Peritoneal microdialysis as a tool for detecting anastomotic leakage in patients after left-side colon and rectal resection. A systematic review.
The objective was to evaluate peritoneal microdialysis in the detection of clinical anastomotic leakage after left-sided colon and rectal resection through a systematic review. A systematic review (PRISMA guidelines) based on a systematic search through PubMed, Cochrane Library, and EMBASE (1 February 2017) was performed. Methodological index of non-randomised studies score was selected to assess the methodological quality. Patient demographics and raw data for intraperitoneal microdialysis concentrations of glucose, lactate, glycerol and pyruvate for 5 d postoperative were obtained from the respective study groups. Ten studies with a total of 128 patients were included. Thirty (23%) patients developed clinical anastomotic leakage. The area under the curve for intraperitoneal lactate concentration was significant higher in patients with anastomotic leakage (58.2; 95% CI 39.2, 77.2) compared with the no leakage group (41.0; 95% CI 35.2, 46.1; p = .007). Receiver operating characteristic curve analysis of the maximum measured lactate concentration demonstrated 25% sensitivity, 88% specificity and 74% accuracy for AL at a cut-off value of 9.8 mmol/L. The odds ratio for a 5 mmol/L increase in lactate in relation to the risk of AL was 2.9 (CI 1.1, 8.0). Increased intraperitoneal lactate concentration within the first 5 d postoperative was significantly associated with clinical anastomotic leakage, but with low predictive values. The microdialysis method is not yet ready for clinical implication before large prospective studies have defined cut off values for a biologic marker in the setting of a clear definitions of leakage.


Complex 5d magnetism in a novel S = 1/2 trimer system, the 12L hexagonal perovskite Ba4BiIr3O12.
The 12L hexagonal perovskite Ba4BiIr3O12 has been synthesized for the first time and characterized using high-resolution neutron and synchrotron X-ray diffraction as well as physical properties measurements. The structure contains Ir3O12 linear face-sharing octahedral trimer units, bridged by corner-sharing BiO6 octahedra. The average electronic configurations of Ir and Bi are shown to be +4(d(5)) and +4(s(1)), respectively, the same as for the S = 1/2 dimer system Ba3BiIr2O9, which undergoes a spin-gap opening with a strong magnetoelastic effect at T* = 74 K. Anomalies in magnetic susceptibility, heat capacity, electrical resistivity, and unit cell parameters indeed reveal an analogous effect at T* ≈ 215 K in Ba4BiIr3O12. However, the transition is not accompanied by the opening of a gap in spin excitation spectrum, because antiferromagnetic coupling among S = 1/2 Ir(4+) (d(5)) cations leads to the formation of a S = 1/2 doublet within the trimers, vs S = 0 singlets within dimers. The change in magnetic state of the trimers at T* leads to a structural distortion, the energy of which is overcompensated for by the formation of S = 1/2 doublets. Extending this insight to the dimer system Ba3BiIr2O9 sheds new light on the more pronounced low-temperature anomalies observed for that compound.


Large-scale analysis of the Alu Ya5 and Yb8 subfamilies and their contribution to human genomic diversity.
We have utilized computational biology to screen GenBank for the presence of recently integrated Ya5 and Yb8 Alu family members. Our analysis identified 2640 Ya5 Alu family members and 1852 Yb8 Alu family members from the draft sequence of the human genome. We selected a set of 475 of these elements for detailed analyses. Analysis of the DNA sequences from the individual Alu elements revealed a low level of random mutations within both subfamilies consistent with the recent origin of these elements within the human genome. Polymerase chain reaction assays were used to determine the phylogenetic distribution and human genomic variation associated with each Alu repeat. Over 99 % of the Ya5 and Yb8 Alu family members were restricted to the human genome and absent from orthologous positions within the genomes of several non-human primates, confirming the recent origin of these Alu subfamilies in the human genome. Approximately 1 % of the analyzed Ya5 and Yb8 Alu family members had integrated into previously undefined repeated regions of the human genome. Analysis of mosaic Yb8 elements suggests gene conversion played an important role in generating sequence diversity among these elements. Of the 475 evaluated elements, a total of 106 of the Ya5 and Yb8 Alu family members were polymorphic for insertion presence/absence within the genomes of a diverse array of human populations. The newly identified Alu insertion polymorphisms will be useful tools for the study of human genomic diversity.


'Click' functionalised polymer resins: a new approach to the synthesis of surface attached bipyridinium and naphthalene diimide [2]rotaxanes.
Herein we describe the design and synthesis of a series of solid-tethered [2]rotaxanes utilising crown ether-naphthalene diimide or crown ether-bipyridinium host guest interactions. TentaGel polystyrene resins were initially modified in a two-stage procedure to azide functionalised beads before the target supramolecular architectures were attached using a copper catalysed "click" procedure. The final assembly was examined using IR spectroscopy and gel-phase (1)H High Resolution Magic Angle Spinning (HR MAS) NMR spectroscopy. The HR MAS technique enabled a direct comparison between the solid-tethered architectures and the synthesis and characterisation of analogous solution-based [2]rotaxanes to be made.


Relation of major volcanic center concentration on venus to global tectonic patterns.
Global analysis of NASA Magellan image data indicates that a major concentration of volcanic centers covering approximately 40 percent of the surface of Venus occurs between the Beta, Atla, and Themis regiones. Associated with this enhanced concentration are geological characteristics commonly interpreted as rifting and mantle upwelling. Interconnected low plains in an annulus around this concentration are characterized by crustal shortening and infrequent volcanic centers that may represent sites of mantle return flow and net down-welling. Together, these observations suggest the existence of relatively simple, largescale patterns of mantle circulation similar to those associated with concentrations of intraplate volcanism on Earth.


To reduce technology prior discharge from intensive care--important but difficult? A grounded theory.
The aim of this study was to provide a deeper understanding of the experience of intensive care staff regarding the reduction in the use of medical technology prior to patients' transfer from the ICU. The goal of ICU transitional care, provided for intensive care patients before, during and after the transfer from the ICU to another care unit, is to ensure minimal disruption and optimal continuity of care for the patient. To smooth this transition, there is a need to prepare for a less technological environment and therefore also a need for a gradual reduction in the use of monitoring equipment. Group interviews and individual interviews, together with participant observations, were conducted with ICU staff in two hospitals in Sweden. The data were analysed using classic grounded theory. The main concern was the ICU staff's ambiguity regarding whether and how to reduce the use of medical technology devices. Insecurity about weaning patients from medical equipment combined with a lack of standardized routines made it difficult for staff to reduce the technical support. The core category describes how the ambiguity was solved primarily by 'prioritizing control'. However, this often caused the ICU staff to use advanced technology while the patients were in the ICU until the ward staff arrived, even if this should have been handled otherwise. Why and how the ICU staff used the strategy of 'prioritizing control' is further explained in the categories 'being affected by cultural/contextual aspects', 'searching for guidance and a shared understanding' and 'weighing advantages with more v s less technology'. It is important to consider ICU staff ambiguity concerning the reduction in technology and to establish strategies for a safe and structured transitional phase with step-down procedures in which technology and monitoring is gradually reduced prior to transfer from ICU.


[Can ultrasonography be used to measure torsion angle of the humerus? Comparison of ultrasonographic, computerized tomographic and anthropometric methods].
Sonographic measurements on 40 cadavaric humeri of the angle of torsion according to Harland had been performed. The results were referred to different computertomographic and anthropometric methods. Sonographic measurements proofed to be accurate and were easily reproducible. Comparing sonographically detected angles with the anthropometric results there was shown a mean difference of 28.9 degrees +/- 7.8 degrees. We conclude that sonography is able to give accurate and reproducible results that do not exactly reflect the angle of torsion.


Linking elements to biochemicals: effects of nutrient supply ratios and growth rates on fatty acid composition of phytoplankton species.
Three species of marine phytoplankton, Rhodomonas sp., Isochrysis galbana Parke, and Phaeodactylum tricornutum Bohlin, were cultivated in semicontinuous cultures to test biochemical responses (fatty acids; FAs) to five nitrogen (N):phosphorus (P) supply ratios and four growth rates (dilution rates). The characteristic FA profile was observed for each algal species (representing particular algal class), which remained relatively stable across the entire ranges of N:P supply ratios and growth rates. For all species, significant direct effects of N:P supply ratios on FAs were found at lower growth rates. The highest saturated and monounsaturated fatty acid (SFA and MUFA) contents were observed under N deficiency at the lowest growth rate in all three species, while responses of polyunsaturated fatty acids (PUFAs) revealed no consistent pattern. Total FAs (and SFAs and MUFAs) in all species showed significant negative correlations with N cell quota (QN ) under N deficiency, but PUFAs had species-specific correlations with QN . The results show that characteristic FA profiles of algal genus or species (representing particular algal classes) underlie fluctuations according to culture conditions. The significant correlation between FAs and QN under N deficiency suggests that elemental and biochemical limitation of phytoplankton should be considered mutually as determinants of food quality for zooplankton in marine ecosystems.


Elevated urinary N-acetyl-β-D-glucosaminidase is associated with high glycoalbumin-to-hemoglobin A1c ratio in type 1 diabetes patients with early diabetic kidney disease.
Urinary N-acetyl-β-D-glucosaminidase (uNAG) predicted the progression of diabetic kidney disease (DKD) prior to development of albuminuria in diabetes patients. We sought whether uNAG level is associated with glycoalbumin-to-hemoglobin A1c ratio (G/A ratio), a marker of postprandial hyperglycemia and glycemic excursion, independent of albuminuria and kidney function. The association between uNAG excretion and G/A ratio was assessed in 204 consecutive subjects with type 1 diabetes (T1D) (mean age 43.9 years; 49.0% men). uNAG excretion level increased along with older age, hyperglycemia, and degree of albuminuria, but was not correlated with body mass index or estimated glomerular filtration rate (eGFR). Elevated uNAG showed robust association with higher G/A ratio (adjusted β = 0.103, P = 0.020) after adjustment for age, sex, body mass index, duration of diabetes, uACR, angiotensin blockers use, fasting plasma glucose, and hemoglobin level. uNAG showed better discriminatory performance for individuals with high G/A ratio than albuminuria (AUC 0.613 vs. 0.518, P = 0.038). Measurement of uNAG improved AUC for high G/A ratio from 0.699 to 0.748 (P = 0.043) when added to conventional risk factors (cutoff 5.24 U/g creatinine; sensitivity 62.5% and specificity 58.0%). In conclusion, Elevated uNAG was found to be associated with high G/A ratio in patients with T1D with early stage DKD, independent of age and albuminuria.


Molecular cloning and characterization of tetrahydroprotoberberine cis-N-methyltransferase, an enzyme involved in alkaloid biosynthesis in opium poppy.
S-Adenosyl-l-methionine:tetrahydroprotoberberine cis-N-methyltransferase (EC 126.96.36.199) catalyzes the conversion of (S)-stylopine to the quaternary ammonium alkaloid, (S)-cis-N-methylstylopine, as a key step in the biosynthesis of protopine and benzophenanthridine alkaloids in plants. A full-length cDNA encoding a protein exhibiting 45 and 48% amino acid identity with coclaurine N-methyltransferase from Papaver somniferum (opium poppy) and Coptis japonica, respectively, was identified in an elicitor-treated opium poppy cell culture expressed sequence tag data base. Phylogenetic analysis showed that the protein belongs to a unique clade of enzymes that includes coclaurine N-methyltransferase, the predicated translation products of the Arabidopsis thaliana genes, At4g33110 and At4g33120, and bacterial S-adenosyl-L-methionine-dependent cyclopropane fatty acid synthases. Expression of the cDNA in Escherichia coli produced a recombinant enzyme able to convert the protoberberine alkaloids stylopine, canadine, and tetrahydropalmatine to their corresponding N-methylated derivatives. However, the protoberberine alkaloids tetrahydroxyberbine and scoulerine, and simple isoquinoline, benzylisoquinoline, and pavine alkaloids were not accepted as substrates, demonstrating the strict specificity of the enzyme. The apparent K(m) values for (R,S)-stylopine and S-adenosyl-L-methionine were 0.6 and 11.5 microm, respectively. TNMT gene transcripts and enzyme activity were detected in opium poppy seedlings and all mature plant organs and were induced in cultured opium poppy cells after treatment with a fungal elicitor. The enzyme was detected in cell cultures of other members of the Papaveraceae but not in species of related plant families that do not accumulate protopine and benzophenanthridine alkaloids.


Repair of Fractures of the Lateral Aspect of the Humeral Condyle in Skeletally Mature Dogs with Locking and Non-Locking Plates.
The aim of this study was to report the functional outcome and complications following transcondylar screw and epicondylar plate fixation of skeletally mature dogs with fractures of the lateral aspect of the humeral condyle using a string of pearls, limited contact dynamic compression plate (LC-DCP) or locking compression plate (LCP). This study was a retrospective review of clinical records, radiographic images and canine brief pain inventory evaluation with a supplementary non-validated questionnaire. Thirty-one fractures in 29 dogs were included. The majority of dogs were Spaniels (23/29). Thirty fractures had evidence of humeral intracondylar fissure. Sixteen fractures had string of pearls, eight LCP and seven LC-DCP. There were two minor complications and three major complications. Perioperative (median: 6 weeks, range: 6-12) outcome was considered fully functional for 24 fractures and acceptable for 6 fractures. One dog had an unacceptable outcome due to the failure of an LCP that healed following revision surgery. Fractures with radiographic follow-up had a persistent visible intracondylar fracture line in 20/26. Twenty-six of twenty-seven owners perceived an excellent outcome (median: 135 weeks, range: 25-246). Repair of fractures of the lateral aspect of the humeral condyle with a transcondylar screw and epicondylar plate has a good perioperative, mid- and long-term outcome with 10% major complication rate, despite incomplete or non-union of the intracondylar fracture in 20/26 elbows.


Orientational wetting and dynamical correlations toward glass transition on the surface of imidazolium-based ionic liquids.
Surface induces many fascinating physical phenomena, such as dynamic acceleration, surface anchoring, and orientational wetting, and, thus, is of great interest to study. Here, we report classic molecular dynamics simulations on the free-standing surface of imidazolium-based ionic liquids (ILs) [C4mim][PF6] and [C10mim][PF6]. On [C10mim][PF6] surface, a significant orientational wetting is observed, with the wetting strength showing a diverging tendency. Depth of the wetting was captured from the density and orientational order profile by a static length, which remarkably increases below the temperature Tstat upon cooling down. The dynamical correlation length that measures the distance of surface-dynamics acceleration into the bulk was characterized via the spatial-dependent mobility. The translational correlation exhibits a similar drastic increment at Tstat, while the rotational correlation drastically increases at a lower temperature Trot. We connect these results to the dynamics in bulk liquids, by finding Tstat and Trot that correspond to the onset temperatures where the liquids become cooperative for translational and rotational relaxation, respectively. This signifies the importance of collective dynamics in the bulk on the orientational wetting and surface dynamics in the ILs.


Perioperative outcomes of laparoscopic transhiatal inversion esophagectomy compare favorably with those of combined thoracoscopic-laparoscopic esophagectomy.
Wide acceptance of laparoscopic esophagectomy has been hampered by the technical difficulty of the procedure and inconsistent improvements in morbidity and mortality. Most case series have utilized a combined thoracoscopic-laparoscopic approach (TLE), but laparoscopic inversion esophagectomy (LIE), a method of transhiatal esophagectomy, has been proposed as an alternative. Inversion esophagectomy simplifies retraction and improves exposure during the mediastinal dissection; however, no previous studies have directly compared LIE outcomes with those of the combined approach. Between July 2003 and March 2008, 70 consecutive patients underwent minimally invasive esophagectomy by LIE (N = 40) or TLE (N = 30). Data for all patients were collected prospectively and stored in a relational database. Recorded outcome measures included operative time, blood loss, length of hospital stay, intensive care unit stay, and perioperative complications. There were no significant differences in patient age, gender, body mass index (BMI), or American Society of Anesthesiologists (ASA) class between the groups, but LIE patients had lower stage of esophageal cancer, and were less likely to have received induction chemoradiotherapy than TLE patients. Patients undergoing LIE had significantly lower operative time (398 vs. 537 min, p < 0.001), intraoperative blood loss (100 vs. 200 ml, p < 0.001), and overall length of stay (9 vs. 14 days, p = 0.003) compared with TLE patients. LIE yielded a median of 10 lymph nodes removed compared with 13 for TLE (p = 0.016). Atrial arrhythmia and postoperative pneumonia were less common in LIE patients than in TLE patients, occurring in 17.5% vs. 27.1% (p = 0.036), and in 7.5% vs. 15.7% of cases (p = 0.029), respectively. LIE provides safe and effective approach to minimally invasive esophagectomy for patients with early esophageal cancer and high-grade dysplasia. Compared with TLE, inversion esophagectomy requires less operative time and has lower operative blood loss and length of hospital stay. LIE may also result in fewer perioperative cardiac and pulmonary complications compared with TLE. Based on these results, we reserve TLE for more advanced esophageal cancer and those undergoing preoperative radiochemotherapy.


Does dopamine block the spawning of the acroporid coral Acropora tenuis?
Most corals undergo spawning after a particular moon phase, but how moon-related spawning is endogenously regulated in corals remains unknown. The objective of the present study was to evaluate whether dopamine (DA) affects spawning in Acropora tenuis. When pieces of four A. tenuis colonies were reared under a natural photoperiod and water temperature, spawning was observed after the predicted moon phase. After exposure to water containing DA at 0.1 μM, pieces of the same colonies only released 5 to 10 bundles. Co-treatment with DA and pimozide (D1 and D2 receptors antagonist), but not domperidone (D2 receptor antagonist), induced mass release of bundles from the colonies. A cross-experiment revealed high fertilization rates between the control colonies (95%) and between the control and DA-treated colonies (90%), suggesting that gametes developed normally in coral tissue. Therefore, DA appears to have an inhibitory effect on the spawning of A. tenuis.


Effect Modifiers of Low-Dose Tamoxifen in a Randomized Trial in Breast Noninvasive Disease.
Low-dose tamoxifen halved recurrence after surgery in a phase III trial in breast noninvasive disease without increasing adverse events. We explored the effect of low-dose tamoxifen in clinically relevant subgroups, including menopausal status, estradiol levels, smoking, body mass index, and proliferation of baseline lesion. Incidence of invasive breast cancer or ductal carcinoma in situ was the primary endpoint. HRs and interaction terms were estimated using Cox models. A favorable HR and 95% confidence interval (CI) could be demonstrated for postmenopausal status (HR = 0.30; 95% CI, 0.11-0.82 vs. HR = 0.73; 95% CI, 0.30-1.76 in premenopausal women; P interaction = 0.13), women with estradiol less than 15.8 pg/mL, presence of menopausal symptoms at baseline, and never smoking (P interaction = 0.07), although the interaction P value was >0.05 for all characteristics. Efficacy was similar in all body mass index categories. Tumors with Ki-67 above the median level of 10% had a greater benefit (HR = 0.27; 95% CI, 0.09-0.81) than those with Ki-67 ≤10% (HR = 1.58; 95% CI, 0.45-5.60; P interaction = 0.04). The efficacy of low-dose tamoxifen seems to be greater in postmenopausal women and in women with lower estradiol levels. Benefits appear to be larger also in women with menopausal symptoms, never smokers, and tumors with Ki-67 >10%. Our results by menopausal status provide important insight into low-dose tamoxifen personalized treatment, although caution is necessary given their exploratory nature. Observation of an improved response in tumors with Ki-67 >10% is consistent but the use of the marker in this setting is investigational.See related commentary by Fabian, p. 3510.


Phytoestrogens and other botanicals: on the problems of evidence-based evaluation.
Soy is the principal plant that produces phytoestrogens, named so because they bind to the estrogen receptors, however weakly compared to the estrogens. Recent reviews concluded that there is no convincing evidence in favor of alleviation of menopausal symptoms by phytoestrogens. However, some studies suggest their efficacy. The question is discussed here, whether vegetable analogues should be used for replacement therapy instead of physiological hormones, also because phytoestrogen preparations often contain a mixture of different components. There is a controversy: phytoestrogens are used to compensate for estrogen deficiency in menopause; but their estrogenic potential does not prevent from the use of soy in infant formulas and other foodstuffs. Feminizing effect of phytoestrogens and soy products may be subtle, detectable only in large populations. This matter should be clarified by independent research, which can be of importance for the future of soy in agriculture. Furthermore a tendency to present placebos and substances with unproven effects in the guise of evidence-based medications is discussed. In conclusion, research quality and possible influence by the industry should be taken into account defining inclusion criteria for studies into meta-analyses and reviews. The article presented patents discussion relevant to the article.


Expression of cellular FLICE-inhibitory protein and its association with p53 mutation in colon cancer.
To investigate the expression of cellular FLICE (Fas associated death domain-like IL-1beta-converting enzyme)-inhibitory protein (c-FLIP) and its association with p53 mutation in colon cancer. Immunohistochemical staining of c-FLIP and mutant p53 by using specific antibodies was performed by the standard streptavidin-peroxidase technique for 45 colon cancer tissue samples with matched normal tissues. Semi-quantitative reverse transcriptional (RT)-PCR was used to measure c-FLIP mRNA levels. t-test statistical method was used in data analyses. c-FLIP mRNA was expressed in all colon cancer tissues and its level (0.63+/-0.12) was significantly higher than that in normal tissues (0.38+/-0.10, P<0.01). Immuno-histochemically, c-FLIP protein was also expressed in all colon cancers (45/45) and 71.1% (32/45) showed an intense immunostaining, in contrast, 93.3% (42/45) of normal colonic mucosa showed positive staining and none of them immunostained intensely. The quantity of c-FLIP protein was significantly higher in cancer tissues than in normal mucosa (7.04+/-1.20 vs 5.21+/-0.86, P<0.01). Positive staining of mutant p53 protein was found in 60% (27/45) colon cancers. c-FLIP mRNA level was decreased in p53 positive group compared with p53 negative cancer tissues (0.59+/-0.13 vs 0.69+/-0.14, P<0.01), but c-FLIP protein had a significantly higher level in p53 positive cancer tissues than in negative ones (7.57+/-1.30 vs 6.25+/-1.27, P<0.01). c-FLIP is specially overexpressed in colon cancers and it might contribute to carcinogenesis of normal colonic mucosa. p53 may exert transcriptional upregulation effects on c-FLIP gene and more potent effects on promoting the degradation of c-FLIP protein.


Carotid Ultrasound in Assessing Fluid Responsiveness in Patients with Hypotension and Suspected Sepsis.
We sought to assess whether ultrasound (US) measurements of carotid flow time (CFTc) and carotid blood flow (CBF) predict fluid responsiveness in patients with suspected sepsis. This was a prospective observational study of hypotensive (systolic blood pressure < 90) patients "at risk" for sepsis receiving intravenous fluids (IVF) in the emergency department. US measurements of CFTc and CBF were performed at time zero and upon completion of IVF. All US measurements were repeated after a passive leg raise (PLR) maneuver. Fluid responsiveness was defined as normalization of blood pressure without persistent hypotension or need for vasopressors. A convenience sample of 69 patients was enrolled. The mean age was 65; 49% were female. Fluid responders comprised 52% of the cohort. CFTc values increased significantly with both PLR (P = 0.047) and IVF administration (P = 0.003), but CBF values did not (P = 0.924 and P = 0.064 respectively). Neither absolute CFTc or CBF measures, nor changes in these values with PLR or IVF bolus, predicted fluid responsiveness, mortality, or the need for intensive care unit admission. In patients with suspected sepsis, a fluid challenge resulted in a significant change in CFTc, but not CBF. Neither absolute measurement nor delta measurements with fluid challenge predicted clinical outcomes.


Feedforward Inhibition Conveys Time-Varying Stimulus Information in a Collision Detection Circuit.
Feedforward inhibition is ubiquitous as a motif in the organization of neuronal circuits. During sensory information processing, it is traditionally thought to sharpen the responses and temporal tuning of feedforward excitation onto principal neurons. As it often exhibits complex time-varying activation properties, feedforward inhibition could also convey information used by single neurons to implement dendritic computations on sensory stimulus variables. We investigated this possibility in a collision-detecting neuron of the locust optic lobe that receives both feedforward excitation and inhibition. We identified a small population of neurons mediating feedforward inhibition, with wide visual receptive fields and whose responses depend both on the size and speed of moving stimuli. By studying responses to simulated objects approaching on a collision course, we determined that they jointly encode the angular size of expansion of the stimulus. Feedforward excitation, on the other hand, encodes a function of the angular velocity of expansion and the targeted collision-detecting neuron combines these two variables non-linearly in its firing output. Thus, feedforward inhibition actively contributes to the detailed firing-rate time course of this collision-detecting neuron, a feature critical to the appropriate execution of escape behaviors. These results suggest that feedforward inhibition could similarly convey time-varying stimulus information in other neuronal circuits.


Beyond Structural Bioinformatics for Genomics with Dynamics Characterization of an Expanded KRAS Mutational Landscape.
Current capabilities in genomic sequencing outpace functional interpretations. Our previous work showed that 3D protein structure calculations enhance mechanistic understanding of genetic variation in sequenced tumors and patients with rare diseases. The KRAS GTPase is among the critical genetic factors driving cancer and germline conditions. Because KRAS-altered tumors frequently harbor one of three classic hotspot mutations, nearly all studies have focused on these mutations, leaving significant functional ambiguity across the broader KRAS genomic landscape observed in cancer and non-cancer diseases. Herein, we extend structural bioinformatics with molecular simulations to study an expanded landscape of 86 KRAS mutations. We identify multiple coordinated changes strongly associated with experimentally established KRAS biophysical and biochemical properties. The patterns we observe span hotspot and non-hotspot alterations, which can all dysregulate Switch regions, producing mutation-restricted conformations with different effector binding propensities. We experimentally measured mutation thermostability and identified shared and distinct patterns with simulations. Our results indicate mutation-specific conformations which show potential for future research into how these alterations reverberate into different molecular and cellular functions. The data we present is not predictable using current genomic tools, demonstrating the added functional information derived from molecular simulations for interpreting human genetic variation.


Effect of vasopressin on atrial natriuretic peptide release and renal function in dogs.
To assess the effect of arginine vasopressin (AVP) and 1-deamino-8-D-AVP (DDAVP) on atrial natriuretic peptide (ANP) release, renal water and electrolyte excretion, and cardiovascular function, AVP and DDAVP were administered at a dose of 10 ng.kg-1.min-1 iv for 30 min into anesthetized dogs receiving saline infusion at a rate of 1 ml.kg-1.min-1 (n = 12). In the control study, saline was infused alone (n = 6). AVP potentiated the plasma ANP response to an increase in plasma volume produced by saline infusion, increased mean arterial blood pressure (MAP), and exaggerated the natriuresis and kaliuresis. DDAVP did not potentiate the increase in plasma ANP but enhanced the natriuresis without any rise in MAP. Saline alone increased plasma ANP as well as sodium and potassium excretion with no changes in MAP. Inulin and p-aminohippuric acid clearances did not change during these studies. The results suggest that in hydrated dogs, AVP may increase ANP release and arterial blood pressure via the vasopressor activity of AVP and potentiate the natriuresis and kaliuresis, but the increased ANP may play little role in the natriuresis.


Bait attractiveness changes community metrics in dung beetles (Coleoptera: Scarabaeidae: Scarabaeinae).
Species relative abundance (SRA) is an essential attribute of biotic communities, which can provide an accurate description of community structure. However, the sampling method used may have a direct influence on SRA quantification, since the use of attractants (e.g., baits, light, and pheromones) can introduce additional sources of variation in trap performance. We tested how sampling aided by baits affect community data and therefore alter derived metrics. We tested our hypothesis on dung beetles using data from flight interception traps (FITs) as a baseline to evaluate baited pitfall trap performance. Our objective was to assess the effect of bait attractiveness on estimates of SRA and assemblage metrics when sampled by pitfall traps baited with human feces.Dung beetles were sampled at three terra firme primary forest sites in the Brazilian Amazon. To achieve our objective, we (i) identified species with variable levels of attraction to pitfall baited with human feces; (ii) assessed differences in SRA; and (iii) assessed the effect of bait on the most commonly used diversity metrics derived from relative abundance (Shannon and Simpson indices). We identified species less and highly attracted to the baits used, because most attracted species showed greater relative abundances within baited pitfall traps samples compared with our baseline. Assemblages sampled by baited pitfall traps tend to show lower diversity and higher dominance than those sampled by unbaited FITs. Our findings suggest that for ecological questions focused on species relative abundance, baited pitfall traps may lead to inaccurate conclusions regarding assemblage structure. Although tested on dung beetles, we suggest that the same effect could be observed for other insect taxa that are also sampled with baited traps. We highlight a need for further studies on other groups to elucidate any potential effects of using baits.


Anterior chest wall resection and sternal body wedge for primary chest wall tumour: reconstruction technique with biological meshes and titanium plates.
Chest wall tumours are heterogeneous neoplasms, either primary or metastatic, with a malignancy rate of 50%. Surgical resection is one of the mainstays of the treatment, however, chest wall resections can be particularly challenging depending onto the resection size, site and patient habitus. The surgical strategy should be carefully analysed preoperatively, keeping in mind the need of an oncological radical resection (R0) in accordance to the reconstruction principles elicited by le Roux and Sherma since 1983, which include restoring the chest wall rigidity, preserving pulmonary mechanics, protect the intrathoracic organs, avoiding paradox movements of the chest cavity and, possibly, to reduce the thoracic deformity. In this context, we herewith report our surgical reconstruction technique following an anterior chest wall resection and sternal body wedge for a primary chest wall tumour (chondrosarcoma).


The shaker and shaking-B genes specify elements in the processing of gustatory information in Drosophila melanogaster.
Mutations that affect the physiological properties of neurones or the development of neuronal circuits are likely to have profound effects on sensory and motor pathways. We have examined the effects of mutations in two loci--the Shaker complex and shaking-B-on the taste pathway of Drosophila melanogaster Meigen at the behavioural and electrophysiological levels. The Shaker locus encodes a variety of A-type potassium channels that are likely to be tissue- and stage-specific. Flies containing the different Shaker alleles examined exhibit a variety of defects in their gustatory responses to sucrose, NaCl and KCl. The firing patterns of the labellar chemosensory neurones in response to these stimuli are normal. This suggests that the channels encoded by the Shaker locus are probably not involved in taste transduction, but affect central gustatory circuits. The shaking-B locus affects neuronal connectivity, though its molecular nature is unknown. Mutants at this locus show increased thresholds for detection of sucrose and fructose and lack the attraction response to 0.1 mol l-1 sodium chloride that is exhibited by the wild-type fly.


Comparison of thermosensitive alleles of the CDC25 gene involved in the cAMP metabolism of Saccharomyces cerevisiae.
The CDC25 gene from Saccharomyces cerevisiae is an essential component of the RAS-adenylate cyclase pathway. Genetic and biochemical evidence has led to the proposal that the gene product may act upstream of RAS, possibly as a guanine nucleotide exchange factor. We report here the cloning, sequencing and characterization of four mutations in the CDC25 gene. All four are missense mutations which reside within the carboxy-terminal quarter of the single open reading frame found within the gene. Three of the four are missense mutations in the same amino acid codon. A search of protein data bases reveals that the carboxy terminus of the putative CDC25 gene product is similar to that of LTE1, a gene required for growth at low temperature and SCD25, a suppressor of cdc25. Taken together these data indicate that the carboxy terminus of CDC25 plays a critical role in the function of the CDC25 gene product and that other proteins, such as LTE1 or SCD25, may have related activities.


Rapid confirmation of Listeria monocytogenes isolated from foods by a colony blot assay using a digoxigenin-labeled synthetic oligonucleotide probe.
An oligodeoxyribonucleotide probe based on the sequence of a 321-bp internal fragment of the msp gene encoding a major secreted polypeptide of Listeria monocytogenes was labeled with digoxigenin by using terminal deoxynucleotidyl transferase. The specificity of the digoxigenin-labeled probe was determined by dot blot assays. The probe reacted with all strains of L. monocytogenes tested (12 of 12 strains representing five serotypes). The probe did not react with any other Listeria species or with other gram-positive bacteria (Brochothrix, Erysipelothrix, Corynebacterium, Rhodococcus, Lactobacillus, Leuconostoc, Bacillus, Staphylococcus, and Streptococcus). The probe was used to develop a colony blot assay for the rapid confirmation of L. monocytogenes on Listeria-selective agars which had been streaked with food enrichment cultures. Forty-eight food samples were tested by conventional culture and DNA colony blot assay. The sensitivity and specificity of the DNA colony blot were 100 and 97%, respectively.


Metalloprotein structures at ambient conditions and in real-time: biological crystallography and spectroscopy using X-ray free electron lasers.
Although the structure of enzymes and the chemistry at the catalytic sites have been studied intensively, an understanding of the atomic-scale chemistry requires a new approach beyond steady state X-ray crystallography and X-ray spectroscopy at cryogenic temperatures. Following the dynamic changes in the geometric and electronic structure of metallo-enzymes at ambient conditions, while overcoming the severe X-ray-induced changes to the redox active catalytic center, is key for deriving reaction mechanisms. Such studies become possible by the intense and ultra-short femtosecond (fs) X-ray pulses from an X-ray free electron laser (XFEL) by acquiring a signal before the sample is destroyed. This review describes the recent and pioneering uses of XFELs to study the protein structure and dynamics of metallo-enzymes using crystallography and scattering, as well as the chemical structure and dynamics of the catalytic complexes (charge, spin, and covalency) using spectroscopy during the reaction to understand the electron-transfer processes and elucidate the mechanism.


Effects of HIV infection on pregnancy. A clinical and immunologic evaluation.
To understand better the effects of HIV infection on pregnancy, a group of 16 HIV seropositive pregnant patients was evaluated prospectively. Complications and results of pregnancy in these individuals were compared with those of 1,906 births at Charity Hospital, New Orleans during the time of the study. A subgroup of five patients underwent immunologic testing prenatally and postpartum. Thirty-one percent of HIV seropositive subjects had premature labor and low birth weight infants. Only 3.1% of the controls had premature labor and low birth weight infants (p less than or equal to .05). The percentage of C-sections was lower in the study group (12.5% versus 31.3%, p less than .10). All patients remained free of HIV-related disease through the postpartum period. Mean CD4 percentages were lower postpartum (21.9 +/- 3.4) than prepartum (45.1 +/- 6.7) in all five patients studied, although proliferative responses to the mitogens phytohemagglutinin and pokeweed were unchanged during the study period. One of the five seropositive patients who underwent immunologic testing with a CD4:CD8 ratio of less than 1.0 prepartum, had a progressive fall in CD4 percentages from 30% to 11% during the course of the study. HIV-I core antigen was not detected in the serum of any of the five patients during pregnancy or postpartum. Thus, HIV-I infection influenced both the course and outcome of pregnancy in the population studied. Further studies are indicated to assess the effects of pregnancy on the progression of HIV-related immunodeficiency in this population.


Uncovering high-strain rate protection mechanism in nacre.
Under high-strain-rate compression (strain rate approximately 10(3) s(-1)), nacre (mother-of-pearl) exhibits surprisingly high fracture strength vis-à-vis under quasi-static loading (strain rate 10(-3) s(-1)). Nevertheless, the underlying mechanism responsible for such sharply different behaviors in these two loading modes remains completely unknown. Here we report a new deformation mechanism, adopted by nacre, the best-ever natural armor material, to protect itself against predatory penetrating impacts. It involves the emission of partial dislocations and the onset of deformation twinning that operate in a well-concerted manner to contribute to the increased high-strain-rate fracture strength of nacre. Our findings unveil that Mother Nature delicately uses an ingenious strain-rate-dependent stiffening mechanism with a purpose to fight against foreign attacks. These findings should serve as critical design guidelines for developing engineered body armor materials.


Retinoic acid receptor/retinoid X receptor heterodimers can be activated through both subunits providing a basis for synergistic transactivation and cellular differentiation.
The receptor for 9-cis-retinoic acid, retinoid X receptor (RXR), forms heterodimers with several nuclear receptors, including the receptor for all-trans-retinoic acid, RAR. Previous studies have shown that retinoic acid receptor can be activated in RAR/RXR heterodimers, whereas RXR is believed to be a silent co-factor. In this report we show that efficient growth arrest and differentiation of the human monocytic cell line U-937 require activation of both RAR and RXR. Also, we demonstrate that the allosteric inhibition of RXR is not obligatory and that RXR can be activated in the RAR/RXR heterodimer in the presence of RAR ligands. Remarkably, RXR inhibition by RAR can also be relieved by an RAR antagonist. Moreover, the dose response of RXR agonists differ between RXR homodimers and RAR/RXR heterodimers, indicating that these complexes are pharmacologically distinct. Finally, the AF2 activation domain of both subunits contribute to activation even if only one of the receptors is associated with ligand. Our data emphasize the importance of signaling through both subunits of a heterodimer in the physiological response to retinoids and show that the activity of RXR is dependent on both the identity and the ligand binding state of its partner.


The Antibiotic Resistome: A Guide for the Discovery of Natural Products as Antimicrobial Agents.
The use of life-saving antibiotics has long been plagued by the ability of pathogenic bacteria to acquire and develop an array of antibiotic resistance mechanisms. The sum of these resistance mechanisms, the antibiotic resistome, is a formidable threat to antibiotic discovery, development, and use. The study and understanding of the molecular mechanisms in the resistome provide the basis for traditional approaches to combat resistance, including semisynthetic modification of naturally occurring antibiotic scaffolds, the development of adjuvant therapies that overcome resistance mechanisms, and the total synthesis of new antibiotics and their analogues. Using two major classes of antibiotics, the aminoglycosides and tetracyclines as case studies, we review the success and limitations of these strategies when used to combat the many forms of resistance that have emerged toward natural product-based antibiotics specifically. Furthermore, we discuss the use of the resistome as a guide for the genomics-driven discovery of novel antimicrobials, which are essential to combat the growing number of emerging pathogens that are resistant to even the newest approved therapies.


Sparse multi-shell diffusion imaging.
Diffusion magnetic resonance imaging (dMRI) is an important tool that allows non-invasive investigation of neural architecture of the brain. The data obtained from these in-vivo scans provides important information about the integrity and connectivity of neural fiber bundles in the brain. A multi-shell imaging (MSI) scan can be of great value in the study of several psychiatric and neurological disorders, yet its usability has been limited due to the long acquisition times required. A typical MSI scan involves acquiring a large number of gradient directions for the 2 (or more) spherical shells (several b-values), making the acquisition time significantly long for clinical application. In this work, we propose to use results from the theory of compressive sampling and determine the minimum number of gradient directions required to attain signal reconstruction similar to a traditional MSI scan. In particular, we propose a generalization of the single shell spherical ridgelets basis for sparse representation of multi shell signals. We demonstrate its efficacy on several synthetic and in-vivo data sets and perform quantitative comparisons with solid spherical harmonics based representation. Our preliminary results show that around 20-24 directions per shell are enough for robustly recovering the diffusion propagator.


Intermediate filaments in alpha-keratins.
Previous x-ray diffraction studies on the alpha-keratins of hair and wool have revealed that the intermediate filaments (IF) have a helical structure rendered imperfect by a precisely defined dislocation. It has also been possible to deduce a surface lattice for the IF and to determine the number of IF molecules associated with each lattice point. In this work this information is combined with data on the ionic interactions between the coiled-coil rope segments of the IF molecules to provide a plausible model for the pattern of interactions that stabilize the framework of the IF in the "hard" alpha-keratins. Similar interaction studies of the proteins from the IF in the so-called "soft" alpha-keratin from the stratum corneum layer of the skin suggest that they are likely to have an essentially similar pattern.


Attenuated nuclear shrinkage in neurons with nuclear aggregates--a morphometric study on pontine neurons of Machado-Joseph disease brains.
Nuclear aggregates (NAs) and neurodegeneration in brains from patients with Machado-Joseph disease (MJD) are both triggered by pathological expansion of CAG/polyglutamine repeat in ataxin-3, but it remains to be clarified whether NA formation is associated with accelerated neurodegeneration. In an attempt to clarify a possible influence of NAs on neurons in human brains, we quantified the size and deformity of neuronal nuclei (those with or without NAs, separately) cross-sectioned on pontine preparations of autopsied brains from four patients with MJD and five controls. Nuclear shrinkage and deformity were more marked in MJD brains than in controls, and these changes were attenuated in neurons harboring NAs. NAs of MJD are presumably linked to a mechanism that attenuates rather than accelerates nuclear shrinkage and deformity. This finding leads us to consider that NAs are not necessarily toxic to neurons in diseased human brains.


Antibiotics and nephrostomy tube care: preliminary observations. Part I. Bacteriuria.
The authors sought to establish whether bacteriuria or candiduria would develop in patients with a noninfected, obstructed renal collecting system after placement of a percutaneous nephrostomy tube. Tubes were placed in 27 sterile systems in 25 patients. Urine was cultured at 7-10-day intervals, and urinalysis was performed. Bacteriuria and pyuria were defined on the basis of laboratory values of greater than or equal to 10(4) colony-forming units per milliliter and greater than or equal to five white blood cells per high-powered field, respectively. Twenty-two systems were followed until all demonstrated multiple organisms. No clinical symptoms were related to bacteriuria.


A Ferrous Iron Exporter Mediates Iron Resistance in Shewanella oneidensis MR-1.
Shewanella oneidensis strain MR-1 is a dissimilatory metal-reducing bacterium frequently found in aquatic sediments. In the absence of oxygen, S. oneidensis can respire extracellular, insoluble oxidized metals, such as iron (hydr)oxides, making it intimately involved in environmental metal and nutrient cycling. The reduction of ferric iron (Fe(3+)) results in the production of ferrous iron (Fe(2+)) ions, which remain soluble under certain conditions and are toxic to cells at higher concentrations. We have identified an inner membrane protein in S. oneidensis, encoded by the gene SO_4475 and here called FeoE, which is important for survival during anaerobic iron respiration. FeoE, a member of the cation diffusion facilitator (CDF) protein family, functions to export excess Fe(2+) from the MR-1 cytoplasm. Mutants lacking feoE exhibit an increased sensitivity to Fe(2+). The export function of FeoE is specific for Fe(2+), as an feoE mutant is equally sensitive to other metal ions known to be substrates of other CDF proteins (Cd(2+), Co(2+), Cu(2+), Mn(2+), Ni(2+), or Zn(2+)). The substrate specificity of FeoE differs from that of FieF, the Escherichia coli homolog of FeoE, which has been reported to be a Cd(2+)/Zn(2+) or Fe(2+)/Zn(2+) exporter. A complemented feoE mutant has an increased growth rate in the presence of excess Fe(2+) compared to that of the ΔfeoE mutant complemented with fieF. It is possible that FeoE has evolved to become an efficient and specific Fe(2+) exporter in response to the high levels of iron often present in the types of environmental niches in which Shewanella species can be found.


[The MB1 family of repeats in clones from the genomes of mammals].
Members of MB1 family repeats are revealed in genomes of many mammals (cow, rabbit, opossum, horse, ...). The MB1 repeats from cow and rabbit genomes are mirror-reflected about the SINE families repeats from cow and rabbit genomes. The life time of MB1 repeats are no less than 100 million years. Classification of MB1 repeats from human genome using the information similarity was performed. This classification has revealed two subfamily MB1 repeats in human genome. Possible processes of creation of MB1 family repeats common for many mammals are discussed.


Current concepts in olfaction.
Because of the location of the olfactory epithelium and bulb, the processes involved in olfaction have been difficult to evaluate. However, the recent application of comprehensive nasal endoscopy, reproducible olfactory testing, and molecular biologic techniques to the study of the transduction of olfactory information has elucidated some of the events in odorant perception.


Static in vitro digestion model adapted to the general older adult population: an INFOGEST international consensus.
Understanding the mechanisms of food digestion is of paramount importance to determine the effect foods have on human health. Significant knowledge on the fate of food during digestion has been generated in healthy adults due to the development of physiologically-relevant in vitro digestion models. However, it appears that the performance of the oro-gastrointestinal tract is affected by ageing and that a model simulating the digestive conditions found in a younger adult (<65 years) is not relevant for an older adult (>65 years). The objectives of the present paper were: (1) to conduct an exhaustive literature search to find data on the physiological parameters of the older adult oro-gastrointestinal tract, (2) to define the parameters of an in vitro digestion model adapted to the older adult. International experts have discussed all the parameters during a dedicated workshop organized within the INFOGEST network. Data on food bolus properties collected in the older adult were gathered, including food particle size found in older adult boluses. In the stomach and small intestine, data suggest that significant physiological changes are observed between younger and older adults. In the latter, the rate of gastric emptying is slowed down, the pH of the stomach content is higher, the amount of secretions and thus the hydrolytic activities of gastric and intestinal digestive enzymes are reduced and the concentration of bile salts lower. The consensus in vitro digestion model of the older adult proposed here will allow significant progress to be made in understanding the fate of food in this specific population, facilitating the development of foods adapted to their nutritional needs. Nevertheless, better foundational data when available and further refinement of the parameters will be needed to implement the proposed model in the future.


Removal and transformation of dissolved organic matter (DOM) during the treatment of partially stabilized leachate in membrane bioreactor.
The removal of dissolved organic matter (DOM) during municipal solid waste leachate treatment in a membrane bioreactor (MBR) was investigated. The MBR was operated with fouled and cleaned membranes, at a time. DOM, mixed liquor, and effluent were characterized by fractionation and fluorescence excitation-emission matrix (EEM) spectroscopy. Results showed DOM removals of 82% during the MBR operation with the fouled membrane. The DOM characterization indicated that most of the hydrophilic compounds and protein-like substances could be significantly removed by microbial activities in the treatment processes. Furthermore partial removal or transformation of organic substances was observed during membrane filtration. The fouled membrane improved DOM rejection by only about 2% when compared to the cleaned membrane, which demonstrated the significance of irreversible foulants on DOM rejection. There were insignificant differences in DOM fractionation and EEM fluorescence spectra between water samples filtered through the fouled and cleaned membranes.


The stomatal fates: Understanding initiation and enforcement of stomatal cell fate transitions.
In the stomatal lineage, repeated arcs of initiation, stem-cell proliferation, and terminal cell fate commitment are displayed on the surface of aerial organs. Over the past two decades, the core transcription and signaling elements that guide cell divisions, patterning, and fate transitions were defined. Here we highlight recent work that extends the core using a variety of cutting-edge techniques in different plant species. New work has discovered transcriptional circuits that initiate and reinforce stomatal fate transitions, while also enabling the lineage to interpret and respond to environmental inputs. Recent developments show that some key stomatal factors are more flexible or potentially even interchangeable, opening up avenues to explore stomatal fates and regulatory networks.


Localized immunoglobulin light chain amyloidosis: Novel insights including prognostic factors for local progression.
In localized light chain amyloidosis (locAL), amyloidogenic light chains (aLC) are produced and deposited locally by a B-cell clone. We present 293 patients with immunohistochemically confirmed locAL. Lung (nodular pulmonary) with 63 patients was the most involved organ. The aLC was λ in 217 cases (κ:λ ratio 1:3). A local B-cell clone was identified in 30% of cases. Sixty-one (21%) had a concomitant autoimmune disorder (cAD). A monoclonal component (MC) were present in 101 (34%) patients and were more frequent in subjects with cAD (51% vs 34%; P = .03). Cigarette smoking was more prevalent in lung locAL (54% vs 37%; P = .018). After a median follow-up of 44 months, 16 patients died and 5- and 10-years locAL progression-free survival (PFS) were 62% and 44%. Interestingly, locAL-PFS was shorter among patients with an identified clonal infiltrate at amyloid deposition site (40 vs 109 months; P = .02) and multinuclear giant cells and/or an inflammatory infiltrate resulted in longer locAL-PFS in lung involvement (65 vs 42 months; P = .01). However, no differences in locAL PFS were observed in patients with cAD, a MC and involved organ site. Treatment was administered in 163 (54%) patients and was surgical in 135 (46%). Median locAL-PFS after first treatment was 56 months. Responders had longer locAL-PFS (78 vs 17 months; P < .001). Three patients with lung locAL and a MC were diagnosed as systemic AL amyloidosis at follow-up. In summary, locAL pathogenesis seems to be heterogeneous and the clonal infiltrate leads local progression.


Ensuring Access to Safe, Legal Abortion in an Increasingly Complex Regulatory Environment.
Restrictions on access to abortion in the United States have reached proportions unprecedented since the nationwide legalization of abortion in 1973. Although some restrictions aim to discourage women from having abortions, many others impede access by affecting the timeliness, affordability, or availability of services. Evidence indicates that these restrictions do not increase abortion safety; rather, they create logistic barriers for women seeking abortion, and they have the greatest effect on women with the fewest resources. In this commentary, we recall the important role that obstetrician-gynecologists (ob-gyns) have played, both before and after Roe v. Wade, in facilitating access to safe abortion care. Using the literature on abortion safety and access as a foundation, we propose several practical ideas about what we as ob-gyns can do to address the current threat to abortion access, whether or not we provide abortion services in practice. We hope that this commentary will encourage discourse within our profession and prompt other suggestions. As ob-gyns who are dedicated to addressing health disparities and promoting the health and well-being of our patients, we can make a difference.


Determinants of condom use among youth in Madagascar.
The objective of this study was to identify the key determinants of condom use with regular and casual partners among youth in Madagascar. Data stem from a reproductive health survey conducted in October-December 2000 among a representative sample of 2440 youth aged 15-24 living in Toamasina province. Following theoretical models of behaviour change, logistic regression was used to assess the effect of AIDS awareness,personal risk perception, condom access, perceived condom effectiveness,self-efficacy and social support on condom use. Among sexually experienced youth, only about four in ten males and two in ten females have ever used condoms. Fewer than 15% of youth used a condom in last intercourse with their regular partner. Whether youth will try condoms appears to depend largely on the perceived effectiveness of condoms for family planning, access to a nearby condom source, parental support for condom use, and patterns of risky sexual behaviour. Young males' likelihood of using a condom with a regular partner increases significantly if they perceive condoms to be effective for family planning (OR=11.4; p=0.019). For females, it increases with level of self-efficacy (OR=2 1; p=0.042) and having discussed HIV prevention with someone in the last year (OR=2.8; p=0.022). Among males,condom use with casual partners is significantly higher among those who perceive themselves to be at high risk of sexually transmitted infections(OR=2.3; p=0.014), who believe condoms are effective for family planning(OR=2.8; p=0.048), who have good access to condoms (OR=2.9; p=0.002)and who perceive their parents support condom use (OR=1-7; p=0.048). In conclusion, very few youth in Toamasina are using condoms, highlighting the need to continue and expand adolescent reproductive health interventions. In this low HIV prevalence setting, it is important for these programmes to emphasize that condoms are effective for both pregnancy prevention and STI/HIV prevention.


Enriching the care of patients with dementia in acute settings? The Dementia Champions Programme in Scotland.
Admission to hospital has been found to have a negative impact on people with dementia. The Scottish Dementia Champions programme was developed to prepare health and social service Dementia Champions working in acute settings as Change Agents. The programme was initially delivered to a cohort of 100 health professionals via blended learning, and comprised five study days, a half day spent in a local community setting, and e-learning. In order to complete the programme and graduate, participants were required to complete and submit reports relating to three work-based activities. The evaluation of the project adopted a two-pronged approach: Impact on programme participants was assessed by scores derived from the Approaches to Dementia Questionnaire (ADQ) (Lintern, 1996) completed at Study Days 1 and 5, and analysis of qualitative data derived from the three written assignments. Participants were asked to evaluate course materials and input for each of the five study days, as well as satisfaction with delivery. Analysis of data derived from the ADQ and 100 reflective reports of the community experience indicate that participants' perceptions of people with dementia shifted significantly during the Programme. Participants identified a range of issues which should be addressed with a view to improving the experiences of people with dementia in acute settings, and put in place actions to bring about change. The format of the programme provided a cost effective means to prepare NHS and Social Service Dementia Champions as Change Agents for practice within a relatively short period of time, and would be transferrable to other staff groups as well as different organisational structures in other countries.


TGF-β/NF1/Smad4-mediated suppression of ANT2 contributes to oxidative stress in cellular senescence.
Oxidative stress and persistent activation of DNA damage response (DDR) are causally involved in the development of cellular senescence, a phenomenon implicated in fundamental (patho)physiological processes such as aging, fetal development and tumorigenesis. Here, we report that adenine nucleotide translocase-2 (ANT2) is consistently down-regulated in all three major forms of cellular senescence: replicative, oncogene-induced and drug-induced, in both normal and cancerous human cells. We previously reported formation of novel NF1/Smad transcription repressor complexes in growth-arrested fibroblasts. Here we show that such complexes form in senescent cells. Mechanistically, binding of the NF1/Smad complexes to the NF1-dependent repressor elements in the ANT2 gene promoter repressed ANT2 expression. Etoposide-induced formation of these complexes and repression of ANT2 were relatively late events co-incident with production and secretion of, and dependent on, TGF-β. siRNA-mediated knock-down of ANT2 in proliferating cells resulted in increased levels of reactive oxygen species (ROS) and activation of the DDR. Knock-down of ANT2, together with etoposide treatment, further intensified ROS production and DNA damage signaling, leading to enhanced apoptosis. Together, our data show that TGF-β-mediated suppression of ANT2 through NF1/Smad4 complexes contributes to oxidative stress and DNA damage during induction of cellular senescence.


New human microvascular endothelial cell lines with specific adhesion molecules phenotypes.
Vascular endothelial cells recognize blood-borne circulating cells and allow them to extravasate in a tissue-specific manner. Because this property determines the selectivity of lymphocyte homing, it is fundamental in physiological as well as pathological processes (inflammation, autoimmune diseases, metastasis). As a tool to assess the molecular basis of endothelium selectivity, microvascular endothelial cell lines of distinct tissue origin were established. Endothelial cells, isolated from lymphoid tissues (lymph nodes and appendix) and from nonlymphoid immune sites--intestine, lung, and skin--were immortalized in vitro. Their general endothelial characteristics, such as the presence of von Willebrand factor (wWf), angiotensin-converting enzyme (ACE), VE-cadherin, and the intracellular E-selectin, were preserved. This article shows that these cell lines display phenotypic characteristics related to their tissue origin. Hence, endothelial cells from lymph nodes expressed peripheral lymph node addressins (PNAds). Endothelial cells from nonlymphoid tissues were ICAM-1 (intercellular adhesion molecule-1) and CD49e positive, whereas P-selectin was not equally distributed among the cell lines. Endothelial cells from mucosal sites reacted with antibody against human MAdCAM-1 (mucosal addressin cell adhesion molecule). In the adhesion test, lymphoid and myeloid cells adhere to endothelial cell lines in a distinct manner. These lines could be useful to study molecular mechanisms involved in tissue-specific cell-cell interaction.


De novo whole-genome assembly in Chrysanthemum seticuspe, a model species of Chrysanthemums, and its application to genetic and gene discovery analysis.
Cultivated chrysanthemum (Chrysanthemum morifolium Ramat.) is one of the most economically important ornamental crops grown worldwide. It has a complex hexaploid genome (2n = 6x = 54) and large genome size. The diploid Chrysanthemum seticuspe is often used as a model of cultivated chrysanthemum, since the two species are closely related. To expand our knowledge of the cultivated chrysanthemum, we here performed de novo whole-genome assembly in C. seticuspe using the Illumina sequencing platform. XMRS10, a C. seticuspe accession developed by five generations of self-crossing from a self-compatible strain, AEV2, was used for genome sequencing. The 2.72 Gb of assembled sequences (CSE_r1.0), consisting of 354,212 scaffolds, covered 89.0% of the 3.06 Gb C. seticuspe genome estimated by k-mer analysis. The N50 length of scaffolds was 44,741 bp. For protein-encoding genes, 71,057 annotated genes were deduced (CSE_r1.1_cds). Next, based on the assembled genome sequences, we performed linkage map construction, gene discovery and comparative analyses for C. seticuspe and cultivated chrysanthemum. The generated C. seticuspe linkage map revealed skewed regions in segregation on the AEV2 genome. In gene discovery analysis, candidate flowering-related genes were newly found in CSE_r1.1_cds. Moreover, single nucleotide polymorphism identification and annotation on the C. × morifolium genome showed that the C. seticuspe genome was applicable to genetic analysis in cultivated chrysanthemums. The genome sequences assembled herein are expected to contribute to future chrysanthemum studies. In addition, our approach demonstrated the usefulness of short-read genome assembly and the importance of choosing an appropriate next genome sequencing technology based on the purpose of the post-genome analysis.


Prostatic localization of spontaneous early invasive carcinoma in Lobund-Wistar rats.
Animal models of human prostate cancer are very limited in number but are of obvious importance to develop. Dr. Morris Pollard (M. Pollard, J. Natl. Cancer Inst., 51: 1235-1241, 1973) has reported that Lobund-Wistar rats develop spontaneous metastatic prostatic cancer when they become old (approximately 25% incidence after 25 months). A chemically induced form of the disease has also been described in Lobund-Wistar rats. However, recent reports suggest that most of the chemically induced adenocarcinomas are not prostatic in origin, with most arising in the seminal vesicle, and thereby raise questions about the origin of the spontaneous cancers. We herein report cancer spontaneously arising in the lateral lobes of the prostates in Lobund-Wistar rats. One of 8 rats killed at 16 months of age showed prostatic carcinoma in situ. Two of 39 rats killed at 20 months displayed early invasive adenocarcinomas with no signs of metastases. Because sectioning of the prostates in this study was limited to face sections from a single block for each rat, it is highly probable that the true incidence of dysplasias and carcinomas is underestimated by these data. Dysplastic or neoplastic changes were not seen in either the seminal vesicles or other portions of the prostatic complex. The nuclei of adenocarcinoma cells showed less labeling with antibody to the androgen hormone receptor than did normal cells. These data strongly support the validity of the Pollard model of spontaneous prostate cancer in Lobund-Wistar rats.


The physician's response to climate change.
Climate change will have an effect on the health and well-being of the populations cared for by practicing physicians. The anticipated medical effects include heat- and cold-related deaths, cardiovascular illnesses, injuries and mental harms from extreme weather events, respiratory illnesses caused by poor air quality, infectious diseases that emanate from contaminated food, water, or spread of disease vectors, the injuries caused by natural disasters, and the mental harm associated with social disruption. Within several years, such medical problems are likely to reach the doorsteps of many physicians. In the face of this reality, physicians should assume their traditional roles as medical professionals, health educators, and community leaders. Clinicians provide individual health services to patients, some of whom will be especially vulnerable to the emerging health consequences of global warming. Physicians also work in academic medical institutions and hospitals that educate and provide continuing medical education to students, residents, and practitioners. The institutions also produce a measurable carbon footprint. Societies of physicians at national, state, and local levels can choose to use their well-developed avenues of communication to raise awareness of the key issues that are raised by climate change as well as other environmental concerns that have profound implications for human health and well-being.


Effects of intron length on differential processing of mouse mu heavy-chain mRNA.
Production of membrane-bound and secreted forms of mouse mu heavy-chain mRNA is controlled by differential processing in a developmental-stage-specific manner. We have analyzed the effects of various deletions and insertions in the C4-M1 intron of the mouse mu gene on the differential processing of mu mRNA. We show that there is a correlation between the length of the C4-M1 intron and the molar ratio of membrane-bound to secreted mu mRNAs, i.e., the shorter the C4-M1 intron, the higher the ratio. Since the poly(A) addition signal in the C4-M1 intron seems to be intact in the mutant mu genes, it is likely that the efficiency of splicing of the C4-M1 intron is affected by changes in the intron length.


Secondary organic aerosol formation from in-use motor vehicle emissions using a potential aerosol mass reactor.
Secondary organic aerosol (SOA) formation from in-use vehicle emissions was investigated using a potential aerosol mass (PAM) flow reactor deployed in a highway tunnel in Pittsburgh, Pennsylvania. Experiments consisted of passing exhaust-dominated tunnel air through a PAM reactor over integrated hydroxyl radical (OH) exposures ranging from ∼ 0.3 to 9.3 days of equivalent atmospheric oxidation. Experiments were performed during heavy traffic periods when the fleet was at least 80% light-duty gasoline vehicles on a fuel-consumption basis. The peak SOA production occurred after 2-3 days of equivalent atmospheric oxidation. Additional OH exposure decreased the SOA production presumably due to a shift from functionalization to fragmentation dominated reaction mechanisms. Photo-oxidation also produced substantial ammonium nitrate, often exceeding the mass of SOA. Analysis with an SOA model highlight that unspeciated organics (i.e., unresolved complex mixture) are a very important class of precursors and that multigenerational processing of both gases and particles is important at longer time scales. The chemical evolution of the organic aerosol inside the PAM reactor appears to be similar to that observed in the atmosphere. The mass spectrum of the unoxidized primary organic aerosol closely resembles ambient hydrocarbon-like organic aerosol (HOA). After aging the exhaust equivalent to a few hours of atmospheric oxidation, the organic aerosol most closely resembles semivolatile oxygenated organic aerosol (SV-OOA) and then low-volatility organic aerosol (LV-OOA) at higher OH exposures. Scaling the data suggests that mobile sources contribute ∼ 2.9 ± 1.6 Tg SOA yr(-1) in the United States, which is a factor of 6 greater than all mobile source particulate matter emissions reported by the National Emissions Inventory. This highlights the important contribution of SOA formation from vehicle exhaust to ambient particulate matter concentrations in urban areas.


Working through mass incarceration: gender and the politics of prison labor from east to west.
This article explores the politics and practices of labor in two penal institutions for women: a maximum security facility for women in Hungary and a community‐based facility for women in California. Diverging from other accounts of imprisonment that tend to operate at either the individual or macroeconomic level, this article analyzes the concrete institutional relations of prison and complicates the assumption that they simply reflect the logic of the prison‐industrial complex. Based on years of ethnographic work in two very different penal systems, I describe variation in how prisons institute labor within and across institutions and cultures: the Hungarian facility positioned wage labor as a right and an obligation that formed the basis of women's social relationships and ties to others, while the U.S. prison excluded wage labor from women's lives so they could get on with the work of self‐improvement and personal healing. From the comparison, I reveal how prisons can both draw on and subvert broader social meanings assigned to women's work, making it difficult to view prison labor as wholly exploitative or abusive. I also argue that refusing to allow female inmates to engage in wage labor can be a more profound form of punishment than requiring it of them. By juxtaposing the discourses and practices of work in two very different penal contexts, this article offers a critical reflection on the political economy of prison labor from the ground up.


Daily treatment with diazepam differentially modifies sensitivity to the effects of gamma-aminobutyric acid(A) modulators on schedule-controlled responding in rhesus monkeys.
The present study examined how daily treatment with the benzodiazepine (BZ) diazepam modifies the effects of positive modulators acting at different sites on the gamma-aminobutyric acid(A) (GABA(A)) receptor complex and negative modulators acting at BZ sites on the receptor complex. GABA(A) modulators were administered alone or in combination with acute or chronic diazepam to rhesus monkeys (n = 4) responding under a multiple fixed ratio (FR/FR) schedule of food presentation and stimulus-shock termination (SST). There was mutual antagonism between the rate-decreasing effects of diazepam (5.6 mg/kg, p.o.) and high efficacy BZ site negative modulators [ethyl beta-carboline-3-carboxylate (beta-CCE), methyl beta-carboline-3-carboxylate (beta-CCM) and methyl-6,7-dimethoxyl-4-ethyl-beta-carboline-3-carboxylate (DMCM)]. Antagonism of beta-CCE, beta-CCM, and DMCM by diazepam was markedly reduced in monkeys receiving diazepam daily. In contrast, daily diazepam treatment enhanced the rate-decreasing effects of Ro 15-4513 (ethyl 8-azido-6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5-alpha]-[1,4]benzodiazepine-3-carboxylate) and flumazenil. Chronic diazepam elicited cross-tolerance to the BZ triazolam and not to the barbiturate pentobarbital or the neuroactive steroid pregnanolone. These results suggest that tolerance to the rate-decreasing effects of BZs is not accompanied by cross-tolerance to positive GABA(A) modulators acting at other sites on the receptor complex. Moreover, changes in sensitivity to negative GABA(A) modulators during chronic diazepam treatment appeared to be related to negative efficacy and not clearly related to the precipitation of withdrawal for all drugs. These results indicate that changes in sensitivity to the behavioral effects of drugs that act at different sites on the GABA(A) receptor complex might be especially useful for identifying and characterizing the functional consequences of GABA(A) receptor heterogeneity.


Correlates of sedentary behavior in middle-aged and old age people with mild cognitive impairment: a multinational study.
ABSTRACTObjective:Despite the benefits of being active for people with mild cognitive impairment (MCI) on cognition and the acknowledgement that MCI is a critical period for intervening to prevent dementia, little is known about the actual sedentary levels in people with MCI. This study investigates correlates of sedentary behavior (SB) in people with MCI. This was a cross-sectional study. Data from the World Health Organization's Study on Global Ageing and Adult Health were analyzed. Individuals aged ≥50 years with MCI were included. SB was assessed by the Global Physical Activity Questionnaire. Associations between SB levels and the correlates were examined using multivariable linear and logistic regressions. 4,082 individuals aged ≥50 years with MCI (64.4 ± 17.0 years; 55.1% female) were included. The prevalence of high SB (i.e., ≥8 hours/day) was 14.0% (95%CI = 12.2%-16.0%), while the time spent sedentary was 262 ± 290 minutes/day. Correlates significantly associated with being sedentary ≥8 hours/day and time spent sedentary per day were, older age, being unemployed, depression, sleep problems, obesity (vs. normal weight), diabetes, stroke, poor self-rated health, and lower levels of social cohesion. Future research exploring interventions to reduce SB in people with MCI should target the identified sociodemographic and mental and physical health correlates, while the promotion of social cohesion may have the potential to increase the efficacy of future public health initiatives.


Gestational diabetes mellitus in India.
Glucose intolerance during pregnancy predisposes the offspring for increased risk of developing glucose intolerance in the future. This vicious cycle is likely to influence and perpetuate the incidence and prevalence of glucose intolerance in any population. No data is available about the prevalence of glucose intolerance during pregnancy in our country and hence a study was undertaken on this aspect. This study was performed in the antenatal clinic of Government Maternity Hospital, Chennai, India. As a pregnant woman in second or third trimester checks into the antenatal clinic, she was given 50 gm oral glucose load and blood sample was collected after one hour. This test was performed on 1251 pregnant women. They were requested to come after 72 hours for the 75 gm OGTT recommended by WHO. Among the 1251 women, 891 responded. The blood sample was taken in the fasting state and at 2 hours after 75 gm of oral glucose. Diagnosis was based on the WHO criteria for gestational diabetes mellitus (GDM). The mean age of these pregnant women was 23+/-4 years. There was a significant increase in the prevalence of GDM in relation to gravida. The effect of BMI did not quite reach statistical significance (chi2 (df=1) = 3.659, P = 0.055), but a model of linear trend was significant. Of the 1251 women who underwent the 50 gm oral glucose challenge test, 670 (53.55%) had one hour plasma glucose > or = 130 mg/dl. Among the 891 pregnant women who had 75 gms OGTT, 168 (18.9%) were diagnosed as GDM, taking both FPG > or = 126 mg/dl and/or 2 hr PPG > or = 140 mg/dl as cut-off values. Taking only 2 hr plasma glucose for analysis, 144 (16.2%) had a value > or = 140 mg/dl. A similar study was conducted in different parts of the country taking only the 2 hr 75 gm post-glucose value of > or = 140 mg/dl as diagnostic criteria for GDM. Of the total number of pregnant women (n = 3674) screened, 16.55% of them found to have GDM. Our study has documented the increased prevalence of GDM in our population necessitating universal screening for glucose intolerance in pregnancy. Using 2 hr plasma glucose > or = 140 mg/dl as a one step procedure is simple and economical, particularly for the countries ethnically more prone to high prevalence of diabetes.


Value of brominated fluorocarbons for the radiographic diagnosis of small-bowel obstruction: comparison with other contrast agents in rats.
The disadvantages of water-soluble gastrointestinal contrast agents include high osmolality, contrast dilution, and severe toxicity if aspirated. Perfluorocarbons are nontoxic in the lung and peritoneal cavity. Because perfluorocarbons are immiscible with water, they have no osmotic effect and cannot be diluted. Because these properties offer theoretical advantages over traditional gastrointestinal contrast agents, we compared two perfluorocarbons with barium and ionic and nonionic iodinated contrast material in a rat model of small-bowel obstruction. Twelve groups of six rats each had ligation of the terminal ileum (obstruction model) or of the terminal ileum and mesenteric artery (obstruction with ischemia model). Each rat received 3 ml of barium, meglumine sodium diatrizoate, iohexol, neat perfluorooctyl bromide, neat perfluorohexyl bromide, or saline (control animals). Contrast media were given at the recommended concentrations, and their progression was evaluated on serial radiographs by an observer who was not aware of the model or the contrast medium given. When one contrast material reached the point of obstruction, all rats in the group were sacrificed and a final radiograph was obtained. Three radiologists, who were not aware of the contrast medium given, on two separate occasions independently reviewed the radiographs and ranked the contrast agents for their relative radiopacity, mucosal definition, speed of transit, gastric retention, and bowel distension. When data from both models were combined, perfluorocarbons were judged on the final image to be the most radiodense, to provide the sharpest mucosal detail, to have the least gastric retention, and to have faster progression than barium. Whereas meglumine sodium diatrizoate and iohexol reached the point of obstruction more rapidly than the perfluorocarbons and barium, they had the greatest gastric retention, caused the most bowel distension, and were the least radiopaque. Our results show that the radiopaque perfluorocarbons are suitable as gastrointestinal contrast agents and have favorable radiographic characteristics in this animal model. When these results are combined with the low-toxicity profile of perflubron, clinical evaluation of this agent for the radiographic assessment of bowel obstruction is warranted.


[Correlation between Borg's scale and spirometer in asthmatic patients].
Spirometry is the ideal evidence to diagnose and evaluate the degree of bronchial obstruction which is present in asthma; however, it is not easy for physicians to have access to it during the first contact due to its high cost. The Borg's scale is an analogous visual scale which has an economical and simple use; it evaluates in a subjective way the respiratory difficulty in a patient in a scale from 0 to 10. It is necessary to have the right tools to know the degree of pulmonary obstruction that asthmatic patients have and to be able to give them the right medical treatment. The use of the Borg's scale to evaluate the severity of the asthmatic patients causes controversy. To determine the correlation between the spirometry and the Borg's scale in adult asthmatic patients. One hundred and one cases of male and female asthmatic patients were studied (GINA2002) from the consultation of specialties in Allergy and Clinic Immunology. A spirometry was done to all patients (Multispiro 2000) and the Borg's scale method was applied to them. The project was approved by the local research committee. The SPSS program version 12.0 was used and the Pearson's correlation test was applied. The correlation between the spirometry and the Borg's scale was r = 0.345, with p < 0.05. There is a scarce correlation between the spirometry and the Borg's scale. The result obtained in this study does not support the use of the Borg's scale to appreciate the bronchial obstruction in adult asthmatic patients.


Kainic Acid Induces mTORC1-Dependent Expression of Elmo1 in Hippocampal Neurons.
Epileptogenesis is a process triggered by initial environmental or genetic factors that result in epilepsy and may continue during disease progression. Important parts of this process include changes in transcriptome and the pathological rewiring of neuronal circuits that involves changes in neuronal morphology. Mammalian/mechanistic target of rapamycin (mTOR) is upregulated by proconvulsive drugs, e.g., kainic acid, and is needed for progression of epileptogenesis, but molecular aspects of its contribution are not fully understood. Since mTOR can modulate transcription, we tested if rapamycin, an mTOR complex 1 inhibitor, affects kainic acid-evoked transcriptome changes. Using microarray technology, we showed that rapamycin inhibits the kainic acid-induced expression of multiple functionally heterogeneous genes. We further focused on engulfment and cell motility 1 (Elmo1), which is a modulator of actin dynamics and therefore could contribute to pathological rewiring of neuronal circuits during epileptogenesis. We showed that prolonged overexpression of Elmo1 in cultured hippocampal neurons increased axonal growth, decreased dendritic spine density, and affected their shape. In conclusion, data presented herein show that increased mTORC1 activity in response to kainic acid has no global effect on gene expression. Instead, our findings suggest that mTORC1 inhibition may affect development of epilepsy, by modulating expression of specific subset of genes, including Elmo1, and point to a potential role for Elmo1 in morphological changes that accompany epileptogenesis.


Cystic fibrosis transmembrane conductance regulator regulates epithelial cell response to Aspergillus and resultant pulmonary inflammation.
Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) alter epithelial cell (EC) interactions with multiple microbes, such that dysregulated inflammation and injury occur with airway colonization in people with cystic fibrosis (CF). Aspergillus fumigatus frequently colonizes CF airways, but it has been assumed to be an innocent saprophyte; its potential role as a cause of lung disease is controversial. To study the interactions between Aspergillus and EC, and the role of the fungus in evoking inflammatory responses. A. fumigatus expressing green fluorescent protein was developed for in vitro and in vivo models, which used cell lines and mouse tracheal EC. Fungal spores (conidia) are rapidly ingested by ECs derived from bronchial cell lines and murine tracheas, supporting a role for EC in early airway clearance. Bronchial ECs harboring CFTR mutations (ΔF508) or deletion demonstrate impaired uptake and killing of conidia, and ECs with CFTR mutation undergo more conidial-induced apoptosis. Germinated (hyphal) forms of the fungus evoke secretion of inflammatory mediators, with CFTR mutation resulting in increased airway levels of macrophage inflammatory protein 2 and KC, and higher lung monocyte chemotactic protein-1. After A. fumigatus inhalation, CFTR(-/-) mice develop exaggerated lymphocytic inflammation, mucin accumulation, and lung injury. Data demonstrate a critical role for CFTR in mediating EC responses to A. fumigatus. Results suggest that the fungus elicits aberrant pulmonary inflammation in the setting of CFTR mutation, supporting the potential role of antifungals to halt progressive CF lung disease.


Visuomotor integration deficits are common to familial and sporadic preclinical Alzheimer's disease.
We investigated whether subtle visuomotor deficits were detectable in familial and sporadic preclinical Alzheimer's disease. A circle-tracing task-with direct and indirect visual feedback, and dual-task subtraction-was completed by 31 individuals at 50% risk of familial Alzheimer's disease (19 presymptomatic mutation carriers; 12 non-carriers) and 390 cognitively normal older adults (members of the British 1946 Birth Cohort, all born during the same week; age range at assessment = 69-71 years), who also underwent β-amyloid-PET/MRI to derive amyloid status (positive/negative), whole-brain volume and white matter hyperintensity volume. We compared preclinical Alzheimer's groups against controls cross-sectionally (mutation carriers versus non-carriers; amyloid-positive versus amyloid-negative) on speed and accuracy of circle-tracing and subtraction. Mutation carriers (mean 7 years before expected onset) and amyloid-positive older adults traced disproportionately less accurately than controls when visual feedback was indirect, and were slower at dual-task subtraction. In the older adults, the same pattern of associations was found when considering amyloid burden as a continuous variable (Standardized Uptake Value Ratio). The effect of amyloid was independent of white matter hyperintensity and brain volumes, which themselves were associated with different aspects of performance: greater white matter hyperintensity volume was also associated with disproportionately poorer tracing accuracy when visual feedback was indirect, whereas larger brain volume was associated with faster tracing and faster subtraction. Mutation carriers also showed evidence of poorer tracing accuracy when visual feedback was direct. This study provides the first evidence of visuomotor integration deficits common to familial and sporadic preclinical Alzheimer's disease, which may precede the onset of clinical symptoms by several years.


Hospitalization risk in patients with schizophrenia treated with dose-equivalent antipsychotics.
This study compared the risk of hospitalization among adults with schizophrenia being treated with equivalent dose ranges of lurasidone versus aripiprazole, olanzapine, quetiapine, or risperidone. Administrative claims data for this analysis came from the IBM MarketScan Commercial, Medicare Supplemental, and Multi-State Medicaid databases between January 2011 and June 2017. The study included adults with schizophrenia who initiated treatment with an antipsychotic and were continuously enrolled for 360 days prior to and following the date of the initial antipsychotic prescription. Risk of all-cause and schizophrenia-related hospitalization among patients who received lurasidone monotherapy versus aripiprazole, olanzapine, quetiapine, or risperidone in equivalent dose ranges were assessed. Marginal structural models that accounted for preindex characteristics, changes in antipsychotic treatment, and time-varying covariates assessed differences between lurasidone and other second-generation antipsychotics on all-cause and schizophrenia-related hospitalizations. A sensitivity analysis was conducted without the dose-equivalence requirement. A total of 20,212 patients met the study inclusion criteria. Compared with those treated with lurasidone monotherapy, the adjusted risk of all-cause hospitalization was significantly higher among patients treated with olanzapine (adjusted rate ratio [aRR], 1.49; P = .04), quetiapine (aRR, 1.64; P = .01), or risperidone (aRR, 1.47; P = .04), but not aripiprazole (aRR, 1.24; P = .28). A similar, non-statistically significant pattern of adjusted risks of schizophrenia-related hospitalizations was observed. A sensitivity analysis without the dose-equivalence requirement produced consistent results. As hospitalization is a major cost driver of direct healthcare cost, lurasidone may be a cost-saving treatment option for patients with schizophrenia.


Level of cytosolic free calcium during acetaminophen toxicity in mouse hepatocytes.
It has been suggested that elevated cytosolic free calcium plays a key role in acetaminophen-induced cell death. The present study has examined the effect of a toxic concentration of acetaminophen on cytosolic free calcium in single mouse hepatocytes, using the dye fura-2 and video imaging fluorescence microscopy. Cytosolic free calcium was calculated from the ratio of emitted fluorescence at greater than 475 nm produced by excitation at 340 and 380 nm, using a double-intensified silicon target camera and digital image processing. In the presence of 5 mM acetaminophen, cell death did not occur for 2 hr, but the toxic lesion that ultimately killed the cells occurred as early as 1 hr. If cytosolic free calcium plays an important role in these toxic events, it would be expected to increase during this period. However, during a 2-hr exposure, cytosolic free calcium concentration in cells exposed to acetaminophen was not different from control. In hepatocytes incubated for longer than 2 hr, the calcium concentration increased shortly before loss of cell viability (i.e., as a late event), consistent with an influx of calcium through a damaged cell membrane. This late increase in calcium occurred well after the appearance of cell surface blebs. The data suggest that there is no sustained change in cytosolic free calcium in acetaminophen injury either before or during the time when irreversible toxic events occur in hepatocytes.


Innovative approaches for enhancing cancer gene therapy.
Gene therapy provides a novel platform for therapeutic intervention of several genetic and non-genetic disorders. With the recent developments in the field, a wide variety of viral and non-viral vectors have emerged that can deliver genetic payloads to target cells. However, non-targeted delivery of transgenes often results in undesirable effects, low tumor transduction, and reduced therapeutic index. In this review, we focus on some of the novel approaches that can be used to meet the present challenges in the field and translate the potential of cancer gene therapy from 'bench to bedside' in the near future.


Contextual Flexibility in Pseudomonas aeruginosa Central Carbon Metabolism during Growth in Single Carbon Sources.
Pseudomonas aeruginosa is an opportunistic human pathogen, particularly noted for causing infections in the lungs of people with cystic fibrosis (CF). Previous studies have shown that the gene expression profile of P. aeruginosa appears to converge toward a common metabolic program as the organism adapts to the CF airway environment. However, we still have only a limited understanding of how these transcriptional changes impact metabolic flux at the systems level. To address this, we analyzed the transcriptome, proteome, and fluxome of P. aeruginosa grown on glycerol or acetate. These carbon sources were chosen because they are the primary breakdown products of an airway surfactant, phosphatidylcholine, which is known to be a major carbon source for P. aeruginosa in CF airways. We show that the fluxes of carbon throughout central metabolism are radically different among carbon sources. For example, the newly recognized "EDEMP cycle" (which incorporates elements of the Entner-Doudoroff [ED] pathway, the Embden-Meyerhof-Parnas [EMP] pathway, and the pentose phosphate [PP] pathway) plays an important role in supplying NADPH during growth on glycerol. In contrast, the EDEMP cycle is attenuated during growth on acetate, and instead, NADPH is primarily supplied by the reaction catalyzed by isocitrate dehydrogenase(s). Perhaps more importantly, our proteomic and transcriptomic analyses revealed a global remodeling of gene expression during growth on the different carbon sources, with unanticipated impacts on aerobic denitrification, electron transport chain architecture, and the redox economy of the cell. Collectively, these data highlight the remarkable metabolic plasticity of P. aeruginosa; that plasticity allows the organism to seamlessly segue between different carbon sources, maximizing the energetic yield from each.IMPORTANCEPseudomonas aeruginosa is an opportunistic human pathogen that is well known for causing infections in the airways of people with cystic fibrosis. Although it is clear that P. aeruginosa is metabolically well adapted to life in the CF lung, little is currently known about how the organism metabolizes the nutrients available in the airways. In this work, we used a combination of gene expression and isotope tracer ("fluxomic") analyses to find out exactly where the input carbon goes during growth on two CF-relevant carbon sources, acetate and glycerol (derived from the breakdown of lung surfactant). We found that carbon is routed ("fluxed") through very different pathways during growth on these substrates and that this is accompanied by an unexpected remodeling of the cell's electron transfer pathways. Having access to this "blueprint" is important because the metabolism of P. aeruginosa is increasingly being recognized as a target for the development of much-needed antimicrobial agents.