Adjuvant therapy of the very young woman.
Women under 35 or 40 with primary breast cancer have a poor prognosis independent of other factors [Albain K, Allred C, Clark G. Breast cancer outcome and predictors of outcome: are there age differentials? J Natl Cancer Inst Monogr 1994;35-42]. In some recent studies, however, age is not independent in multivariate analyses, which include gene signatures [Van De Vijver M, He YD, Van'T Veer L, et al. A gene-expression signature as a predictor of survival in breast cancer. N Engl J Med 2002;347:1999-2009.(132)]. Dissection of such molecular signatures may identify mechanisms, which can be targeted. Today, positive estrogen receptors identify women who require endocrine therapy, and HER2/neu positivity those who require herceptin and also benefit most from anthracyclines. Locoregional recurrences are also more common in younger women. Radiation boost therapy can reduce in-breast recurrence [Bartelink H, Horiot JC, Poortmans PM, Struikmans H, et al. Impact of radiation dose on local control, fibrosis and survival after breast conserving treatment: 10 year results of the EORTC trial 22881-10882. Br Cancer Res Treat 2006;100:S8-10]. There are also particular quality of life issues in young women, for whom fertility concerns and symptoms of premature menopause loom large. Some young women with lower risk may be candidates for endocrine therapy alone but it may be difficult to identify these with current prognostic and predictive factors. In the future more sophisticated molecular factors may identify those who require hormones alone, chemotherapy alone, newer biologic therapies, or combinations of these approaches.


Predicting NMR relaxation of proteins from molecular dynamics simulations with accurate methyl rotation barriers.
The internal dynamics of proteins occurring on time scales from picoseconds to nanoseconds can be sensitively probed by nuclear magnetic resonance (NMR) spin relaxation experiments, as well as by molecular dynamics (MD) simulations. This complementarity offers unique opportunities, provided that the two methods are compared at a suitable level. Recently, several groups have used MD simulations to compute the spectral density of backbone and side chain molecular motions and to predict NMR relaxation rates from these. Unfortunately, in the case of methyl groups in protein side chains, inaccurate energy barriers to methyl rotation were responsible for a systematic discrepancy in the computed relaxation rates, as demonstrated for the AMBER ff99SB*-ILDN force field (and related parameter sets), impairing quantitative agreement between simulations and experiments. However, correspondence could be regained by emending the MD force field with accurate coupled cluster quantum chemical calculations. Spurred by this positive result, we tested whether this approach could be generally applicable, in spite of the fact that different MD force fields employ different water models. Improved methyl group rotation barriers for the CHARMM36 and AMBER ff15ipq protein force fields were derived, such that the NMR relaxation data obtained from the MD simulations even now display very good agreement with the experiment. Results herein showcase the performance of present-day MD force fields and manifest their refined ability to accurately describe internal protein dynamics.


Risk of serious upper gastrointestinal and cardiovascular thromboembolic complications with meloxicam.
To assess the risk of serious gastrointestinal and thromboembolic complications with approved doses of meloxicam. We pooled data from clinical trials of meloxicam at doses of 7.5 or 15 mg/d. A blinded gastrointestinal adjudication committee used prespecified criteria to identify gastric or duodenal perforation, gastric outlet obstruction, or hemodynamically important upper gastrointestinal bleeding. For analysis of thromboembolic complications, investigator-reported events were analyzed without adjudication. We analyzed data from 24,196 patients from 28 trials, most of whom had been followed for up to 60 days. Of these patients, 13,118 received meloxicam (10,158 received a daily dose of 7.5 mg and 2960 received 15 mg), 5283 were treated with diclofenac 100 mg, 181 received diclofenac 150 mg, 5371 were treated with piroxicam 20 mg, and 243 received naproxen 500 mg twice daily. Patients who received 7.5 mg of meloxicam daily had a 0.03% risk of serious upper gastrointestinal events, which was significantly lower than the risk in those who received diclofenac, naproxen, or piroxicam (P <0.02). With the 15 mg daily dose of meloxicam, this risk was significantly different only when compared with piroxicam (P = 0.03). The risk of thromboembolic events in patients treated with meloxicam at either dose was lower than with diclofenac, but similar to that observed with piroxicam and naproxen. This pooled analysis of 24,196 patients demonstrates that meloxicam has a favorable gastrointestinal and thromboembolic safety profile. However, only a small number of patients were followed for more than 60 days, and meaningful comparisons were not possible in this subgroup.


Self-assembly of β-alanine homotetramer: formation of nanovesicles for drug delivery.
The present paper describes the fabrication of nanovesicles using the stirring induced self-assembly of a β-alanine homotetramer (H2N-βAla-βAla-βAla-βAla-CONH2) in an aqueous medium. Dynamic light scattering (DLS) studies revealed the formation of nanostructures with an average particle size of ∼463 nm. The morphology and size of the nanostructures was also confirmed by electron microscopy. The formation of well-defined spherical nanovesicles in the size range of ∼100 nm to 250 nm is displayed. Secondary structure estimations using circular dichroism (CD) and infrared spectroscopic (IR) studies suggest an intramolecularly hydrogen bonded conformation for the tetrapeptide. CD and IR studies conducted in an aqueous medium before and after self-assembly depict a significant change in structure and hydrogen bonding patterns. Supportive evidence regarding the conformation of the tetrapeptide is provided by the conformational search using computational methods. The theoretically predicted minimum energy structure depicts an 8-helix type of backbone folding with the formation of two 8-membered intramolecularly hydrogen bonded rings. Four of the short helical type strands of the tetrapeptide were then subjected to a geometry optimization that resulted in the formation of an assembly with a central cavity, completely in agreement with experimental findings. Finally these nanovesicles were evaluated for their potential use as carriers of the potent anti-Parkinson's drug l-DOPA. We have observed that in aqueous media these nanovesicles are able to encapsulate the l-DOPA molecules, indicated by DLS measurements and also by absorption and emission spectroscopic methods. By lowering the pH to 6.2 these loaded vesicles are able to release the drug slowly. These assemblies thus can be considered as potential delivery vehicles for l-DOPA which normally suffers from poor administration.


Plant Polyphenols as Chemopreventive Agents for Lung Cancer.
Lung cancer may be prevented by a diet rich in fruits and vegetables as they are enriched with dietary antioxidant polyphenols, such as flavonoids, proanthocyanidins, lignans, stilbenes, and phenolic acids. Dietary polyphenols exert a wide range of beneficial biological functions beyond their antioxidative properties and are involved in regulation of cell survival pathways leading to anticarcinogenic and antimutagenic functions. There are sufficient evidence from in vitro, in vivo, and epidemiological studies to suggest that the dietary intervention of polyphenols in cancer prevention, including the chemopreventive ability of dietary polyphenols, act against lung carcinogens. Cohort and epidemiological studies in selected risk populations have evaluated clinical effects of polyphenols. Polyphenols have demonstrated three major actions: antioxidative activity, regulation of phase I and II enzymes, and regulation of cell survival pathways against lung carcinogenesis. They have also shown an inverse association of lung cancer occurrences among high risk populations who consumed considerable amounts of fruits and vegetables in their daily diet. In in vitro cell culture experimental models, polyphenols bind with electrophilic metabolites from carcinogens, inactivate cellular oxygen radicals, prevent membrane lipid peroxidation and DNA oxidative damage, and adduct formation. Further, polyphenols enhance the detoxifying enzymes such as the phase II enzymes, glutathione transferases and glucuronosyl transferases.


Osteonectin/SPARC secreted by RPE and localized to the outer plexiform layer of the monkey retina.
Osteonectin/SPARC is a secreted protein that has been implicated in ocular disease. Deletion of osteonectin/SPARC causes age-onset cataract in mice and the cataractous human lens has increased expression of osteonectin/SPARC. In this study, the expression and localization of osteonectin/SPARC in the monkey retina were determined as was secretion by cultured human retinal pigment epithelial (RPE) cells. Adult Rhesus monkey eyes (Macaca mulatta) were dissected, and 5-mm macula and peripheral retina punches were obtained. Supernatants were collected from cultured human RPE cells. Subcellular fractionation of whole monkey retina was also performed. Osteonectin/SPARC expression and/or secretion was monitored by Northern and Western blot analyses, and localization was determined by immunocytochemistry. Outside of the retina osteonectin/SPARC mRNA is broadly expressed in many human tissues. Northern blot analysis shows that in the retina osteonectin/SPARC is expressed almost exclusively by the macular RPE/choroid. Western blot analysis revealed osteonectin/SPARC in both the macula and the peripheral neural retina but only in trace amounts in the RPE/choroid. In subcellular fractions of the whole retina, osteonectin/SPARC was detected, mainly in the soluble fraction but also in the membrane and nuclear fractions. Immunohistochemical analysis localized osteonectin/SPARC specifically to the outer plexiform layer. Western blot analysis of conditioned medium from human RPE cells cultured on porous substrates indicated that osteonectin/SPARC is secreted in large amounts from both the apical and basal sides of the RPE. Collectively these data provide evidence that osteonectin/SPARC is synthesized in the macular RPE, secreted, and subsequently transported to the outer plexiform layer. The expression pattern of osteonectin/SPARC in the subcellular retinal fractions is consistent with a soluble protein that is transported and internalized.


Bone mineral density and fracture rate in response to intravenous and oral bisphosphonates in adult osteogenesis imperfecta.
The effect of bisphosphonate treatment on bone mineral density (BMD) and fracture rates was assessed in adults with osteogenesis imperfecta (OI). This observational nonrandomized study included 90 OI adults treated with intravenous pamidronate (n = 28), oral alendronate (n = 10), or oral residronate (n = 17) or not treated (n = 35). There were 63 type I, 15 type III, and 12 type IV OI patients. BMD results were observed for up to 161 months and an average of 52 months of treatment. For type I and grouped type III/IV patients, treatment with pamidronate showed an increasing rate in L1-L4 BMD from baseline (0.006 [P = 0.03] and 0.016 [P < 0.001] gm/cm(2)/year, respectively); oral bisphosphonate treatment showed a significant increasing rate in L1-L4 BMD (0.004 gm/cm(2)/year [P = 0.047]) for type I patients. Pamidronate-treated type III/IV and oral bisphosphonate-treated type I patients showed significant increases in total-hip BMD (0.006 [P = 0.003] and 0.011 [P = 0.046] gm/cm(2)/year, respectively). Bisphosphonate effect on fracture rate was assessed for 5-year periods before and after treatment in 51 treated and 22 nontreated individuals matched for age at which bisphosphonate was first administered to the treated group. Bisphosphonate treatment did not decrease fracture rate in type I OI patients. Fracture rate decreased in type III/IV patients following pamidronate but not following oral bisphosphonate treatment. These results underscore a need to consider whether bisphosphonate treatment is appropriate for all adults with OI.


Coagulation profile in healthy Nigerian neonates.
A study of some coagulation factors were carried out in preterm and term infants on the first day of life. Screening coagulation tests--prothrombin time (PT), partial thromboplastin time with kaolin (PTTK) and the concentrations of Factors VIII: C and fibrinogen were determined in 100 normal newborn infants classified into three groups according to their gestational ages: 28 to 30 weeks, 31 to 36 weeks, 37 to 42 weeks. The respective values were compared with those of six-month-old infants as well as adults. The mean values of the screening coagulation tests, PT and PTTK, Factor VIII: C and fibrinogen were significantly different in all the three gestational age groups of nonates when compared with those of six-month-old infants and adult Nigerians. This suggests that a relative hypocagulable state exists among newborn infants and could be responsible for increased bleeding tendencies in these groups of infants. This is the first documented report of coagulation profile in Nigerian neonates, and the range obtained in this study can thus be regarded as standard for healthy Nigerian neonates. The body of data should therefore provide a basis for evaluating newborn noenates with bleeding problems.


[A marathon runner with rhabdomyolysis].
Serious complications after running a marathon are relatively rare. We saw a patient at the emergency department (A&E) who developed acute renal failure as a result of rhabdomyolysis after excessive exertion. A 31-year-old, healthy male was referred to the A&E by the GP service two days after running a marathon because of a CK level of 131,900 U/l. At the A&E, he was experiencing severe muscle pain, was continuously vomiting and experiencing reduced diuresis. Lab tests revealed a serum creatinine level of 705 µmol/l, indicating acute renal failure. In order to improve renal perfusion, the patient was treated with aggressive fluid administration and was monitored. After a hospital stay of 17 days, the patient was discharged in good condition. Acute renal failure caused by rhabdomyolysis after exertion is caused by a combination of adverse environmental factors, risk factors and genetic predisposition. We recommend referring runners who have a body temperature of > 40°C and are experiencing changed awareness after a marathon to the hospital for assessment.


Perceptual anchoring in preschool children: not adultlike, but there.
Recent studies suggest that human auditory perception follows a prolonged developmental trajectory, sometimes continuing well into adolescence. Whereas both sensory and cognitive accounts have been proposed, the development of the ability to base current perceptual decisions on prior information, an ability that strongly benefits adult perception, has not been directly explored. Here we ask whether the auditory frequency discrimination of preschool children also improves when given the opportunity to use previously presented standard stimuli as perceptual anchors, and whether the magnitude of this anchoring effect undergoes developmental changes. Frequency discrimination was tested using two adaptive same/different protocols. In one protocol (with-reference), a repeated 1-kHz standard tone was presented repeatedly across trials. In the other (no-reference), no such repetitions occurred. Verbal memory and early reading skills were also evaluated to determine if the pattern of correlations between frequency discrimination, memory and literacy is similar to that previously reported in older children and adults. Preschool children were significantly more sensitive in the with-reference than in the no-reference condition, but the magnitude of this anchoring effect was smaller than that observed in adults. The pattern of correlations among discrimination thresholds, memory and literacy replicated previous reports in older children. The processes allowing the use of context to form perceptual anchors are already functional among preschool children, albeit to a lesser extent than in adults. Nevertheless, immature anchoring cannot fully account for the poorer frequency discrimination abilities of young children. That anchoring is present among the majority of typically developing preschool children suggests that the anchoring deficits observed among individuals with dyslexia represent a true deficit rather than a developmental delay.


The cotton WRKY transcription factor GhWRKY17 functions in drought and salt stress in transgenic Nicotiana benthamiana through ABA signaling and the modulation of reactive oxygen species production.
Drought and high salinity are two major environmental factors that significantly limit the productivity of agricultural crops worldwide. WRKY transcription factors play essential roles in the adaptation of plants to abiotic stresses. However, WRKY genes involved in drought and salt tolerance in cotton (Gossypium hirsutum) are largely unknown. Here, a group IId WRKY gene, GhWRKY17, was isolated and characterized. GhWRKY17 was found to be induced after exposure to drought, salt, H2O2 and ABA. The constitutive expression of GhWRKY17 in Nicotiana benthamiana remarkably reduced plant tolerance to drought and salt stress, as determined through physiological analyses of the germination rate, root growth, survival rate, leaf water loss and Chl content. GhWRKY17 transgenic plants were observed to be more sensitive to ABA-mediated seed germination and root growth. However, overexpressing GhWRKY17 in N. benthamiana impaired ABA-induced stomatal closure. Furthermore, we found that GhWRKY17 modulated the increased sensitivity of plants to drought by reducing the level of ABA, and transcript levels of ABA-inducible genes, including AREB, DREB, NCED, ERD and LEA, were clearly repressed under drought and salt stress conditions. Consistent with the accumulation of reactive oxygen species (ROS), reduced proline contents and enzyme activities, elevated electrolyte leakage and malondialdehyde, and lower expression of ROS-scavenging genes, including APX, CAT and SOD, the GhWRKY17 transgenic plants exhibited reduced tolerance to oxidative stress compared with wild-type plants. These results therefore indicate that GhWRKY17 responds to drought and salt stress through ABA signaling and the regulation of cellular ROS production in plants.


Cell Patterning Technology on Polymethyl Methacrylate through Controlled Physicochemical and Biochemical Functionalization.
In recent years, innovative cell-based biosensing systems have been developed, showing impact in healthcare and life science research. Now, there is a need to design mass-production processes to enable their commercialization and reach society. However, current protocols for their fabrication employ materials that are not optimal for industrial production, and their preparation requires several chemical coating steps, resulting in cumbersome protocols. We have developed a simplified two-step method for generating controlled cell patterns on PMMA, a durable and transparent material frequently employed in the mass manufacturing of microfluidic devices. It involves air plasma and microcontact printing. This approach allows the formation of well-defined cell arrays on PMMA without the need for blocking agents to define the patterns. Patterns of various adherent cell types in dozens of individual cell cultures, allowing the regulation of cell-material and cell-cell interactions, were developed. These cell patterns were integrated into a microfluidic device, and their viability for more than 20 h under controlled flow conditions was demonstrated. This work demonstrated the potential to adapt polymeric cytophobic materials to simple fabrication protocols of cell-based microsystems, leveraging the possibilities for commercialization.


Patent Foramen Ovale and Ascending Aortic Dilatation Causing Platypnea-Orthodeoxia Syndrome.
Platypnea-orthodeoxia syndrome (POS) is an underdiagnosed clinical syndrome characterized by dyspnea (platypnea) and hypoxemia (orthodeoxia) in the upright position that resolves when recumbent. POS is often due to an underlying right-to-left shunt. Four broad mechanisms for the shunt have been described: intracardiac shunts, intrapulmonary shunts, hepatopulmonary syndrome, and pulmonary ventilation-perfusion mismatch. A 68-year-old male with a past medical history of chronic obstructive pulmonary disease (COPD), obstructive sleep apnea, ascending aortic dilation (3.9 cm), myelofibrosis, and status post stem cell transplant complicated by graft versus host disease was found hypoxemic (oxygen saturation: 82%) on routine visit prompting hospitalization. Hypoxemia initially responded to 40% FiO2 but subsequently progressed to refractory hypoxemia on 100% FiO2. A chest computed tomography (CT) scan showed evidence of multiple segmental pulmonary emboli with patent central pulmonary arteries. Hypoxemia out of proportion to pulmonary embolism clot burden and examination findings consistent with orthodeoxia prompted further investigations. Nuclear medicine scan showed radiotracer activity in both brain and kidneys consistent with a small right-to-left shunt (5.9%). Transesophageal echocardiography (TEE) revealed a patent foramen ovale (PFO) with a right-to-left shunt across the atrial septum, with a maximum opening of 3.5 mm and tunnel length of 25 mm. Right heart catheterization (RHC) is consistent with the right-to-left shunt and normal right heart pressures. The degree of the shunt was not significant enough to explain the degree of hypoxemia, but all the diagnostic studies were performed in a supine position, possibly underestimating the degree of the shunt. PFO closure with transcatheter 30-mm Gore device (GORE® CARDIOFORM, Arizona, USA) decreased supplemental oxygen requirement from 75% high-flow nasal cannula (NC) to room air (RA) immediately after the procedure. The patient was subsequently discharged home on a baseline oxygen requirement of 2 L NC at nighttime. POS should be suspected when a patient develops severe hypoxemia after changing from a recumbent position to a sitting or standing position. The identification and correction of the shunting or mismatch often allow complete resolution of POS. Transthoracic echocardiography with agitated saline, TEE, and RHC are the diagnosis modalities of choice. Left heart cardiac catheterization remains the gold standard, which would demonstrate a mismatch in oxygen saturation between the pulmonary vein and the aorta. Our patient's PFO was successfully closed by a percutaneous transcatheter closure device leading to the complete resolution of hypoxemia immediately.


Effect of phenolic glycosides on Agrobacterium tumefaciens virH gene induction and plant transformation.
O-Aryl-d-glucoside (4-7) and d-xyloside (8-10) derivatives were synthesized and tested on Agrobacterium virH gene induction and plant transformation. alpha- or beta-Glycosides enhanced vir activity at concentrations above 250 micromicro. The highest vir activity was observed with beta-glucoside derivative 4 at 10 mM. A marked difference between phenol glucoside derivative 4 and the corresponding free phenol on the growth of transformants was observed. The regenerated transgenic tissues, after transformation on medium containing acetosyringyl beta-glucoside 4, grew at twice the rate of those on medium containing only free acetosyringone (AS). Compound 4 was less toxic for tobacco explants compared to the corresponding free phenol. However, the xyloside derivatives tested (8-10) were less effective for gene induction compared with corresponding free phenols.


Influence of dietary cis- and trans-fat on 1,2-dimethylhydrazine-induced colon tumors and fecal steroid excretion in Fischer 344 rats.
To investigate the effects of the difference in the geometry of dietary fatty acids on colon tumorigenesis, male rats were fed semipurified diets containing either partially hydrogenated corn oil (trans-monoene fat) or olive oil (cis-monoene fat) at the 10% level and received a single oral dosage of 1,2-dimethylhydrazine (DMH). The difference in the fatty acid composition of dietary fats was confined essentially to the geometrical isomerism of octadecenoate, and the linoleic acid content was made equivalent (2% of total energy). After about 15 mo of feeding, colon tumor incidence in DMH treated rats was nearly the same in both fat groups. Fecal neutral steroid excretion was higher, while the transformation of cholesterol to coprostanol was lower in rats given the trans-fat. There were no marked differences in the excretion and composition of fecal bile acids between two fat groups. Serum cholesterol and tocopherol levels of rats given trans-fat diets tended to be low. The results suggested that the trans-monoene behaves much like the cis-monoene in the incidence of DMH-induced colon tumors, although there were characteristic differences in metabolic events in the intestine.


Characterization of structural features controlling the receptiveness of empty class II MHC molecules.
MHC class II molecules (MHC II) play a pivotal role in the cell-surface presentation of antigens for surveillance by T cells. Antigen loading takes place inside the cell in endosomal compartments and loss of the peptide ligand rapidly leads to the formation of a non-receptive state of the MHC molecule. Non-receptiveness hinders the efficient loading of new antigens onto the empty MHC II. However, the mechanisms driving the formation of the peptide inaccessible state are not well understood. Here, a combined approach of experimental site-directed mutagenesis and computational modeling is used to reveal structural features underlying "non-receptiveness." Molecular dynamics simulations of the human MHC II HLA-DR1 suggest a straightening of the α-helix of the β1 domain during the transition from the open to the non-receptive state. The movement is mostly confined to a hinge region conserved in all known MHC molecules. This shift causes a narrowing of the two helices flanking the binding site and results in a closure, which is further stabilized by the formation of a critical hydrogen bond between residues αQ9 and βN82. Mutagenesis experiments confirmed that replacement of either one of the two residues by alanine renders the protein highly susceptible. Notably, loading enhancement was also observed when the mutated MHC II molecules were expressed on the surface of fibroblast cells. Altogether, structural features underlying the non-receptive state of empty HLA-DR1 identified by theoretical means and experiments revealed highly conserved residues critically involved in the receptiveness of MHC II. The atomic details of rearrangements of the peptide-binding groove upon peptide loss provide insight into structure and dynamics of empty MHC II molecules and may foster rational approaches to interfere with non-receptiveness. Manipulation of peptide loading efficiency for improved peptide vaccination strategies could be one of the applications profiting from the structural knowledge provided by this study.


The Role of HMGB1 in Radioresistance of Bladder Cancer.
Although radical cystectomy surgery is the standard-of-care for muscle-invasive bladder cancer, it entails complete removal of the bladder and surrounding organs which leads to substantial loss in the quality-of-life of patients. Radiotherapy, which spares the bladder, would be a more appropriate treatment modality if we can utilize molecular markers to select patients with better response to radiation. In this study, we investigate a protein called high mobility group box protein 1 (HMGB1) as a predictive marker for radiotherapy response in bladder cancer. Our in vitro results indicate a positive correlation between higher levels of HMGB1 protein and resistance to radiation in various cell lines. Upon HMGB1 protein knockdown, highly significant (>1.5-fold) sensitization to radiotherapy was achieved. We saw that loss of HMGB1 was associated with at least two times higher (P < 0.001) DNA damage in cell lines postradiation. Our results also depicted that autophagy was inhibited more than 3-fold (P < 0.001) upon HMGB1 knockdown, implicating its role in autophagy as another cause of bladder cancer radioresistance. Further validation was done in vivo by conducting mouse tumor xenograft experiments, where HMGB1 knockdown tumors showed a significantly better (P < 0.001) response to radiotherapy and decreased autophagy (shown by P62 staining) as compared with controls. The cumulative findings of our in vitro and in vivo studies highlight the significance of HMGB1 as a radiation response marker as well as its utility in radiosensitization of bladder cancer.


Octopine and nopaline metabolism in Agrobacterium tumefaciens and crown gall tumor cells: role of plasmid genes.
Crown gall tumors produced octopine or nopaline or neither compound, depending on the bacterial strain that incited the tumor. The genes specifying production of octopine or nopaline by the tumor were transferred to recipient bacterial strains when the large plasmid associated with virulence was transferred by either conjugation or deoxyribonucleic acid-mediated transformation. Our results, which confirm the work of others (Bomhoff et al., 1976; Goldman et al., 1968; Petit et al., 1970), indicate that, in general, the strains that utilize octopine induce tumors that synthesize octopine, and those that utilize nopaline induce tumors that synthesize nopaline. However, there were several notable exceptions. One class utilized both octopine and nopaline, but the tumors induced by these strains produced only nopaline. Another class utilized nopaline, but their tumors synthesized neither nopaline nor octopine. Mutants were isolated from a number of either octopine- or nopaline-utilizing strains that no longer could utilize the relevant guanido amino acid. These strains, which were mutant in the gene specifying octopine or nopaline oxidase, still retained the permease for these amino acids as well as virulence. Tumors induced by these mutants still synthesized approximately the same levels of octopine and nopaline as tumors induced by their parents. These results suggest that the plasmid gene that determines production of octopine or nopaline by the tumor is distinct from the plasmid gene that determines their catabolism by the bacteria.


Axotomy-dependent and -independent synapse elimination in organ cultures of Wld(s) mutant mouse skeletal muscle.
Progressive "dying back" neurodegenerative diseases are debilitating due to loss of connectivity after nerve terminal and axonal withdrawal, which impairs peripheral nerve function and leads ultimately to neuronal cell death. The mutant mouse (Wallerian degeneration slow; Wld(s)) provides an accessible model system to understand orthograde and retrograde degeneration, because in these mice axotomy induces slow, progressive withdrawal of nerve terminals from motor endplates. Axon degeneration itself is about 10 times slower than in wild-type mice. We describe an organ culture paradigm that permits direct observation of the progressive changes in morphology of neuromuscular junctions in Wld(s) mutant mice. Normal nerve terminal and motor endplate morphology were maintained at most Wld(s) neuromuscular junctions for up to 72 hr in vitro. At others, synaptic boutons were removed from postsynaptic junctional folds in piecemeal fashion, as observed in adults in vivo. By contrast, nerve terminals degenerated rapidly and synchronously in wild-type muscle cultures, resembling Wallerian degeneration in vivo. These observations confirm that in Wld(s) mice, axotomy triggers a mechanism of nerve-terminal withdrawal that seems qualitatively different from that in wild-type animals. The piecemeal dismantling of presynaptic terminals resembles that occurring during neonatal synapse elimination. Organ cultures of neonatal Wld(s) muscle maintained for 1-2 days in vitro also showed no evidence of synaptic terminal degeneration, but elimination of polyneuronal innervation progressed in vitro at approximately the same rate as in vivo. Taken together, the data suggest that both natural and axotomy-induced forms of synapse withdrawal may be accessible to continuous observation and analysis, in organ-cultures of Wld(S) mouse muscles. This offers several advantages over repeated visualization of synaptic remodeling that has thus far been possible only in vivo.


Familial correlation of bone mineral density, birth data and lifestyle factors among adolescent daughters, mothers and grandmothers.
This study aimed to clarify the relationship between skeletal or lifestyle factors among Japanese daughter-mother, mother-grandmother and daughter-grandmother pairs. We performed a cross-sectional study in a cohort of Japanese adolescent daughters (12-18 years of age), their mothers (339 pairs) and grandmothers on their mothers' side (34 pairs). Gestational age, birth weight, age at menarche and presence of menarche or menopause were surveyed in the participants. Height, body weight and lumbar 2-4 bone mineral density (BMD) were measured. Dietary intake and current physical activity were assessed by using questionnaires. Gestational age and age at menarche were significantly correlated among daughters, mothers and grandmothers (P < 0.001). BMD was significantly correlated between daughters and mothers (P < 0.001), while it was not significantly correlated between daughters and grandmothers or between mothers and grandmothers. Dietary intake of calcium and vitamin D, and the frequency, duration and intensity of current physical activity were significantly correlated between daughters and mothers (P < 0.05), although no significant correlation was found between daughters and grandmothers, or between mothers and grandmothers. The parameters for exercise indicated a positive correlation for BMD in the daughters and the mothers, but not in the grandmothers. The results suggested that estrogen deficiency decreases familial correlation for BMD after menopause. Achieving high BMD through exercise may be important for prevention of postmenopausal osteoporosis in premenopausal low-height mothers.


EditR: A Method to Quantify Base Editing from Sanger Sequencing.
CRISPR-Cas9-Cytidine deaminase fusion enzymes-termed "base editors"-allow targeted editing of genomic deoxycytidine to deoxythymidine (C:G→T:A) without the need for double-stranded break induction. Base editors represent a paradigm shift in gene editing technology due to their unprecedented efficiency to mediate targeted, single-base conversion. However, current analysis of base editing outcomes rely on methods that are either imprecise or expensive and time-consuming. To overcome these limitations, we developed a simple, cost-effective, and accurate program to measure base editing efficiency from fluorescence-based Sanger sequencing, termed "EditR." We provide EditR as a free online tool or downloadable desktop application requiring a single Sanger sequencing file and guide RNA sequence. EditR is more accurate than enzymatic assays, and provides added insight to the position, type, and efficiency of base editing. Furthermore, EditR is likely amenable to quantify base editing from the recently developed adenosine deaminase base editors that act on either DNA (adenosine deaminase base editors [ABEs]) or RNA (REPAIRs) (catalyzes A:T→G:C). Collectively, we demonstrate that EditR is a robust, inexpensive tool that will facilitate the broad application of base editing technology, thereby fostering further innovation in this burgeoning field.


Identifying Policy Implications and Future Research Directions Regarding Military Community Support and Child Psychosocial Adjustment.
As former U.S. Army Surgeon General Horoho points out, a large fraction of what determines the health and readiness of our military families does not occur during appointments with professionals, but rather within the "Lifespace-where health really happens...." Indeed, when children of military families experience psychosocial difficulties, such stress impacts the service members' personal well-being and ability to focus at work, impairing their capacity to attend to the mission. As such, the Department of Defense (DoD) has instituted a family readiness system to bolster resiliency within military families, including children, e.g., by linking families with support networks. Bolstering military family resiliency, including the prevention of and effective intervention for child psychosocial problems, is an important issue at all levels of the DoD. Service members, leaders, and policy makers have a vested interest in promoting mission readiness and a healthy force. Research can play an important role in shaping decision-making by consolidating what is currently known and not known about a particular expertise area. To date, there has been no consolidation of research regarding outcomes associated with military community support and the programs that currently exist to bolster child and family resiliency. Given the importance of military families to mission readiness, a review of the relevant research is warranted. This commentary article reviews the literature on community support for military children, provides an overview of currently available resources, discusses concerns with the current provision of support services to military families, and offers recommendations for future research, policy, and implementation of military community support programs. Although there is a dearth of research on available support programs, there appears to be no lack of services available to military families. However, several steps could be taken to make these resources into a more supportive system. Family members must be able to identify what support services exist, distinguish which service is most suitable for their needs, and be able to readily access these services in a resource-conserving manner. Considerable overlap in support services seems to suggest limited coordination between organizations and service providers, particularly in regard to the government/civilian interface, which inherently lies outside of DoD control. This overlap suggests a redundancy, which may not be efficient economically or in regard to accessing support. There also may be some confusion over which support service is most suitable for the consumer's need. Although some overlap is useful, such as the provision of different services to different populations (e.g., having separate programs for the Army, Navy, Air Force, and Marine subcultures), limited organization and parsimonious provision of services makes it particularly difficult for spouses and family members to navigate resources. Initial attempts are being made to organize and consolidate resources in both the government and civilian sectors. However, it is clear that these initiatives have not completely solved problems related to resource access, redundancy, and lack of research-supported efficacy.


Soliton formation from a pulse passing the zero-dispersion point in a nonlinear Schrodinger equation
We consider in detail the self-trapping of a soliton from a wave pulse that passes from a defocusing region into a focusing one in a spatially inhomogeneous nonlinear waveguide, described by a nonlinear Schrodinger equation in which the dispersion coefficient changes its sign from normal to anomalous. The model has direct applications to dispersion-decreasing nonlinear optical fibers, and to natural waveguides for internal waves in the ocean. It is found that, depending on the (conserved) energy and (nonconserved) "mass" of the initial pulse, four qualitatively different outcomes of the pulse transformation are possible: decay into radiation; self-trapping into a single soliton; formation of a breather; and formation of a pair of counterpropagating solitons. A corresponding chart is drawn on a parametric plane, which demonstrates some unexpected features. In particular, it is found that any kind of soliton(s) (including the breather and counterpropagating pair) eventually decays into pure radiation with an increase of energy, the initial "mass" being kept constant. It is also noteworthy that a virtually direct transition from a single soliton into a pair of symmetric counterpropagating ones seems possible. An explanation for these features is proposed. In two cases when analytical approximations apply, viz., a simple perturbation theory for broad initial pulses and the variational approximation for narrow ones, comparison with direct simulations shows reasonable agreement.


Hypo-angiogenesis: a possible pathological factor in the development of dry age-related macular degeneration and a novel therapeutic target.
Angiogenesis causes severe vision loss in patients with exudative or wet forms of age-related macular degeneration (AMD). The pathogenesis involves upregulation of several proangiogenic factors, particularly the vascular endothelial growth factor (VEGF). Contrary to the pathogenesis of exudative AMD, molecular events leading to the development of dry AMD remain unclear. Dry AMD is characterized by loss of the retinal pigment epithelium (RPE). The mechanism that triggers RPE cell loss remains unclear. Choriocapillaris development is absent in mice with RPE-specific deletion of VEGF. Moreover, in later life, background VEGF secretion promotes the survival of the RPE and maintains choriocapillaris integrity. We hypothesized that reduced synthesis of VEGF (hypo-angiogenesis) or abnormalities in its receptors, VEGF receptor-1 (VEGFR1) and VEGFR2, may be involved in the pathogenesis of non-exudative AMD or dry AMD. If the concept of hypo-angiogenesis as a driver for dry AMD is proven, treatment with VEGF or induction of angiogenesis could be considered. Similar attempts at therapeutic angiogenesis have been actively investigated in cardiac and limb ischemia. The reasons for a patient developing exudative AMD or dry AMD remain poorly understood. Nevertheless, targeting increased VEGF production in patients with exudative AMD using anti-VEGF drugs is highly efficacious in preserving vision. Similarly, dry AMD may be a manifestation of reduced VEGF synthesis (hypo-angiogenesis) and subsequent decreased RPE cell survival. Experimental studies exploring the possibility of reduced VEGF secretion and/or increased receptor resistance/abnormality could pave the way for clinical trials of angiogenesis to treat dry AMD.


Hantavirus infections in Finland.
Hantaviruses cause haemorrhagic fever with renal syndrome (HFRS) and hantaviruspulmonary syndrome (HPS), and occur throughout the world. The viruses aretransmitted to humans in aerosols of rodent secretions. About 200 000 people,mainly in Eurasia, develop.


Impacts of water and nitrogen supplies on the physiology, leaf demography and nitrogen dynamics of Betula pendula.
We determined the response of Betula pendula Roth. trees to a restricted water supply, and quantified the interactions between tree N and water status on leaf demography and internal N cycling. In April 1993, 3-year-old trees were planted in sand culture and four treatments applied: high-N supply (56 mg tree(-1) week(-1)) with either 2 dm(3) water week(-1) (HN+) or 0.9 dm(3) water week(-1) (HN-), or low-N supply (14 mg tree(-1) week(-1)) with 2 dm(3) (LN+) or 0.9 dm(3) (LN-) water week(-1). Until 1994, the N supplied to trees was enriched with (15)N to 5.4 atom %. During 1993, there were few differences in the growth or leaf demography of trees in the LN+ and LN- treatments, but the high-N treatment increased tree growth. Leaf mass and area were initially similar in trees in the HN+ and HN- treatments, but the trees in the HN- treatment had a smaller root system. Net assimilation rate under saturating light was higher in trees in the HN+ treatment than in trees in the LN+ treatment. There was an N x water supply interaction as a result of trees in the HN- treatment closing their stomata by the beginning of August. However, there was no difference in gas exchange characteristics of leaves in the LN+ and LN- treatments. Although leaf senescence and abscission started in the HN- treatment by mid-August and continued for about 90 days, whereas leaf abscission in the other treatments did not start until the beginning of October and only lasted 25-30 days, the trees in the HN+ and HN- treatments remobilized similar amounts of (15)N for leaf growth in the spring of 1994. There were no differences in predawn water potential among treatments and no evidence of osmotic adjustment. We conclude that B. pendula trees avoid rather than tolerate drought. The interaction between the effects of nitrogen and water supplies on leaf demography and internal cycling of N are discussed.


RNase T affects Escherichia coli growth and recovery from metabolic stress.
To determine the essentiality and role of RNase T in RNA metabolism, we constructed an Escherichia coli chromosomal rnt::kan mutation by using gene replacement with a disrupted, plasmid-borne copy of the rnt gene. Cell extracts of a strain with mutations in RNases BN, D, II, and I and an interuppted rnt gene were devoid of RNase T activity, although they retained a low level (less than 10%) of exonucleolytic activity on tRNA-C-C-[14C]A due to two other unidentified RNases. A mutant lacking tRNA nucleotidyltransferase in addition to the aforementioned RNases accumulated only about 5% as much defective tRNA as did RNase T-positive cells, indicating that this RNase is responsible for essentially all tRNA end turnover in E. coli. tRNA from rnt::kan strains displayed a slightly reduced capacity to be aminoacylated, raising the possibility that RNase T may also participate in tRNA processing. Strains devoid of RNase T displayed slower growth rates than did the wild type, and this phenotype was accentuated by the absence of the other exoribonucleases. A strain lacking RNase T and other RNases displayed a normal response to UV irradiation and to the growth of bacteriophages but was severely affected in its ability to recover from a starvation regimen. The data demonstrate that the absence of RNase T affects the normal functioning of E. coli, but it can be compensated for to some degree by the presence of other RNases. Possible roles of RNase T in RNA metabolism are discussed.


Insights into degradation mechanism of N-end rule substrates by p62/SQSTM1 autophagy adapter.
p62/SQSTM1 is the key autophagy adapter protein and the hub of multi-cellular signaling. It was recently reported that autophagy and N-end rule pathways are linked via p62. However, the exact recognition mode of degrading substrates and regulation of p62 in the autophagic pathway remain unknown. Here, we present the complex structures between the ZZ-domain of p62 and various type-1 and type-2 N-degrons. The binding mode employed in the interaction of the ZZ-domain with N-degrons differs from that employed by classic N-recognins. It was also determined that oligomerization via the PB1 domain can control functional affinity to the R-BiP substrate. Unexpectedly, we found that self-oligomerization and disassembly of p62 are pH-dependent. These findings broaden our understanding of the functional repertoire of the N-end rule pathway and provide an insight into the regulation of p62 during the autophagic pathway.


Toxicity evaluations of various carbon nanomaterials.
After the discovery of fullerene and carbon nanotubes, various carbon nanomaterials were discovered or synthesized. The carbon nanomaterials have remarkable properties, different from bulk materials with the same chemical composition, and are therefore useful for industrial applications. However, the toxicity of nanomaterials may also differ from that of the bulk materials; this difference poses a concern. The physical similarity of nanomaterials to asbestos has led to evaluations for toxicity by many researchers using various methods. In this review, we compile and compare the toxicity evaluations of each carbon nanomaterial.


Localized in vivo hyperpolarization transfer sequences.
In vivo localized and fully adiabatic homonuclear and heteronuclear polarization transfer experiments were designed and performed in the rat brain at 9.4 T after infusion of hyperpolarized sodium [1,2-(13)C(2)] and sodium [1-(13)C] acetate. The method presented herein leads to highly enhanced in vivo detection of short-T(1) (13)C as well as attached protons. This indirect detection scheme allows for probing additional molecular sites in hyperpolarized substrates and their metabolites and can thus lead to improved spectral resolution such as in the case of (13)C-acetate metabolism.


Pleiotropic effects of statins: evidence against benefits beyond LDL-cholesterol lowering.
3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) are extremely effective at reducing low-density lipoprotein (LDL) cholesterol and have been demonstrated to reduce mortality and the risk of major cardiovascular events in a number of large primary and secondary prevention studies. The linear relationship between LDL-cholesterol and cardiovascular risk suggests statins work solely by reducing LDL-cholesterol, and that ancillary properties do not contribute to cardiovascular risk reduction. In recent years, however, a number of additional non-lipid-lowering, or 'pleiotropic', effects of statins have been suggested to contribute to their efficacy in cardiovascular disease. The first data to suggest that statins may have benefits beyond lipid lowering came from the Heart Protection Study, in which simvastatin reduced mortality and morbidity even in patients with 'normal' LDL-cholesterol levels (2.6 mmol/L or 100 mg/dL). It has since been demonstrated, however, that cardiovascular risk remains high at this LDL concentration, but is substantially reduced in those achieving levels below 2.0 mmol/L (77 mg/dL). Evidence for the pleiotropic effects of statins in heart failure comes largely from retrospective and subgroup analyses of large studies. When statin therapy is compared with placebo, or when high-dose statin therapy is compared with low-dose treatment, a lower incidence of heart failure or hospitalization is observed. Despite promising retrospective data, however, two prospective studies of rosuvastatin in the treatment of patients with New York Heart Association class II-IV heart failure showed no impact on the primary endpoint and only one of the studies showed a lower rate of hospitalization favouring rosuvastatin. A number of small studies has shown evidence for mechanisms of action of statins outside of LDL-cholesterol lowering, including improvements in endothelial function, halting or retardation of atheroma development, reduction in inflammation and antithrombotic effects. The linear relationship between LDL-cholesterol lowering and reduction in coronary heart disease risk, as well as a lack of conclusive evidence for other mechanisms of action raise the question of whether any cholesterol-lowering agent is equally effective for reducing cardiovascular risk, but recent data from the torcetrapib clinical trial programme suggest this is not the case. Future cholesterol-lowering modalities must be able to demonstrate efficacy and good tolerability in large-scale clinical trials.


Central administration of [Phe1psi(CH2-NH)Gly2]nociceptin(1-13)-NH2 and orphanin FQ/nociceptin (OFQ/N) produce similar cardiovascular and renal responses in conscious rats.
In vitro studies have shown that [Phe1Psi(CH2-NH)Gly2]OFQ/N(1-13)-NH2 (referred to as [FG]OFQ/N(1-13)-NH2) is the first selective antagonist to prevent the binding of the endogenous ligand orphanin FQ/Nociceptin (OFQ/N) at the orphan opioid-like receptor. In the present study, we examined the potential changes in cardiovascular and renal function produced by the i.c.v. injection of [FG]OFQ/N(1-13)-NH2 in conscious Sprague-Dawley rats. In conscious rats, i.c.v. injection of [FG]OFQ/N(1-13)-NH2 produced a marked and sustained decrease in heart rate, mean arterial pressure, and urinary sodium excretion and a profound increase in urine flow rate (i.e., a water diuresis). The cardiovascular and renal excretory responses produced by i.c.v. [FG]OFQ/N(1-13)-NH2 were dose dependent and were similar in pattern but of longer duration than responses evoked by i.c.v. OFQ/N. In other animals, the i.c.v. injection of OFQ/N(1-13)-NH2, a potential metabolite of [FG]OFQ/N(1-13)-NH2, produced changes in cardiovascular and renal function that were comparable to those evoked by i.c.v. [FG]OFQ/N(1-13)-NH2. In contrast, OFQ/N(2-17), a fragment of OFQ/N [OFQ/N(1-17)], was inactive when administered centrally. Finally, studies were performed to determine whether [FG]OFQ/N(1-13)-NH2 may be an antagonist at the orphan opioid-like receptor receptor when administered centrally at a dose that alone was inactive. In these studies, i.c.v. pretreatment of animals with low-dose [FG]OFQ/N(1-13)-NH2 failed to prevent the cardiovascular and renal excretory response to i.c.v. OFQ/N. Although [FG]OFQ/N(1-13)-NH2 is reported to be an antagonist of the OFQ/N receptor in vitro, these findings indicate that this compound has agonist activity similar to that of the endogenous ligand OFQ/N when administered centrally in vivo.


Strip gingival autograft used to correct mucogingival problems around implants.
This case report describes the use of a strip gingival autograft to transplant narrow strips of keratinized gingiva around dental implants. Replacement of unattached, nonkeratinized mucosa with keratinized gingiva resulted in firmly attached gingiva and an improved seal around implants that was healthier and more resistant to inflammation. The strip gingival autograft technique is a simple surgery that results in less discomfort for the patient and provides predictable results.


Adenylate cyclase agonist properties of CGP-12177A in brown fat: evidence for atypical beta-adrenergic receptors.
Thermogenesis in brown adipose tissue (BAT) is stimulated by catecholamine activation of adenylate cyclase through the beta-adrenergic receptor. Recently it was reported that the beta-adrenergic antagonist CGP-12177A stimulates oxygen consumption in BAT. To investigate the mechanism of action of CGP-12177A in BAT, we assessed the inhibitory and stimulatory affects of CGP-12177A on the adenylate cyclase system in myocardial and BAT membranes from rats. CGP-1277A inhibited isoproterenol-stimulated adenylate cyclase activity in a dose-dependent manner, with an inhibitory constant (Ki) of 1.94 +/- 0.18 microM in BAT and 0.49 +/- 0.11 microM in the heart. However, in the absence of isoproterenol, CGP-12177A stimulated adenylate cyclase in BAT with two components of activation, and half-maximal stimulation occurred at 1 microM and 1.5 mM. In contrast, CGP-12177A did not stimulate adenylate cyclase activity in heart membranes. Propranolol inhibited the isoproterenol-stimulated activity with a potency that was one log less in BAT compared with heart. Propranolol fully blocked the high-affinity component but only weakly blocked the low-affinity component of CGP-12177A-stimulated activity in BAT. Pindolol was also less potent in BAT but inhibited the CGP-12177A-stimulated activity in a manner similar to the inhibition of the isoproterenol-stimulated activity, suggesting the CGP-12177A activation was beta-receptor mediated. Binding curves of [125I]iodocyanopindolol ([125I]ICYP) in competition with CGP-12177A demonstrated a shift to lower affinity in the presence of beta,gamma-imidoguanosine 5'-triphosphate, indicating that CGP-12177A has agonist properties with respect to the [125I]ICYP binding site.(ABSTRACT TRUNCATED AT 250 WORDS)


Clinical features and the risk factors of infantile exotropia recurrence.
To evaluate the clinical features and the risk factors of recurrence in infantile exotropia. Retrospective, observational case series. The clinical records of 20 infantile exotropia patients who underwent exotropia surgery were reviewed retrospectively. Patients were divided into 2 groups according to the amount of exodeviation present at 1 year after surgery. Preoperative patient characteristics and postoperative alignments were evaluated at 1 day and 1, 3, 6, and 12 months after surgery. The successful group was composed of 12 patients with an alignment of less than 10 prism diopters (PD) of exodeviation, and the recurrent group was composed of 8 patients with an alignment of 10 PD or more of exodeviation (range, 10 to 18 PD) at 1 year after surgery. The incidences of dissociated vertical deviation, inferior oblique overaction, and rates of constancy before operation were not significantly different in the 2 groups (P = .603, P = .158, and P = .347, respectively), and postoperative deviations were not significantly different at 1 day after surgery. However, postoperative deviations were significantly different in the 2 groups at 1 month (P = .069) and 3 months (P < .001) after surgery, and this difference was maintained at 12 months after surgery. Furthermore, correlation between initial alignments at 1 month after surgery and final alignments was significant in successful group (P = .012). No single factor affecting the surgical outcome was identified in infantile exotropia, but the recurrence of infantile exotropia was apparent from the first postoperative month.


Numerical Simulation of a Biventricular Assist Device with Fixed Right Outflow Cannula Banding During Pulmonary Hypertension.
As a left ventricular assist device is designed to pump against the systemic vascular resistance (SVR), pulmonary congestion may occur when using such device for right ventricular support. The present study evaluates the efficacy of using a fixed right outflow banding in patients receiving biventricular assist device support under various circulatory conditions, including variations in the SVR, pulmonary vascular resistance (PVR), total blood volume (BV), as well as ventricular contractility. Effect of speed variation on the hemodynamics was also evaluated at varying degrees of PVR. Pulmonary congestion was observed at high SVR and BV. A reduction in right ventricular assist device (RVAD) speed was required to restore pulmonary pressures. Meanwhile, at a high PVR, the risk of ventricular suction was prevalent during systemic hypotension due to low SVR and BV. This could be compensated by increasing RVAD speed. Isolated right heart recovery may aggravate pulmonary congestion, as the failing left ventricle cannot accommodate the resultant increase in the right-sided flow. Compared to partial assistance, the sensitivity of the hemodynamics to changes in VAD speed increased during full assistance. In conclusion, our results demonstrated that the introduction of a banding graft with a 5 mm diameter guaranteed sufficient reserve of the pump speed spectrum for the regulation of acceptable hemodynamics over different clinical scenarios, except under critical conditions where drug administration or volume management is required.


How Retroviruses Escape the Nonsense-Mediated mRNA Decay.
Many posttranscriptional processes are known to regulate gene expression and some of them can act as an antiviral barrier. The nonsense-mediated mRNA decay (NMD) was first identified as an mRNA quality control pathway that triggers rapid decay of mRNA containing premature stop codons due to mutations. NMD is now considered as a general posttranscriptional regulation pathway controlling the expression of a large set of cellular genes. In addition to premature stop codons, many other features including alternative splicing, 5' uORF, long 3' UTR, selenocystein codons, and frameshift are able to promote NMD. Interestingly, many viral mRNAs exhibit some of these features suggesting that virus expression and replication might be sensitive to NMD. Several studies, including recent ones, have shown that this is the case for retroviruses; however, it also appears that retroviruses have developed strategies to overcome NMD in order to protect their genome and ensure a true expression of their genes. As a consequence of NMD inhibition, these viruses also affect the expression of host genes that are prone to NMD, and therefore can potentially trigger pathological effects on infected cells. Here, we review recent studies supporting this newly uncovered function of the NMD pathway as a defense barrier that viruses must overcome in order to replicate.


Rayleigh Instability-Driven Coaxial Spinning of Knotted Cell-Laden Alginate Fibers as Artificial Lymph Vessels.
Constructing artificial lymph vessels, which play a key role in the immune response, can provide new insights into immunology and disease pathologies. An immune tissue is a highly complex network that consists of lymph vessels, with a "beads-on-a-string" knotted structure. Herein, we present the facile and rapid fabrication of beads-on-a-string knotted cell-laden fibers using coaxial spinning of alginate by exploiting the Plateau-Rayleigh instability. It is shown how alterations in the flow rate and alginate concentration greatly affect the beads-on-a-string structure, rooted in the Plateau-Rayleigh instability theory. Biocompatibility was confirmed by the lactate dehydrogenase (LDH) assay and live/dead staining of the encapsulated human white blood cells. Finally, the encapsulated white blood cells were still functional as indicated by their anti-CD3 activation to secrete interleukin 2. The rapid fabrication of a cell-laden beads-on-a-string three-dimensional (3D) culture platform enables a crude mimicry of the lymph vessel structure. With joint expertise in immunology, microfluidics, and bioreactors, the technology may contribute to the mechanistic assay of human immune response in vitro and functional replacement.


Phosphorus-31 brain MR spectroscopy in women during and after pregnancy compared with nonpregnant control subjects.
A reversible decrease in brain size has been demonstrated during normal pregnancy that is maximal at term and returns to normal after many months. The purpose of this longitudinal study was to use phosphorus-31 MR spectroscopy to determine if metabolic changes explain this physiologic event. Pregnant women (n = 12) were examined at term and up to 6 months after delivery. Nonpregnant control subjects (n = 7) were imaged twice (a month apart) to exclude hormone effects. Brain (31)P MR spectra were acquired at 1.5 T, and intracellular pH was calculated from the chemical shift between phosphocreatine and inorganic phosphate resonances. Statistical analysis was performed by using an analysis of variance. We found no statistically significant differences in the relative levels of metabolite associated with cerebral bioenergetics and cell membrane metabolism between pregnant women and nonpregnant women. However, a significant increase in cerebral pH was observed in pregnant women at 6 weeks after delivery compared with control subjects (7.074 +/- 0.063 vs 7.017 +/- 0.041; P < .05). pH returned to normal by 6 months after delivery (7.014 +/- 0.010). Changes in brain size associated with pregnancy appear to be associated with an increase in intracellular pH after delivery. The observed alkalosis may reflect altered cellular metabolism. These persistent brain perturbations associated with pregnancy indicate that, when postpartum physiologic and pharmacologic changes are measured, long-term effects may be expected in central nervous system processing.


A Björling representation for Jacobi fields on minimal surfaces and soap film instabilities.
We develop a general framework for the description of instabilities on soap films using the Björling representation of minimal surfaces. The construction is naturally geometric and the instability has the interpretation as being specified by its amplitude and transverse gradient along any curve lying in the minimal surface. When the amplitude vanishes, the curve forms part of the boundary to a critically stable domain, while when the gradient vanishes the Jacobi field is maximal along the curve. In the latter case, we show that the Jacobi field is maximally localized if its amplitude is taken to be the lowest eigenfunction of a one-dimensional Schrödinger operator. We present examples for the helicoid, catenoid, circular helicoids and planar Enneper minimal surfaces, and emphasize that the geometric nature of the Björling representation allows direct connection with instabilities observed in soap films.


Idiopathic hypereosinophilic syndrome in 3 Rottweilers.
Three Rottweilers with marked peripheral eosinophilia and infiltration of the liver, spleen, lungs, and bone marrow with eosinophils were diagnosed with idiopathic hypereosinophilic syndrome (IHES). Mean serum immunoglobulin E concentrations were markedly high. On cytogenetic analysis, no evidence of karyotypic abnormalities was found in bone marrow aspirates. Despite an extensive search, no underlying cause for the eosinophilia could be identified. In this study, cytogenetic analysis and measurement of serum IgE concentrations were used to differentiate IHES and eosinophilic leukemia.


[Development of the duodenal glands in rats on a high cellulose diet].
Wistar weaned rats were kept on cellulose-rich diet for 3 months. They revealed the duodenal gland area of increased length, as compared to the control group kept on standard pellets. Glandular-duodenal index, that is a relative indication of gland development, also rose. Cellulose-fed rats showed a lower pH of the stomach contents. The correlation between pH and gland development has been established. Possible reasons for a more intensive gland enlargement in herbivorous animals are being discussed.


Phylogenetic relationships of the genera Acetobacterium and Eubacterium sensu stricto and reclassification of Eubacterium alactolyticum as Pseudoramibacter alactolyticus gen. nov., comb. nov.
16S rRNA gene sequences of the type strains of the seven previously described Acetobacterium species were determined. The Acetobacterium species were found to form a tight phylogenetic cluster within the Clostridium subphylum of the gram-positive bacteria. Within this subphylum these organisms belong to cluster XV as defined by Collins et al. (M.D. Collins, P.A. Lawson, A. Willems, J.J. Cordoba, J. Fernandez-Garayzabal, P. Garcia, J. Cai, H. Hippe, and J. A. E. Farrow, Int. J. Syst. Bacteriol. 44:812-826, 1994) together with Eubacterium alactolyticum barkeri, Eubacterium callanderi, and Eubacterium limosum. Our data indicate that Clostridium cluster XV consists of at least the following three genera: the genus Acetobacterium, the genus Eubacterium sensu stricto (comprising E. limosum, E. barkeri, and E. callanderi), and the genus Pseudoramibacter gen. nov., which is created for E. alactolyticum, which we reclassify as Pseudoramibacter alactolyticus comb. nov.


Transcriptional repression of type I IFN genes.
Transcriptional regulation is a consequence of the combination of both activation and repression for establishing specific patterns of eukaryotic gene expression. The regulation of the expression of type I interferon (IFN-A and IFN-B) multigene family is controlled primarily at the transcriptional level and has been widely studied as a model for understanding the mechanisms of stable repression, transient virus induction and postinduction repression of the genes. The positive and negative regulatory elements required for this on/off switch have been defined within a complex 5' upstream region of their transcription start site. The differential expression pattern of type I IFN genes is thought to involve both substitutions in the virus responsive element (VRE) and presence or absence of negatively acting sequences surrounding the VRE. In this review we discuss several mechanisms of negative regulation due to the existence of common or specific elements in the IFN-B and IFN-A genes and we summarize recent studies on transcriptional repressors that bind to these promoters.


Tracing nitrates and sulphates in river basins using isotope techniques.
The objective of this paper is to outline how stable isotope techniques can contribute to the elucidation of the sources and the fate of riverine nitrate and sulphate in watershed studies. The example used is the Oldman River Basin (OMRB), located in southern Alberta (Canada). Increasing sulphate concentrations and decreasing delta(34)S values along the flowpath of the Oldman River indicate that oxidation of pyrite in tills is a major source of riverine sulphate in the agriculturally used portion of the OMRB. Chemical and isotopic data showed that manure-derived nitrogen contributes significantly to the increase in nitrate concentrations in the Oldman River and its tributaries draining agricultural land. It is suggested that hydrological conditions control agricultural return flows to the surface water bodies in southern Alberta and impart significant seasonal variations on concentrations and isotopic compositions of riverine nitrate. Combining isotopic, chemical, and hydrometric data permitted us to estimate the relative contribution of major sources to the total solute fluxes. Hence, we submit that isotopic measurements can make an important contribution to the identification of nutrient and pollutant sources and to river basin management.


A genomic approach reveals a novel mitotic pathway in papillomavirus carcinogenesis.
More than 90% of cervical carcinomas are associated with human papillomavirus (HPV) infection. The two viral oncogenes E6 and E7 play a major role in transforming the cells by disrupting p53- and pRb-dependent cell cycle checkpoints. A hallmark of HPV-associated cervical carcinoma is loss of the expression of the viral E2 protein, often by disruption of E2-encoding gene. We showed previously that reintroduction of E2 in HPV18-associated cervical carcinoma cells induces cell cycle arrest in G(1) because of the transcriptional repression of the viral oncogenes E6 and E7 and concomitant reactivation of the p53 and pRb pathways. Here we describe global gene profiling of HeLa cells expressing different HPV18 E2 mutants to study the effects of repression of the viral oncogenes. We identified 128 genes transcriptionally regulated by the viral oncogenes in cervical carcinoma. Surprisingly, E2 repressed a subset of E2F-regulated mitotic genes in an E6/E7-dependent pathway. This was corroborated by the observation that E2 delayed mitotic progression, suggesting the involvement of a mitotic pathway in HPV carcinogenesis. These mitotic genes constitute an as yet unrecognized set of genes, which were also found deregulated in other HPV-associated cervical carcinoma cell lines and therefore represent new targets for both diagnosis and therapeutic approaches in cervical cancer.


Surface presentation of functional peptides in solution determines cell internalization efficiency of TAT conjugated nanoparticles.
Functionalizing nanoparticles with cell-penetrating peptides is a popular choice for cellular delivery. We investigated the effects of TAT peptide concentration and arrangement in solution on functionalized nanoparticles' efficacy for membrane permeation. We found that cell internalization correlates with the positive charge distribution achieved prior to nanoparticle encountering interactions with membrane. We identified a combination of solution based properties required to maximize the internalization efficacy of TAT-functionalized nanoparticles.


Comparison of morbidity and function after colectomy with ileorectal anastomosis or restorative proctocolectomy for familial adenomatous polyposis.
Restorative proctocolectomy with an ileal reservoir (RPC) should prevent colorectal cancer in patients with familial adenomatous polyposis. Until this is confirmed its role compared with total colectomy and ileorectal anastomosis (IRA) will depend on the relative morbidity and postoperative bowel function after the two procedures. This was analysed in 99 patients (37 RPC, 62 IRA) operated on between 1977 and 1989. Morbidity was greater after RPC with subsequent ileostomy closure (median hospital stay, 24 versus 11 days; complications, 60 versus 21 per cent; reoperation, 29 versus 3 per cent; return to normal activity; 31 versus 14 weeks). There was little difference in bowel function; after IRA median frequency was 3/24 h and urgency (unable to wait 15 min) occurred in 50 per cent, compared with 4.5/24h and 17 per cent after RPC. Night evacuation occurred in 10 and 43 per cent respectively. IRA was performed in younger patients (median 19 versus 31 years) who had fewer bowel motions before operation (2 versus 5/24 h). The greater morbidity of RPC suggests that it should be restricted to patients at higher risk of developing later rectal cancer, including those unavailable for follow-up and those with large or confluent rectal polyps or with curable colon cancer at the initial colectomy.


Gamma Knife irradiation-induced histopathological changes in the trigeminal nerves of rhesus monkeys.
The authors' goal was to observe histopathological changes in the trigeminal nerve after Gamma Knife surgery (GKS) in rhesus monkeys, and to investigate the radiobiological mechanism of GKS for primary trigeminal neuralgia. The nerve length-dosage effect of irradiation is also discussed. One of 5 rhesus monkeys randomly served as a control, and the other 4 monkeys were randomly administered a target radiation dose of 60, 70, 80, or 100 Gy (a different dose in each animal). The size of the collimator was 4 mm, and the target point was the trigeminal nerve root. In each experimental monkey, one side was exposed to single-target-point irradiation, and the contralateral side was exposed to double-target-point irradiation. After 6 months, the trigeminal nerve root was examined using light microscopy, transmission electron microscopy, and immunohistochemistry. At each radiation dose, the damage to the nerve tissue by single-target-point irradiation was identical to that caused by double-target-point irradiation. In the trigeminal nerve tissues of the monkeys irradiated with 60 and 70 Gy, there was limited nerve demyelination and degeneration, fragmentation, or loss of axons. In the trigeminal nerve tissue of the monkey irradiated with 80 Gy, the nerve tissue showed a disordered structure. In the trigeminal nerve tissue of the monkey irradiated with 100 Gy, there was severe derangement in the structure of the nerve tissue, and extensive demyelination, fragmentation, and loss of axons. The target doses of 60 and 70 Gy have very little impact on the structure of the trigeminal nerve. Irradiation at 80 Gy can cause partial degeneration and loss of axons and demyelination. A 100-Gy dose can cause some necrosis of neurons. Comparing the single-target-point with the double-target-point irradiation, the extent of damage to the nerve tissue is identical, and no difference in the nerve length-dosage effect was found.


Collections from the mesophytic zone off Bermuda reveal three species of Kallymeniaceae (Gigartinales, Rhodophyta) in genera with transoceanic distributions.
A molecular survey of red algae collected by technical divers and submersibles from 90 m in the mesophotic zone off the coast of Bermuda revealed three species assignable to the Kallymeniaceae. Two of the species are representative of recently described genera centered in the western Pacific in Australia and New Zealand, Austrokallymenia and Psaromenia and the third from the Mediterranean Sea and the eastern Atlantic, Nothokallymenia. A phylogenetic analysis of concatenated mitochondrial (COI-5P) and chloroplast (rbcL) genes, as well as morphological characteristics, revealed that two are shown to be new species with distant closest relatives (N. erosa and Psaromenia septentrionalis), while the third represents a deep water western Atlantic species now moved to an Australasian genus (A. westii).


Comparative proteomic and metabolomic profiling of citrus fruit with enhancement of disease resistance by postharvest heat treatment.
From field harvest to the consumer's table, fresh citrus fruit spends a considerable amount of time in shipment and storage. During these processes, physiological disorders and pathological diseases are the main causes of fruit loss. Heat treatment (HT) has been widely used to maintain fruit quality during postharvest storage; however, limited molecular information related to this treatment is currently available at a systemic biological level. Mature 'Kamei' Satsuma mandarin (Citrus unshiu Marc.) fruits were selected for exploring the disease resistance mechanisms induced by HT during postharvest storage. Proteomic analyses based on two-dimensional gel electrophoresis (2-DE), and metabolomic research based on gas chromatography coupled to mass spectrometry (GC-MS), and liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QToF-MS) were conducted. The results show resistance associated proteins were up-regulated in heat treated pericarp, such as beta-1, 3-glucanase, Class III chitinase, 17.7 kDa heat shock protein and low molecular weight heat-shock protein. Also, redox metabolism enzymes were down-regulated in heat treated pericarp, including isoflavone reductase, oxidoreductase and superoxide dismutase. Primary metabolic profiling revealed organic acids and amino acids were down-regulated in heat treated pericarp; but significant accumulation of metabolites, including tetradecanoic acid, oleic acid, ornithine, 2-keto-d-gluconic acid, succinic acid, turanose, sucrose, galactose, myo-inositol, glucose and fructose were detected. Noticeably, H2O2 content decreased, while, lignin content increased in heat treated pericarp compared to the control, which might increase fruit resistibility in response to external stress. Also, flavonoids, substances which are well-known to be effective in reducing external stress, were up-regulated in heat treated pericarp. This study provides a broad picture of differential accumulation of proteins and metabolites in postharvest citrus fruit, and gives new insights into HT improved fruit disease resistance during subsequent storage of 'Kamei' Satsuma mandarin. Interpretation of the data for the proteins and metabolites revealed reactive oxygen species (ROS) and lignin play important roles in heat treatment induced fruit resistance to pathogens and physiological disorders.


The Portrayal of Premium Cigar Selling Propositions in Lifestyle Magazines: A Content Analysis.
Although lifestyle magazines are an important marketing tool for premium cigars, little is known about their recent portrayal of the products. We expand on research conducted for the National Academies of Sciences, Engineering, and Medicine's Committee on Patterns of Use and Health Effects of "Premium Cigars" and Priority Research. A content analysis of magazine covers, articles, and advertisements published in 2021 from Cigar Aficionado and Cigar Snob (five issues each) and Cigar Journal (three issues) assessed magazine themes and mentions of lower harm. Magazine covers (n = 14), advertisements (n = 105), and every fifth article (n = 45) were coded. In total, 92% of the magazine covers, 72.4% of advertisements, and 62.2% of articles had premium cigar content. Celebrities (e.g. musicians and actors) appeared on 92.9% of covers. The most common themes in the advertisements and articles were "high quality." Regarding article content, 80% portrayed the premium cigars' quality (e.g. superior materials), and the taste of featured products (e.g. "tones," "aromas," and "notes") was portrayed in 42.4%. While no articles described their health risks, 6.7% described the positive health effects of premium cigar use (e.g. stress relief and clear cognition). Additionally, a digital search of Cigar Snob and Cigar Aficionado for terms related to lower harm (e.g. "organic," "healthy," "clean," "pure," and "natural") found 7.7 mentions of lower-harm words per issue. Our findings indicate that lifestyle magazines are an important marketing strategy that promotes and normalizes premium cigar use as a high-quality product that can have positive health effects for users. Our manuscript characterized the premium cigar companies' use of selling propositions, including promoting the products' features, safety, taste, and flavors, as a part of their advertising promotion practices. Premium cigar companies used digital and print lifestyle magazines as marketing tools to promote and normalize the use of their products by emphasizing their high quality and positive health benefits (e.g. reducing stress). Premium cigars were promoted as a symbol of power and success, featured in advertisements of upscale social events (e.g. exclusive trade shows, social clubs, and lounges), and often juxtaposed with expensive alcohol, food, and other luxurious goods. Future research should assess if exposure to premium cigar content increases consumer appeal and detracts from the products' potential adverse health outcomes.


Linked sets of antinuclear antibodies: what do they mean?
Among the many antinuclear antibodies found among patients with systemic lupus erythematosus, a subset predominates in terms of frequency of occurrence and titer. These antibodies include anti-DNA, antihistone, anti-U1 RNP, anti-Sm and anti-Ro and they tend to occur in linked sets which correspond to their autoantigenic epitopes on the nucleosome, the U1 snRNP, and the Ro particle. This pattern of occurrence suggests that these nucleoprotein structures elicit at least some of the autoimmune responses in this disease.


Management of castration-resistant prostate cancer: a global approach.
Treatment options for castration-resistant prostate cancer (crpc) have evolved since the start of the 2000s, with most of the new effective therapies appearing since 2010. In 2004, docetaxel was the first chemotherapeutic agent to improve survival in crpc, but little else was available once patients recurred. Since 2010, four new options have been shown to improve survival in patients with refractory or recurring disease after docetaxel. In the management of bone metastases, two bone-targeted therapies have been shown to reduce the risk of bone complications, and they are part of the overall management strategy in crpc patients. Therapeutic options before chemotherapy have shown promising results and may soon become available in Canada. The present article reviews the treatment options that have shown to be effective in crpc and also some of the ongoing work in the field.


[Autophagy and brain: the case of neurodegenerative diseases].
The autophagy-lysosome system is an essential pathway to get rid of unwanted cellular components (proteins and organelles). The brain, and specifically neurons, are very sensitive to abnormalities of the proteome because altered proteins or damaged organelles cannot be diluted by cell division that does not occur in these cells. Most neurodegenerative disorders are characterized by accumulation of undegraded misfolded proteins and are currently associated with autophagy-lysosome dysfunctions. Recent studies have highlighted the modulation of this complex pathway as a putative therapeutic strategy. This review provides an update on the brain-related specificities and dysfunctions of this pathway and discusses the autophagy-based therapies couteracting neurodegeneration.


A retrospective review of the use of thromboprophylaxis in patients who subsequently developed a venous thromboembolism after discharge from hospital.
Thromboprophylaxis is highly effective, safe and cost-effective. However, data from international studies have shown that many at-risk hospitalised patients do not receive appropriate prophylaxis, leaving them at risk of venous thromboembolism (VTE) and its consequences. To assess whether patients readmitted with a VTE to the North Shore Hospital, Auckland, received appropriate thromboprophylaxis during their index admission. Discharge coding was used to identify all patients admitted for VTE-related events between 1 July 2005 to 30 June 2007. Further analysis identified those patients hospitalised for any cause in the 3 months prior to the VTE event. Data were collected on patient characteristics, risk factors for VTE and the thromboprophylaxis regimen prescribed. The seventh American College of Chest Physicians (ACCP) guidelines on the prevention of venous thromboembolism were used to assess VTE risk and to determine whether patients received appropriate prophylaxis. Of the 962 patients admitted with a VTE event, 243 (25%) had been hospitalised within the preceding 3 months. 108 patients were eligible for inclusion in the study; 58 were categorized as medical admissions and 50 as surgical. Using the ACCP-defined criteria, 29 (50%) medical and 38 (76%) surgical patients were identified as being at increased risk of VTE. Only one third of at-risk patients received ACCP-recommended prophylaxis during their index hospital admission. More surgical patients received recommended prophylaxis than medical patients, although this was not statistically significant (37% vs 24%, p=0.3, Fisher's exact test). A significant number of at-risk patients, who subsequently developed VTE after discharge from hospital, failed to receive appropriate thromboprophylaxis during their index admission. Implementing national and local strategies, to improve the use of thromboprophylaxis in hospitalised patients, may help to reduce the burden of VTE.


Rhodomicrobium lacus sp. nov., an alkalitolerent bacterium isolated from Umiam lake, Shillong, India.
A Gram-stain-negative, motile, alkali-tolerant, swollen-rod shaped, reddish brown coloured, phototrophic bacterium designated as strain JA980T, was isolated from freshwater sampled at Umiam lake, Shillong, India. Strain JA980T grew well up to pH 9.0. Respiratory quinones were ubiquinone 10 and rhodoquinone 10. The major fatty acid was C18: 1ω7c/C18:1ω6c with minor amounts of C18:0, C16:0, C18:0 3-OH and C16:0 3-OH. Strain JA980T contained bacteriochlorophyll-a and carotenoids of the spirilloxanthin series. The polar lipids of strain JA980T comprised phosphatidylethanolamine, phosphatidylcholine, diphosphatidylglycerol, an unidentified phospholipid, unidentified amino lipids (AL1,3,4,5) and an unidentified lipid (L1). Strain JA980T had the highest (99.57 %) 16S rRNA gene sequence similarity to the type strains of Rhodomicrobium vannielii ATCC17100T and Rhodomicrobium udaipurense JA643T. The genome of strain JA980T was 3.88 Mbp with a DNA G+C content of 62.4 mol%. Based on the results of phylogenetic analyses, low in silico DNA-DNA hybridization values (33 %), low (87 %) average nucleotide identity results, chemotaxonomic characteristics and differential physiological properties, strain JA980T could not be classified into either of the two recognized species of the genus Rhodomicrobium, suggesting that it represents a novel species, for which the name Rhodomicrobium lacus sp. nov. is proposed. The type strain is JA980T (=KCTC 15697T= MCC 3714T= NBRC 113803T).


The Multifactorial Nature of Early Numeracy and Its Stability.
Early numeracy is a robust predictor of later mathematical abilities. So far, early numeracy has typically been presented as a unitary or two-factorial construct. Nevertheless, there is recent evidence suggesting that it may also be reflected by more basic numerical competences. However, the structure and stability of such a multifactorial model of early numeracy over time has not been investigated yet. In the present study, we used data from a large, longitudinal sample (N = 1292) in the United States with assessments of math ability in prekindergarten and kindergarten to evaluate both the factorial structure of early numeracy and its stability over time. Confirmatory factor analysis identified four distinct basic numerical competences making up early numeracy in prekindergarten: patterning/geometry, number sense, arithmetic, and data analysis/statistics. Stability as tested by means of measurement invariance indicated configural invariance of these four factors from prekindergarten to kindergarten. This reflected that early numeracy in kindergarten was made up by the same four basic numerical competences as in prekindergarten and thus seemed rather stable over the course of preschool. These findings may not only have implications for research on numerical cognition but particularly for diagnostic processes or the development of interventions in educational practice.


Clinical assessment of venous thromboembolic risk in surgical patients.
Prophylaxis against postoperative venous thromboembolism should be tailored according to the patient's level of risk. However, risk assessment is not yet in widespread use in surgical practice. In this study, 538 general surgical patients were prospectively assessed based on a scoring system containing 20 risk factors. Depending on the total risk factor score, the patients were grouped into low (0 to 1, 34.5%), moderate (2 to 4, 48.5%), or high risk (more than 4, 17.2%) categories. Statistically significant (p less than 0.0001) correlation was found between our results and those of three existing risk assessment systems. Overall, of the 538 patients, 37.2% received prophylaxis; 10%, 42.1%, and 76% received prophylaxis in the low-, moderate-, and high-risk categories, respectively. Mechanical prophylactic modalities (graduated elastic compression and sequential intermittent compression) were preferred over pharmacologic modalities. These results suggest that implementation of prophylaxis remains underutilized despite published reports, including NIH guidelines. Our results indicate that the majority of surgical patients seen in this suburban hospital have two or more risk factors for developing venous thromboembolism.


Intramolecular heme ligation of the cytochrome P450 2C9 R108H mutant demonstrates pronounced conformational flexibility of the B-C loop region: implications for substrate binding.
A previous study [Dickmann, L., et al. (2004) Mol. Pharmacol. 65, 842-850] revealed some unusual properties of the R108H mutant of cytochrome P450 2C9 (CYP2C9), including elevated thermostability relative to that of CYP2C9, as well as a UV-visible absorbance spectrum that was indicative of nitrogenous ligation to the heme iron. In our study, size-exclusion chromatography and UV-visible absorbance spectroscopy of CYP2C9 R108H monomers demonstrated that nitrogen ligation is indeed intramolecular. Pulsed electron paramagnetic resonance of CYP2C9 R108H monomers showed that a histidine is most likely bound to the heme as previously hypothesized. An energy-minimized model of the R108H mutant maintained a CYP fold, despite substantial movement of several loop regions of the mutant, and, therefore, represents an extreme example of a closed conformation of the enzyme. Molecular dynamics (MD) simulations of CYP2C9 were performed to study the range of energetically accessible CYP2C9 conformations. These in silico studies showed that the B-C loop region of CYP2C9 moves away from the heme to a position resembling the putative open conformation described for rabbit CYP2B4. A model involving the movement of the B-C loop region and R108 between the open and closed conformations of CYP2C9 is presented, which helps to explain the enzyme's ability to regio- and stereospecifically metabolize some ligands while allosterically activating others.


The distal location of the iron responsive region of the hepcidin promoter.
The response of hepcidin transcription to iron has been repeatedly documented in living mice, but it is difficult to demonstrate the response in ex vivo systems. We have hydrodynamically transfected mice with plasmid constructs composed of a murine hepcidin 1 promoter and fragments of the promoter fused to a firefly luciferase reporter. This method enabled us to quantitate the response of the hepcidin promoter to short-term feeding of a high-iron diet to mice that have been maintained on an iron-deficient diet. We show that the region of the promoter between 1.6 Kb and 1.8 Kb upstream from the start of translation is essential for the response to iron. The promoter region between -260 bp and -1.6 Kb is not essential for the iron responsiveness of hepcidin promoter. The iron-responsive region that we have mapped is the same region required for the in vitro response of HepG2 cells to stimulation with bone morphogenetic proteins and differs from the LPS/IL-6 responsive area.


Establishment of an External Ventricular Drain Best Practice Guideline: The Quest for a Comprehensive, Universal Standard for External Ventricular Drain Care.
External ventricular drains (EVDs) are commonly used to facilitate removal of cerebrospinal fluid in patients with neurologic dysfunction. Despite a high risk for infection (upward of 45%), many hospitals lack strict protocols for EVD placement and maintenance. In addition, EVD infections are typically not tracked with the same diligence as central-line catheter infections, because there are no widely accepted standards for routine management of EVDs. The purpose of this review is to provide a guide for the development of a standardized, best practice EVD protocol for catheter insertion, care, and maintenance to reduce ventriculostomy-related infections. A secondary goal of this review is to provide support for the future development of guidelines for the consistent tracking of EVD insertion and maintenance practices.At an academic medical center, an interdisciplinary team of nurses, advanced practice nurses, and neurointensivists reviewed recent medical and nursing literature as well as research-based institutional protocols on EVD insertion and maintenance from the United States and abroad to determine global best practices. The goal of this literature review was to identify key areas of focus in EVD insertion and maintenance as well as to identify recent studies that have shown success in managing EVDs with low rates of infection. The following terms were used in this search: EVD, externalized ventricular drains, EVD infections, EVD insertion, EVD Care and Maintenance, Nursing and EVDS. The following databases were utilized by each member of the interdisciplinary team to establish a state of the science on EVD management: American Association of Neurosurgical Surgeons, CINAHL, Cochrane, National Guidelines Clearinghouse, and PubMed. The following common EVD themes were identified: preinsertion hair removal and skin preparation, aseptic technique, catheter selection, monitoring of EVD insertion technique using a "bundle" approach, postinsertion dressing type and frequency of dressing changes, techniques for maintenance and cerebrospinal fluid sampling, duration of catheter placement, staff education/competence, and surveillance.


The efficacy of atomoxetine for the treatment of children and adolescents with attention-deficit/hyperactivity disorder: a comprehensive review of over a decade of clinical research.
Atomoxetine was first licensed to treat attention-deficit/hyperactivity disorder (ADHD) in children and adolescents in the US in 2002. The aim of this paper is to comprehensively review subsequent publications addressing the efficacy of atomoxetine in 6- to 18-year-olds with ADHD. We identified 125 eligible papers using a predefined search strategy. Overall, these papers demonstrate that atomoxetine is an effective treatment for the core ADHD symptoms (effect sizes 0.6-1.3, vs. placebo, at 6-18 weeks), and improves functional outcomes and quality of life, in various pediatric populations with ADHD (i.e., males/females, patients with co-morbidities, children/adolescents, and with/without prior exposure to other ADHD medications). Initial responses to atomoxetine may be apparent within 1 week of treatment, but can take longer (median 23 days in a 6-week study; n=72). Responses often build gradually over time, and may not be robust until after 3 months. A pooled analysis of six randomized placebo-controlled trials (n=618) indicated that responses at 4 weeks may predict response at 6-9 weeks, although another pooled analysis of open-label data (n=338) suggests that the probability of a robust response to atomoxetine [≥40% decrease in ADHD-Rating Scale (ADHD-RS) scores] may continue to increase beyond 6-9 weeks. Atomoxetine may demonstrate similar efficacy to methylphenidate, particularly immediate-release methylphenidate, although randomized controlled trials are generally limited by short durations (3-12 weeks). In conclusion, notwithstanding these positive findings, before initiating treatment with atomoxetine, it is important that the clinician sets appropriate expectations for the patient and their family with regard to the likelihood of a gradual response, which often builds over time.


Effects of early phase of preconditioning on rat testicular ischemia.
Brief episodes of ischemia followed by periods of reperfusion generate a powerful protective mechanism in cell, tissue or organ, which increase the resistance to further ischemic damage. This is known as ischemic preconditioning, and has not been investigated in testis. The present experiments were undertaken to determine whether early phase of ischemic preconditioning is evident in rat testis. Surgery was conducted under thiopental (60 mg/kg, intraperitoneal) anesthesia in male Wistar rats. Surgical procedures were performed through a midline incision. Group 1 was designed as a sham group. In group 2, which served as the ischemia group, the animals were subjected to unilateral testicular torsion by rotating the left testis 720 degrees in a clockwise direction. Then, this testis was maintained in the torsion position by fixing with a silk suture to the scrotal wall for 90 min. In groups 3 and 4, 5 or 10 min ischemia followed by 10 min reperfusion was introduced, respectively, to induce single cycle ischemic preconditioning. In group 5, which served as the multiple cycle preconditioning group, 3 cycles of 10 min ischemia and 10 min reperfusion were applied prior to 90 min ischemia. Both ipsilateral and contralateral testes were removed from the rats at the end of the experimental periods, and tissue malondialdehyde (MDA), nitric oxide (NO) levels, xanthine oxidase (XO), myeloperoxidase (MPO) and superoxide dismutase (SOD) activities were measured. Both testes were also evaluated histologically, assessing interstitial edema, congestion, hemorrhages, rupture of tubules and Leydig cell proliferation. 90 min ischemia produced a marked increase in MDA level in left testis. However, all ischemic preconditioning protocols used in this study did not show any significant modification in MDA, NO levels or XO, MPO and SOD activities. Histological grading scale was also similar in ischemia and preconditioning groups. These results suggest that there are no protective effects with ischemic preconditioning in rat testis as showed by biochemical analysis and histological examinations.