Effectiveness and Functional Mechanism of a Multicomponent Sanitizer against Biofilms Formed by Escherichia coli O157:H7 and Five Salmonella Serotypes Prevalent in the Meat Industry.
Biofilm formation by Escherichia coli O157:H7 and Salmonella enterica at meat processing plants poses a potential risk of meat product contamination. Many common sanitizers are unable to completely eradicate biofilms formed by these foodborne pathogens because of the three-dimensional biofilm structure and the presence of bacterial extracellular polymeric substances (EPSs). A novel multifaceted approach combining multiple chemical reagents with various functional mechanisms was used to enhance the effectiveness of biofilm control. We tested a multicomponent sanitizer consisting of a quaternary ammonium compound (QAC), hydrogen peroxide, and the accelerator diacetin for its effectiveness in inactivating and removing Escherichia coli O157:H7 and Salmonella enterica biofilms under meat processing conditions. E. coli O157:H7 and Salmonella biofilms on common contact surfaces were treated with 10, 20, or 100% concentrations of the multicomponent sanitizer solution for 10 min, 1 h, or 6 h, and log reductions in biofilm mass were measured. Scanning electron microscopy (SEM) was used to directly observe the effect of sanitizer treatment on biofilm removal and bacterial morphology. After treatment with the multicomponent sanitizer, viable E. coli O157:H7 and Salmonella biofilm cells were below the limit of detection, and the prevalence of both pathogens was low. After treatment with a QAC-based control sanitizer, surviving bacterial cells were countable, and pathogen prevalence was higher. SEM analysis of water-treated control samples revealed the three-dimensional biofilm structure with a strong EPS matrix connecting bacteria and the contact surface. Treatment with 20% multicomponent sanitizer for 10 min significantly reduced biofilm mass and weakened the EPS connection. The majority of the bacterial cells had altered morphology and compromised membrane integrity. Treatment with 100% multicomponent sanitizer for 10 min dissolved the EPS matrix, and no intact biofilm structure was observed; instead, scattered clusters of bacterial aggregates were detected, indicating the loss of cell viability and biofilm removal. These results indicate that the multicomponent sanitizer is effective, even after short exposure with dilute concentrations, against E. coli O157:H7 and S. enterica biofilms.


Regional differences in hamstring muscle damage after a marathon.
Previous studies suggest that marathon running induces lower extremity muscle damage. This study aimed to examine inter- and intramuscular differences in hamstring muscle damage after a marathon using transverse relaxation time (T2)-weighted magnetic resonance images (MRI). 20 healthy collegiate marathon runners (15 males) were recruited for this study. T2-MRI was performed before (PRE) and at 1 (D1), 3 (D3), and 8 days (D8) after marathon, and the T2 values of each hamstring muscle at the distal, middle, and proximal sites were calculated. Results indicated that no significant intermuscular differences in T2 changes were observed and that, regardless of muscle, the T2 values of the distal and middle sites increased significantly at D1 and D3 and recovered at D8, although those values of the proximal site remained constant. T2 significantly increased at distal and middle sites of the biceps femoris long head on D1 (p = 0.030 and p = 0.004, respectively) and D3 (p = 0.007 and p = 0.041, respectively), distal biceps femoris short head on D1 (p = 0.036), distal semitendinosus on D1 (p = 0.047) and D3 (p = 0.010), middle semitendinosus on D1 (p = 0.005), and distal and middle sites of the semimembranosus on D1 (p = 0.008 and p = 0.040, respectively) and D3 (p = 0.002 and p = 0.018, respectively). These results suggest that the distal and middle sites of the hamstring muscles are more susceptible to damage induced by running a full marathon. Conditioning that focuses on the distal and middle sites of the hamstring muscles may be more useful in improving recovery strategies after prolonged running.


Anticipatory pleasure in current psychosis: Cognitive and emotional correlates.
Anticipation of pleasure - a key aspect of hedonic experience - is a motivating factor for engaging in activities. Low levels of anticipatory pleasure and activity are found in individuals with psychosis. Cognitive factors (e.g., working memory and IQ) have been a focus of explanation for anticipation of pleasure in psychosis. However, cognitive factors do not fully account for such difficulties. It is plausible that emotional factors (e.g., depression, self-beliefs) also contribute. We examined anticipatory pleasure in relation to cognitive and emotional processes in patients with current psychosis. 128 patients with persecutory delusions in the context of non-affective psychosis completed assessments of anticipatory pleasure, cognitive functioning, emotional processes, and activity. Lower anticipatory pleasure was significantly associated with depression, insomnia, negative-self beliefs, suicidal ideation, poorer psychological wellbeing, and paranoia-related avoidance. There were no significant associations with working memory, physical activity, or meaningful activity. Emotional factors may play a more significant role than cognitive difficulties in the experience of anhedonia in psychosis. However, the cross-sectional design precludes causal inferences. Future research should examine whether, for example, improving self-concept or reducing paranoia-related avoidance leads to improvement in anticipatory pleasure in patients with psychosis.


Rhabdomyosarcoma of the breast - a rare malignancy.
Female, 60 FINAL DIAGNOSIS: Rhabdomyosarcoma of the breast Symptoms: Lump in axilla Medication: - Clinical Procedure: Mastectomy Specialty: Oncology. Rare disease. Primary nonepithelial malignancies of the breast include primary breast sarcomas, therapy-related breast sarcomas, the phyllodes tumors, and primary breast lymphomas. They account for less than 5% of all breast neoplasms. We report the case of a 60-year-old postmenopausal female diagnosed with rhabdomyosarcoma with infiltrating duct carcinoma. She was treated with modified radical mastectomy with axillary clearance and postoperative chemotherapy. Primary rhabdomyosarcoma of the breast in adults is extremely rare. Rhabdomyosarcomas in adults account for less than 3% of all adult primary soft-tissue sarcomas. Primary breast sarcomas usually present as large painless breast lumps with no associated skin and nipple changes or axillary lymphadenopathy; they are more aggressive and have more rapid growth than epithelial malignancies or benign breast lesions. The tumor can grow to large size, around 5.8 cm. Affected patients are typically women in their 50 s (ranging from 17 to 89 years), but it is also seen in men. The treatment of primary breast sarcomas requires a multidisciplinary approach. Surgery remains the mainstay of therapy. Chemotherapy has no clearly defined role in primary breast or soft-tissue sarcomas. The prognosis of primary breast sarcomas depends on the histologic grade and size of the tumor. They spread locally and hematogenously, but they are not usually associated with axillary lymphadenopathy.


Management of zygomatic complex residual deformity due to war in Iraq.
Since March 20, 2003, maxillofacial surgeons in Iraq encounter several trauma cases, part of which are zygomatic residual deformities that result from conventional war, civil unrest, crimes, and car explosions. This study aimed to determine the relationship between etiological factors and severity of posttraumatic zygomatic residual deformity and to investigate methods of surgical repairs, treatment modalities, and any complications that may have occurred during this critical time in Iraq. There were a total of 40 patients with posttraumatic deformity of the zygoma included in this study. Records of these patients (29 men and 11 women) who have been treated in the maxillofacial unit of a specialized surgery hospital (Medical City Hospital, Baghdad) between 2006 and 2011 were collected. Of all patients, 28 (70%) had injuries from missile whereas 12 patients (30%) had injuries from other causes (aggressive social behavior). According to the causes of delayed treatment, 30 patients (60%) were untreated previously (15 patients [50%] of them had delay because severe comminution, severe damage, or loss of the overlying soft tissue; 3 patients [10%] were undiagnosed; and 12 patients [40%] had treatment delayed as a result of major damage to other body parts or medical condition), 3 patients (30%) had improper reduction, and 7 patients (70%) had improper fixation. Our definitive treatment was as follows: 13 patients (32.5%) were treated by osteotomy, 10 patients (25%) were treated by onlay bone graft, 14 patients (35 %) were treated by both methods, and 3 patients (7.5%) were untreated. [corrected] We recommend that every surgeon who deals with residual deformity should clinically evaluate the residual deformity.


A novel metric for quantification of homogeneous and heterogeneous tumors in PET for enhanced clinical outcome prediction.
Oncologic PET images provide valuable information that can enable enhanced prognosis of disease. Nonetheless, such information is simplified significantly in routine clinical assessment to meet workflow constraints. Examples of typical FDG PET metrics include: (i) SUVmax, (2) total lesion glycolysis (TLG), and (3) metabolic tumor volume (MTV). We have derived and implemented a novel metric for tumor quantification, inspired in essence by a model of generalized equivalent uniform dose as used in radiation therapy. The proposed metric, denoted generalized effective total uptake (gETU), is attractive as it encompasses the abovementioned commonly invoked metrics, and generalizes them, for both homogeneous and heterogeneous tumors, using a single parameter a. We evaluated this new metric for improved overall survival (OS) prediction on two different baseline FDG PET/CT datasets: (a) 113 patients with squamous cell cancer of the oropharynx, and (b) 72 patients with locally advanced pancreatic adenocarcinoma. Kaplan-Meier survival analysis was performed, where the subjects were subdivided into two groups using the median threshold, from which the hazard ratios (HR) were computed in Cox proportional hazards regression. For the oropharyngeal cancer dataset, MTV, TLG, SUVmax, SUVmean and SUVpeak produced HR values of 1.86, 3.02, 1.34, 1.36 and 1.62, while the proposed gETU metric for a = 0.25 (greater emphasis on volume information) enabled significantly enhanced OS prediction with HR = 3.94. For the pancreatic cancer dataset, MTV, TLG, SUVmax, SUVmean and SUVpeak resulted in HR values of 1.05, 1.25, 1.42, 1.45 and 1.52, while gETU at a = 3.2 (greater emphasis on SUV information) arrived at an improved HR value of 1.61. Overall, the proposed methodology allows placement of differing degrees of emphasis on tumor volume versus uptake for different types of tumors to enable enhanced clinical outcome prediction.


Genetic analysis reveals the wild ancestors of the llama and the alpaca.
The origins of South America's domestic alpaca and llama remain controversial due to hybridization, near extirpation during the Spanish conquest and difficulties in archaeological interpretation. Traditionally, the ancestry of both forms is attributed to the guanaco, while the vicuña is assumed never to have been domesticated. Recent research has, however, linked the alpaca to the vicuña, dating domestication to 6000-7000 years before present in the Peruvian Andes. Here, we examine in detail the genetic relationships between the South American camelids in order to determine the origins of the domestic forms, using mitochondrial (mt) and microsatellite DNA. MtDNA analysis places 80% of llama and alpaca sequences in the guanaco lineage, with those possessing vicuña mtDNA being nearly all alpaca or alpaca-vicuña hybrids. We also examined four microsatellites in wild known-provenance vicuña and guanaco, including two loci with non-overlapping allele size ranges in the wild species. In contrast to the mtDNA, these markers show high genetic similarity between alpaca and vicuña, and between llama and guanaco, although bidirectional hybridization is also revealed. Finally, combined marker analysis on a subset of samples confirms the microsatellite interpretation and suggests that the alpaca is descended from the vicuña, and should be reclassified as Vicugna pacos. This result has major implications for the future management of wild and domestic camelids in South America.


Allelic variants of the NLR protein Rpi-chc1 differentially recognize members of the Phytophthora infestans PexRD12/31 effector superfamily through the leucine-rich repeat domain.
Phytophthora infestans is a pathogenic oomycete that causes the infamous potato late blight disease. Resistance (R) genes from diverse Solanum species encode intracellular receptors that trigger effective defense responses upon the recognition of cognate RXLR avirulence (Avr) effector proteins. To deploy these R genes in a durable fashion in agriculture, we need to understand the mechanism of effector recognition and the way the pathogen evades recognition. In this study, we cloned 16 allelic variants of the Rpi-chc1 gene from Solanum chacoense and other Solanum species, and identified the cognate P. infestans RXLR effectors. These tools were used to study effector recognition and co-evolution. Functional and non-functional alleles of Rpi-chc1 encode coiled-coil nucleotide-binding leucine-rich repeat (CNL) proteins, being the first described representatives of the CNL16 family. These alleles have distinct patterns of RXLR effector recognition. While Rpi-chc1.1 recognized multiple PexRD12 (Avrchc1.1) proteins, Rpi-chc1.2 recognized multiple PexRD31 (Avrchc1.2) proteins, both belonging to the PexRD12/31 effector superfamily. Domain swaps between Rpi-chc1.1 and Rpi-chc1.2 revealed that overlapping subdomains in the leucine-rich repeat (LRR) domain are responsible for the difference in effector recognition. This study showed that Rpi-chc1.1 and Rpi-chc1.2 evolved to recognize distinct members of the same PexRD12/31 effector family via the LRR domain. The biased distribution of polymorphisms suggests that exchange of LRRs during host-pathogen co-evolution can lead to novel recognition specificities. These insights will guide future strategies to breed durable resistant varieties.


Nutritional Keys for Intestinal Barrier Modulation.
The intestinal tract represents the largest interface between the external environment and the human body. Nutrient uptake mostly happens in the intestinal tract, where the epithelial surface is constantly exposed to dietary antigens. Since inflammatory response toward these antigens may be deleterious for the host, a plethora of protective mechanisms take place to avoid or attenuate local damage. For instance, the intestinal barrier is able to elicit a dynamic response that either promotes or impairs luminal antigens adhesion and crossing. Regulation of intestinal barrier is crucial to control intestinal permeability whose increase is associated with chronic inflammatory conditions. The cross talk among bacteria, immune, and dietary factors is able to modulate the mucosal barrier function, as well as the intestinal permeability. Several nutritional products have recently been proposed as regulators of the epithelial barrier, even if their effects are in part contradictory. At the same time, the metabolic function of the microbiota generates new products with different effects based on the dietary content. Besides conventional treatments, novel therapies based on complementary nutrients are now growing. Fecal therapy has been recently used for the clinical treatment of refractory Clostridium difficile infection instead of the classical antibiotic therapy. In the present review, we will outline the epithelial response to nutritional components derived from dietary intake and microbial fermentation focusing on the consequent effects on the integrity of the epithelial barrier.


The genetic basis of cerebral palsy.
Although prematurity and hypoxic-ischaemic injury are well-recognized contributors to the pathogenesis of cerebral palsy (CP), as many as one-third of children with CP may lack traditional risk factors. For many of these children, a genetic basis to their condition is suspected. Recent findings have implicated copy number variants and mutations in single genes in children with CP. Current studies are limited by relatively small patient numbers, the underlying genetic heterogeneity identified, and the paucity of validation studies that have been performed. However, several genes mapping to intersecting pathways controlling neurodevelopment and neuronal connectivity have been identified. Analogous to other neurodevelopmental disorders such as autism and intellectual disability, the genomic architecture of CP is likely to be highly complex. Although we are just beginning to understand genetic contributions to CP, new insights are anticipated to serve as a unique window into the neurobiology of CP and suggest new targets for intervention.


Cross-sectional survey of treatment practices for urethritis at pharmacies, private clinics and government health facilities in coastal Kenya: many missed opportunities for HIV prevention.
While bacterial sexually transmitted infections (STIs) are important cofactors for HIV transmission, STI control has received little attention in recent years. The aim of this study was to assess STI treatment and HIV testing referral practices among health providers in Kenya. In 2011 we assessed quality of case management for male urethritis at pharmacies, private clinics and government health facilities in coastal Kenya using simulated visits at pharmacies and interviews at pharmacies and health facilities. Quality was assessed using Ministry of Health guidelines. Twenty (77%) of 26 pharmacies, 20 (91%) of 22 private clinics and all four government facilities in the study area took part. The median (IQR) number of adult urethritis cases per week was 5 (2-10) at pharmacies, 3 (1-3) at private clinics and 5 (2-17) at government facilities. During simulated visits, 10% of pharmacies prescribed recommended antibiotics at recommended dosages and durations and, during interviews, 28% of pharmacies and 27% of health facilities prescribed recommended antibiotics at recommended dosages and durations. Most regimens were quinolone-based. HIV testing was recommended during 10% of simulated visits, 20% of pharmacy interviews and 25% of health facility interviews. In an area of high STI burden, most men with urethritis seek care at pharmacies and private clinics. Most providers do not comply with national guidelines and very few recommend HIV testing. In order to reduce the STI burden and mitigate HIV transmission, there is an urgent need for innovative dissemination of up-to-date guidelines and inclusion of all health providers in HIV/STI programmes.


RStrucFam: a web server to associate structure and cognate RNA for RNA-binding proteins from sequence information.
RNA-binding proteins (RBPs) interact with their cognate RNA(s) to form large biomolecular assemblies. They are versatile in their functionality and are involved in a myriad of processes inside the cell. RBPs with similar structural features and common biological functions are grouped together into families and superfamilies. It will be useful to obtain an early understanding and association of RNA-binding property of sequences of gene products. Here, we report a web server, RStrucFam, to predict the structure, type of cognate RNA(s) and function(s) of proteins, where possible, from mere sequence information. The web server employs Hidden Markov Model scan (hmmscan) to enable association to a back-end database of structural and sequence families. The database (HMMRBP) comprises of 437 HMMs of RBP families of known structure that have been generated using structure-based sequence alignments and 746 sequence-centric RBP family HMMs. The input protein sequence is associated with structural or sequence domain families, if structure or sequence signatures exist. In case of association of the protein with a family of known structures, output features like, multiple structure-based sequence alignment (MSSA) of the query with all others members of that family is provided. Further, cognate RNA partner(s) for that protein, Gene Ontology (GO) annotations, if any and a homology model of the protein can be obtained. The users can also browse through the database for details pertaining to each family, protein or RNA and their related information based on keyword search or RNA motif search. RStrucFam is a web server that exploits structurally conserved features of RBPs, derived from known family members and imprinted in mathematical profiles, to predict putative RBPs from sequence information. Proteins that fail to associate with such structure-centric families are further queried against the sequence-centric RBP family HMMs in the HMMRBP database. Further, all other essential information pertaining to an RBP, like overall function annotations, are provided. The web server can be accessed at the following link: http://caps.ncbs.res.in/rstrucfam .


Verification in referral-based crowdsourcing.
Online social networks offer unprecedented potential for rallying a large number of people to accomplish a given task. Here we focus on information gathering tasks where rare information is sought through "referral-based crowdsourcing": the information request is propagated recursively through invitations among members of a social network. Whereas previous work analyzed incentives for the referral process in a setting with only correct reports, misreporting is known to be both pervasive in crowdsourcing applications, and difficult/costly to filter out. A motivating example for our work is the DARPA Red Balloon Challenge where the level of misreporting was very high. In order to undertake a formal study of verification, we introduce a model where agents can exert costly effort to perform verification and false reports can be penalized. This is the first model of verification and it provides many directions for future research, which we point out. Our main theoretical result is the compensation scheme that minimizes the cost of retrieving the correct answer. Notably, this optimal compensation scheme coincides with the winning strategy of the Red Balloon Challenge.


Automated Microscopic Analysis of Metal Sulfide Colonization by Acidophilic Microorganisms.
Industrial biomining processes are currently focused on metal sulfides and their dissolution, which is catalyzed by acidophilic iron(II)- and/or sulfur-oxidizing microorganisms. Cell attachment on metal sulfides is important for this process. Biofilm formation is necessary for seeding and persistence of the active microbial community in industrial biomining heaps and tank reactors, and it enhances metal release. In this study, we used a method for direct quantification of the mineral-attached cell population on pyrite or chalcopyrite particles in bioleaching experiments by coupling high-throughput, automated epifluorescence microscopy imaging of mineral particles with algorithms for image analysis and cell quantification, thus avoiding human bias in cell counting. The method was validated by quantifying cell attachment on pyrite and chalcopyrite surfaces with axenic cultures of Acidithiobacillus caldus, Leptospirillum ferriphilum, and Sulfobacillus thermosulfidooxidans. The method confirmed the high affinity of L. ferriphilum cells to colonize pyrite and chalcopyrite surfaces and indicated that biofilm dispersal occurs in mature pyrite batch cultures of this species. Deep neural networks were also applied to analyze biofilms of different microbial consortia. Recent analysis of the L. ferriphilum genome revealed the presence of a diffusible soluble factor (DSF) family quorum sensing system. The respective signal compounds are known as biofilm dispersal agents. Biofilm dispersal was confirmed to occur in batch cultures of L. ferriphilum and S. thermosulfidooxidans upon the addition of DSF family signal compounds.IMPORTANCE The presented method for the assessment of mineral colonization allows accurate relative comparisons of the microbial colonization of metal sulfide concentrate particles in a time-resolved manner. Quantitative assessment of the mineral colonization development is important for the compilation of improved mathematical models for metal sulfide dissolution. In addition, deep-learning algorithms proved that axenic or mixed cultures of the three species exhibited characteristic biofilm patterns and predicted the biofilm species composition. The method may be extended to the assessment of microbial colonization on other solid particles and may serve in the optimization of bioleaching processes in laboratory scale experiments with industrially relevant metal sulfide concentrates. Furthermore, the method was used to demonstrate that DSF quorum sensing signals directly influence colonization and dissolution of metal sulfides by mineral-oxidizing bacteria, such as L. ferriphilum and S. thermosulfidooxidans.


Mechanisms for similarity matching in disparity measurement.
Early neural mechanisms for the measurement of binocular disparity appear to operate in a manner consistent with cross-correlation-like processes. Consequently, cross-correlation, or cross-correlation-like procedures have been used in a range of models of disparity measurement. Using such procedures as the basis for disparity measurement creates a preference for correspondence solutions that maximize the similarity between local left and right eye image regions. Here, we examine how observers' perception of depth in an ambiguous stereogram is affected by manipulations of luminance and orientation-based image similarity. Results show a strong effect of coarse-scale luminance similarity manipulations, but a relatively weak effect of finer-scale manipulations of orientation similarity. This is in contrast to the measurements of depth obtained from a standard cross-correlation model. This model shows strong effects of orientation similarity manipulations and weaker effects of luminance similarity. In order to account for these discrepancies, the standard cross-correlation approach may be modified to include an initial spatial frequency filtering stage. The performance of this adjusted model most closely matches human psychophysical data when spatial frequency filtering favors coarser scales. This is consistent with the operation of disparity measurement processes where spatial frequency and disparity tuning are correlated, or where disparity measurement operates in a coarse-to-fine manner.


Identification of adducts formed in the reaction of 5'-acetoxy-N'-nitrosonornicotine with deoxyguanosine and DNA.
N'-Nitrosonornicotine (NNN) is believed to play an important role as a cause of cancer in people who use tobacco products and is considered to be a human carcinogen. NNN requires metabolism to form DNA adducts, which are absolutely critical to its carcinogenic properties. Previous studies have identified cytochrome P450-catalyzed 2'- and 5'-hydroxylation of NNN as potential DNA adduct forming metabolic pathways. 5'-Hydroxylation is the more prevalent of these in monkeys and humans and is known to generate mutagenic intermediates, but the DNA adducts formed by this pathway have never been characterized. In this study, we used 5'-acetoxyNNN as a stable precursor to 5'-hydroxyNNN and investigated its esterase-catalyzed reactions with deoxyguanosine (dGuo) and DNA. Adducts resulting from carbocation and oxonium ion intermediates, produced by the spontaneous decomposition of 5'-hydroxyNNN, were identified. The carbocation pathway resulted in the formation of 2-[2-hydroxy-5-(3-pyridyl)pyrrolidin-1-yl]deoxyinosine (12) which was characterized by comparison to an independently synthesized standard. Treatment of 12 with NaBH(3)CN produced two diastereomers of 2-[2-(3-pyridyl)pyrrolidin-1-yl]deoxyinosine (14), and their absolute configurations at the 2-position were determined by comparison to synthetic standards. The oxonium ion pathway produced diastereomers of N(2)[5-(3-pyridyl)tetrahydrofuran-2-yl]dGuo (16), identified by comparison to synthetic standards. The absolute configuration at the 5-position was determined by establishing the stereochemistry of the enantiomers of 5-(3-pyridyl)-2-hydroxytetrahydrofuran at the 5-position and allowing these to react individually with dGuo. Treatment of 16 with NaBH(3)CN produced N(2)[4-hydroxy-4-(3-pyridyl)but-1-yl]dGuo (18) which was also synthesized independently. Using liquid chromatography-electrospray ionization-tandem mass spectrometry with selected reaction monitoring, we identified adducts 12 and 16 as products of the reactions of 5'-acetoxyNNN with dGuo. Similarly, adducts 14 and 18 were identified as products of the reaction of 5'-acetoxyNNN with DNA followed by NaBH(3)CN treatment and enzymatic hydrolysis. These results provide the first structural characterization of DNA adducts that can be formed by 5'-hydroxylation of NNN.


Functional kinomics identifies candidate therapeutic targets in head and neck cancer.
To identify novel therapeutic drug targets for p53-mutant head and neck squamous cell carcinoma (HNSCC). RNAi kinome viability screens were performed on HNSCC cells, including autologous pairs from primary tumor and recurrent/metastatic lesions, and in parallel on murine squamous cell carcinoma (MSCC) cells derived from tumors of inbred mice bearing germline mutations in Trp53, and p53 regulatory genes: Atm, Prkdc, and p19(Arf). Cross-species analysis of cell lines stratified by p53 mutational status and metastatic phenotype was used to select 38 kinase targets. Both primary and secondary RNAi validation assays were performed on additional HNSCC cell lines to credential these kinase targets using multiple phenotypic endpoints. Kinase targets were also examined via chemical inhibition using a panel of kinase inhibitors. A preclinical study was conducted on the WEE1 kinase inhibitor, MK-1775. Our functional kinomics approach identified novel survival kinases in HNSCC involved in G2-M cell-cycle checkpoint, SFK, PI3K, and FAK pathways. RNAi-mediated knockdown and chemical inhibition of the WEE1 kinase with a specific inhibitor, MK-1775, had a significant effect on both viability and apoptosis. Sensitivity to the MK-1775 kinase inhibitor is in part determined by p53 mutational status, and due to unscheduled mitotic entry. MK-1775 displays single-agent activity and potentiates the efficacy of cisplatin in a p53-mutant HNSCC xenograft model. WEE1 kinase is a potential therapeutic drug target for HNSCC. This study supports the application of a functional kinomics strategy to identify novel therapeutic targets for cancer.


Gap geometry dictates epithelial closure efficiency.
Closure of wounds and gaps in tissues is fundamental for the correct development and physiology of multicellular organisms and, when misregulated, may lead to inflammation and tumorigenesis. To re-establish tissue integrity, epithelial cells exhibit coordinated motion into the void by active crawling on the substrate and by constricting a supracellular actomyosin cable. Coexistence of these two mechanisms strongly depends on the environment. However, the nature of their coupling remains elusive because of the complexity of the overall process. Here we demonstrate that epithelial gap geometry in both in vitro and in vivo regulates these collective mechanisms. In addition, the mechanical coupling between actomyosin cable contraction and cell crawling acts as a large-scale regulator to control the dynamics of gap closure. Finally, our computational modelling clarifies the respective roles of the two mechanisms during this process, providing a robust and universal mechanism to explain how epithelial tissues restore their integrity.


Assessing risk of bias in studies that evaluate health care interventions: recommendations in the misinformation age.
Methods to assess the risk of bias in a way that is reliable, reproducible. and transparent to readers, have evolved over time. Viswanathan et al. recently provided updated recommendations for assessing risk of bias in systematic reviews of health care interventions. We comment on their recommendations and discuss new tools in development that we, as co-convenors and coordinators of the Cochrane Bias Methods Group, are leading, which complement the methods recommended.


Cost-effectiveness of water quality interventions for preventing diarrhoeal disease in developing countries.
Using effectiveness data from a recent systematic review and cost data from programme implementers and World Health Organization (WHO) databases, we conducted a cost-effectiveness analysis to compare non-piped in source- (dug well, borehole and communal stand post) and four types of household- (chlorination, filtration, solar disinfection, flocculation/disinfection) based interventions to improve the microbial quality of water for preventing diarrhoeal disease. Results are reported for two WHO epidemiological sub-regions, Afr-E (sub-Saharan African countries with very high adult and child mortality) and Sear-D (South East Asian countries with high adult and child mortality) at 50% intervention coverage. Measured against international benchmarks, source- and household-based interventions were generally cost effective or highly cost effective even before the estimated saving in health costs that would offset the cost of implementation. Household-based chlorination was the most cost-effective where resources are limited; household filtration yields additional health gains at higher budget levels. Flocculation/disinfection was strongly dominated by all other interventions; solar disinfection was weakly dominated by chlorination. In addition to cost-effectiveness, choices among water quality interventions must be guided by local conditions, user preferences, potential for cost recovery from beneficiaries and other factors.


Addition of clonidine to bupivacaine in transversus abdominis plane block prolongs postoperative analgesia after cesarean section.
The aim was to compare duration of postoperative analgesia with addition of clonidine to bupivacaine in bilateral transversus abdominis plane (TAP) block after lower segment cesarean section (LSCS). One hundred American Society of Anesthesiologists (ASA) grade I and II pregnant patients undergoing LSCS under spinal anesthesia were randomly divided to receive either 20 ml bupivacaine 0.25% (Group B; n = 50) or 20 ml bupivacaine+1ug/kg clonidine bilaterally (Group BC; n = 50) in TAP block in a double-blind fashion. The total duration of analgesia, patient satisfaction score, total requirement of analgesics in the first 24 h, and the side effects of clonidine such as sedation, dryness of mouth, hypotension, and bradycardia were observed. P < 0.05 was taken as significant. In 99 patients analyzed, TAP block failed in five patients. Duration of analgesia was significantly longer in Group BC (17.8 ± 3.7 h) compared to Group B (7.3 ± 1.2 h; P < 0.01). Mean consumption of diclofenac was 150 mg and 65.4 mg in Groups B and BC (P < 0.01), respectively. All patients in Group BC were extremely satisfied (P < 0.01) while those in Group B were satisfied. Thirteen patients (28%) in Group BC were sedated but arousable (P = 0.01) compared to none in Group B. In Group BC, 19 patients complained of dry mouth compared to 13 in Group B (P = 0.121). None of the patients experienced hypotension or bradycardia. Addition of clonidine 1 μg/kg to 20 ml bupivacaine 0.25% in TAP block bilaterally for cesarean section significantly increases the duration of postoperative analgesia, decreases postoperative analgesic requirement, and increases maternal comfort compared to 20 ml of bupivacaine 0.25% alone.


Loss of inner retinal neurons after retinal ischemia in rats.
Ischemia is a risk factor for eye diseases like ocular vein occlusion or glaucoma. To investigate effects of ischemia-reperfusion (I/R) a lot of different animal models are used, studying one or two different cell types, which creates heterogeneity of data. The aim of this study was to investigate the function and morphology of the whole retina and different retinal cell types in an I/R model. I/R was induced by elevating the intraocular pressure in the right eyes of rats. Twenty-one days after ischemia, electroretinogram measurements were performed. Changes in layer thickness were investigated. Changes of RGC, amacrine-, rod bipolar-, and glia cells as well as presence of apoptosis were analyzed immunohistologically. A-wave- and b-wave amplitudes were decreased; histology showed a reduction of RGC- and inner plexiform layer thickness and a 29% loss of RGCs occurred in ischemic eyes (P = 0.016). An increase of apoptotic cells was detected in the GCL and INL of ischemic retinas (P < 0.05). Also, a loss of cholinergic amacrine cells (control: 11 ± 1 cells/mm, I/R: 4 ± 1 cells/mm, P < 0.001), but no change in rod bipolar cell numbers was noted. Our study allowed a comparison of the effects of I/R for different retinal cell types. Cells in the outer retina seemed to be more resistant to ischemic damage compared with cells of the inner retina. We hypothesize that a degenerative process, like a secondary wave of apoptosis, occurs 21 days after I/R, causing progressive damage in the retina.


Tailoring of Atlantic salmon (Salmo salar L.) flesh lipid composition and sensory quality by replacing fish oil with a vegetable oil blend.
Atlantic salmon (Salmo salar L.) juveniles were fed either 100% fish oil (FO), 75% vegetable oil (VO), or 100% VO throughout their life cycle to harvest weight followed by a finishing diet period when all groups were fed 100% FO. The two experimental VO diets were tested at two different locations (Scotland and Norway) against the same control diet (100% FO). The VO blend was composed of rapeseed oil, palm oil, and linseed oil using capelin oil as a control for fatty acid class compositions. Flesh fatty acid profiles were measured regularly throughout the experiment, with the times of sampling determined by changes in pellet size/lipid content and fish life stage. Growth and mortality rates were not significantly affected by dietary fatty acid compositions throughout the life cycle, except during the seawater winter period in Norway when both growth and protein utilization were increased in salmon fed 100% VO compared to 100% FO. Flesh fatty acid composition was highly influenced by that of the diet, and after the finishing diet period the weekly intake recommendations of very long chain n-3 polyunsaturated fatty acid (VLCn-3 PUFA) for human health were 80 and 56% satisfied by a 200 g meal of 75% VO and 100% VO flesh, respectively. No effect on flesh astaxanthin levels was observed in relation to changing dietary oil sources. Sensory evaluation showed only minor differences between salmon flesh from the dietary groups, although prior to the finishing diet period, flesh from 100% VO had less rancid and marine characteristics and was preferred over flesh from the other dietary groups by a trained taste panel. After the finishing diet period, the levels of typical vegetable oil fatty acids in flesh were reduced, whereas those of VLCn-3 PUFA increased to levels comparable with a 100% FO fed salmon. No differences in any of the sensory characteristics were observed between dietary groups. By blending VOs to provide balanced levels of dietary fatty acids, up to 100% of the fish oil can be replaced by the VO blend without compromising growth or flesh quality. At the same time, 75% of the dietary fish oil can be replaced without compromising flesh VLCn-3 PUFA content, thereby providing a beneficial nutritional profile for human consumption.


The ontogeny of alpha-fetoprotein gene expression in the mouse gastrointestinal tract.
The ontogeny of alpha-fetoprotein (AFP) gene expression has been examined in the fetal and adult mouse gastrointestinal tract. AFP mRNA constitutes approximately 0.1% of total mRNA in the fetal gut. The transcripts were localized by in situ hybridization to the epithelial cells lining the villi of the fetal gut. At birth, AFP mRNA declines rapidly to achieve low adult basal levels, which are not affected by different alleles of raf, a gene that determines the adult basal level of AFP mRNA in the liver. The basal level in the adult gut is the consequence of continued AFP transcription in a small number of enteroendocrine cells that are distributed infrequently on the villi. These cells were identified by double antibody staining with antibodies to chromogranin A, an enteroendocrine cell marker and AFP. Previous studies resulted in the generation of a line of transgenic mice containing an internally deleted AFP gene that was greatly overexpressed in the fetal gut. The basis for the inappropriately high level expression of the transgene was shown to be the consequence of very high levels of transcription in the epithelial cells of the villi rather than to expression in inappropriate cell types. The cis-acting DNA sequences required for expression of the AFP gene in the gut were investigated using Caco-2 cells, a human colon adenocarcinoma cell line. These experiments indicated that, with one exception, the regulatory elements required in both the promoter and enhancer regions of the gene coincided with those that are necessary for high level expression in the liver. The one exception was enhancer II, located 5 kbp of DNA upstream of the gene, which exhibited no activity in Caco-2 cells.


Spectral Processing for Denoising and Compression of 3D Meshes Using Dynamic Orthogonal Iterations.
Recently, spectral methods have been extensively used in the processing of 3D meshes. They usually take advantage of some unique properties that the eigenvalues and the eigenvectors of the decomposed Laplacian matrix have. However, despite their superior behavior and performance, they suffer from computational complexity, especially while the number of vertices of the model increases. In this work, we suggest the use of a fast and efficient spectral processing approach applied to dense static and dynamic 3D meshes, which can be ideally suited for real-time denoising and compression applications. To increase the computational efficiency of the method, we exploit potential spectral coherence between adjacent parts of a mesh and then we apply an orthogonal iteration approach for the tracking of the graph Laplacian eigenspaces. Additionally, we present a dynamic version that automatically identifies the optimal subspace size that satisfies a given reconstruction quality threshold. In this way, we overcome the problem of the perceptual distortions, due to the fixed number of subspace sizes that is used for all the separated parts individually. Extensive simulations carried out using different 3D models in different use cases (i.e., compression and denoising), showed that the proposed approach is very fast, especially in comparison with the SVD based spectral processing approaches, while at the same time the quality of the reconstructed models is of similar or even better reconstruction quality. The experimental analysis also showed that the proposed approach could also be used by other denoising methods as a preprocessing step, in order to optimize the reconstruction quality of their results and decrease their computational complexity since they need fewer iterations to converge.


Insertion element IS987 from Mycobacterium bovis BCG is located in a hot-spot integration region for insertion elements in Mycobacterium tuberculosis complex strains.
Most strains of the Mycobacterium tuberculosis complex carry multiple copies of an IS3-like element, and these strains are highly polymorphic with regard to the site of integration in the chromosome. In contrast, Mycobacterium bovis BCG contains a single copy of the insertion element, and in all strains this copy is integrated at the same site in the chromosome. In this study, we determined the sequence of the single-copy insertion element from M. bovis BCG, IS987, and its flanking regions. The analysis of IS987 revealed that this element was virtually identical to the sequence of IS986 from M. tuberculosis. IS987 is located in a region containing direct repeats (DRs). The cloned flanking regions contained 20 virtually identical DRs of 36 bp, each separated by 35 to 41 bp of spacer DNA. Analysis of chromosomal DNA by the polymerase chain reaction revealed the presence of a cluster of 49 DRs, and IS987 is inserted in the 30th DR. Furthermore, the DR sequences were found to occur only in species of the M. tuberculosis complex and not in nine other mycobacterial species tested. Analysis of 14 M. tuberculosis strains revealed the presence of one insertion sequence element in the DR-containing region of eight strains, two insertion sequence elements were located in the DR region of five strains, and one strain did not contain an insertion sequence element in this region. Additionally, the DR-containing regions of these 14 M. tuberculosis strains were polymorphic in length and composition. We conclude that the DR cluster is a specific, hot-spot region for integration of insertion elements in the chromosome of M. tuberculosis complex strains.


Methylation profile of genes involved in inflammation, in the blood from pregnancies with maternal preeclampsia due to untreated gestational diabetes mellitus.
To investigate DNA methylation changes in peripheral blood from patients with gestational diabetes mellitus (GDM) and preeclampsia (PE) due to poorly treated GDM. Eighteen pregnant women participated in the study: 6 with GDM, 6 with PE, and 6 healthy controls. The promoter methylation status of genes was profiled using the Human Inflammatory Response and Autoimmunity EpiTect Methyl II Signature PCR Array profiles. The results were validated with quantitative real-time polymerase chain reaction (qRT-PCR). Fewer inflammation-related genes were significantly hypomethylated in PE cases compared to healthy subjects than in GDM cases. Some of the examined genes show different methylation patterns between GDM and PE. The epigenetic changes observed in this study indicate that GDM and PE exhibit specific DNA methylation profiles, with possible clinical applications.


The diversity of prokaryotic DDE transposases of the mutator superfamily, insertion specificity, and association with conjugation machineries.
Transposable elements (TEs) are major components of both prokaryotic and eukaryotic genomes and play a significant role in their evolution. In this study, we have identified new prokaryotic DDE transposase families related to the eukaryotic Mutator-like transposases. These genes were retrieved by cascade PSI-Blast using as initial query the transposase of the streptococcal integrative and conjugative element (ICE) TnGBS2. By combining secondary structure predictions and protein sequence alignments, we predicted the DDE catalytic triad and the DNA-binding domain recognizing the terminal inverted repeats. Furthermore, we systematically characterized the organization and the insertion specificity of the TEs relying on these prokaryotic Mutator-like transposases (p-MULT) for their mobility. Strikingly, two distant TE families target their integration upstream σA dependent promoters. This allowed us to identify a transposase sequence signature associated with this unique insertion specificity and to show that the dissymmetry between the two inverted repeats is responsible for the orientation of the insertion. Surprisingly, while DDE transposases are generally associated with small and simple transposons such as insertion sequences (ISs), p-MULT encoding TEs show an unprecedented diversity with several families of IS, transposons, and ICEs ranging in size from 1.1 to 52 kb.


Acute renal failure: nephrosonographic findings in asphyxiated neonates.
To determine the incidence of acute renal failure (ARF) and nephrosonographic findings among asphyxiated neonates, and to correlate this with uric acid levels and the severity of hypoxic encephalopathy, we studied 80 full-term appropriate-for-date singleton neonates with perinatal asphyxia, and 30 healthy full-term neonates as controls from March 2006 to February 2007. A detailed history, thorough clinical examination along with investigations, including urine examination, 24-h urine collection, ultrasonography of abdomen and cranium, serum electrolytes, blood urea nitrogen, serum creatinine, and serum uric acid were obtained. ARF developed in 45% (36/80) of the asphyxiated neonates. Forty-eight (60%) neonates showed significant elevation of blood urea and 41 (51.3%) neonates had significant elevation of serum creatinine than the control group (P < 0.001). Sixty-two (77.5%) neonates developed significant elevation of serum uric acid levels, and nephrosonography revealed hyperechogenicity in all of them, while only two among the healthy neonates showed the raised uric acid levels (P < 0.001). Nonoliguric renal failure was seen 28/36 (77.8%) of the neonates with ARF, whereas eight (22.2%) neonates had oliguric renal failure. Eight (27.8%) patients among ARF patients maintained abnormal biochemical parameters after 2 weeks, and of whom four patients died after variable lengths of time with a mortality rate of 11.11%. Kidneys are the most common organs involved in perinatal asphyxia, and uric acid might be a causative factor for failure in addition to hypoxic insult. Routine use of kidney function test, along with abdominal ultrasonography form an important screening tool to detect any additional morbidity in these patients.


Predictors of noncompliance in patients with schizophrenia.
Around 50% of patients with schizophrenia do not fully comply with treatment, and noncompliance is linked to relapse, rehospitalization, poor outcome, and high economic costs. The health belief model views noncompliance as a decision made by the patient, arrived at after weighing the perceived risks and benefits of treatment. A MEDLINE search for the years 1980-2002 using combinations of the keywords schizophrenia, compliance, adherence, antipsychotics, tolerability, and side effects was used to identify articles investigating the factors influencing compliance in schizophrenia. Many factors influence compliance, including those that affect patients' beliefs about their illness and the benefits of treatment (e.g., insight into illness, belief that medication can ameliorate symptoms), perceived costs of treatment (e.g., medication side effects), and barriers to treatment (e.g., ease of access to treatment, degree of family or social support). Medication side effects that are distressing to patients and linked to noncompliance include extrapyramidal side effects, neuroleptic dysphoria, akathisia, sexual dysfunction, and weight gain. Compliance can be improved by cognitive-behavioral therapies, such as compliance therapy, and other psychosocial interventions associated with improved social functioning and a lower risk of rehospitalization. Treatment adherence may also be improved by use of atypical antipsychotics with few perceived side effects. By considering the factors leading to noncompliance and adopting a comprehensive strategy for improving compliance, encompassing psychosocial intervention and optimum choice of medication, the management of schizophrenia could be greatly improved.


General prognostic scores in outcome prediction for cancer patients admitted to the intensive care unit.
Intensivists and nursing staff are often reluctant to admit patients with cancer to the intensive care unit even though these patients' survival rate has improved since the 1980s. To identify factors associated with mortality in cancer patients admitted to the intensive care unit and to assess and compare the effectiveness of 3 general prognostic models: the Acute Physiology and Chronic Health Evaluation (APACHE) II, the Simplified Acute Physiology Score (SAPS II), and the Sequential Organ Failure Assessment (SOFA). A prospective observational cohort study was performed in 2 general intensive care units. Discrimination was assessed by using area under the receiver operating characteristic curves, and calibration was evaluated by using Hosmer-Lemeshow goodness-of-fit tests. A total of 126 patients were included during a 3-year period. The observed mortality was 46.8%. All 3 general models showed excellent discrimination (area under the curve >0.8) and good calibration (P = .17, .14, and .22 for APACHE II, SAPS II, and SOFA, respectively). However, discrimination was significantly better with APACHE II scores than with SOFA scores (P = .02). Multivariate analyses indicated that independent of the 3 severity-of-illness scores, unfavorable risk factors for mortality included a patient's preadmission performance status, source of admission (internal medicine vs surgery department), and the presence of septic shock, infection, or anemia. Combining SOFA and SAPS II scores with these variables created prognostic models with improved calibration and discrimination. The general prognostic models seem fairly accurate in the prediction of mortality in critically ill cancer patients in the intensive care unit.


Pregnancy-associated Cushing's disease? An exploratory retrospective study.
In most clinical series of Cushing's disease (CD), over 80% of patients are women, many of whom are of reproductive age. The year following pregnancy may be a common time to develop CD. We sought to establish the incidence of CD onset associated with pregnancy. A retrospective review was conducted for patients with biochemically-proven CD. Demographics, clinical history, biochemistry, imaging, pathology, and outcomes were reviewed. Pregnancy-associated CD was defined as symptom onset within 1 year of childbirth. Over 10 years, 77 patients including 64 women (84%), with CD underwent endonasal surgery. Of the 64 women, 64% were of reproductive age (15-45 years) at the time of diagnosis, and 11 (27%) met criteria for pregnancy-associated CD. Of these 11 women, median number of pregnancies prior to onset of CD was 2 (range 1-4) compared to zero (range 0-7) for 30 other women with CD onset during reproductive age (p = 0.0024). With an average follow-up of 47 ± 34 months, sustained surgical remission rates for woman with pregnancy-associated CD, other women of reproductive age, and women not of reproductive age were 91%, 80% and 83%, respectively. The average lag-time from symptom onset to diagnosis for women with pregnancy-associated CD was 4 ± 2 years. In this exploratory study, over one quarter of women of reproductive age with CD appeared to have symptomatic disease onset within 1 year of childbirth. This relatively high rate of pregnancy-associated CD suggests a possible causal relationship related to the stress of pregnancy and pituitary corticotroph hyperactivity in the peripartum period. This possible association suggests a heightened degree of clinical suspicion and biochemical testing for CD may be warranted after childbirth. Further study of this possible link between pregnancy and CD is warranted.


A Markov random field model for network-based analysis of genomic data.
A central problem in genomic research is the identification of genes and pathways involved in diseases and other biological processes. The genes identified or the univariate test statistics are often linked to known biological pathways through gene set enrichment analysis in order to identify the pathways involved. However, most of the procedures for identifying differentially expressed (DE) genes do not utilize the known pathway information in the phase of identifying such genes. In this article, we develop a Markov random field (MRF)-based method for identifying genes and subnetworks that are related to diseases. Such a procedure models the dependency of the DE patterns of genes on the networks using a local discrete MRF model. Simulation studies indicated that the method is quite effective in identifying genes and subnetworks that are related to disease and has higher sensitivity and lower false discovery rates than the commonly used procedures that do not use the pathway structure information. Applications to two breast cancer microarray gene expression datasets identified several subnetworks on several of the KEGG transcriptional pathways that are related to breast cancer recurrence or survival due to breast cancer. The proposed MRF-based model efficiently utilizes the known pathway structures in identifying the DE genes and the subnetworks that might be related to phenotype. As more biological networks are identified and documented in databases, the proposed method should find more applications in identifying the subnetworks that are related to diseases and other biological processes.


Modified "Allele-Specific qPCR" Method for SNP Genotyping Based on FRET.
The proposed method is a modified and improved version of the existing "Allele-specific q-PCR" (ASQ) method for genotyping of single nucleotide polymorphism (SNP) based on fluorescence resonance energy transfer (FRET). This method is similar to frequently used techniques like Amplifluor and Kompetitive allele specific PCR (KASP), as well as others employing common universal probes (UPs) for SNP analyses. In the proposed ASQ method, the fluorophores and quencher are located in separate complementary oligonucleotides. The ASQ method is based on the simultaneous presence in PCR of the following two components: an allele-specific mixture (allele-specific and common primers) and a template-independent detector mixture that contains two or more (up to four) universal probes (UP-1 to 4) and a single universal quencher oligonucleotide (Uni-Q). The SNP site is positioned preferably at a penultimate base in each allele-specific primer, which increases the reaction specificity and allele discrimination. The proposed ASQ method is advanced in providing a very clear and effective measurement of the fluorescence emitted, with very low signal background-noise, and simple procedures convenient for customized modifications and adjustments. Importantly, this ASQ method is estimated as two- to ten-fold cheaper than Amplifluor and KASP, and much cheaper than all those methods that rely on dual-labeled probes without universal components, like TaqMan and Molecular Beacons. Results for SNP genotyping in the barley genes HvSAP16 and HvSAP8, in which stress-associated proteins are controlled, are presented as proven and validated examples. This method is suitable for bi-allelic uniplex reactions but it can potentially be used for 3- or 4-allelic variants or different SNPs in a multiplex format in a range of applications including medical, forensic, or others involving SNP genotyping.


The adeno-associated virus rh10 vector is an effective gene transfer system for chronic spinal cord injury.
Treatment options for chronic spinal cord injury (SCI) remain limited due to unfavourable changes in the microenvironment. Gene therapy can overcome these barriers through continuous delivery of therapeutic gene products to the target tissue. In particular, adeno-associated virus (AAV) vectors are potential candidates for use in chronic SCI, considering their safety and stable gene expression in vivo. Given that different AAV serotypes display different cellular tropisms, it is extremely important to select an optimal serotype for establishing a gene transfer system during the chronic phase of SCI. Therefore, we generated multiple AAV serotypes expressing ffLuc-cp156, a fusion protein of firefly luciferase and Venus, a variant of yellow fluorescent protein with fast and efficient maturation, as a reporter, and we performed intraparenchymal injection in a chronic SCI mouse model. Among the various serotypes tested, AAVrh10 displayed the highest photon count on bioluminescence imaging. Immunohistological analysis revealed that AAVrh10 showed favourable tropism for neurons, astrocytes, and oligodendrocytes. Additionally, with AAVrh10, the area expressing Venus was larger in the injury epicentre and extended to the surrounding tissue. Furthermore, the fluorescence intensity was significantly higher with AAVrh10 than with the other vectors. These results indicate that AAVrh10 may be an appropriate serotype for gene delivery to the chronically injured spinal cord. This promising tool may be applied for research and development related to the treatment of chronic SCI.


Can gastric irrigation prevent infection during NOTES mesh placement?
Natural orifice transluminal endoscopic surgery (NOTES) ventral hernia repair could avoid abdominal wall incisions. The infectious risk for mesh placement is of concern. We compared NOTES with laparoscopic mesh placement. Thirty-seven swine were randomized to abdominal wall polypropylene mesh placement via NOTES or laparoscopy or NOTES control. All animals received antibiotics and gastric irrigation; the laparoscopy group also received preoperative acid suppression. In the NOTES mesh group, the 2-cm(2) polypropylene mesh was placed using a transgastric transportation device and clipped to the anterior abdominal wall. The control animals underwent endoscopy (no gastrotomy) followed by laparoscopic mesh placement or NOTES only without mesh placement. Necropsy was performed at 14 days. One NOTES mesh placement was incomplete (endoscope failure). All mesh animals survived to 14 days. At necropsy, significantly more mesh infections were noted in the NOTES mesh versus laparoscopy group (4:11 vs 0:14; p = 0.03). Gastric irrigation reduced the bacterial load significantly in all groups (p < 0.001). Infection was independent of gastric bacterial load. No difference between acid suppressed and non-suppressed animals was seen. The mesh placement via NOTES is technically feasible but has a high infection rate despite irrigation. Sterile conduits are needed to enable NOTES-type hernia repair with mesh.


Tuberculosis among individuals with community-acquired pneumonia presenting to emergency in Gaborone, Botswana.
Delays in diagnosing Tuberculosis (TB) are associated with increased transmission. TB may present as a clinical syndrome that mimics community-acquired pneumonia (CAP). The aim of this paper was to determine frequency of TB among patients with CAP at a referral hospital in Gaborone, Botswana. We performed a retrospective study of adults presenting with CAP from April 2010-October 2011 to the Emergency Department (ED); we matched this cohort to the National Botswana Tuberculosis Registry (NBTR) to identify individuals subsequently diagnosed with TB. We assessed demographics, time to TB diagnosis, clinical outcomes and performed logistic regressions to identify factors associated with TB diagnosis. We identified 1305 individuals presenting with CAP; TB was subsequently diagnosed in 68 (5.2%). The median time to TB diagnosis was 9.5 days. Forty percent were AFB sputum smear positive and 87% were identified as being HIV-positive. Subsequent diagnosis of TB is common among individuals with CAP at our ED, suggesting that TB may be present at the time of CAP presentation. Given the lack of distinguishing clinical factors between pulmonary TB and CAP, adults presenting with CAP should be evaluated for active TB in Botswana.


Severe neurotoxicity following 5-fluorouracil-based chemotherapy in a patient with dihydropyrimidine dehydrogenase deficiency.
Patients with decreased dihydropyrimidine dehydrogenase (DPD) activity are at increased risk for experiencing serious adverse reactions following 5-fluorouracil (5-FU)-based chemotherapy. Symptoms include severe and potentially life-threatening gastrointestinal toxicity, myelosuppression, and neurological toxicity. In the present study, we describe a 50-year-old Caucasian man who developed severe encephalopathy during his second cycle of 5-FU chemotherapy. The patient remained in a comatose state for 4 days but then showed dramatic improvement in his neurological status following continuous i.v. infusion of thymidine at 8 g/m2/day. Laboratory studies revealed the patient to be severely DPD deficient, as demonstrated by DPD enzyme activity from peripheral blood mononuclear cells being below the lower limit of the 95th percentile of a control population and by Western immunoblot analysis showing undetectable levels of DPD protein. Additional studies revealed a significant defect in pyrimidine catabolism with a 3.3- and 365-fold increase in the levels of uracil in plasma and urine, respectively, compared to normal subjects. Family studies suggest that the inheritance pattern of this syndrome is complex and most consistent with an autosomal recessive trait. This study demonstrates that cancer patients with DPD deficiency are at increased risk for developing severe neurological toxicity secondary to 5-FU chemotherapy, and that infusional thymidine should be considered as a potential rescue agent against this particular host toxicity.


Electrochemical quantification of transmitter concentration in single nanoscale vesicles isolated from PC12 cells.
We use an electrochemical platform, nanoparticle tracking analysis, and differential centrifugation of single catecholamine vesicles to study the properties of nanometer transmitter vesicles, including the number of molecules, size, and catecholamine concentration inside. Vesicle impact electrochemical cytometry (VIEC) was used to quantify the catecholamine content of single vesicles in different batches isolated from pheochromocytoma (PC12) cells with different ultracentrifugation speeds. We show that, vesicles containing less catecholamine are obtained at subsequent centrifugation steps with higher speed (force). Important to quantification, the cumulative content after subsequent centrifugation steps is equivalent to that of one-step centrifugation at the highest speed, 70 000g. Moreover, as we count molecules in the vesicles, we compared molecular numbers from VIEC, flow VIEC, and intracellular VIEC to corresponding vesicle size measured by nanoparticle tracking analysis to evaluate catecholamine concentration in vesicles. The data suggest that vesicular catecholamine concentration is relatively constant and independent of the vesicular size, indicating vesicular transmitter content as a main factor regulating the vesicle size.


Effect of Thyroperoxidase and Deiodinase Inhibition on Anterior Swim Bladder Inflation in the Zebrafish.
A set of adverse outcome pathways (AOPs) linking inhibition of thyroperoxidase and deiodinase to impaired swim bladder inflation in fish has recently been developed. These AOPs help to establish links between these thyroid hormone (TH) disrupting molecular events and adverse outcomes relevant to aquatic ecological risk assessment. Until now, very little data on the effects of TH disruption on inflation of the anterior chamber (AC) of the swim bladder were available. The present study used zebrafish exposure experiments with three model compounds with distinct thyroperoxidase and deiodinase inhibition potencies (methimazole, iopanoic acid, and propylthiouracil) to evaluate this linkage. Exposure to all three chemicals decreased whole body triiodothyronine (T3) concentrations, either through inhibition of thyroxine (T4) synthesis or through inhibition of Dio mediated conversion of T4 to T3. A quantitative relationship between reduced T3 and reduced AC inflation was established, a critical key event relationship linking impaired swim bladder inflation to TH disruption. Reduced inflation of the AC was directly linked to reductions in swimming distance compared to controls as well as to chemical-exposed fish whose ACs inflated. Together the data provide compelling support for AOPs linking TH disruption to impaired AC inflation in fish.


Modifications of default-mode network connectivity in patients with cerebral glioma.
The aim of the study was to evaluate connectivity modifications in the Default Mode Network (DMN) in patients with cerebral glioma, and to correlate these modifications to tumor characteristics. Twenty-four patients with a left-hemisphere cerebral tumor (14 grade II and 10 grade IV gliomas) and 14 healthy age-matched right-hand volunteers were enrolled in the study. Subjects underwent fMRI while performing language tasks for presurgical mapping. Data was analyzed with independent component analysis in order to identify the DMN. DMN group maps were produced by random-effect analysis (p<0.001, FDR-corrected). An analysis of variance across the three groups (p<0.05) and post-hoc t-test contrasts between pairs of groups were calculated (p<0.05, FDR-corrected). All three groups showed typical DMN areas. However, reduced DMN connectivity was detected in tumor patients with respect to controls. A significantly increased and reduced integration of DMN areas was observed in the hippocampal and prefrontal regions, respectively. Modifications were closely related to tumor grading. Moreover, the DMN lateralized to the hemisphere contralateral to tumor in the low-grade, but not in the high-grade tumor patients. Modifications of DMN connectivity were induced by gliomas and differed for high and low grade tumors.


Varicella vaccine effectiveness during an outbreak in a partially vaccinated population in Spain.
An outbreak of varicella occurred between December 2004 and April 2005 in the primary school and day-care center of a town of 5430 inhabitants in Spain. Although the varicella vaccine is not included in the infant vaccination program, some children had been vaccinated before the outbreak. The aim of this study was to estimate varicella vaccine effectiveness during an outbreak that took place in a partially vaccinated population of children. A cohort study was carried out. Cases were identified through notification by doctors and active search. Information was gathered on the current disease, history of varicella and previous vaccinations together with age, course, school year and other sociodemographic factors. The relative risk (RR) of varicella and the contribution of the previously mentioned factors to the probability of contracting the disease was estimated for vaccinated and unvaccinated children. Participation reached 96.5% (387 of 401 children) in the school and 91.2% (31 of 34 children) in the day-care center. Of 269 children with no history of varicella and with a documented vaccination record, 96 (35.7%) had been previously vaccinated. During the outbreak, 148 cases of varicella were observed. The overall attack rate was 54.4%, 22.9% in vaccinated and 72.8% in unvaccinated children. RR of varicella in vaccinated children was 0.31 (95% confidence interval [CI] = 0.21-0.46). The overall adjusted vaccine effectiveness against varicella was 69.5% (95% CI = 50.5-81.5%) and 96.9% (95% CI = 77.5-99.6%) against mild and severe forms. Only time since vaccination was associated with vaccine failure. In the varicella outbreak studied, we conclude that vaccine was effective in the prevention of the disease, particularly in its moderate and severe forms, but because the proportion of vaccinated children was low, an outbreak still occurred.


Laparoscopic Liver Resection Difficulty Score-a Validation Study.
The technical complexity of laparoscopic liver resection (LLR) poses unique challenges distinct from open surgery. An objective scoring system was developed that preoperatively quantifies the difficulty of LRR to help guide surgeon decision-making regarding the feasibility and safety of minimally invasive approaches. The aim of this multiinstitutional study was to externally validate this scoring system. Patients who underwent LLR at two institutions were reviewed. LLR difficulty score (LDS) was calculated based on patient, tumor, and anatomic characteristics by two independent, blinded hepatobiliary surgeons. Surrogates of case complexity (e.g., conversion rate, operative time) were used for validation of this index. From 2006 to 2016, 444 LLR were scored as low (n = 94), intermediate (n = 98), and high difficulty (n = 152) with respective conversion rates of 5.3%, 15.7%, and 25%. Cases of higher LDS correlated with larger mean blood loss (203 ml vs. 331 ml vs. 635 ml). Mean operative and Pringle maneuver used were associated with increasing LDS (155 min vs. 202 min vs. 315 min and 14.4% vs. 29.7% vs. 45.1% respectively). These operative surrogates of difficulty correlated significantly with the LDS (all p < 0.0001). This comprehensive external validation of the LDS is robust and applicable in diverse patient populations. This LDS serves as a useful objective predictor of technical difficulty for LLR to help surgeons in selecting patients according to their individual operative experience and is valuable for preoperative risk estimation and stratification in randomized trials.


Management of a 3-year-old child with ambiguous genitalia.
True hermaphrodites have characteristics tissue both sex in the form of presence of vagina uterus, fallopian tubes and ovaries as in female and presence of testis seminiferous tubules but penis as in penoscrotal hypospadias and bifid scrotal sac confuses as well developed labia majora. Determination of sex as per parents view in our social set up. The case was provisionally diagnosed as intersex. Total abdominal hysterectomy, bilateral salphingo-oophorectomy and colpectomy were conducted. A 3-year-old child referred by a pediatrician to Nalanda Medical College, child surgical department for investigation and management of his ambiguous genitalia having following features like, urine was not passing from the tip of penis; penis along with apparently looking female genitalia was taken under treatment. Examination under general anesthesia showed a presence of both male/female internal genitalia and gonads with features of external genitalia of both sexes. After total abdominal hysterectomy and human chorionic gonadotrophin treatment growth of phallus and testicle was noted. The child was made male after extirpation of female genitals. This case illustrates that gender correction can be made as per patients consent in adult but at this stage social customs and parents consent dictated the treatment.


A New Edible Film to Produce In Vitro Meat.
In vitro meat is a novel concept of food science and biotechnology. Methods to produce in vitro meat employ muscle cells cultivated on a scaffold in a serum-free medium using a bioreactor. The microstructure of the scaffold is a key factor, because muscle cells must be oriented to generate parallel alignments of fibers. This work aimed to develop a new scaffold (microstructured film) to grow muscle fibers. The microstructured edible films were made using micromolding technology. A micromold was tailor-made using a laser cutting machine to obtain parallel fibers with a diameter in the range of 70-90 µm. Edible films were made by means of solvent casting using non-mammalian biopolymers. Myoblasts were cultured on flat and microstructured films at three cell densities. Cells on the microstructured films grew with a muscle fiber morphology, but in the case of using the flat film, they only produced unorganized cell proliferation. Myogenic markers were assessed using quantitative polymerase chain reaction. After 14 days, the expression of desmin, myogenin, and myosin heavy chain were significantly higher in microstructured films compared to the flat films. The formation of fiber morphology and the high expression of myogenic markers indicated that a microstructured edible film can be used for the production of in vitro meat.


Variation in male effects on female fecundity in Drosophila melanogaster.
In many species, males have the capacity to directly influence (either positively or negatively) the fitness of their mates and offspring, not only via parental care contributions and/or precopulatory resource provisioning, but also via the post-copulatory activity of those substances passed on to their mates in their ejaculates. Here, we examine how an individual male's identity may be related to phenotypic variation in short-term female fecundity in the model species, Drosophila melanogaster. The effect of male identity on short-term fecundity stimulation of females was repeatable across time and accounted for over a fifth of the total observed phenotypic variation in fecundity in two independent populations. The functional explanations for these results and the implications for our understanding of the factors that contribute to the adaptive significance of mating preferences and/or sexual conflict are discussed.


Biomechanics of the thoracolumbar vertebral column of dogs during lateral bending.
To study biomechanical characteristics of the normal and surgically altered canine thoracolumbar vertebral column to determine the effects of surgery and trauma on lateral stability. The T13-L1 vertebral motion units of 48 mixed-breed dogs were dissected free of surrounding musculature and prepared for biomechanical testing by cross-pinning the vertebral bodies and mounting in polymethylmethacrylate. Normal and surgically altered spinal specimens were subjected to lateral bending. The mean slope of the bending moment versus angular displacement curve and the load to failure were compared between treatment groups and significance was determined by the method of least squares (P < 0.05). Specimens were surgically altered by facetectomy, lateral fenestration, diskectomy, and combinations of these procedures. Each specimen was subjected to lateral bending to failure at a rate of 2.5 cm/min in a swing arm bending jig designed to simulate 4-point bending and subject the specimen to pure bending. Only specimens undergoing diskectomy had a significant decrease in slope and load at failure. Unilateral and bilateral facetectomies and fenestration induced a nonsignificant decrease in stiffness, compared with control specimens. Fenestrations and facetectomies do not appear to increase the risk of injury to the canine thora-columbar spinal cord during lateral bending. Fenestrations and facetectomies, as used in routine laminectomies, may be performed without concern for significant destabilization of the spine in lateral bending; however, it is possible that thoracolumbar spinal fractures involving only the vertebral body may significantly destabilize the spine in all modes of bending.


Jaw movement tremor as a predictor of chewing performance.
The purpose of this study was to investigate normal physiologic tremor in jaw movement as a factor that may influence chewing performance more directly than either muscle activity or jaw displacement. Chewing performance was defined in terms of the reduction in food particle size after 15 chewing strokes. Data on chewing particle size and electromyographic activity were available for 24 asymptomatic adults from an earlier study. Jaw movements during chewing were recorded using electrognathography, and velocity and acceleration in three planes were determined. Power spectrum for acceleration was calculated during opening and closing phases of the chewing cycle. The frequency of the peak amplitude in the power spectrum represented physiologic tremor of the jaw. Tremor frequencies during both opening and closing phases of the chewing cycle were strong predictors of chewing performance. A multivariate model composed of variables derived from acceleration, together with electromyographic and jaw movement variables, produced a multivariate model that was able to predict chewing performance with an adjusted R2 value of .78.


Proteomic Analysis of Exosomes and Exosome-Free Conditioned Media From Human Osteosarcoma Cell Lines Reveals Secretion of Proteins Related to Tumor Progression.
Osteosarcomas are the most prevalent bone tumors in pediatric patients, but can also occur later in life. Bone tumors have the potential to metastasize to lung and occasionally other vital organs. To understand how osteosarcoma cells interact with their micro-environment to support bone tumor progression and metastasis, we analyzed secreted proteins and exosomes from three human osteosarcoma cell lines. Exosome isolation was validated by transmission electron microscopy (TEM) and immuno-blotting for characteristic biomarkers (CD63, CD9, and CD81). Exosomal and soluble proteins (less than 100 kDa) were identified by mass spectrometry analysis using nanoLC-MS/MS and classified by functional gene ontology clustering. We identified a secretome set of >3,000 proteins for both fractions, and detected proteins that are either common or unique among the three osteosarcoma cell lines. Protein ontology comparison of proteomes from exosomes and exosome-free fractions revealed differences in the enrichment of functional categories associated with different biological processes, including those related to tumor progression (i.e., angiogenesis, cell adhesion, and cell migration). The secretome characteristics of osteosarcoma cells are consistent with the pathological properties of tumor cells with metastatic potential. J. Cell. Biochem. 118: 351-360, 2017. © 2016 Wiley Periodicals, Inc.


Primary men alcoholics with histories of suicide attempts.
A structured questionnaire administered to 913 primary alcoholics entering an alcoholism treatment program revealed that 155 (17%) had attempted suicide. Compared to other primary alcoholics, those with histories of suicide attempts were more likely to have had early life difficulties with police, school and parents; reported more misuse of drugs; and were significantly more likely to have an alcoholic or depressed close relative.


Complications of carpal tunnel surgery.
During a 12-year period, the authors treated 25 patients with 26 complications of previous carpal tunnel surgery. Twenty-four of these patients were referred following initial surgery elsewhere. The most frequent complication identified was neuroma of the palmar cutaneous branch of the median nerve in 14 of the cases. Other complications were hypertrophic scars, dysesthesias after multiple procedures to release the carpal tunnel, joint stiffness, failure to relieve symptoms, and neuromas of the dorsal sensory branch of the radial nerve. All of these complications are potentially preventable. With a properly placed incision, exposure carried out under magnification, and surgery under direct vision the majority of these complications may be prevented. It is further noted that the technique of transverse incision at the wrist for release of the carpal tunnel is potentially dangerous and should be abandoned.


Intragenic deletions and a deep intronic mutation affecting pre-mRNA splicing in the dihydropyrimidine dehydrogenase gene as novel mechanisms causing 5-fluorouracil toxicity.
Dihydropyrimidine dehydrogenase (DPD) is the initial enzyme acting in the catabolism of the widely used antineoplastic agent 5-fluorouracil (5FU). DPD deficiency is known to cause a potentially lethal toxicity following administration of 5FU. Here, we report novel genetic mechanisms underlying DPD deficiency in patients presenting with grade III/IV 5FU-associated toxicity. In one patient a genomic DPYD deletion of exons 21-23 was observed. In five patients a deep intronic mutation c.1129-5923C>G was identified creating a cryptic splice donor site. As a consequence, a 44 bp fragment corresponding to nucleotides c.1129-5967 to c.1129-5924 of intron 10 was inserted in the mature DPD mRNA. The deleterious c.1129-5923C>G mutation proved to be in cis with three intronic polymorphisms (c.483 + 18G>A, c.959-51T>G, c.680 + 139G>A) and the synonymous mutation c.1236G>A of a previously identified haplotype. Retrospective analysis of 203 cancer patients showed that the c.1129-5923C>G mutation was significantly enriched in patients with severe 5FU-associated toxicity (9.1%) compared to patients without toxicity (2.2%). In addition, a high prevalence was observed for the c.1129-5923C>G mutation in the normal Dutch (2.6%) and German (3.3%) population. Our study demonstrates that a genomic deletion affecting DPYD and a deep intronic mutation affecting pre-mRNA splicing can cause severe 5FU-associated toxicity. We conclude that screening for DPD deficiency should include a search for genomic rearrangements and aberrant splicing.


Changing trends in the treatment of dry-eye disease.
Dry eye is a visually disabling disease encountered in many countries with a wide variation of treatment practices all over the world. On that front, the 2007 Report of the International Dry Eye WorkShop (DEWS) reviewed the current knowledge on all aspects of dry-eye disease (DED), in an evidence-based manner, and outlined the trends and recommendations in the treatment of DED on the basis of disease severity. This review mainly focuses on treatments for DED based on severity as recommended in the DEWS report, particularly artificial eye drops, hyaluronate sodium eye drops, autologous serum, anti-inflammatory eye drops including cyclosporine and steroids, and mucin secretagogues. New dry-eye treatment modalities in current trials outlined on the clinicaltrial.gov site are also outlined. Further investigations into the mechanism of action of the new mucin and tear secretagogues which have been suggested to have anti-inflammatory properties will enrich our understanding in relation to relevant ocular surface responses after treatment with these new agents.


Prefrontal cortical mechanisms underlying individual differences in cognitive flexibility and stability.
The pFC is critical for cognitive flexibility (i.e., our ability to flexibly adjust behavior to changing environmental demands), but also for cognitive stability (i.e., our ability to follow behavioral plans in the face of distraction). Behavioral research suggests that individuals differ in their cognitive flexibility and stability, and neurocomputational theories of working memory relate this variability to the concept of attractor stability in recurrently connected neural networks. We introduce a novel task paradigm to simultaneously assess flexible switching between task rules (cognitive flexibility) and task performance in the presence of irrelevant distractors (cognitive stability) and to furthermore assess the individual "spontaneous switching rate" in response to ambiguous stimuli to quantify the individual dispositional cognitive flexibility in a theoretically motivated way (i.e., as a proxy for attractor stability). Using fMRI in healthy human participants, a common network consisting of parietal and frontal areas was found for task switching and distractor inhibition. More flexible persons showed reduced activation and reduced functional coupling in frontal areas, including the inferior frontal junction, during task switching. Most importantly, the individual spontaneous switching rate antagonistically affected the functional coupling between inferior frontal junction and the superior frontal gyrus during task switching and distractor inhibition, respectively, indicating that individual differences in cognitive flexibility and stability are indeed related to a common prefrontal neural mechanism. We suggest that the concept of attractor stability of prefrontal working memory networks is a meaningful model for individual differences in cognitive stability versus flexibility.


Sox2 conditional mutation in mouse causes ataxic symptoms, cerebellar vermis hypoplasia, and postnatal defects of Bergmann glia.
Sox2 is a transcription factor active in the nervous system, within different cell types, ranging from radial glia neural stem cells to a few specific types of differentiated glia and neurons. Mutations in the human SOX2 transcription factor gene cause various central nervous system (CNS) abnormalities, involving hippocampus and eye defects, as well as ataxia. Conditional Sox2 mutation in mouse, with different Cre transgenes, previously recapitulated different essential features of the disease, such as hippocampus and eye defects. In the cerebellum, Sox2 is active from early embryogenesis in the neural progenitors of the cerebellar primordium; Sox2 expression is maintained, postnatally, within Bergmann glia (BG), a differentiated cell type essential for Purkinje neurons functionality and correct motor control. By performing Sox2 Cre-mediated ablation in the developing and postnatal mouse cerebellum, we reproduced ataxia features. Embryonic Sox2 deletion (with Wnt1Cre) leads to reduction of the cerebellar vermis, known to be commonly related to ataxia, preceded by deregulation of Otx2 and Gbx2, critical regulators of vermis development. Postnatally, BG is progressively disorganized, mislocalized, and reduced in mutants. Sox2 postnatal deletion, specifically induced in glia (with GLAST-CreERT2), reproduces the BG defect, and causes (milder) ataxic features. Our results define a role for Sox2 in cerebellar function and development, and identify a functional requirement for Sox2 within postnatal BG, of potential relevance for ataxia in mouse mutants, and in human patients.


The effect of motivation on movement: a study of bradykinesia in Parkinson's disease.
Bradykinesia is a cardinal feature of Parkinson's disease (PD). Despite its disabling impact, the precise cause of this symptom remains elusive. Recent thinking suggests that bradykinesia may be more than simply a manifestation of motor slowness, and may in part reflect a specific deficit in the operation of motivational vigour in the striatum. In this paper we test the hypothesis that movement time in PD can be modulated by the specific nature of the motivational salience of possible action-outcomes. We developed a novel movement time paradigm involving winnable rewards and avoidable painful electrical stimuli. The faster the subjects performed an action the more likely they were to win money (in appetitive blocks) or to avoid a painful shock (in aversive blocks). We compared PD patients when OFF dopaminergic medication with controls. Our key finding is that PD patients OFF dopaminergic medication move faster to avoid aversive outcomes (painful electric shocks) than to reap rewarding outcomes (winning money) and, unlike controls, do not speed up in the current trial having failed to win money in the previous one. We also demonstrate that sensitivity to distracting stimuli is valence specific. We suggest this pattern of results can be explained in terms of low dopamine levels in the Parkinsonian state leading to an insensitivity to appetitive outcomes, and thus an inability to modulate movement speed in the face of rewards. By comparison, sensitivity to aversive stimuli is relatively spared. Our findings point to a rarely described property of bradykinesia in PD, namely its selective regulation by everyday outcomes.


The System for Coding Interactions and Family Functioning (SCIFF) in low-income and urban adolescents.
Reliable, brief, and cost-effective methods to assess parenting are critical for advancing etiological research and translational efforts within parenting science. In the current study, we adapted the System for Coding Interactions and Family Functioning (SCIFF) for use among a sample of mostly racial minority adolescents aged 15 years old, growing up in a low-income urban setting. A multiethnic team coded videotapes of a family interaction task designed to elicit conflict. First, we assessed the reliability of SCIFF codes (N = 187; 54% female; 77% African American). Second, we tested whether SCIFF codes assessing harsh parenting, positive parenting, dyadic conflict, and dyadic closeness converged with parent-child reports of the same constructs. Third we explored links between observed harsh and positive parenting in early childhood (ages 3 and 5) and SCIFF codes at age 15. Our training and SCIFF coding protocols produced high interrater reliability. In support of convergent validity, we found specificity in the associations between negative aspects of parenting across methods: the SCIFF harsh parenting and dyadic conflict codes uniquely converged with concurrent parent-child reports of the same constructs. There was a longitudinal cross-construct association between more observed harshness in early childhood and lower dyadic closeness at age 15. Finally, the convergence of the SCIFF codes with other parenting measures was similar by gender and for families living below or above 200% of the poverty line. A modified version of the SCIFF can be used with reliability in low-income urban samples with variation in gender and race. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Potential link between exposure to fungicides chlorothalonil and mancozeb and haemic neoplasia development in the soft-shell clam Mya arenaria: a laboratory experiment.
The aetiology of haemic neoplasia (HN) is unknown, so far but many causative factors are suggested such as viral, pollution and genetics. The aim of this study was to determine if, under chronic exposure, two major pesticides (chlorothalonil and mancozeb) which are used in potato production could induce HN in soft-shell clams (Mya arenaria). Short-term experiments with acute exposure were also performed. Clams were collected from an epizootic site (North River, PEI) and from a site free of the disease (Magdalen Islands, Quebec). The tetraploid level of haemocytes was assessed by flow cytometry for each clam to determine the HN status. The bioaccumulation of pesticides in tissues was quantified by gas chromatography/mass spectrometry (GC/MS) for chlorothalonil while mancozeb and manganese were quantified by inductively coupled plasma-mass spectrometer (ICP/MS). Long term exposure to fungicide Bravo 500((R)) did not induce high tetraploid levels on negative calm from North River and the analysis of the digestive gland and the mantle did not reveal any detectable level of chlorothalonil. In the Manzate 200 DF((R)), some clams revealed high level of tetraploid cells but no difference were observed between the treatments and the control. The analysis of the digestive gland and the mantle for manganese did not highlight any significant difference in tissue concentration (p=0.05). For the acute exposure, chlorothalonil analysis showed that the active ingredient is distributed between four chlorinated compounds: 99.5% for chlorothalonil isomers, 0.4% for pentachlorothalonil and 0.1% for trichlorothalonil isomers. For a 72 h experiment, the accumulation was within 4h; the higher tissue concentration of chlorothalonil was 59.2 microg g(-1) in the mantle after 48 h, following by a decrease to an undetectable level at the end. For the manganese, the accumulation was detected after 4h; the higher tissue concentration was 48.8 microg g(-1) in the mantle after 24h and, over the following 48 h, the accumulation decreased until the end of the trial. Based on the data, the accumulation of these fungicides seems to be transitory. Chlorothalonil and mancozeb are both oxidative-stress promoters and could have induced cell dysfunction while in the tissue. Study on the effect of these fungicides on the p53 protein system is an example of strategy that would provide information on cellular events promoting neoplasia.


Craniosynostosis: novel insights into pathogenesis and treatment.
The identification in craniosynostosis syndromes of mutations in genes belonging to the fibroblast growth factor signalling pathway and the transcriptional regulator MSX2 provides important clues to the pathogenesis of these disorders. Although surgery continues to be the mainstay of treatment, new animal models and improved uncerstanding of cranial suture biology and pathology may lead to complementary therapies.


Anatomical Study of Temporal Fat Compartments and its Clinical Application for Temporal Fat Grafting.
Low satisfaction rates and severe complications are two major limitations for temporal hollowing augmentation using autologous fat grafting. Despite fat compartments in temporal region have been reported, its clinical applied anatomy for fat grafting have not been the subject of studies that show its benefits objectively and statistically. To investigate temporal fat compartments and relative neurovascular structures in cadavers, developing a safe and effective fat grafting technique for temporal hollowing augmentation. The study was conducted on 8 cadavers (16 temples). The tissue layers, fat compartments, ligaments, and neurovascular structures in the temporal region were analysed. The variables were the number and location of sentinel veins, perforator vessels of the middle temporal vein. Measurements were taken with a digital calliper. Two separate fat compartments, the lateral temporal-cheek fat compartment and lateral orbital fat compartment, were found in the subcutaneous layer, and two separate septum compartments, the upper and lower temporal compartment, were found in the loose areolar tissue layer. One sentinel vein and 1 to 6 perforator vessels were found to travel through the subcutaneous tissue layer, traverse the overlapping tissue layers in the lower temporal septum region, and finally join in the middle temporal vein. The four fat compartments in the temporal region are ideal receipt sites for fat grafting. The medial border of the junction of the hairline and temporal line is a safe and effective cannula entry site for temporal fat grafting. The anterior half of the lower temporal compartment is a "zone of caution" for temporal fat grafting.


[Cross-cultural adaptation to Brazil of the instrument Caregiver Abuse Screen (CASE) for detection of abuse of the elderly by caregivers].
This first of two papers focuses on the first part in the cross-cultural adaptation of the Portuguese-language version of Caregiver Abuse Screen (CASE), a brief instrument for detecting domestic violence against the elderly. CASE was originally developed in Canada and used to screen violence against the elderly by interviewing their caregivers. Besides a broad literature review, the evaluation of conceptual and item equivalences involved expert discussion groups. Semantic equivalence included the following steps: two translations and respective back-translations; an evaluation of referential and general (connotative) equivalence between the original instrument and each version; further discussions with experts in order to define the final version; and pre-testing the latter in 40 caregivers of elderly subjects in an outpatient geriatric clinic. It was possible to establish high-quality conceptual, item, and semantic equivalence for the Portuguese-language version. Although the results shown here were encouraging, they should be reevaluated in light of a forthcoming psychometric analysis (measurement equivalence) to be performed by the research group.