kinase C and the MAPK/ERK pathways in K562 cell proliferation are independent of cholinergic activation.


Transcription factor EB overexpression prevents neurodegeneration in experimental synucleinopathies.
The synucleinopathies Parkinson's disease (PD) and Multiple system atrophy (MSA) - characterized by α-synuclein intracytoplasmic inclusions into, respectively, neurons and oligodendrocytes - are associated with impairment of the autophagy-lysosomal pathways (ALP). Increased expression of the master regulator of ALP, transcription factor EB (TFEB), is hypothesized to promote the clearance of WT α-synuclein and survival of dopaminergic neurons. Here, we explore the efficacy of targeted TFEB overexpression either in neurons or oligodendrocytes to reduce the pathological burden of α-synuclein in a PD rat model and a MSA mouse model. While TFEB neuronal expression was sufficient to prevent neurodegeneration in the PD model, we show that only TFEB oligodendroglial overexpression leads to neuroprotective effects in the MSA model. These beneficial effects were associated with a decreased accumulation of α-synuclein into oligodendrocytes through recovery of the ALP machinery. Our study demonstrates that the cell type where α-synuclein aggregates dictates the target of TFEB overexpression in order to be protective, paving the way for adapted therapies.


Pharmacokinetics and Tissue Distribution of Isovitexin-2''-O-β-D-glucopyranoside (IVG) in Sprague-Dawley Rats.
Isovitexin-2"-O-D-glucopyranoside (IVG) has been known to exhibit sedative and hypnotic effects. However, there is little understanding of the in vivo pharmacokinetics and tissue distribution of IVG. This study aimed to investigate the pharmacokinetics and tissue distribution of IVG. The study employed an HPLC-ESI-MS/MS method to analyze the pharmacokinetics and tissue distribution of IVG. Under mass spectrometry, IVG and internal standard (IS) showed strong negative ionization signals. MRM analysis chose ion transitions m/z 593.3 → 293.0 (IVG) and m/z 579.8 → 271.4 (IS). Method validation indicated high precision, accuracy, and reliability with a quantitation limit under 20 ng/mL. After intravenously administering 5.0 mg/kg of IVG, rapid clearance from rat plasma was observed, with a half-life (t1/2) of 3.49 ± 0.99 h and a clearance rate of 54.53 ± 11.90 mL/kg/h. The area under the curve (AUC0-12h) of 37.79 ± 7.65 μg·h/mL indicated a brisk metabolic rate. Evaluating the tissue distribution, the highest accumulation was seen in the liver (30.32 ± 3.06 μg/g), followed by the kidney (20.58 ± 2.12 μg/g) and intestine (6.69 ± 0.93 μg/g), suggesting a propensity for IVG to concentrate in these tissues. Importantly, the presence of IVG in the brain underlines its potential to traverse the blood-brain barrier. These findings revealed that following intravenous administration, IVG was swiftly and broadly distributed throughout various rat tissues. This study provides valuable information on the pharmacokinetics and tissue distribution of IVG, implicating its potential as a novel and effective drug candidate for sedative and anxiolytic treatment.


Enantioselective enzymes by computational design and in silico screening.
Computational enzyme design holds great promise for providing new biocatalysts for synthetic chemistry. A strategy to design small mutant libraries of complementary enantioselective epoxide hydrolase variants for the production of highly enantioenriched (S,S)-diols and (R,R)-diols is developed. Key features of this strategy (CASCO, catalytic selectivity by computational design) are the design of mutations that favor binding of the substrate in a predefined orientation, the introduction of steric hindrance to prevent unwanted substrate binding modes, and ranking of designs by high-throughput molecular dynamics simulations. Using this strategy we obtained highly stereoselective mutants of limonene epoxide hydrolase after experimental screening of only 37 variants. The results indicate that computational methods can replace a substantial amount of laboratory work when developing enantioselective enzymes.


Practice patterns in classic bladder exstrophy: A global perspective.
While evaluation and management options for classic bladder exstrophy (CBE) patients are numerous and varied, little is known regarding the relative utilization of these different methods throughout the world. A large group of exstrophy surgeons practicing globally was surveyed, seeking to document their methods of care. A list of international exstrophy surgeons' email addresses was compiled using professional contacts and referral networking. An online survey was sent to each email address. Surgeons who had not performed a CBE closure within the previous 5 years were excluded. Survey questions queried the respondents' surgical practice type, years since training, and their preferred methods of preoperative evaluation, operative management, and postoperative management. Survey invitations were sent out starting in December 2014 and responses were collected for approximately 6 months. A total of 1152 valid email addresses were invited, resulting in 293 respondents (25%) from 39 countries and every American Urological Association (AUA) section. Seventy-six were excluded, leaving 217 respondents (Table). Respondents reported a median of 17 years since finishing their surgical training (IQR 8-25 years). Practice types included pediatric urology (n = 209), general urology (n = 9), pediatric surgery (n = 59), and other practice makeup (n = 3). On subgroup analyses, there were no significant regional practice differences, with the exception of complete primary repair of exstrophy (CPRE) and oral opioid prescribing being significantly higher in North America compared to other regions. Findings indicate that there may be diversity in CBE practice patterns globally. While most responding surgeons from regions outside of North America indicated modern staged repair of exstrophy (MSRE) as their preferred closure technique, a relatively equal distribution of respondents from North America selected CPRE and MSRE. A majority of North American surgeons chose performing osteotomies for both newborn and delayed closures, while an appreciable number of respondents from other regions selected never using osteotomies in their closures. Limitations to this study include a low survey response rate, particularly from surgeons outside of the United States, which may have significantly impacted the ability to draw meaningful global comparisons. Global variation among practices of surgeons performing CBE closures may exist. The wide range of methods demonstrated by this survey suggests the need for more conclusive comparative studies to elucidate whether an optimal standard exists. Local social factors, access to surgical expertise and transportation to referral centers, and finances play a role in what constitutes the best operative approach.


Reduction in targeted potentially inappropriate medication use in elderly inpatients: a pragmatic randomized controlled trial.
The use of potentially inappropriate medications (PIMs) in hospitalized older adults is a complex problem, but the use of computerized alert systems (CAS) has shown some potential. The study's objective is to assess the change in PIM use with a CAS-based pharmacist-physician intervention model compared to usual clinical care. Pragmatic single-site randomized controlled trial was conducted at a university teaching hospital. Hospitalizations identified with selected Beers or STOPP criteria were randomized to usual clinical care or to the CAS-based pharmacist-physician intervention. The primary outcome was PIM drug cessation or dosage decrease. Clinical relevance of the CAS alerts was assessed. Analyses included 231 patients who had 128 and 126 hospitalizations in the control and intervention groups, respectively. Patients had a mean age of 81, and 60% were female. In the intervention compared to the control group, drug cessation or dosage decrease were more frequent at 48 h post-alert (45.8 vs 15.9%; absolute difference 30.0%; 95%CI 13.8 to 46.1%) and at discharge from the hospital (48.1 vs 27.3%; absolute difference 20.8%; 95%CI 4.6 to 37.0%). In a post hoc analysis of all alerts, regardless of their clinical relevance, the absolute difference in drug cessation or dosage decrease between the intervention and control groups was 16.2% (95%CI 2.9 to 29.6%) at 48 h and 8.0% (95%CI -4.0 to 20.0%) at discharge from the hospital. In hospitalized older adults, a CAS-based pharmacist-physician intervention, compared to usual clinical care, resulted in significant higher number of drug cessation and dosage reductions for targeted PIMs.


A theoretical study of the effect of zero-crossing points within the dispersion map upon a longhaul RZ-DPSK system.
A theoretical study focusing on the effect of zero-crossing points in the dispersion map for the long-haul return-to-zero differential phase shift keying (RZ-DPSK) transmission system had been conducted. Difference of the transmission performance was characterized with regard to number of zero-crossing points within the dispersion map. It was observed that increasing the number of zero-crossing points in the dispersion map combined with the self phase modulation (SPM) caused the performance degradation near the system zero dispersion wavelength.


[Structural characteristics of paramagnetic analogs of enzyme-substrate complexes of phosphorylation coupling factors].
Mn2+-ion is linked to isolated chloroplast coupling factor CF-1 via the ATP bridge in the catalytically competent ternary complex as deduced from water proton relaxation rate measurements. Two essential SH-groups in CF-1 protein were modified with nitroxyl mercuric derivative as spin label. The substrate complex Ca2+-ATP is shown to induce the structural transition near the active site to the state with a stronger immobilized spin label. The distances between the paramagnetic metal ions and nitroxyl bound to the protein SH-group were evaluated as being in the range of 5-8,5 A for Cu2+ and 14-22 A for Mn2+.


ClC-7 expression levels critically regulate bone turnover, but not gastric acid secretion.
Mutations in the 2Cl(-)/1H(+)-exchanger ClC-7 impair osteoclast function and cause different types of osteoclast-rich osteopetrosis. However, it is unknown to what extent ClC-7 function has to be reduced to become rate-limiting for bone resorption. In osteoclasts from osteopetrosis patients expression of the mutated ClC-7 protein did not correlate with disease severity and resorption impairment. Therefore, a series of transgenic mice expressing ClC-7 in osteoclasts at different levels was generated. Crossing of these mice with Clcn7(-/-) mutants rescued the osteopetrotic phenotype to variable degrees. One resulting double transgenic line mimicked human autosomal dominant osteopetrosis. The trabecular bone of these mice showed a reduction of osteoblast numbers, osteoid, and osteoblast marker gene expression indicative of reduced osteoblast function. In osteoclasts from these mutants ClC-7 expression levels were 20 to 30% of wildtype levels. These reduced levels not only impaired resorptive activity, but also increased numbers, size and nucleus numbers of osteoclasts differentiated in vitro. Although ClC-7 was expressed in the stomach and PTH levels were high in Clcn7(-/-) mutants loss of ClC-7 did not entail a relevant elevation of gastric pH. In conclusion, we show that in our model a reduction of ClC-7 function by approximately 70% is sufficient to increase bone mass, but does not necessarily enhance bone formation. ClC-7 does not appear to be crucially involved in gastric acid secretion, which explains the absence of an osteopetrorickets phenotype in CLCN7-related osteopetrosis.