Isolation of genes by complementation in yeast: molecular cloning of a cell-cycle gene.
cdc28, one of several genes required for cell division in the yeast Saccharomyces cerevisiae, has been isolated on recombinant plasmids. A recombinant plasmid pool containing the entire yeast genome was constructed by partial digestion of yeast DNA with the four-base recognition restriction endonuclease Sau3A to give the equivalent of random fragments, size selection on sucrose gradients, and introduction of the fragments into the yeast vector YRp7 by use of the homology of Sau3A ends with those generated in the vector by cleavage with BamHI. Recombinant plasmids capable of complementing cdc28 mutations were isolated by transformation of a cdc28ts strain and selection for clones capable of growth at the restrictive temperature. Plasmids responsible for complementing the cdc28ts phenotype were shown to recombine specifically with the chromosomal cdc28 locus, confirming the identity of the cloned sequences. In addition, one of the recombinant plasmids was capable of complementing a mutation in tyr1, a gene genetically linked to cdc28. This method of gene isolation and identification should be applicable to all yeast genes for which there are readily scorable mutants.


Identification of a residue responsible for UDP-sugar donor selectivity of a dihydroxybenzoic acid glycosyltransferase from Arabidopsis natural accessions.
UDP-glycosyltransferase (UGT) plays a major role in the diversity and reactivity of plant specialized metabolites by catalyzing the transfer of the sugar moiety from activated UDP-sugars to various acceptors. Arabidopsis UGT89A2 was previously identified from a genome-wide association study as a key factor that affects the differential accumulation of dihydroxybenzoic acid (DHBA) glycosides in distinct Arabidopsis natural accessions, including Col-0 and C24. The in vitro enzyme assays indicate that these distinct metabolic phenotypes reflect the divergence of UGT89A2 enzyme properties in the Col-0 and C24 accessions. UGT89A2 from Col-0 is highly selective toward UDP-xylose as the sugar donor, and the isoform from C24 can utilize both UDP-glucose and UDP-xylose but with a higher affinity to the glucose donor. The sequences of the two isozymes only differ at six amino acid residues. Examination of these amino acid residues in more natural accessions revealed a strong correlation between the amino acid polymorphism at position 153 and the DHBA glycoside accumulation pattern. Site-directed mutagenesis that swapped residue 153 between UGT89A2 from Col-0 and C24 reversed the UDP-sugar preferences, indicating that residue 153 plays an important role in determining sugar donor specificity of UGT89A2. This study provides insight into the key amino acid changes that confer sugar donor selectivity on UGTs, and demonstrates the usefulness of natural variation in understanding the structure-function relationship of enzymes involved in specialized metabolism.


Multi-stepping saccadic sequences in humans.
A number of paradigms are reported in which multi-stepping saccadic sequences (MSS) are reliably elicited in normal subjects. It was observed that MSS are consistently associated with a prolongation of the initial saccade latency. Further experiments show that this phenomenon is validated across a number of independent eye movement tasks. However, the probability of MSS occurring is reduced when temporal constraints are imposed on the latency of the response. An interpretation of these task-elicited MSS in terms of the neural mechanisms controlling voluntary saccades is proposed.


A comparative study of formalin-ethyl acetate sedimentation technique and Mini Parasep® solvent-free method in the rapid diagnosis of intestinal parasites.
Stool microscopy and concentration techniques are the two most important and necessary aspects of diagnostic parasitology. In an era when there is increased disease burden due to intestinal parasites, an early and appropriate diagnosis is warranted. Direct microscopy is usually labor intensive and tedious. Thirty-two fresh fecal specimens from patients presenting with eosinophilia and/or anemia (hemoglobin levels <10 g%), HIV-positive patients, and in patients clinically suspected of harboring parasites, were collected for the study. All the positive samples were processed by both the standard methodology, i.e., formalin-ethyl acetate sedimentation technique and Mini Parasep® SF method, by the standard operating procedure of our laboratory and the manufacturer's instruction. Stool pellet concentrates were subjected to saline/iodine wet mount, modified acid fast staining for intestinal coccidian parasites and trichrome staining for Blastocystis hominis. The average number of organisms counted in 0.5 ml of pellet was used for comparison of the two techniques. The morphology of eggs was maintained in both the techniques; however, the wet mount prepared from the sedimentation technique had more background fecal debris in comparison to the Parasep® technique. The parasite yield was equal for both the techniques while Mini parasep had the advantage of less distortion of parasite morphology. We found that Parasep® offered a better parasitic yield, a better workflow capacity, and a reduced turnaround time, which would further benefit resource-restrained laboratories and those with a high sample turnover.


The alignment of the knee joint in relationship to age and osteoarthritis: the Copenhagen Osteoarthritis Study.
The aim of the present study was to describe the changes in the axis of the knee joint in both radiologically osteoarthritic and non-osteoarthritic knees, on the basis of angles measurable in standardized clinical short knee radiographs, in a cross sectional study of an epidemiological cohort. From the third inclusion of the Copenhagen City Heart Study, 4,151 subjects were selected for standardized radiography of the knees. After censuring the inclusion, the resulting cohort was comprised of 3,488 individuals. Images were analyzed for radiological knee joint osteoarthritis (OA) and the anatomical femorotibial axis of the knee joint was measured. The prevalence of knee joint OA in males was 27.9% and 27.5%, for the left and right knees respectively. In females this was 32.8% and 36.4%. The mean knee joint angles were 4.11° in males; and 5.45° in females. A difference of 1.3° was found between the genders. In non-osteoarthritic knees the increase in valgus orientation in relationship to increasing age was found to be 0.03° and 0.04° per year, respectively, for males and females. Likewise, Kellgren and Lawrence found that OA was seen to influence a shift towards varus of 0.55°-0.76° per level of OA. Stratification in accordance with morphological severity of OA documented a clear tendency for the axis of the diseased knees to depart from the mean, primarily in the direction of varus. In knees exhibiting no signs of radiographic osteoarthritis we found a significant relationship between increasing age and a shift in the anatomical axis in the direction of valgus.


Retention in care and connection to care among HIV-infected patients on antiretroviral therapy in Africa: estimation via a sampling-based approach.
Current estimates of retention among HIV-infected patients on antiretroviral therapy (ART) in Africa consider patients who are lost to follow-up (LTF) as well as those who die shortly after their last clinic visit to be no longer in care and to represent limitations in access to care. Yet many lost patients may have "silently" transferred and deaths shortly after the last clinic visit more likely represent limitations in clinical care rather than access to care after initial linkage. We evaluated HIV-infected adults initiating ART from 1/1/2004 to 9/30/2007 at a clinic in rural Uganda. A representative sample of lost patients was tracked in the community to obtain updated information about care at other ART sites. Updated outcomes were incorporated with probability weights to obtain "corrected" estimates of retention for the entire clinic population. We used the competing risks approach to estimate "connection to care"--the percentage of patients accessing care over time (including those who died while in care). Among 3,628 patients, 829 became lost, 128 were tracked and in 111, updated information was obtained. Of 111, 79 (71%) were alive and 35/48 (73%) of patients interviewed in person were in care and on ART. Patient retention for the clinic population assuming lost patients were not in care was 82.3%, 68.9%, and 60.1% at 1, 2 and 3 years. Incorporating updated care information from the sample of lost patients increased estimates of patient retention to 85.8% to 90.9%, 78.9% to 86.2% and 75.8% to 84.7% at the same time points. Accounting for "silent transfers" and early deaths increased estimates of patient retention and connection to care substantially. Deaths soon after the last clinic visit (potentially reflecting limitations in clinical effectiveness) and disconnection from care among patient who were alive each accounted for approximately half of failures of retention.


Bead-Based Multiplex Assay for the Simultaneous Detection of Antibodies to African Swine Fever Virus and Classical Swine Fever Virus.
African swine fever (ASF) and Classical swine fever (CSF) are both highly contagious diseases of domestic pigs and wild boar. In the last years, several cases of both diseases have been reported in the Caucasus, Russian Federation and Eastern Europe. Thus, the probability of encountering these two viruses in the same area is increasing. Since differentiation by clinical or post-mortem examination is not possible, laboratory tools for differential diagnosis are required. In the present work, we have developed a triplex bead-based assay using some of the most immunogenic antigens of each virus, for the simultaneous detection of antibodies; i.e. the VP72 and VP30 of ASF virus (ASFV) and the E2 protein of CSF virus (CSFV). The assay was firstly set up and optimized using well characterized reference serum samples specific for each pathogen. Then, a panel of 352 sera from experimentally infected animals with either ASFV or CSFV were analyzed in the multiplex assay. A collection of 253 field negative sera was also included in the study. The results of the multiplex analysis were compared to those obtained by two commercially available ELISAs for detection of antibodies against ASFV or CSFV, and considered in this study as the reference techniques. The data obtained showed values of 97.3% sensitivity and 98.3% specificity for detection of antibodies to ASFV and 95.7% of sensitivity and 99.8% specificity for detection of antibodies to CSFV. This multiplex assay allows the simultaneous and differential detection of antibodies against ASFV and CSFV, providing a valuable tool for surveillance studies. Moreover, this method is rather versatile, offering the possibility of increasing the panel of antigens from other swine diseases that could be of interest for a differential diagnosis along with ASF and CSF.


Structural observation of the primary isomerization in vision with femtosecond-stimulated Raman.
The primary event that initiates vision is the light-induced 11-cis to all-trans isomerization of retinal in the visual pigment rhodopsin. Despite decades of study with the traditional tools of chemical reaction dynamics, both the timing and nature of the atomic motions that lead to photoproduct production remain unknown. We used femtosecond-stimulated Raman spectroscopy to obtain time-resolved vibrational spectra of the molecular structures formed along the reaction coordinate. The spectral evolution of the vibrational features from 200 femtoseconds to 1 picosecond after photon absorption reveals the temporal sequencing of the geometric changes in the retinal backbone that activate this receptor.


Sensory basis for detection of benthic prey in two Lake Malawi cichlids.
The adaptive radiations of African cichlids resulted in a diversity of feeding morphologies and strategies, but the role of sensory biology in prey detection and feeding ecology remains largely unexplored. Two endemic Lake Malawi cichlid genera, Tramitichromis and Aulonocara, feed on benthic invertebrates, but differ in lateral line morphology (narrow and widened lateral line canals, respectively) and foraging strategy. The hypothesis that they use their lateral line systems differently was tested by looking at the relative contribution of the lateral line system and vision in prey detection by Tramitichromis sp. and comparing results to those from a complementary study using Aulonocara stuartgranti (Schwalbe et al., 2012). First, behavioral trials were used to assess the ability of Tramitichromis sp. to detect live (mobile) and dead (immobile) benthic prey under light and dark conditions. Second, trials were run before, immediately after, and several weeks after chemical ablation of the lateral line system to determine its role in feeding behavior. Results show that Tramitichromis sp. is a visual predator that neither locates prey in the dark nor depends on lateral line input for prey detection and is thus distinct from A. stuartgranti, which uses its lateral line or a combination of vision and lateral line to detect prey depending on light condition. Investigating how functionally distinctive differences in sensory morphology are correlated with feeding behavior in the laboratory and determining the role of sensory systems in feeding ecology will provide insights into how sensory capabilities may contribute to trophic niche segregation.


The Berry phase and the Aharonov-Bohm effect on optical activity.
The helical crystal structure in optically active media acts as the natural micro-solenoids for the electromagnetic waves passing through them, producing the longitudinal magnetic field in the direction of the axis of helices. Magnetic flux through the helical structure is quantized. The Berry phase is induced by rotation of the electrons around the helical structure. Optical rotation is related to the difference in the accumulative Berry phase between the right-, and the left-circularly polarized waves, which is proportional to the magnetic flux through the helical structure, according to the Aharonov-Bohm effect. The optical activity is the natural Faraday effect and the natural Aharonov-Bohm effect.


Transcriptomic buffering of cryptic genetic variation contributes to meningococcal virulence.
Commensal bacteria like Neisseria meningitidis sometimes cause serious disease. However, genomic comparison of hyperinvasive and apathogenic lineages did not reveal unambiguous hints towards indispensable virulence factors. Here, in a systems biological approach we compared gene expression of the invasive strain MC58 and the carriage strain α522 under different ex vivo conditions mimicking commensal and virulence compartments to assess the strain-specific impact of gene regulation on meningococcal virulence. Despite indistinguishable ex vivo phenotypes, both strains differed in the expression of over 500 genes under infection mimicking conditions. These differences comprised in particular metabolic and information processing genes as well as genes known to be involved in host-damage such as the nitrite reductase and numerous LOS biosynthesis genes. A model based analysis of the transcriptomic differences in human blood suggested ensuing metabolic flux differences in energy, glutamine and cysteine metabolic pathways along with differences in the activation of the stringent response in both strains. In support of the computational findings, experimental analyses revealed differences in cysteine and glutamine auxotrophy in both strains as well as a strain and condition dependent essentiality of the (p)ppGpp synthetase gene relA and of a short non-coding AT-rich repeat element in its promoter region. Our data suggest that meningococcal virulence is linked to transcriptional buffering of cryptic genetic variation in metabolic genes including global stress responses. They further highlight the role of regulatory elements for bacterial virulence and the limitations of model strain approaches when studying such genetically diverse species as N. meningitidis.


Assembly of human small nuclear RNA gene-specific transcription factor IIIB complex de novo on and off promoter.
In humans, transcription factor IIIB (TFIIIB)-alpha governs basal transcription from small nuclear RNA genes by RNA polymerase III (pol III). One of the components of this complex, BRFU/TFIIIB50, is specific for these promoters, whereas TATA-binding protein (TBP) and hB" are required for pol III transcription from both gene external and internal promoters. We show that hB" is specifically recruited to a promoter-bound TBP.BRFU complex, which we have previously demonstrated as forming on TATA-containing templates. The N-terminal region of BRFU, containing a zinc ribbon domain, acts as a damper of hB" binding. TBP deactivates this negative mechanism through protein-protein contacts with both BRFU and hB", which may then promote their cooperative binding to form TFIIIB-alpha. In addition, we have identified a GC-rich sequence downstream from the TATA box (the BURE) which, depending on the strength of TATA box, can either enhance BRFU binding to the TBP.DNA complex or hB" association with the BRFU.TBP.DNA complex, and subsequently stimulate pol III transcription. Moreover, mutation of the BURE reduces pol III transcription and induces transcription by RNA polymerase II from the U2 gene promoter carrying a minimal TATA box.


Efficacy of sublingual fentanyl vs. oral morphine for cancer-related breakthrough pain.
Breakthrough cancer pain (BTcP) is recognized as a clinically significant complication of chronic cancer pain with most BTcP episodes peaking in intensity within a few minutes and lasting for approximately 30 min. Although a number of rapid-onset fentanyl preparations have been developed in the last decade, BTcP is still typically managed through the use of rescue doses of oral morphine but a comparative study of sublingual fentanyl and oral morphine is still lacking. The aim of this study was to determine the efficacy, tolerability, and patient satisfaction of sublingual fentanyl citrate (SLF) and oral morphine solution (OM) in the treatment of BTcP. In this prospective, longitudinal, controlled-study, 40 patients with BTcP were allocated to receive oral morphine (OM) or sublingual fentanyl (SLF). Pain intensity level on a 0-10 numerical rating visual analog scale (VAS), frequency of BTcP throughout the day, onset of relief (0-5, 6-10, 11-15, or over 16 min), time required for dose titration, patient satisfaction and adverse effects were assessed at 3, 7, 15, and 30 days after starting the treatment. Mean doses of opioids for BTcP were 235 ± 23.4 μg (SLF) and 38 ± 5.2 mg (OM). The mean pain intensity levels were significantly lower with SLF than OM at 3 days (6.0 vs. 6.95; p = 0,001), 7 days (4.15 vs. 6.25, p < 0.001), 15 days (3.45 vs. 5.35, p < 0.001), and 30 days (3.05 vs. 4.45, p < 0.001). SLF provided significantly faster relief for BTcP than OM (p < 0.001) with a shorter dose titration period (mean 6.6 ± 3.3 vs. 13.3 ± 4.9 days; p < 0.001) and better satisfaction scores and with a very good safety profile. Administration of SLF might provide a more effective treatment option than oral morphine for BTcP.


Molecular analysis of the human vitamin D binding protein (group specific component, Gc) in tuberous sclerosis complex (TSC).
Group specific component (Gc) is an abundant plasma protein whose functional role is not clearly established. Gc protein is synthesised in the liver and is known to bind vitamin D, vitamin D metabolites, and G actin; Gc protein is also implicated in macrophage activation. Several polymorphic electrophoretic variants of Gc protein are found in all human populations; the most common alleles are Gc-1f, Gc-1s, and Gc-2. In previous studies, Gc allele frequencies, determined using isoelectric focusing or immunofixation or both, were significantly different in patients with tuberous sclerosis complex (TSC) from matched controls, with an excess of Gc-2 in patients. Linkage association between Gc and TSC is unlikely since the Gc locus maps to chromosome 4q12, whereas the two common forms of TSC map to 9q34 and 16p13.1, respectively. However, a direct cause and effect relationship between Gc protein and TSC symptoms is possible. To investigate further the relationship between the Gc locus and TSC, two Gc restriction site polymorphisms, HaeIII and StyI, were typed in 43 unrelated white subjects with TSC. The frequencies of the restriction site polymorphisms in the TSC patients did not differ from those in control populations. Therefore a direct association between Gc type and TSC is unlikely. The previously reported association was either spurious or the result of typing errors in plasma from subjects with underlying abnormalities in plasma proteins.


Characterization of human mineralocorticosteroid receptor expressed in the baculovirus system.
To investigate the structure and function of the mineralocorticosteroid receptor (MR), one has to circumvent the major difficulty related to its very low abundance. For this purpose, the full-length human MR (hMR) has been produced using the efficient baculovirus system. The recombinant protein is overexpressed in Sf9 insect cells at a concentration of approximately 2 pmol/mg of protein, which is 50-100 times more than the concentration in aldosterone target tissues. It binds aldosterone with high affinity (Kd approximately 1 nM) and clearly displays a mineralocorticosteroid specificity as evidenced by competition studies with steroid ligands and by the monoclonal anti-idiotypic antibody H10E interacting with the steroid-binding domain of MR. After [35S]methionine labeling, a single polypeptide band at approximately 120 kDa is detected and further identified as hMR by immunoblotting with A4, an anti-peptide antibody. Sedimentation analyses show that the native form of MR is recognized by A4 and B174, an antibody to the 90-kDa heat shock protein, since they both induce a shift of the receptor from 9S to 11S sedimentation coefficient. These results clearly demonstrate that MR is a heterooligomer containing the insect equivalent of the 90-kDa heat shock protein. This 9S receptor complex is converted to a 4S form during high-salt gradient ultracentrifugation, suggesting that MR can undergo a complete in vitro transformation. Immunofluorescence studies indicate that hMR, which is almost exclusively a cytoplasmic protein in Sf9 cells, is translocated to the nucleus after aldosterone exposure. Therefore, the recombinant hMR seems to behave as the native receptor. Given the possibility of large-scale protein production, the baculovirus system should prove useful in studies of the molecular basis of MR function as a transcription factor.


A comparison of four culture methods for diagnosing infection in continuous ambulatory peritoneal dialysis.
Dialysate effluent from 41 patients presenting with 54 episodes of CAPD peritonitis was examined by four culture techniques, three employing methods for the lysis of peritoneal leucocytes. The most efficient method employed a distilled water lysis-centrifugation technique (81% of episodes culture-positive). Filtration (without leucocyte lysis) was less effective (74% culture-positive). The results of a mechanical leucocyte lysis technique were less satisfactory still, the culture positive rate of 74% being compromised by the recovery of the infecting organism in low numbers, and by an association with a high incidence of plate contamination. The results of a bile-salt lysis technique were the least satisfactory (67% culture-positive).


Predictors of Treatment Outcome in Group or Individual Cognitive Processing Therapy for Posttraumatic Stress Disorder Among Active Duty Military.
The purpose of this study was to examine demographic, psychological, military, and deployment variables that might predict posttraumatic stress disorder (PTSD) symptom improvement in a sample of active duty service members who received either group or individual cognitive processing therapy (CPT). Data were analyzed from 165 active duty service members with pre- and posttreatment data participating in a randomized controlled trial comparing group with individual CPT. Pretreatment variables were examined as predictors of change in PTSD severity from baseline to posttreatment, assessed using the PTSD Symptom Scale-Interview Version (PSS-I). Predictors of PSS-I change were first evaluated using Pearson correlations, followed by partial and multiple correlations to clarify which associations remained when effects of other predictors were controlled. Multiple regression analyses were used to test for interactions between pretreatment variables and treatment format. Only age was a significant predictor of PTSD symptom change after controlling for other variables and statisitically correcting for testing multiple variables. There was also an interaction between age and treatment format. Younger participants had greater symptom improvement, particularly if they received individual treatment. Other pretreatment variables did not predict outcome. CPT appears to be robust across most pretreatment variables, such that comorbid disorders, baseline symptom severity, and suicidal ideation do not interfere with application of CPT. However, individual CPT may be a better option particularly for younger service members.


Yoga management of breast cancer-related lymphoedema: a randomised controlled pilot-trial.
Secondary arm lymphoedema continues to affect at least 20% of women after treatment for breast cancer requiring lifelong professional treatment and self-management. The holistic practice of yoga may offer benefits as an adjunct self-management option. The aim of this small pilot trial was to gain preliminary data to determine the effect of yoga on women with stage one breast cancer-related lymphoedema (BCRL). This paper reports the results for the primary and secondary outcomes. Participants were randomised, after baseline testing, to receive either an 8-week yoga intervention (n = 15), consisting of a weekly 90-minute teacher-led class and a 40-minute daily session delivered by DVD, or to a usual care wait-listed control group (n = 13). Primary outcome measures were: arm volume of lymphoedema measured by circumference and extra-cellular fluid measured by bioimpedance spectroscopy. Secondary outcome measures were: tissue induration measured by tonometry; levels of sensations, pain, fatigue, and their limiting effects all measured by a visual analogue scale (VAS) and quality of life based on the Lymphoedema Quality of Life Tool (LYMQOL). Measurements were conducted at baseline, week 8 (post-intervention) and week 12 (four weeks after cessation of the intervention). At week 8, the intervention group had a greater decrease in tissue induration of the affected upper arm compared to the control group (p = 0.050), as well as a greater reduction in the symptom sub-scale for QOL (p = 0.038). There was no difference in arm volume of lymphoedema or extra-cellular fluid between groups at week 8; however, at week 12, arm volume increased more for the intervention group than the control group (p = 0.032). An 8-week yoga intervention reduced tissue induration of the affected upper arm and decreased the QOL sub-scale of symptoms. Arm volume of lymphoedema and extra-cellular fluid did not increase. These benefits did not last on cessation of the intervention when arm volume of lymphoedema increased. Further research trials with a longer duration, higher levels of lymphoedema and larger numbers are warranted before definitive conclusions can be made.


Spatial configuration processing in visual word form perception and reading abilities in Chinese.
The analysis of spatial relations among components of a character is important in identifying visual word forms. We investigated such spatial configuration processing for Chinese characters in dyslexic children (aged 9-12 years) and two groups of typically developing children, matched according to chronological age or reading level. In this study, we used real characters and noncharacters to manipulate the spatial configuration between character components while preserving their local features. Both characters were displayed either in an upright or inverted position. Participants were asked to quickly determine whether the two presented stimuli were identical. A significant interaction between character type and character orientation showed children's matching performance was better in upright real characters than in the inverted version, but such effect was absent in noncharacters. This indicated that regardless of reading skills, children developed the ability to use local configuration information to visually identify words. Dyslexic children performed poorly in both real-character and noncharacter conditions. Furthermore, compared with control groups, dyslexic children performed worse in real-character conditions, which were close to the noncharacter performance of other groups. This reflected an impairment of global visual word-form processing (i.e., spatial configuration between components) in dyslexia.


Interdigitated microelectronic bandage augments hemostasis and clot formation at low applied voltage in vitro and in vivo.
Hemorrhage or uncontrolled bleeding can arise either due to a medical condition or from a traumatic injury and are typically controlled with the application of a hemostatic agent. Hemostatic agents are currently derived from animal or human products, which carry risks of blood borne infections and immune dysregulation. Therefore, the need exists for novel biomedical therapies not derived from animal or human products to achieve hemostasis. Accordingly, we created an interdigitated microelectronic bandage that applies low voltage electrical stimulation to an injury site, resulting in faster clot formation without excessive heating, accelerated fibrin formation, and hemostasis overall. Our interdigitated microelectronic bandage found fibrin formed 1.5× faster in vitro. In vivo, total cessation of bleeding was 2.5× faster, resulting in 2× less blood loss. Electricity has been used in medical applications such as defibrillation, cauterization, and electrosurgery, but scant research has focused on hemostasis. Here we report a novel surface treatment using an interdigitated microelectronic device that creates rapid hemostasis in both in vitro and in vivo bleeding models with low applied voltages, representing a new and novel class of hemostatic agents that are electrically-based.


Frequency of polymorphisms pro198leu in GPX-1 gene and ile58thr in MnSOD gene in the altitude Ecuadorian population with bladder cancer.
Bladder cancer represents 8% of all malignancies diagnosed in men and 3% in women. The risk factors for developing bladder cancer, including the incidence rate, morbidity, and mortality, vary according to the ethnic group, exposition rate at work, age, gender, and tobacco consumption. Moreover, there is a risk of developing this carcinoma due to dietary conditions, demonstrating that certain enzymes neutralize oxidative compound derivative of carcinogens, which if not degraded, accumulate in the body and destroy epithelial cells of the bladder, causing an increase in the risk of developing this disease. The detoxifying enzymes inactivate dangerous chemical compounds and anions for the cell; that is, why it is important to know if the polymorphisms pro198leu in GPX-1 and ile58thr in MnSOD are associated with bladder cancer. In this study, 120 individuals were analyzed as controls and 97 individuals with previously diagnosed bladder cancer. In the case of polymorphism pro198leu, highly significant differences were observed and individuals with this polymorphism presented a probability of developing bladder cancer 3.8 times greater than controls (OR = 3.8; 95% CI 2.16-6.78; p < 0.001). No significant differences in polymorphism ile58thr of MnSOD gene occurred when we compared the study population (OR = 2.1; 95% CI 1.26-3.49; p > 0.05). The results indicate that polymorphism of GPX-1 gene influences the risk of developing bladder cancer in the Ecuadorian population, suggesting that more research on detoxifying genes in bladder cancer should be conducted.


The Mycobacterium tuberculosis regulatory network and hypoxia.
We have taken the first steps towards a complete reconstruction of the Mycobacterium tuberculosis regulatory network based on ChIP-Seq and combined this reconstruction with system-wide profiling of messenger RNAs, proteins, metabolites and lipids during hypoxia and re-aeration. Adaptations to hypoxia are thought to have a prominent role in M. tuberculosis pathogenesis. Using ChIP-Seq combined with expression data from the induction of the same factors, we have reconstructed a draft regulatory network based on 50 transcription factors. This network model revealed a direct interconnection between the hypoxic response, lipid catabolism, lipid anabolism and the production of cell wall lipids. As a validation of this model, in response to oxygen availability we observe substantial alterations in lipid content and changes in gene expression and metabolites in corresponding metabolic pathways. The regulatory network reveals transcription factors underlying these changes, allows us to computationally predict expression changes, and indicates that Rv0081 is a regulatory hub.


Dual blockade of HER2 in HER2-overexpressing tumor cells does not completely eliminate HER3 function.
Dual blockade of HER2 with trastuzumab and lapatinib or with pertuzumab is a superior treatment approach compared with single-agent HER2 inhibitors. However, many HER2-overexpressing breast cancers still escape from this combinatorial approach. Inhibition of HER2 and downstream phosphoinositide 3-kinase (PI3K)/AKT causes a transcriptional and posttranslational upregulation of HER3 which, in turn, counteracts the antitumor action of the HER2-directed therapies. We hypothesized that suppression of HER3 would synergize with dual blockade of HER2 in breast cancer cells sensitive and refractory to HER2 antagonists. Inhibition of HER2/HER3 in HER2(+) breast cancer cell lines was evaluated by Western blotting. We analyzed drug-induced apoptosis and two- and three-dimensional growth in vitro. Growth inhibition of PI3K was examined in vivo in xenografts treated with combinations of trastuzumab, lapatinib, and the HER3-neutralizing monoclonal antibody U3-1287. Treatment with U3-1287 blocked the upregulation of total and phosphorylated HER3 that followed treatment with lapatinib and trastuzumab and, in turn, enhanced the antitumor action of the combination against trastuzumab-sensitive and -resistant cells. Mice bearing HER2(+) xenografts treated with lapatinib, trastuzumab, and U3-1287 exhibited fewer recurrences and better survival than mice treated with lapatinib and trastuzumab. Dual blockade of HER2 with trastuzumab and lapatinib does not eliminate the compensatory upregulation of HER3. Therapeutic inhibitors of HER3 should be considered as part of multidrug combinations aimed at completely and rapidly disabling the HER2 network in HER2-overexpressing breast cancers.


Morphology of the ampullae of Lorenzini in juvenile freshwater Carcharhinus leucas.
Ampullae of Lorenzini were examined from juvenile Carcharhinus leucas (831-1,045 mm total length) captured from freshwater regions of the Brisbane River. The ampullary organ structure differs from all other previously described ampullae in the canal wall structure, the general shape of the ampullary canal, and the apically nucleated supportive cells. Ampullary pores of 140-205 µm in diameter are distributed over the surface of the head region with 2,681 and 2,913 pores present in two sharks that were studied in detail. The primary variation of the ampullary organs appears in the canal epithelial cells which occur as either flattened squamous epithelial cells or a second form of pseudostratified contour-ridged epithelial cells; both cell types appear to release material into the ampullary lumen. Secondarily, this ampullary canal varies due to involuted walls that form a clover-like canal wall structure. At the proximal end of the canal, contour-ridged cells abut a narrow region of cuboidal epithelial cells that verge on the constant, six alveolar sacs of the ampulla. The alveolar sacs contain numerous receptor and supportive cells bound by tight junctions and desmosomes. Pear-shaped receptor cells that possess a single apical kinocilium are connected basally by unmyelinated neural boutons. Opposed to previously described ampullae of Lorenzini, the supportive cells have an apical nucleus, possess a low number of microvilli, and form a unique, jagged alveolar wall. A centrally positioned centrum cap of cuboidal epithelial cells overlies a primary afferent lateral line nerve.


Effect of replacing grass silage with red clover silage on nutrient digestion, nitrogen metabolism, and milk fat composition in lactating cows fed diets containing a 60:40 forage-to-concentrate ratio.
Diets based on red clover silage (RCS) typically increase the concentration of polyunsaturated fatty acids (PUFA) in ruminant meat and milk and lower the efficiency of N utilization compared with grass silages (GS). Four multiparous Finnish Ayrshire cows (108 d postpartum) fitted with rumen cannulas were used in a 4 × 4 Latin square design with 21-d periods to evaluate the effect of incremental replacement of GS with RCS on milk production, nutrient digestion, whole-body N metabolism, and milk fatty acid composition. Treatments comprised total mixed rations offered ad libitum, containing 600 g of forage/kg of diet dry matter (DM), with RCS replacing GS in ratios of 0:100, 33:67, 67:33, and 100:0 on a DM basis. Intake of DM and milk yield tended to be higher when RCS and GS were offered as a mixture than when fed alone. Forage species had no influence on the concentration or secretion of total milk fat, whereas replacing GS with RCS tended to decrease milk protein concentration and yield. Substitution of GS with RCS decreased linearly whole-tract apparent organic matter, fiber, and N digestion. Forage species had no effect on total nonammonia N at the omasum, whereas the flow of most AA at the omasum was higher for diets based on a mixture of forages. Replacing GS with RCS progressively lowered protein degradation in the rumen, increased linearly ruminal escape of dietary protein, and decreased linearly microbial protein synthesis. Incremental inclusion of RCS in the diet tended to lower whole-body N balance, increased linearly the proportion of dietary N excreted in feces and urine, and decreased linearly the utilization of dietary N for milk protein synthesis. Furthermore, replacing GS with RCS decreased linearly milk fat 4:0 to 8:0, 14:0, and 16:0 concentrations and increased linearly 18:2n-6 and 18:3n-3 concentrations, in the absence of changes in cis-9 18:1, cis-9, trans-11 18:2, or total trans fatty acid concentration. Inclusion of RCS in the diet progressively increased the apparent transfer of 18-carbon PUFA from the diet into milk, but had no effect on the amount of 18:2n-6 or 18:3n-3 at the omasum recovered in milk. In conclusion, forage species modified ruminal N metabolism, the flow of AA at the omasum, and whole-body N partitioning. A lower efficiency of N utilization for milk protein synthesis with RCS relative to GS was associated with decreased availability of AA for absorption, with some evidence of an imbalance in the supply of AA relative to requirements. Higher enrichment of PUFA in milk for diets based on RCS was related to an increased supply for absorption, with no indication that forage species substantially altered PUFA bioavailability.


A case for a negative-strand coding sequence in a group of positive-sense RNA viruses.
Positive-sense single-stranded RNA viruses form the largest and most diverse group of eukaryote-infecting viruses. Their genomes comprise one or more segments of coding-sense RNA that function directly as messenger RNAs upon release into the cytoplasm of infected cells. Positive-sense RNA viruses are generally accepted to encode proteins solely on the positive strand. However, we previously identified a surprisingly long (∼1,000-codon) open reading frame (ORF) on the negative strand of some members of the family Narnaviridae which, together with RNA bacteriophages of the family Leviviridae, form a sister group to all other positive-sense RNA viruses. Here, we completed the genomes of three mosquito-associated narnaviruses, all of which have the long reverse-frame ORF. We systematically identified narnaviral sequences in public data sets from a wide range of sources, including arthropod, fungal, and plant transcriptomic data sets. Long reverse-frame ORFs are widespread in one clade of narnaviruses, where they frequently occupy >95 per cent of the genome. The reverse-frame ORFs correspond to a specific avoidance of CUA, UUA, and UCA codons (i.e. stop codon reverse complements) in the forward-frame RNA-dependent RNA polymerase ORF. However, absence of these codons cannot be explained by other factors such as inability to decode these codons or GC3 bias. Together with other analyses, we provide the strongest evidence yet of coding capacity on the negative strand of a positive-sense RNA virus. As these ORFs comprise some of the longest known overlapping genes, their study may be of broad relevance to understanding overlapping gene evolution and de novo origin of genes.


Acetone reactions over the surfaces of polycrystalline UO2: a kinetic and spectroscopic study.
The reaction of acetone is studied on the surfaces of polycrystalline UO2, prepared by hydrogen reduction of U3O8 at 770 K. The study is conducted by in situ Fourier transform infrared (FTIR) and temperature-programmed desorption (TPD). Acetone adsorption does not fit the simple Langmuir model, and adsorbate-adsorbate interactions are found to be significant. Acetone adsorbs molecularly on UO2 as evidenced by the nuCO of the eta1(O) mode at 1686 cm(-1). Part of acetone is reduced to the isopropoxide species ((CH3)2HC-O-U4+) upon heating (nu(CC), rho(CH3) at 1167 cm(-1) and nu(CO), rho(CH3) at 980 cm(-1)), and upon further heating, acetates (CH3COO(a), (a) for adsorbed) are observed. Detailed TPD studies indicated that the main reaction of acetone on UO2 is the deoxygenation to propene, driven by the oxophilic nature of UO2. Other reactions were also observed to a lesser extent, and these included reductive coupling to 2,3-dimethylbutene and condensation to mesityl oxide. An attempt to extract kinetic parameters from TPD data was conducted. Three models were studied: variation of heating rate, leading edge analysis (Habenschaden-Kuppers method), and complete analysis. The complete analysis provided the most plausible results, in particular, at low coverage. With this method, at nearly zero coverage the activation energy, Ed, for desorption was found to be close to 140 kJ/mol with a prefactor of 10(13) s(-1). Ed dropped sharply with increasing coverage, theta, to ca. 35 kJ/mol at theta=0.15 with a prefactor of 10(11) s(-1). The activation energy for the desorption of acetone on UO2(111) single crystals, at saturation coverage, was previously found to be equal to 65 kJ/mol using the leading edge analysis.


Changes in 24-Hour Domain-Specific Movement Behaviors and Their Associations With Children's Psychosocial Health During the Transition From Primary to Secondary School: A Compositional Data Analysis.
Little is known about the influence of 24-hour movement behaviors on children's psychosocial health when transitioning from primary to secondary school. This study described changes in 24-hour domain-specific movement behavior composition and explored their associations with changes in psychosocial health during this transition. Data were drawn from the Longitudinal Study of Australian Children. The analytical sample (n = 909) included children who were enrolled in primary school at baseline (2010) and in secondary school at follow-up (2012). Time spent in 8 domains of movement behaviors was derived from the child-completed time-use diaries. Psychosocial health was examined using the self-report version of the Strengths and Difficulties Questionnaires. Analyses included repeated-measures multivariate analysis of variance and compositional regression. Children reported engaging in more social activities and sleeping less over the transition period. Increased time spent in social activities (βilr = -0.06, P = .014) and recreational screen use (βilr = -0.17, P = .003) (relative to other domains) were associated with decreased prosocial behavior in boys. Changes in movement behavior composition were not associated with changes in girls' psychosocial health. This study found considerable changes in children's 24-hour movement behavior composition, but a lack of consistent association with changes in psychosocial health during the primary to secondary school transition.


Osteogenic capillaries orchestrate growth plate-independent ossification of the malleus.
Endochondral ossification is a developmental process by which cartilage is replaced by bone. Terminally differentiated hypertrophic chondrocytes are calcified, vascularized, and removed by chondroclasts before bone matrix is laid down by osteoblasts. In mammals, the malleus is one of three auditory ossicles that transmit vibrations of the tympanic membrane to the inner ear. The malleus is formed from a cartilaginous precursor without growth plate involvement, but little is known about how bones of this type undergo endochondral ossification. Here, we demonstrate that in the processus brevis of the malleus, clusters of osteoblasts surrounding the capillary loop produce bone matrix, causing the volume of the capillary lumen to decrease rapidly in post-weaning mice. Synchrotron X-ray tomographic microscopy revealed a concentric, cylindrical arrangement of osteocyte lacunae along capillaries, indicative of pericapillary bone formation. Moreover, we report that overexpression of Fosl1, which encodes a component of the AP-1 transcription factor complex, in osteoblasts significantly blocked malleal capillary narrowing. These data suggest that osteoblast/endothelial cell interactions control growth plate-free endochondral ossification through 'osteogenic capillaries' in a Fosl1-regulated manner.


Effects of Concentration and Ionization Degree of Anchoring Cationic Polymers on the Lateral Heterogeneity of Anionic Lipid Monolayers.
We employed coarse-grained Monte Carlo simulations to investigate a system composed of cationic polymers and a phosphatidyl-choline membrane monolayer, doped with univalent anionic phosphatidylserine (PS) and tetravalent anionic phosphatidylinositol 4,5-bisphosphate (PIP2) lipid molecules. For this system, we consider the conditions under which multiple cationic polymers can anchor onto the monolayer and explore how the concentration and ionization degree of the polymers affect the lateral rearrangement and fluidity of the negatively charged lipids. Our work shows that the anchoring cationic polymers predominantly bind the tetravalent anionic PIP2 lipids and drag the PIP2 clusters to migrate on the monolayer. The polymer/PIP2 binding is found to be drastically enhanced by increasing the polymer ionization fraction, which causes the PIP2 lipids to form into larger clusters and reduces the mobility of the polymer/PIP2 complexes. As expected, stronger competition effects between anchoring polymers occur at higher polymer concentrations, for which each anchoring polymer partially dissociates from the monolayer and hence sequesters a smaller PIP2 cluster. The desorbed segments of the anchored polymers exhibit a faster mobility on the membrane, whereas the PIP2 clusters are closely restrained by the limited adhering cationic segments of anchoring polymers. We further demonstrate that the PIP2 molecules display a hierarchical mobility in the PIP2 clusters, which is regulated by the synergistic effect between the cationic segments of the polymers. The PS lipids sequester in the vicinity of the polymer/PIP2 complexes if the tetravalent PIP2 lipids cannot sufficiently neutralize the cationic polymers. Finally, we illustrate that the increase in the ionic concentration of the solution weakens the lateral clustering and the mobility heterogeneity of the charged lipids. Our work thus provides a better understanding of the fundamental biophysical mechanism of the concentration gradients and the hierarchical mobility of the anionic lipids in the membrane caused by the cationic polymer anchoring on length and time scales that are generally inaccessible by atomistic models. It also offers insight into the development and design of novel biological applications on the basis of the modulation of signaling lipids.


A hydrodynamic water quality model for propagation of pollutants in rivers.
Numerical modelling can be a useful tool to assess a receiving water body's quality state. Indeed, the use of mathematical models in river water quality management has become a common practice to show the cause-effect relationship between emissions and water body quality and to design as well as assess the effectiveness of mitigation measures. In the present study, a hydrodynamic river water quality model is presented. The model consists of a quantity and a quality sub-model. The quantity sub-model is based on the Saint Venant equations. The solution of the Saint Venant equations is obtained by means of an explicit scheme based on space-time conservation. The method considers the unification of space and time and the enforcement of flux conservation in both space and time. On the other hand, the quality sub-model is based on the advection dispersion equation. Particularly, the principle of upstream weighting applied to finite difference methods is employed. This method enable us to reduce the numerical dispersion avoiding oscillation phenomena. The optimal weighting coefficient was calculated on the basis of the mesh Peclet number. Regarding the quality processes, the model takes into account the main physical/chemical processes; these are degradation of dissolved carbonaceous substances, ammonium oxidation, algal uptake and denitrification, dissolved oxygen balance, including depletion by degradation processes and supply by physical reaeration and photosynthetic production. To properly simulate the river water quality, four state variables were considered: DO, BOD, NH4, and NO. The model was applied to the Savena River (Italy), which is the focus of a European-financed project for which quantity and quality data were gathered. A sensitivity analysis of the model output compared to the model input or parameters was carried out.


Treatment options for third ventricular colloid cysts: comparison of open microsurgical versus endoscopic resection.
We retrospectively reviewed our experience treating third ventricular colloid cysts to compare the efficacy of endoscopic and transcallosal approaches. Between September 1994 and March 2004, 55 patients underwent third ventricular colloid cyst resection. The transcallosal approach was used in 27 patients; the endoscopic approach was used in 28 patients. Age, sex, cyst diameter, and presence of hydrocephalus were similar between the two groups. The operating time and hospital stay were significantly longer in the transcallosal craniotomy group compared with the endoscopic group. Both approaches led to reoperations in three patients. The endoscopic group had two subsequent craniotomies for residual cysts and one repeat endoscopic procedure because of equipment malfunction. The transcallosal craniotomy group had two reoperations for fractured drainage catheters and one operation for epidural hematoma evacuation. The transcallosal craniotomy group had a higher rate of patients requiring a ventriculoperitoneal shunt (five versus two) and a higher infection rate (five versus none). Intermediate follow-up demonstrated more small residual cysts in the endoscopic group than in the transcallosal craniotomy group (seven versus one). Overall neurological outcomes, however, were similar in the two groups. Compared with transcallosal craniotomy, neuroendoscopy is a safe and effective approach for removal of colloid cysts in the third ventricle. The endoscope can be considered a first-line treatment for these lesions, with the understanding that a small number of these patients may need an open craniotomy to remove residual cysts.


Femoral loading mechanics in the Virginia opossum, Didelphis virginiana: torsion and mediolateral bending in mammalian locomotion.
Studies of limb bone loading in terrestrial mammals have typically found anteroposterior bending to be the primary loading regime, with torsion contributing minimally. However, previous studies have focused on large, cursorial eutherian species in which the limbs are held essentially upright. Recent in vivo strain data from the Virginia opossum (Didelphis virginiana), a marsupial that uses a crouched rather than an upright limb posture, have indicated that its femur experiences appreciable torsion during locomotion as well as strong mediolateral bending. The elevated femoral torsion and strong mediolateral bending observed in D. virginiana might result from external forces such as a medial inclination of the ground reaction force (GRF), internal forces deriving from a crouched limb posture, or a combination of these factors. To evaluate the mechanism underlying the loading regime of opossum femora, we filmed D. virginiana running over a force platform, allowing us to measure the magnitude of the GRF and its three-dimensional orientation relative to the limb, facilitating estimates of limb bone stresses. This three-dimensional analysis also allows evaluations of muscular forces, particularly those of hip adductor muscles, in the appropriate anatomical plane to a greater degree than previous two-dimensional analyses. At peak GRF and stress magnitudes, the GRF is oriented nearly vertically, inducing a strong abductor moment at the hip that is countered by adductor muscles on the medial aspect of the femur that place this surface in compression and induce mediolateral bending, corroborating and explaining loading patterns that were identified in strain analyses. The crouched orientation of the femur during stance in opossums also contributes to levels of femoral torsion as high as those seen in many reptilian taxa. Femoral safety factors were as high as those of non-avian reptiles and greater than those of upright, cursorial mammals, primarily because the load magnitudes experienced by opossums are lower than those of most mammals. Thus, the evolutionary transition from crouched to upright posture in mammalian ancestors may have been accompanied by an increase in limb bone load magnitudes.


Hominins on Flores, Indonesia, by one million years ago.
Previous excavations at Mata Menge and Boa Lesa in the Soa Basin of Flores, Indonesia, recovered stone artefacts in association with fossilized remains of the large-bodied Stegodon florensis florensis. Zircon fission-track ages from these sites indicated that hominins had colonized the island by 0.88 +/- 0.07 million years (Myr) ago. Here we describe the contents, context and age of Wolo Sege, a recently discovered archaeological site in the Soa Basin that has in situ stone artefacts and that lies stratigraphically below Mata Menge and immediately above the basement breccias of the basin. We show using (40)Ar/(39)Ar dating that an ignimbrite overlying the artefact layers at Wolo Sege was erupted 1.02 +/- 0.02 Myr ago, providing a new minimum age for hominins on Flores. This predates the disappearance from the Soa Basin of 'pygmy' Stegodon sondaari and Geochelone spp. (giant tortoise), as evident at the nearby site of Tangi Talo, which has been dated to 0.90 +/- 0.07 Myr ago. It now seems that this extirpation or possible extinction event and the associated faunal turnover were the result of natural processes rather than the arrival of hominins. It also appears that the volcanic and fluvio-lacustrine deposits infilling the Soa Basin may not be old enough to register the initial arrival of hominins on the island.


The Golgi zone of the rat spermatid and its role in the formation of cytoplasmic vesicles.
ELECTRON MICROSCOPIC STUDY OF THE GOLGI ZONE OF THE RAT SPERMATIDS REVEALED TWO MAIN TYPES OF VESICULAR STRUCTURES: flat vesicles generally accumulated in stacks and spheroidal vesicles of various sizes. The Golgi zone of young spermatids showed a predominance of flat vesicles and small spherical vesicles, while the "residual" Golgi zone of the maturing spermatids displayed an increased number of large spherical vesicles and thus became increasingly vacuolated in appearance. The images presented were suggestive of a transformation of the flat vesicles into spherical vesicles by fragmentation and dilatation and also of a passage of the spherical vesicles from the Golgi zone to the surrounding cytoplasm.


Invasive pneumococcal disease caused by mucoid serotype 3 Streptococcus pneumoniae: a case report and literature review.
Among the different serotypes of Streptococcus pneumoniae, serotype 3 has received global attention. We report the fatal case of a 76-year-old Japanese man who had an invasive pneumococcal disease associated with pneumonia caused by serotype 3 S. pneumoniae. The patient had a history of hypertension, laryngeal cancer, chronic obstructive pulmonary disease, and type 2 diabetes mellitus. Following a cerebral arteriovenous malformation hemorrhage, he underwent surgery to remove the hematoma and began rehabilitation. On day 66 of hospitalization, he suddenly developed a fever, and coarse crackles and wheezes were heard in his right lung. A diagnosis of hospital-acquired aspiration pneumonia was made, and initial treatment with piperacillin/tazobactam was started. Teicoplanin was added after S. pneumoniae was isolated from the blood culture, however, the patient died 5 days later. The S. pneumoniae detected in the sputum smear was serotype 3, showed mucoid colonies and susceptibility to penicillins, cephalosporins, carbapenems, and levofloxacin, but resistance to erythromycin. We experienced a fatal case of pneumonia caused by mucoid serotype 3 S. pneumoniae with a thick capsule. Serotype 3-associated pneumonia may develop a wider pulmonary infiltrative shadow, a prolonged therapeutic or hospitalization course, and a poor outcome. Careful observation and intervention are required, and the use of additional antibiotics or intravenous immunoglobulins should be considered in such cases. Pneumococcal immunization is also an important public health measure to minimize the development of severe infections caused by serotype 3 strains.


Silicon infrared diffuser for wireless communication.
We show what we believe to be a novel way to use silicon in infrared radio communication as a suitable material for the realization of optical diffusers in the range of 850-1600 nm. A crystalline silicon wafer is made porous by means of electrochemical etching. The porous silicon produced is optically characterized, and measurements report a high reflectance in the band of interest. We also study the angular distribution of diffused radiation by the porous silicon surface at different angles of incident radiation. Measurements show that radiation diffuses in a quasi-Lambertian manner, confirming the good performance of this material as an incident radiation diffuser.


Probabilistic Modeling of Imaging, Genetics and Diagnosis.
We propose a unified Bayesian framework for detecting genetic variants associated with disease by exploiting image-based features as an intermediate phenotype. The use of imaging data for examining genetic associations promises new directions of analysis, but currently the most widely used methods make sub-optimal use of the richness that these data types can offer. Currently, image features are most commonly selected based on their relevance to the disease phenotype. Then, in a separate step, a set of genetic variants is identified to explain the selected features. In contrast, our method performs these tasks simultaneously in order to jointly exploit information in both data types. The analysis yields probabilistic measures of clinical relevance for both imaging and genetic markers. We derive an efficient approximate inference algorithm that handles the high dimensionality of image and genetic data. We evaluate the algorithm on synthetic data and demonstrate that it outperforms traditional models. We also illustrate our method on Alzheimer's Disease Neuroimaging Initiative data.


Programmed activation of cancer cell apoptosis: A tumor-targeted phototherapeutic topoisomerase I inhibitor.
We report here a tumor-targeting masked phototherapeutic agent 1 (PT-1). This system contains SN-38-a prodrug of the topoisomerase I inhibitor irinotecan. Topoisomerase I is a vital enzyme that controls DNA topology during replication, transcription, and recombination. An elevated level of topoisomerase I is found in many carcinomas, making it an attractive target for the development of effective anticancer drugs. In addition, PT-1 contains both a photo-triggered moiety (nitrovanillin) and a cancer targeting unit (biotin). Upon light activation in cancer cells, PT-1 interferes with DNA re-ligation, diminishes the expression of topoisomerase I, and enhances the expression of inter alia mitochondrial apoptotic genes, death receptors, and caspase enzymes, inducing DNA damage and eventually leading to apoptosis. In vitro and in vivo studies showed significant inhibition of cancer growth and the hybrid system PT-1 thus shows promise as a programmed photo-therapeutic ("phototheranostic").


Expanded repeat in canine epilepsy.
Epilepsy afflicts 1% of humans and 5% of dogs. We report a canine epilepsy mutation and evidence for the existence of repeat-expansion disease outside humans. A canid-specific unstable dodecamer repeat in the Epm2b (Nhlrc1) gene recurrently expands, causing a fatal epilepsy and contributing to the high incidence of canine epilepsy. Tracing the repeat origins revealed two successive events, starting 50 million years ago, unique to canid evolution. A genetic test, presented here, will allow carrier and presymptomatic diagnosis and disease eradication. Clinicopathologic characterization establishes affected animals as a model for Lafora disease, the most severe teenage-onset human epilepsy.


From Beyond the Grave: Use of Medical Information from the Deceased to Guide Care of Living Relatives.
In order to inform patients of their genetic risks, access to the medical records and/or stored samples of their relatives is often helpful. We consider some of the obstacles to such access when these relatives are deceased and suggest how they might be navigated. We explore an issue first highlighted in 2004 by Lucassen et al. (Br Med J 328:952-953, 2004) and re-evaluate it in the wake of novel technologies and mainstreaming of genomic medicine. We find that it is still an issue in practice despite professional guidelines advocating access to familial information (Joint Committee on Genomics in Medicine 2019) and that the Human Tissue Act 2004 is often wrongly constructed as a reason to block access. Access is often obstructed by failing to adopt the necessary relational concept of autonomy that applies in genetic medicine as reported by Horton and Lucassen (Curr Genet Med Rep 7:85-91, 2019) and by considering confidentiality to be absolute, even after death. In response to a recent legal case about the confidentiality of genetic test results, and their disclosure to family members (ABC v St George's Healthcare NHS Trust 2020), Dove et al. (J Med Ethics 45:504-507, 2019) suggested that a duty to consider the interests of genetic relatives could co-exist alongside a duty of confidentiality to a patient. In this way, healthcare professionals can use professional judgement about the relative value of genetic information to family members. This is equally relevant in accessing deceased relatives' information. A recent systematic review found a high level of acceptability of postmortem use of genetic data for medical research amongst participants and their relatives, and it is reasonable to assume that this acceptability would extend to clinical practice as reported by Bak et al. (Eur J Hum Genet 28:403-416, 2020). Within clinical practice, access to medical records/samples of deceased relatives is often obstructed unnecessarily, potentially resulting in harm to the living relatives seeking advice. Consent to such access is important but need not be the bureaucratic hurdle that is often imposed.


Risk Factors of Chronic Rhinosinusitis After Functional Endoscopic Sinus Surgery.
BACKGROUND Clinical data of 288 chronic rhinosinusitis patients were retrospectively analyzed to investigate the risk factors of clinical prognosis, aiming to provide clinical evidence for the diagnosis and treatment of chronic rhinosinusitis. MATERIAL AND METHODS A total of 288 patients diagnosed with chronic rhinosinusitis in the Department of Otolaryngology of the First Affiliated Hospital of Xinjiang Medical University were recruited. Among all participants, 177 were male and 111 were female, aged from 22 to 83 years, (52±14) years on average. Subsequent follow-up was conducted to evaluate surgical efficacy. Influencing factors of clinical prognosis were analyzed by univariate and multivariate logistic regression analyses. RESULTS After functional endoscopic sinus surgery by Messerklinger technique, 187 (64.9%) patients were fully recovered, 72 (25.0%) presented with improvement, and 28 (10.1%) were untreated. Univariate logistic regression analysis revealed that 11 variables were correlated with the clinical prognosis of chronic rhinosinusitis. Multivariate logistic regression analysis demonstrated that age, history of allergic rhinitis, severity of dysosmia, history of nasosinusitis surgery, and long-term use of nasal decongestant were the risk factors, whereas comprehensive therapy after surgery was a protective factor. CONCLUSIONS More emphasis should be placed upon the factors associated with the clinical prognosis of patients with chronic rhinosinusitis following undergoing endoscopic sinus surgery, offering consolidated evidence for the prevention and treatment of chronic rhinosinusitis.


Breast MRI in patients after breast conserving surgery with sentinel node procedure using a superparamagnetic tracer.
A procedure for sentinel lymph node biopsy (SLNB) using superparamagnetic iron-oxide (SPIO) nanoparticles and intraoperative sentinel lymph node (SLN) detection was developed to overcome drawbacks associated with the current standard-of-care SLNB. However, residual SPIO nanoparticles can result in void artefacts at follow-up magnetic resonance imaging (MRI) scans. We present a grading protocol to quantitatively assess the severity of these artefacts and offer an option to minimise the impact of SPIO nanoparticles on diagnostic imaging. Follow-up mammography and MRI of two patient groups after a magnetic SLNB were included in the study. They received a 2-mL subareolar dose of SPIO (high-dose, HD) or a 0.1-mL intratumoural dose of SPIO (low-dose, LD). Follow-up mammography and MRI after magnetic SLNB were acquired within 4 years after breast conserving surgery (BCS). Two radiologists with over 10-year experience in breast imaging assessed the images and analysed the void artefacts and their impact on diagnostic follow-up. A total of 19 patients were included (HD, n = 13; LD, n = 6). In the HD group, 9/13 patients displayed an artefact on T1-weighted images up to 3.6 years after the procedure, while no impact of the SPIO remnants was observed in the LD group. SLNB using a 2-mL subareolar dose of magnetic tracer in patients undergoing BCS resulted in residual artefacts in the breast in the majority of patients, which may hamper follow-up MRI. This can be avoided by using a 0.1-mL intratumoural dose.


Behavioral and neurochemical effects of dietary methyl donor deficiency combined with unpredictable chronic mild stress in rats.
Methyl donor deficiencies and chronic stress cause depression independently, but their interaction has never been thoroughly evaluated. In our study, methyl donor deficient diet and chronic stress condition consisted respectively of a B2, B9, B12, and choline-free diet and a chronic mild stress procedure. Rats were randomly assigned to six groups with three "diet" conditions (free-feeding, pair-fed and methyl donor deficient diet) and two "stress" conditions (no-stress and stress) and were evaluated in the open-field, the elevated plus-maze and the forced swimming test. After the behavioral evaluation, corticosterone and homocysteine plasma levels were measured and dopamine, DOPAC, serotonin, 5HIAA concentrations were evaluated in several brain areas. Rats given a methyl donor deficient diet for 11 weeks causing elevated plasma homocysteine levels were compared to pair-fed and free-feeding rats with or without unpredictable chronic mild stress. Regardless of stress environmental conditions, the methyl donor deficient diet decreased plasma corticosterone levels and caused disinhibition in the elevated plus-maze condition relative to both control groups. However, stress potentiated the effects of the deficient regimen on rearing in the open-field and climbing in the forced swim test. The dietary changes involved in behavior and plasma corticosterone could be caused by homocysteine-induced decreases in dopamine and 5-hydroxytryptamine metabolites in selective brain regions and it can be noted that regardless of stress-conditions, methyl donor deficient diet decreases DOPAC/dopamine and 5HIAA/serotonin ratios in striatum and hypothalamus and selectively 5HIAA/serotonin ratio in the sensorimotor cortex. Our experimental data is particularly relevant in the context of neuropsychiatric disorders frequently associated with folate deficiency and hyperhomocysteinemia.


Continuous removal of small nonviable suspended mammalian cells and debris from bioreactors using inertial microfluidics.
Removing nonviable cells from a cell suspension is crucial in biotechnology and biomanufacturing. Label-free microfluidic cell separation devices based on dielectrophoresis, acoustophoresis, and deterministic lateral displacement are used to remove nonviable cells. However, their volumetric throughputs and test cell concentrations are generally too low to be useful in typical bioreactors in biomanufacturing. In this study, we demonstrate the efficient removal of small (<10 μm) nonviable cells from bioreactors while maintaining viable cells using inertial microfluidic cell sorting devices and characterize their performance. Despite the size overlap between viable and nonviable cell populations, the devices demonstrated 3.5-28.0% dead cell removal efficiency with 88.3-83.6% removal purity as well as 97.8-99.8% live cell retention efficiency at 4 million cells per mL with 80% viability. Cascaded and parallel configurations increased the cell concentration capacity (10 million cells per mL) and volumetric throughput (6-8 mL min-1). The system can be used for the removal of small nonviable cells from a cell suspension during continuous perfusion cell culture operations.


Photobiomodulation Therapy in the Treatment of Oral Mucositis, Dysphagia, Oral Dryness, Taste Alteration, and Burning Mouth Sensation Due to Cancer Therapy: A Case Series.
Oral complications of cancer therapy, such as oral dryness, dysphagia, and taste alteration, are associated with a negative impact in the quality of life of the patients. Few supportive care measures are available for such complications. This case series reveals the effectiveness of the photobiomodulation (PBM) therapy when used in a specific protocol and parameters, in the management of oral complications related to cancer therapy. Dysphagia was measured using the functional outcome swallowing scale for staging oropharyngeal dysphagia (FOSS). Oral mucositis was measured according to the National Cancer Institute scale. The quantity of the whole resting and stimulated saliva was measured in order to assess the oral dryness. In addition, the taste alteration was measured according to a protocol suggested by the International Standards organization (ISO). Sensation of burning mouth was measured using a visual analogue scale. These measurements were made before treatment, during, and at the end of the treatment. Diode laser 635 nm was used in 3 J/cm2. Five sessions interleaved with 24 h breaks were conducted for the dysphagia and oral dryness, and 10 sessions were conducted for the taste alteration and burning mouth sensation. Regardless of the limitations of this case series, PBM can be considered safe, time saving, and a promising approach for the management of the oral complications due to cancer therapy and the quality of life of cancer patients.


A randomized assessment of adding the kinase inhibitor lestaurtinib to first-line chemotherapy for FLT3-mutated AML.
The clinical benefit of adding FMS-like tyrosine kinase-3 (FLT3)-directed small molecule therapy to standard first-line treatment of acute myeloid leukemia (AML) has not yet been established. As part of the UK AML15 and AML17 trials, patients with previously untreated AML and confirmed FLT3-activating mutations, mostly younger than 60 years, were randomly assigned either to receive oral lestaurtinib (CEP701) or not after each of 4 cycles of induction and consolidation chemotherapy. Lestaurtinib was commenced 2 days after completing chemotherapy and administered in cycles of up to 28 days. The trials ran consecutively. Primary endpoints were overall survival in AML15 and relapse-free survival in AML17; outcome data were meta-analyzed. Five hundred patients were randomly assigned between lestaurtinib and control: 74% had FLT3-internal tandem duplication mutations, 23% FLT3-tyrosine kinase domain point mutations, and 2% both types. No significant differences were seen in either 5-year overall survival (lestaurtinib 46% vs control 45%; hazard ratio, 0.90; 95% CI 0.70-1.15; P = .3) or 5-year relapse-free survival (40% vs 36%; hazard ratio, 0.88; 95% CI 0.69-1.12; P = .3). Exploratory subgroup analysis suggested survival benefit with lestaurtinib in patients receiving concomitant azole antifungal prophylaxis and gemtuzumab ozogamicin with the first course of chemotherapy. Correlative studies included analysis of in vivo FLT3 inhibition by plasma inhibitory activity assay and indicated improved overall survival and significantly reduced rates of relapse in lestaurtinib-treated patients who achieved sustained greater than 85% FLT3 inhibition. In conclusion, combining lestaurtinib with intensive chemotherapy proved feasible in younger patients with newly diagnosed FLT3-mutated AML, but yielded no overall clinical benefit. The improved clinical outcomes seen in patients achieving sustained FLT3 inhibition encourage continued evaluation of FLT3-directed therapy alongside front-line AML treatment. The UK AML15 and AML17 trials are registered at www.isrctn.com/ISRCTN17161961 and www.isrctn.com/ISRCTN55675535 respectively.


Lateral entry compared with medial and lateral entry pin fixation for completely displaced supracondylar humeral fractures in children. A randomized clinical trial.
Closed reduction and percutaneous pin fixation is the treatment of choice for completely displaced (type-III) extension supracondylar fractures of the humerus in children, although controversy persists regarding the optimal pin-fixation technique. The purpose of this study was to compare the efficacy of lateral entry pin fixation with that of medial and lateral entry pin fixation for the operative treatment of completely displaced extension supracondylar fractures of the humerus in children. This prospective, randomized clinical trial had sufficient power to detect a 10% difference in the rate of loss of reduction between the two groups. The techniques of lateral entry and medial and lateral entry pin fixation were standardized in terms of the pin location, the pin size, the incision and position of the elbow used for medial pin placement, and the postoperative course. The primary study end points were a major loss of reduction and iatrogenic ulnar nerve injury. Secondary study end points included radiographic measurements, clinical alignment, Flynn grade, elbow range of motion, function, and complications. The lateral entry group (twenty-eight patients) and the medial and lateral entry group (twenty-four patients) were similar in terms of mean age, sex distribution, and preoperative displacement, comminution, and associated neurovascular status. No patient in either group had a major loss of reduction. There was no significant difference between the rates of mild loss of reduction, which occurred in six of the twenty-eight patients treated with lateral entry and one of the twenty-four treated with medial and lateral entry (p = 0.107). There were no cases of iatrogenic ulnar nerve injury in either group. There were also no significant differences (p > 0.05) between groups with respect to the Baumann angle, change in the Baumann angle, humerocapitellar angle, change in the humerocapitellar angle, Flynn grade, carrying angle, elbow flexion, elbow extension, total elbow range of motion, return to function, or complications. With use of the specific techniques employed in this study, both lateral entry pin fixation and medial and lateral entry pin fixation are effective in the treatment of completely displaced (type-III) extension supracondylar fractures of the humerus in children. Therapeutic Level I. See Instructions to Authors for a complete description of levels of evidence.


Accurate quantification of transcriptome from RNA-Seq data by effective length normalization.
We propose a novel, efficient and intuitive approach of estimating mRNA abundances from the whole transcriptome shotgun sequencing (RNA-Seq) data. Our method, NEUMA (Normalization by Expected Uniquely Mappable Area), is based on effective length normalization using uniquely mappable areas of gene and mRNA isoform models. Using the known transcriptome sequence model such as RefSeq, NEUMA pre-computes the numbers of all possible gene-wise and isoform-wise informative reads: the former being sequences mapped to all mRNA isoforms of a single gene exclusively and the latter uniquely mapped to a single mRNA isoform. The results are used to estimate the effective length of genes and transcripts, taking experimental distributions of fragment size into consideration. Quantitative RT-PCR based on 27 randomly selected genes in two human cell lines and computer simulation experiments demonstrated superior accuracy of NEUMA over other recently developed methods. NEUMA covers a large proportion of genes and mRNA isoforms and offers a measure of consistency ('consistency coefficient') for each gene between an independently measured gene-wise level and the sum of the isoform levels. NEUMA is applicable to both paired-end and single-end RNA-Seq data. We propose that NEUMA could make a standard method in quantifying gene transcript levels from RNA-Seq data.


Future Directions for Optimizing Clinical Science & Safety: Ecological Momentary Assessments in Suicide/Self-Harm Research.
Mobile technology has facilitated rapid growth in the use of intensive longitudinal methods (ILM), such as ecological momentary assessments (EMA), that help identify proximal indicators of risk in real-time and real-world settings. To realize the potential of ILM for advancing knowledge regarding suicidal and self-injurious thoughts and behaviors (SITB), this article aims to provide a systematic review of safety protocols in published ILM studies of youth SITB, highlight considerations for maximizing safety, and offer an agenda for future research. We conducted a systematic review of risk management strategies in published studies applying ILM to assess SITB in youth. The review indicated diverse safety strategies, with near-universal use of preventive strategies before beginning ILM surveys. Strategies for participant protection during the survey period included automated protective messages to seek support when elevated risk was detected; and staff-led strategies, some of which included active outreach to parents/caregivers when youth responses suggested elevated risk. Studies assessing suicidality all provided staff-led follow-up. There was minimal information on youth reactivity to intensive longitudinal assessments of SITB. Available evidence did not suggest increased suicidal ideation, suicide attempts, self-injurious behavior, or deaths with ILM. Based on the review, we propose a research agenda to inform safety procedures in ILM research and a model for managing risk in future ILM studies of youth SITB. This model begins with a needs assessment and proposes a "goodness of fit" approach for matching safety procedures to the specific needs of each ILM study.


Multi-day low dose ketamine infusion for the treatment of complex regional pain syndrome.
Complex regional pain syndrome (CRPS) is characterized by pain that is out of proportion to the injury and is regional in distribution. A large body of literature supports a dynamic change in the physiology and structure of central pain projecting neurons mediated through the N-methyl-D-aspartate (NMDA) receptor. A critical factor in central sensitization seems to be the release of the magnesium block on the NMDA receptor with influx of calcium and initiation of intracellular cascades. Current literature supports the effectiveness of ketamine in blocking central sensitization through its effects on the NMDA receptor. Recent treatment with anesthetic doses of ketamine in severely ill patients with generalized CRPS prompted our interest in a lower dose therapy. To report on the efficacy of low dose outpatient ketamine infusion for the treatment of CRPS diagnosed by International Association for the Study of Pain (IASP) criteria in patients who have failed conservative treatment. Open label, prospective, pain journal evaluation of a 10-day infusion of intravenous ketamine in the CRPS patient. Patients diagnosed with CRPS by a single neurologist were assigned to receive a 10-day outpatient infusion of ketamine supervised by an Anesthesiologist/Pain Management Specialist. The infusion was administered in a short procedure unit after each patient had been instructed on how to complete a pain questionnaire. Monitoring consisted of continuous ECG, pulse oximetry, and non-invasive blood pressure every 15 minutes. Patients made journal entries each day prior to the infusion of 40-80 mg of ketamine. The subjects were also asked to rate their pain intensity using a verbal analog pain scale of 0-10 and the affective component using a verbal scale of 0-4. There was a significant reduction in pain intensity from initiation of infusion (Day 1) to the 10th day, with a significant reduction in the percentage of patients experiencing pain by Day 10 as well as a reduction in the level of their "worst" pain. The nadirs of pain were lower by Day 10 with a significant reduction in the incidence of "punishing pain." Moreover, there was a significant improvement in the ability to initiate movement by the 10th day. A four-hour ketamine infusion escalated from 40-80 mg over a 10-day period can result in a significant reduction of pain with increased mobility and a tendency to decreased autonomic dysregulation.


14-3-3 proteins are essential for RAS/MAPK cascade signaling during pseudohyphal development in S. cerevisiae.
14-3-3 proteins are highly conserved ubiquitous proteins whose explicit functions have remained elusive. Here, we show that the S. cerevisiae 14-3-3 homologs BMH1 and BMH2 are not essential for viability or mating MAPK cascade signaling, but they are essential for pseudohyphal-development MAPK cascade signaling and other processes. Activated alleles of RAS2 and CDC42 induce pseudohyphal development and FG(TyA)-lacZ signaling in Bmh+ strains but not in ste20 (p65PAK) or bmh1 bmh2 mutant strains. Moreover, Bmh1p and Bmh2p associate with Ste20p in vivo. Three alleles of BMH1 encode proteins defective for FG(TyA)-lacZ signaling and association with Ste20p, yet these alleles complement other 14-3-3 functions. Therefore, the 14-3-3 proteins are specifically required for RAS/MAPK cascade signaling during pseudohyphal development in S. cerevisiae.


When pain is not only pain: inserting needles into the body evokes distinct reward-related brain responses in the context of a treatment.
The aim of this study was to compare behavioral and functional brain responses to the act of inserting needles into the body in two different contexts, treatment and stimulation, and to determine whether the behavioral and functional brain responses to a subsequent pain stimulus were also context dependent. Twenty-four participants were randomly divided into two groups: an acupuncture treatment (AT) group and an acupuncture stimulation (AS) group. Each participant received three different types of stimuli, consisting of tactile, acupuncture, and pain stimuli, and was given behavioral assessments during fMRI scanning. Although the applied stimuli were physically identical in both groups, the verbal instructions differed: participants in the AS group were primed to consider the acupuncture as a painful stimulus, whereas the participants in the AT group were told that the acupuncture was part of therapeutic treatment. Acupuncture yielded greater brain activation in reward-related brain areas (ventral striatum) of the brain in the AT group when compared to the AS group. Brain activation in response to pain stimuli was significantly attenuated in the bilateral secondary somatosensory cortex and the right dorsolateral prefrontal cortex after prior acupuncture needle stimulation in the AT group but not in the AS group. Inserting needles into the body in the context of treatment activated reward circuitries in the brain and modulated pain responses in the pain matrix. Our findings suggest that pain induced by therapeutic tools in the context of a treatment is modulated differently in the brain, demonstrating the power of context in medical practice.


Anthropogenically-Mediated Density Dependence in a Declining Farmland Bird.
Land management intrinsically influences the distribution of animals and can consequently alter the potential for density-dependent processes to act within populations. For declining species, high densities of breeding territories are typically considered to represent productive populations. However, as density-dependent effects of food limitation or predator pressure may occur (especially when species are dependent upon separate nesting and foraging habitats), high territory density may limit per-capita productivity. Here, we use a declining but widespread European farmland bird, the yellowhammer Emberiza citrinella L., as a model system to test whether higher territory densities result in lower fledging success, parental provisioning rates or nestling growth rates compared to lower densities. Organic landscapes held higher territory densities, but nests on organic farms fledged fewer nestlings, translating to a 5 times higher rate of population shrinkage on organic farms compared to conventional. In addition, when parental provisioning behaviour was not restricted by predation risk (i.e., at times of low corvid activity), nestling provisioning rates were higher at lower territory densities, resulting in a much greater increase in nestling mass in low density areas, suggesting that food limitation occurred at high densities. These findings in turn suggest an ecological trap, whereby preferred nesting habitat does not provide sufficient food for rearing nestlings at high population density, creating a population sink. Habitat management for farmland birds should focus not simply on creating a high nesting density, but also on ensuring heterogeneous habitats to provide food resources in close proximity to nesting birds, even if this occurs through potentially restricting overall nest density but increasing population-level breeding success.


Increased yield stability of field-grown winter barley (Hordeum vulgare L.) varietal mixtures through ecological processes.
Crop variety mixtures have the potential to increase yield stability in highly variable and unpredictable environments, yet knowledge of the specific mechanisms underlying enhanced yield stability has been limited. Ecological processes in genetically diverse crops were investigated by conducting field trials with winter barley varieties (Hordeum vulgare), grown as monocultures or as three-way mixtures in fungicide treated and untreated plots at three sites. Mixtures achieved yields comparable to the best performing monocultures whilst enhancing yield stability despite being subject to multiple predicted and unpredicted abiotic and biotic stresses including brown rust (Puccinia hordei) and lodging. There was compensation through competitive release because the most competitive variety overyielded in mixtures thereby compensating for less competitive varieties. Facilitation was also identified as an important ecological process within mixtures by reducing lodging. This study indicates that crop varietal mixtures have the capacity to stabilise productivity even when environmental conditions and stresses are not predicted in advance. Varietal mixtures provide a means of increasing crop genetic diversity without the need for extensive breeding efforts. They may confer enhanced resilience to environmental stresses and thus be a desirable component of future cropping systems for sustainable arable farming.


Unresectable solitary hepatocellular carcinoma not amenable to radiofrequency ablation: multicenter radiology-pathology correlation and survival of radiation segmentectomy.
Resection and radiofrequency ablation (RFA) are treatment options for hepatocellular carcinoma (HCC) <3 cm; there is interest in expanding the role of ablation to 3-5 cm. RFA is considered high-risk when the lesion is in close proximity to critical structures. Combining microcatheter technology and the localized emission properties of Y90, highly selective radioembolization is a possible alternative to RFA in such cases. We assessed the efficacy (response, radiology-pathology correlation, survival) of radiation segmentectomy in solitary HCC not amenable to RFA or resection. Patients with treatment-naïve, unresectable, solitary HCC ≤ 5 cm not amenable to RFA were included in this multicenter study. Administered dose, response rate, time-to-progression (modified Response Evaluation Criteria in Solid Tumors [mRECIST]), radiology-pathology correlation and long-term survival were assessed. In all, 102 patients were included in this study. mRECIST complete response (CR), partial response (PR), and stable disease (SD) were 47/99 (47%), 39/99 (39%), and 12/99 (12%), respectively. Median time-to-disease-progression was 33.1 months. In all, 33/102 (32%) patients were transplanted with a median (interquartile range [IQR]) time-to-transplantation of 6.3 months (3.6-9.7). Pathology revealed 100% and 50-99% necrosis in 17/33 (52%) and 16/33 (48%), respectively. Median overall survival was 53.4 months. Univariate analysis demonstrated a survival benefit for Eastern Cooperative Oncology Group (ECOG) 0 patients. In the multivariate model, age <65, ECOG 0, and Child-Pugh A were characteristics associated with longer survival. Radiation segmentectomy is an effective technique with a favorable risk profile and radiology-pathology outcomes for solitary HCC ≤ 5 cm. This approach may allow for treatment of HCC in difficult locations. Since RFA and resection are not options given tumor location, there appears to be a strong rationale for this technique as second choice.


Amorphous Silica: A New Antioxidant Role for Rapid Critical-Sized Bone Defect Healing.
Traumatic fractures cause structurally unstable sites due to severe bone loss. Such fractures generate a high yield of reactive oxygen species (ROS) that can lead to oxidative stress. Excessive and prolonged ROS activity impedes osteoblast differentiation and instigates long healing times. Stimulation of antioxidants such as superoxide dismutase (SOD1), are crucial to reduce ROS, stimulate osteogenesis, and strengthen collagen and mineral formation. Yet, no current fixative devices have shown an ability to enhance collagen matrix formation through antioxidant expression. This study reports plasma-enhanced chemical vapor deposition based amorphous silicon oxynitride (Si(ON)x) as a potential new fracture healing biomaterial that adheres well to the implant surface, releases Si(+4) to enhance osteogenesis, and forms a surface hydroxyapatite for collagen mineral attachment. These materials provide a sustained release of Si(+4) in physiological environment for extended times. The dissolution rate partially depends on the film chemistry and can be controlled by varying O/N ratio. The presence of Si(+4) enhances SOD1, which stimulates other osteogenic markers downstream and leads to rapid mineral formation. In vivo testing using a rat critical-sized calvarial defect model shows a more rapid bone-regeneration for these biomaterials as compared to control groups, that implies the clinical significance of the presented biomaterial.


Contrastive Learning and Neural Oscillations.
The concept of Contrastive Learning (CL) is developed as a family of possible learning algorithms for neural networks. CL is an extension of Deterministic Boltzmann Machines to more general dynamical systems. During learning, the network oscillates between two phases. One phase has a teacher signal and one phase has no teacher signal. The weights are updated using a learning rule that corresponds to gradient descent on a contrast function that measures the discrepancy between the free network and the network with a teacher signal. The CL approach provides a general unified framework for developing new learning algorithms. It also shows that many different types of clamping and teacher signals are possible. Several examples are given and an analysis of the landscape of the contrast function is proposed with some relevant predictions for the CL curves. An approach that may be suitable for collective analog implementations is described. Simulation results and possible extensions are briefly discussed together with a new conjecture regarding the function of certain oscillations in the brain. In the appendix, we also examine two extensions of contrastive learning to time-dependent trajectories.


American College of Critical Care Medicine Clinical Practice Parameters for Hemodynamic Support of Pediatric and Neonatal Septic Shock.
The American College of Critical Care Medicine provided 2002 and 2007 guidelines for hemodynamic support of newborn and pediatric septic shock. Provide the 2014 update of the 2007 American College of Critical Care Medicine "Clinical Guidelines for Hemodynamic Support of Neonates and Children with Septic Shock." Society of Critical Care Medicine members were identified from general solicitation at Society of Critical Care Medicine Educational and Scientific Symposia (2006-2014). The PubMed/Medline/Embase literature (2006-14) was searched by the Society of Critical Care Medicine librarian using the keywords: sepsis, septicemia, septic shock, endotoxemia, persistent pulmonary hypertension, nitric oxide, extracorporeal membrane oxygenation, and American College of Critical Care Medicine guidelines in the newborn and pediatric age groups. The 2002 and 2007 guidelines were widely disseminated, translated into Spanish and Portuguese, and incorporated into Society of Critical Care Medicine and American Heart Association/Pediatric Advanced Life Support sanctioned recommendations. The review of new literature highlights two tertiary pediatric centers that implemented quality improvement initiatives to improve early septic shock recognition and first-hour compliance to these guidelines. Improved compliance reduced hospital mortality from 4% to 2%. Analysis of Global Sepsis Initiative data in resource rich developed and developing nations further showed improved hospital mortality with compliance to first-hour and stabilization guideline recommendations. The major new recommendation in the 2014 update is consideration of institution-specific use of 1) a "recognition bundle" containing a trigger tool for rapid identification of patients with septic shock, 2) a "resuscitation and stabilization bundle" to help adherence to best practice principles, and 3) a "performance bundle" to identify and overcome perceived barriers to the pursuit of best practice principles.


Resources for functional genomics studies in Drosophila melanogaster.
Drosophila melanogaster has become a system of choice for functional genomic studies. Many resources, including online databases and software tools, are now available to support design or identification of relevant fly stocks and reagents or analysis and mining of existing functional genomic, transcriptomic, proteomic, etc. datasets. These include large community collections of fly stocks and plasmid clones, "meta" information sites like FlyBase and FlyMine, and an increasing number of more specialized reagents, databases, and online tools. Here, we introduce key resources useful to plan large-scale functional genomics studies in Drosophila and to analyze, integrate, and mine the results of those studies in ways that facilitate identification of highest-confidence results and generation of new hypotheses. We also discuss ways in which existing resources can be used and might be improved and suggest a few areas of future development that would further support large- and small-scale studies in Drosophila and facilitate use of Drosophila information by the research community more generally.


Ion mobility tandem mass spectrometry enhances performance of bottom-up proteomics.
One of the limiting factors in determining the sensitivity of tandem mass spectrometry using hybrid quadrupole orthogonal acceleration time-of-flight instruments is the duty cycle of the orthogonal ion injection system. As a consequence, only a fraction of the generated fragment ion beam is collected by the time-of-flight analyzer. Here we describe a method utilizing postfragmentation ion mobility spectrometry of peptide fragment ions in conjunction with mobility time synchronized orthogonal ion injection leading to a substantially improved duty cycle and a concomitant improvement in sensitivity of up to 10-fold for bottom-up proteomic experiments. This enabled the identification of 7500 human proteins within 1 day and 8600 phosphorylation sites within 5 h of LC-MS/MS time. The method also proved powerful for multiplexed quantification experiments using tandem mass tags exemplified by the chemoproteomic interaction analysis of histone deacetylases with Trichostatin A.