Lumican suppresses cell proliferation and aids Fas-Fas ligand mediated apoptosis: implications in the cornea.
Lumican, an extracellular matrix (ECM) keratan sulfate proteoglycan, binds fibrillar collagen and limits collagen fibril diameter in the cornea, skin and tendon. Lumican-deficient mice (Lum(-/-)) develop abnormally thick collagen fibrils, translucent corneas and fragilities of the skin and the tendon. In addition to modulating interstitial ECM structure, here we hypothesized that lumican regulates proliferation and apoptosis of cells residing in the interstitium. Corneal and embryonic fibroblasts from the Lum(-/-) mouse show increased growth in culture. Lum(-/-) mouse embryonic fibroblasts (MEF), compared to their wild type counterparts, display increased rates of proliferation and decreased apoptosis. Ectopic expression of lumican in Lum(-/-) MEF or exogenous recombinant lumican in the culture medium reduces proliferation to rates seen in the Lum(+/+) MEF. We further investigated the implications of lumican's proliferation and apoptosis regulatory role in the cornea where lumican is a major component of the stromal matrix. Stromal keratocytes undergo proliferation and apoptosis during corneal maturation and in the healing of injured cornea. The Lum(-/-) mouse shows increased proliferation and decreased apoptosis of stromal keratocytes during postnatal corneal maturation at the 10-day age. Apoptosis is also significantly down regulated in Lum(-/-) vis-à-vis Lum(+/+) mice during stromal wound healing in the adult 6-week age. Lumican appears to regulate these cellular functions by modulating specific cell growth and apoptosis mediators. Thus, Lum(-/-) MEF have decreased p21(WAF1/CIP1), a universal inhibitor of cyclin-dependent kinases and a consequent increase in cyclins A, D1 and E. Furthermore, the tumor suppressor p53, an upstream regulator of p21 is down regulated in the MEF and the cornea of Lum(-/-) mice. The evidence suggests regulation of p21 by lumican in a p53-dependent way. The MEF and the cornea of Lum(-/-) mice also show a dramatic decrease in Fas (CD95). The Lum(-/-) MEF fail to induce Fas upon treatment with Fas ligand. Fas-Fas ligand interaction is an initiating event in apoptosis and its disruption in lumican-deficiency may partly explain the observed decrease in apoptosis. Fas-Fas ligand mediated apoptosis is critical for maintaining the immune privileged status of the cornea, which implies a new and exciting role for lumican in the cornea.


Prophylactic antibiotics in simple hand lacerations.
A randomized prospective study of 265 patients with hand lacerations was carried out to define the role of prophylactic antibiotics in the management of simple, sutured wounds of the hand. Two hundred sixty-five patients were followed up until suture removal or satisfactory wound healing. There were a total of three infections (1.1%). There was no noticeable differences in the incidence of infection in the antibiotic and placebo treatment groups. It is concluded that prophylactic antibiotics are an unnecessary adjunct in the treatment of simple lacerations of the hand and no replacement for meticulous wound management.


A Methodology for Validating Diversity in Synthetic Time Series Generation.
In order for researchers to deliver robust evaluations of time series models, it often requires high volumes of data to ensure the appropriate level of rigor in testing. However, for many researchers, the lack of time series presents a barrier to a deeper evaluation. While researchers have developed and used synthetic datasets, the development of this data requires a methodological approach to testing the entire dataset against a set of metrics which capture the diversity of the dataset. Unless researchers are confident that their test datasets display a broad set of time series characteristics, it may favor one type of predictive model over another. This can have the effect of undermining the evaluation of new predictive methods. In this paper, we present a new approach to generating and evaluating a high number of time series data. The construction algorithm and validation framework are described in detail, together with an analysis of the level of diversity present in the synthetic dataset.


Synchronised oscillations of the human sensorimotor cortex.
Oscillations are a prominent feature of macroscopic human sensorimotor cortical activity as recorded non-invasively with electroencephalography (EEG) and magnetoencephalography (MEG). The advent of whole-scalp MEG systems allowing rapid non-invasive recording from the entire cortex and accurate localisation of neural sources, and the development of refined signal analysis methods are important factors that led to an increasing interest in studies of sensorimotor oscillations during the last 10 years. Investigations on healthy subjects revealed frequency-specific localisation and modality-specific reactivity of 10 Hz and 20 Hz sensorimotor oscillations. Task-specific coherence between motor cortical and electromyographic oscillations, reflecting cortico-motoneuronal coupling, point towards a functional role of precentral oscillations in the cortical control of voluntary movements. Furthermore, abnormal cortico-motoneuronal coupling may underlie clinical symptoms of motor disorders, such as tremor. Thus, investigation of oscillatory sensorimotor activity proceeds from phenomenology to function and provides an interesting approach to address questions in human motor physiology and pathophysiology.


Assembly of chimeric connexin-aequorin proteins into functional gap junction channels. Reporting intracellular and plasma membrane calcium environments.
Chimeric proteins comprising connexins 26, 32, and 43 and aequorin, a chemiluminescent calcium indicator, were made by fusing the amino terminus of aequorin to the carboxyl terminus of connexins. The retention of function by the chimeric partners was investigated. Connexin 32-aequorin and connexin 43-aequorin retained chemiluminescent activity whereas that of connexin 26-aequorin was negligible. Immunofluorescent staining of COS-7 cells expressing the chimerae showed they were targeted to the plasma membrane. Gap junction intercellular channel formation by the chimerae alone and in combination with wild-type connexins was investigated. Stable HeLa cells expressing connexin 43-aequorin were functional, as demonstrated by Lucifer yellow transfer. Paris of Xenopus oocytes expressing connexin 43-aequorin were electrophysiologically coupled, but those expressing chimeric connexin 26 or 32 showed no detectable levels of coupling. The formation of heteromeric channels constructed of chimeric connexin 32 or connexin 43 and the respective wild-type connexins was inferred from the novel voltage gating properties of the junctional conductance. The results show that the preservation of function by each partner of the chimeric protein is dictated mainly by the nature of the connexin, especially the length of the cytoplasmic carboxyl-terminal domain. The aequorin partner of the connexin 43 chimera reported calcium levels in COS-7 cells in at least two different calcium environments.


Perioperative management of patients on chronic antithrombotic therapy.
Perioperative management of antithrombotic therapy is a situation that occurs frequently and requires consideration of the patient, the procedure, and an expanding array of anticoagulant and antiplatelet agents. Preoperative assessment must address each patient's risk for thromboembolic events balanced against the risk for perioperative bleeding. Procedures can be separated into those with a low bleeding risk, which generally do not require complete reversal of the antithrombotic therapy, and those associated with an intermediate or high bleeding risk. For patients who are receiving warfarin who need interruption of the anticoagulant, consideration must be given to whether simply withholding the anticoagulant is the optimal approach or whether a perioperative "bridge" with an alternative agent, typically a low-molecular-weight heparin, should be used. The new oral anticoagulants dabigatran and rivaroxaban have shorter effective half-lives, but they introduce other concerns for perioperative management, including prolonged drug effect in patients with renal insufficiency, limited experience with clinical laboratory testing to confirm lack of residual anticoagulant effect, and lack of a reversal agent. Antiplatelet agents must also be considered in the perioperative setting, with particular consideration given to the potential risk for thrombotic complications in patients with coronary artery stents who have antiplatelet therapy withheld.


Visuospatial Functioning in the Primary Progressive Aphasias.
The aim of this study was to identify whether the three main primary progressive aphasia (PPA) variants would show differential profiles on measures of visuospatial cognition. We hypothesized that the logopenic variant would have the most difficulty across tasks requiring visuospatial and visual memory abilities. PPA patients (n=156), diagnosed using current criteria, and controls were tested on a battery of tests tapping different aspects of visuospatial cognition. We compared the groups on an overall visuospatial factor; construction, immediate recall, delayed recall, and executive functioning composites; and on individual tests. Cross-sectional and longitudinal comparisons were made, adjusted for disease severity, age, and education. The logopenic variant had significantly lower scores on the visuospatial factor and the most impaired scores on all composites. The nonfluent variant had significant difficulty on all visuospatial composites except the delayed recall, which differentiated them from the logopenic variant. In contrast, the semantic variants performed poorly only on delayed recall of visual information. The logopenic and nonfluent variants showed decline in figure copying performance over time, whereas in the semantic variant, this skill was remarkably preserved. This extensive examination of performance on visuospatial tasks in the PPA variants solidifies some previous findings, for example, delayed recall of visual stimuli adds value in differential diagnosis between logopenic variant PPA and nonfluent variant PPA variants, and illuminates the possibility of common mechanisms that underlie both linguistic and non-linguistic deficits in the variants. Furthermore, this is the first study that has investigated visuospatial functioning over time in the PPA variants. (JINS, 2018, 24, 259-268).


Yin-Yang tongue sign: An imaging clue of lesions involving the skull base segment in the hypoglossal pathway.
To investigate the diagnostic value of the Yin-Yang tongue sign in patients with tongue deviation. According to the presence of the Yin-Yang tongue sign on CT/MR, 107 patients with tongue deviation were divided into a positive group and a negative group. The involvement categories of the hypoglossal canal (HC) in the positive group were evaluated and classified as HC dilation and HC erosion. The correlations between HC involvement categories and the presence of the sign were analysed. There were 55 cases (55/107, 51.4%) in the positive group and 52 cases (52/107, 48.6%) in the negative group. Hypoglossal nerve (HN) involvement mainly occurred in the skull base (61.8%), skull base and carotid space (10.9%), and carotid space segment (12.7%). Neurogenic (50.9%), squamous cell carcinoma (14.5%), and metastases (12.7%) were the predominant aetiologies. The sensitivity, specificity, and accuracy of this sign for suggesting skull base lesions around HC were 72.4%, 80.8%, and 76.6%, respectively. In the positive group, HC dilation was seen in 21 patients (21/55, 38.2%) and 21 cases were all benign. HC erosion were noted in 19 patients (19/55, 34.5%), of whom 12 cases were malignant. The Yin-Yang tongue sign is formed by unilateral tongue atrophy and fat infiltration caused by lesions in the HN pathway, especially compressive or invasive lesions involving the skull base segment.


Beta-very low density lipoprotein is sequestered in surface-connected tubules in mouse peritoneal macrophages.
beta-very low density lipoprotein (VLDL) is a large lipoprotein with multiple apoprotein E (apoE) molecules that bind to the LDL receptors on mouse macrophages. Even though they bind to the same receptor, the endocytic processing of beta-VLDL differs from low density lipoprotein (LDL). LDL is rapidly delivered to perinuclear lysosomes and degraded, but much of the beta-VLDL is retained in peripheral compartments for several minutes. We have investigated the properties of these peripheral compartments. Measurement of the pH was made using FITC-phosphatidylethanolamine incorporated into the beta-VLDL, and we found that the peripheral compartments were near neutral in pH. These peripheral, beta-VLDL containing compartments were poorly accessible to antibodies, but a low molecular weight fluorescence quencher (trypan blue) entered the compartments within a few seconds. Intermediate voltage EM of cells labeled with colloidal-gold-beta-VLDL revealed that the peripheral compartments are tubular, surface-connected invaginations. Kinetic studies with fluorescent beta-VLDL showed that the compartments become fully sealed with a half-time of 6 min, and the beta-VLDL is then delivered rapidly to perinuclear lysosomes. By monitoring fluorescence energy transfer between lipid analogs incorporated into the beta-VLDL, some processing of the lipoprotein in the peripheral tubular compartments is demonstrated. The novel mode of uptake of beta-VLDL may account for the high cholesterol ester accumulation induced by this lipoprotein.


Malnutrition is Associated with Behavioral and Psychiatric Symptoms of Dementia in Older Women with Mild Cognitive Impairment and Early-Stage Alzheimer's Disease.
We examined the nutritional status and its association with behavioral psychiatric symptoms of dementia (BPSD) among 741 memory clinic patients (normal cognition (NC), 152; mild cognitive impairment (MCI), 271; early-stage Alzheimer disease (AD), 318). Nutritional status and BPSD were assessed using the Mini Nutritional Assessment Short-Form (MNA-SF) and the Dementia Behavior Disturbance Scale (DBD), respectively. Compared to subjects with NC, more subjects with MCI and early-stage AD were at risk of malnutrition (MNA-SF, 8-11: NC, 34.2%; MCI, 47.5%; early-stage AD, 53.8%) and were malnourished (MNA-SF, 0-7: NC, 4.6%; MCI, 5.9%; early-stage AD, 8.2%). Among patients with MCI or early-stage AD, those at risk of/with malnutrition showed higher DBD scores than those well-nourished (12.7 ± 9.0 vs. 9.5 ± 7.3; p < 0.001). Moreover, analysis of covariance adjusting for confounders showed that nutritional status was significantly associated with specific BPSD, including "verbal aggressiveness/emotional disinhibition" (F = 5.87, p = 0.016) and "apathy/memory impairment" (F = 15.38, p < 0.001), which were revealed by factor analysis of DBD. Our results suggest that malnutrition is common among older adults with mild cognitive decline, and possibility that nutritional problems are associated with individual BPSD.


Myelination and demyelination processes in the rat cerebellum cell culture: an electron microscopic study.
We observed manifestations of the myelination process in dissociated culture of the cerebellar tissue of newborn rats and modifications of the structure of myelin sheaths after treating the culture with a demyelinating factor, blood serum of patients suffering from multiple sclerosis (MS). On day in vitro (DIV) 26, in the control myelin sheaths of the axons demonstrated the closest resemblance to those observed in vivo, and we selected this term for inducing demyelination. Addition of the serum of MS patients to the culturing medium evoked rapid (in 3-6 hours) dramatic changes in the ultrastructure of myelin sheaths; these were a decrease in the number of the lamellae, their splitting and invagination, formation of vesicles, etc. The serum of MS patients in an acute stage of the disease exerted more intensive demyelination effects than that of patients in a remission stage.


The ARH and Macrodomain Families of α-ADP-ribose-acceptor Hydrolases Catalyze α-NAD+ Hydrolysis.
ADP-ribosyltransferases transfer ADP-ribose from β-NAD+ to acceptors; ADP-ribosylated acceptors are cleaved by ADP-ribosyl-acceptor hydrolases (ARHs) and proteins containing ADP-ribose-binding modules termed macrodomains. On the basis of the ADP-ribosyl-arginine hydrolase 1 (ARH1) stereospecific hydrolysis of α-ADP-ribosyl-arginine and the hypothesis that α-NAD+ is generated as a side product of β-NAD+/ NADH metabolism, we proposed that α-NAD+ was a substrate of ARHs and macrodomain proteins. Here, we report that ARH1, ARH3, and macrodomain proteins (i.e., MacroD1, MacroD2, C6orf130 (TARG1), Af1521, hydrolyzed α-NAD+ but not β-NAD+. ARH3 had the highest α-NADase specific activity. The ARH and macrodomain protein families, in stereospecific reactions, cleave ADP-ribose linkages to N- or O- containing functional groups; anomerization of α- to β-forms (e.g., α-ADP-ribosyl-arginine to β-ADP-ribose- (arginine) protein) may explain partial hydrolysis of ADP-ribosylated acceptors with an increase in content of ADP-ribosylated substrates. Af1521 and ARH3 crystal structures with bound ADP-ribose revealed similar ADP-ribose-binding pockets with the catalytic residues of the ARH and macrodomain protein families in the N-terminal helix and loop. Although the biological roles of the ARHs and macrodomain proteins differ, they share enzymatic and structural properties that may regulate metabolites such as α-NAD+.


Robotic-assisted pelvic lymph node dissection for prostate cancer: frequency of nodal metastases and oncological outcomes.
Limited data are available regarding the oncologic efficacy of pelvic lymph node dissection (PLND) performed during robotic-assisted laparoscopic prostatectomy (RALP) for prostate cancer. We aimed to determine the frequency of pelvic lymph node metastasis and oncological outcomes following RALP with PLND in patients who did not receive adjuvant androgen deprivation therapy (ADT). We retrospectively reviewed the records of 1740 consecutive patients who underwent RALP and extended PLND. The primary endpoint was biochemical recurrence (BCR). The estimated BCR probability was obtained using the Kaplan-Meier method. Cox proportional hazard regression models were used to assess for predictors of BCR. One hundred and eight patients (6 %) with positive LNs were identified. The median number of LNs removed was 17 (IQR 11-24), and median follow-up was 26 months (IQR 14-43). Ninety-one (84 %) patients did not receive adjuvant ADT of whom 60 % had BCR with a median time to recurrence of 8 months. The 1- and 3-year BCR-free probability was 42 and 28 %, respectively. Patients with ≤2 LN+ had significantly better biochemical-free estimated probability compared to those with >2 LN+ (p = 0.002). The total number of LN+ (HR = 1.1; 95 % CI 1.01-1.2, p = 0.04) and Gleason 8-10 (HR = 1.96; 95 % CI 1.1-3.4, p = 0.02) were predictors of BCR on multivariate analysis. Among men with positive lymph nodes at time of robotic prostatectomy, those with two or fewer positive nodes and Gleason <8 exhibited favorable biochemical-free survival without adjuvant therapy.


[Effectivity of different methods for disinfection of applanation tonometer prisms (author's transl)].
In 13,440 bacteriological investigations the effectivity of different methods for desinfection of Goldmann tonometer prisms (merfen, pantasept, sekusept steril, UV sterilizer Sklar) is tested. Sekusept steril and UV sterilizer show the quickest and most sufficient results. They cause no damage to corneal epithelium or to plastic objects. We can recommend them for desinfection of applanation tonometer prims, Goldmann's gonioscopy and fundus contact lenses as routine methods.


Carrot man: a case of excessive beta-carotene ingestion.
In this case report, the authors describe a 48-year-old male who complained to his primary care physician of abdominal discomfort and yellow/orange skin discoloration. Physical examination was normal except for some mild mid-abdominal discomfort (no observed skin color changes). An abdominal CT scan indicated a colon that was full of stool. Laboratory studies indicated elevated liver enzymes. Upon further questioning, the patient reported ingesting 6-7 pounds of carrots per week to facilitate his dieting effort. The patient was diagnosed with constipation, hypercarotinemia, and possible vitamin A toxicity. Following the cessation of excessive carrot ingestion, his liver enzymes normalized within 1 month.


Protein annotation as term categorization in the gene ontology using word proximity networks.
We participated in the BioCreAtIvE Task 2, which addressed the annotation of proteins into the Gene Ontology (GO) based on the text of a given document and the selection of evidence text from the document justifying that annotation. We approached the task utilizing several combinations of two distinct methods: an unsupervised algorithm for expanding words associated with GO nodes, and an annotation methodology which treats annotation as categorization of terms from a protein's document neighborhood into the GO. The evaluation results indicate that the method for expanding words associated with GO nodes is quite powerful; we were able to successfully select appropriate evidence text for a given annotation in 38% of Task 2.1 queries by building on this method. The term categorization methodology achieved a precision of 16% for annotation within the correct extended family in Task 2.2, though we show through subsequent analysis that this can be improved with a different parameter setting. Our architecture proved not to be very successful on the evidence text component of the task, in the configuration used to generate the submitted results. The initial results show promise for both of the methods we explored, and we are planning to integrate the methods more closely to achieve better results overall.


Captive propagation of vervet monkeys (Cercopithecus aethiops) in harems.
A small breeding colony of captive vervet monkeys (Cercopithecus aethiops), consisting of ten single male harem groups, was established in Kenya. Vervets were extremely prone to stress after capture and manipulative procedures led to high mortalities. Six weeks were required for partial habituation, after which routine handling was not too problematic. However, complete adjustment to captivity took a minimum of one year. Establishment of stable breeding groups from adult animals initially proved difficult because of fighting among females. Once harem groups stabilized, reproductive rate was high with just under 90% pregnancies and 85% livebirths annually. Births exhibited a seasonal pattern similar to that reported from wild populations in Kenya.


Key clinical observations after 5-azacytidine and decitabine treatment of myelodysplastic syndromes suggest practical solutions for better outcomes.
Clinical experience with 5-azacytidine and decitabine treatment of myelodysplastic syndromes (MDS), complemented by biological and pharmacological studies, has revealed compelling mechanism of action differences compared with traditional myeloid cancer treatment mainstays such as cytarabine. For example, 5-azacytidine and decitabine produce remissions and better overall survival in MDS with high-risk chromosome abnormalities at a surprisingly high rate, consistent with experimental observations that noncytotoxic DNA methyltransferase depletion by 5-azacytidine/decitabine can trigger cell cycle exit independently of p53, thus circumventing a basis for resistance to apoptosis-based DNA-damaging therapy. That responses cut across the chaotic genomic landscape of MDS highlights common threads in disease, such as high expression in myeloblasts of differentiation-driving transcription factors yet paradoxical epigenetic suppression of proliferation-terminating late-differentiation genes. Less toxic regimens (lower dosages but more frequent administration) of 5-azacytidine/decitabine have been more successful, underscoring the importance of preserving functionally normal stem cells, which are rendered more precious by attrition from age, previous cytotoxic treatments, and the disease process and are needed to relieve cytopenias, the cause of morbidity and mortality. Also emphasized is that there can be no therapeutic benefit, regardless of mutation or cytogenetic subtype, if DNA methyltransferase is not depleted by sufficient overlap between intracellular drug half-lives and S-phase entries of malignant cells. Improved understanding of mechanism-of-action differences demands new approaches, from historic (but not scientific) more-is-better and one-size-fits-all empiricism to pharmacodynamic-based designs and combinations directed not solely at suppressing malignant clones, but at improving therapeutic indices.


Influence of social connectedness, communication and monitoring on adolescent sexual activity in Ghana.
This paper examines connectedness to, communication with and monitoring of unmarried adolescents in Ghana by parents, other adults, friends and key social institutions and the roles these groups play with respect to adolescent sexual activity. The paper draws on 2004 nationally-representative survey data and qualitative evidence from focus group discussions and in-depth interviews with adolescents in 2003. Adolescents show high levels of connectedness to family, adults, friends, school and religious groups. High levels of adult monitoring are also observed, but communication with family about sex-related matters was not as high as with non-family members. The qualitative data highlight gender differences in communication. Multivariate analysis of survey data shows a strong negative relationship between parental monitoring and recent sexual activity for males and females, and limited effects of communication. Creating a supportive environment and showing interest in the welfare of adolescents appear to promote positive sexual and reproductive health outcomes.


Long-term stability of contour augmentation with early implant placement following single tooth extraction in the esthetic zone: a prospective, cross-sectional study in 41 patients with a 5- to 9-year follow-up.
Early implant placement with simultaneous contour augmentation is documented with short- and medium-term studies. The long-term stability of contour augmentation is uncertain. In this prospective, cross-sectional study, 41 patients with an implant-borne single crown were examined twice, in 2006 and 2010. Clinical, radiologic, and esthetic parameters were assessed at both examinations. In addition, a cone beam computed tomographic (CBCT) image was obtained during the second examination to assess the dimensions of the facial bone wall. All 41 implants demonstrated ankylotic stability without signs of peri-implant infection at both examinations. The clinical parameters remained stable over time. Satisfactory esthetic outcomes were noted, as assessed by the pink and white esthetic score (PES/WES) indices. Overall, the PES scores were slightly higher than the WES scores. None of the implants developed mucosal recession over time, as confirmed by values of the distance between implant shoulder and mucosal margin and cast measurements. The periapical radiographs yielded stable peri-implant bone levels, with a mean distance between implant shoulder and first visible bone-implant contact value of 2.18 mm. The CBCT analysis demonstrated a mean thickness of the facial bone wall ≈2.2 mm. In two implants (4.9%) no facial bone wall was detectable radiographically. This prospective cross-sectional study demonstrates stable peri-implant hard and soft tissues for all 41 implants examined and satisfactory esthetic outcomes overall. The follow-up of 5 to 9 years confirmed again that the risk for mucosal recession is low with early implant placement. In addition, contour augmentation with guided bone regeneration was able to establish and maintain a facial bone wall in 95% of patients.


Assessing the removal of inorganic colloids and Cryptosporidium parvum from drinking water.
A new batch device that simulates the conditions in water and wastewater treatment plants and enables the study of low-concentration feeds is described. The application of this apparatus to the monitoring of the concentration of inorganic and/or biological contaminants is demonstrated, using kaolin particles and Cryptosporidium parvum oocysts, respectively. The rate of inorganic particle attachment to the static medium is found to be directly proportional to the initial influent concentration. On the other hand, Cryptosporidium parvum removal is found to be more effective in the presence of additional (kaolin) particles, and this is attributed to electrostatic interactions between oppositely charged layers on the absorbing medium. Accordingly, the addition of humic materials is found to inhibit the removal process, possibly by neutralizing the positive charge on the kaolin particles. The relevance of these results to existing water purification processes in general and to risk assessment of microbiological contamination in particular is discussed.


Feedstock optimization of in-vessel food waste composting systems for inactivation of pathogenic microorganisms.
An optimum composting recipe was investigated to reduce pathogenic microorganisms in a forced-aerated in-vessel system (55 liters). The feedstocks used for in-vessel composting were food waste, cow manure, and bulking materials (wood shavings and mulch hay). A statistical extreme vertices mixture design method was used to design the composting experiments and analyze the collected data. Each mixture (nine total) was replicated randomly three times. Temperature was monitored as an indicator of the efficiency of the composting experiments. The maximum temperature values of the mixtures were used as a response for both extreme vertices mixture design and statistical analyses. Chemical changes (moisture content, carbon/nitrogen ratio, volatile solids, and pH) and reductions of indicator (fecal coliforms and fecal streptococci) and pathogenic microorganisms (Salmonella and Escherichia coli O157:H7) were measured by the most-probable-number method before and after a 12-day composting period. Maximum temperatures for the tested compost mixtures were in the range of 37.0 to 54.7 degrees C. Extreme vertices mixture design analysis of the surface plot suggested an optimum mixture containing 50% food waste, 40% manure, and 10% bulking agents. This optimum mixture achieved maximum temperatures of 54.7 to 56.6 degrees C for about 3.3 days. The total reduction of Salmonella and E. coli O157:H7 were 92.3%, whereas fecal coliforms and fecal streptococci reductions were lower (59.3 and 27.1%, respectively). Future study is neededto evaluate the extreme vertices mixture design method for optimization of large-scale composting.


Challenges to evaluating respiratory syncytial virus mortality in Bangladesh, 2004-2008.
Acute lower respiratory illness is the most common cause of death among children, globally. Data are not available to make accurate estimates on the global mortality from respiratory syncytial virus (RSV), specifically. Respiratory samples collected from children under 5 years of age during 2004 to 2008 as part of population-based respiratory disease surveillance in an urban community in Dhaka, Bangladesh were tested for RSV, human metapneumovirus (HMPV), human parainfluenza virus (PIV) types 1, 2, and 3, influenza and adenovirus by RT-PCR. Verbal autopsy data were used to identify children who died from respiratory illness in a nearby rural community. Significance of the correlation between detections and community respiratory deaths was determined using Spearman's coefficient. RSV activity occurred during defined periods lasting approximately three months but with no clear seasonal pattern. There was no significant correlation between respiratory deaths and detection of any of the respiratory viruses studied. Outbreaks of respiratory viruses may not be associated with deaths in children in the study site; however, the few respiratory deaths observed and community-to-community variation in the timing of outbreaks may have obscured an association. An accurate assessment of respiratory virus-associated deaths will require detections and death data to come from the same location and a larger study population.


Self-Regulatory Strategies in Daily Life: Selection, Optimization, and Compensation and Everyday Memory Problems.
The effective use of self-regulatory strategies, such as selection, optimization, and compensation (SOC) requires resources. However, it is theorized that SOC use is most advantageous for those experiencing losses and diminishing resources. The present study explored this seeming paradox within the context of limitations or constraints due to aging, low cognitive resources, and daily stress in relation to everyday memory problems. We examined whether SOC usage varied by age and level of constraints, and if the relationship between resources and memory problems was mitigated by SOC usage. A daily diary paradigm was used to explore day-to-day fluctuations in these relationships. Participants (n=145, ages 22 to 94) completed a baseline interview and a daily diary for seven consecutive days. Multilevel models examined between- and within-person relationships between daily SOC use, daily stressors, cognitive resources, and everyday memory problems. Middle-aged adults had the highest SOC usage, although older adults also showed high SOC use if they had high cognitive resources. More SOC strategies were used on high stress compared to low stress days. Moreover, the relationship between daily stress and memory problems was buffered by daily SOC use, such that on high-stress days, those who used more SOC strategies reported fewer memory problems than participants who used fewer SOC strategies. The paradox of resources and SOC use can be qualified by the type of resource-limitation. Deficits in global resources were not tied to SOC usage or benefits. Conversely, under daily constraints tied to stress, the use of SOC increased and led to fewer memory problems.


MdMYB66 Is Associated with Anthocyanin Biosynthesis via the Activation of the MdF3H Promoter in the Fruit Skin of an Apple Bud Mutant.
Skin color is an important trait that is mainly determined by the content and composition of anthocyanins in apples. In this study, a new bud mutant (RM) from 'Oregon Spur II' (OS) of Red Delicious apple was obtained to reveal the mechanism underlying red color formation. Results showed that the total anthocyanin content in RM was significantly higher than that in OS with the development of fruit. Through widely-targeted metabolomics, we found that cyanidin-3-O-galactoside was significantly accumulated in the fruit skin of RM. Transcriptome analysis revealed that the structural gene MdF3H and MdMYB66 transcription factor were significantly up-regulated in the mutant. Overexpression of MdMYB66 in apple fruit and apple callus significantly promoted anthocyanin accumulation and significantly increased the expression level of MdMYB66 and structural genes related to anthocyanin synthesis. Y1H and LUC analysis verified that MdMYB66 could specifically bind to the promoter of MdF3H. The results of the double luciferase activity test showed that MdMYB66 activated MdF3H 3.8 times, which led to increased anthocyanin contents. This might explain the phenotype of red color in RM at the early stage. Taken together, these results suggested that MdMYB66 was involved in regulating the anthocyanin metabolic pathways through precise regulation of gene expression. The functional characterization of MdMYB66 provides insight into the biosynthesis and regulation of anthocyanins.


[Effect of inhibitors of cyclooxygenase and lypoxygenase pathway of metabolism of arachidonic acid on the development and suppression of the immune response in mice].
The influence of indomethacin (IM) and nordihydroguaiaretic acid (NDGA) as inhibitors of cyclooxygenase and lypoxygenase pathways of arachidonic acid metabolism, accordingly, on the development of the immune response (IR) to sheep red blood cells (SRBC), as well as on the formation and functional activity of the antigen-induced (by the tolerogenous dose of SRBC) T-suppressors (AITs) was studied. Investigation was carried out on mice of line CBA. The IR was estimated by quantity of antibody-forming cells in the mouse spleen. The functional activity of AITs was determined in the transfer adopting system by the intensity of IR suppression in mice--recipients. The data obtained have demonstrated, that both inhibitors mainly stimulated the IR which was more expressed at NDGA application and depended on a phase of the IR. The stimulation of the IR was related with a suppression of AITs and a decrease in their functional activity.


Biochanin A Modulates Cell Viability, Invasion, and Growth Promoting Signaling Pathways in HER-2-Positive Breast Cancer Cells.
Overexpression of HER-2 receptor is associated with poor prognosis and aggressive forms of breast cancer. Scientific literature indicates a preventive role of isoflavones in cancer. Since activation of HER-2 receptor initiates growth-promoting events in cancer cells, we studied the effect of biochanin A (an isoflavone) on associated signaling events like receptor activation, downstream signaling, and invasive pathways. HER-2-positive SK-BR-3 breast cancer cells, MCF-10A normal breast epithelial cells, and NIH-3T3 normal fibroblast cells were treated with biochanin A (2-100 muM) for 72 hours. Subsequently cell viability assay, western blotting and zymography were carried out. The data indicate that biochanin A inhibits cell viability, signaling pathways, and invasive enzyme expression and activity in SK-BR-3 cancer cells. Biochanin A did not inhibit MCF-10A and NIH-3T3 cell viability. Therefore, biochanin A could be a unique natural anticancer agent which can selectively target cancer cells and inhibit multiple signaling pathways in HER-2-positive breast cancer cells.


Spectral elementary-coherence-function representation for partially coherent light pulses.
A broad class of partially coherent non-stationary fields can be expressed in terms of the recently proposed independent-elementary-pulse model. In this work we first introduce a corresponding dual representation in the frequency domain and then extend this concept by considering shifted and weighted elementary spectral coherence functions. We prove that this method, which closely describes practical optical systems, leads to properly defined correlation functions. As an example, we demonstrate that our new model characterizes, in a natural way, trains of ultra-short pulses, affected by noise and timing jitter, emitted by usual modulators employed in telecom applications.


Effect of a Single Yoga Asana on Blood Glucose Levels in Type 2 Diabetes Mellitus: A Self-Controlled Study.
Diabetes is a metabolic disorder characterized by high blood sugar levels. Yoga has been shown to have positive effects on blood sugar levels in diabetes patients. However, there is limited research on the effects of specific yoga poses on blood sugar levels in patients with type 2 diabetes (T2DM). This study aimed to evaluate the effect of a single yoga asana, Ardha Matsyendrasana, on random blood glucose (RBG) levels in patients with T2DM. Specifically, we aimed to investigate whether a 15-minute practice of Ardha Matsyendrasana could reduce RBG levels in patients with T2DM. This study employed a self-controlled design to evaluate the effect of Ardha Matsyendrasana on blood glucose levels in patients with type 2 diabetes mellitus. 100 patients with type 2 diabetes mellitus (T2DM) were recruited for this study. All participants underwent two sessions: a control session (CS) and an asana session (AS), each lasting 15 minutes. During the CS, participants rested in a sitting pose, while during the AS, they practiced Ardha Matsyendrasana. The order of the sessions was randomized, with half the participants undergoing the CS on day 1, the AS on day 2, and the other half undergoing the sessions in reverse order. We measured participants' random blood glucose (RBG) levels immediately before and after each intervention. The statistical package for the social sciences, version 16, was used to perform a paired t-test to compare RBG levels before and after each intervention. The study demonstrated a significant reduction in random blood glucose (RBG) levels in the Ardha Matsyendrasana session compared to the control session. This trend was observed in both males and females with T2DM. A single session of Ardha Matsyendrasana for 15 minutes can effectively reduce blood glucose levels in patients with T2DM. However, further studies are required to determine the long-term effects of this asana on glycemic control.


Renal response to lithogenic and anti-lithogenic supplement challenges in a stone-free population group.
In South Africa, urolithiasis is extremely rare in the black population, but is common in the white population. The objective of this study was to investigate the individual effects of 5 different dietary and supplemental challenges (high dietary calcium, calcium supplement, vitamin B6 supplement, L-glutamine supplement, and L-cysteine supplement) on the urinary risk factors for calcium oxalate urolithiasis in subjects from both race groups. Complete Latin Square design. University research laboratory. Subjects were recruited from the student cohort of the University of Cape Town (10 male subjects from each race group). Selection criteria were no history of renal or metabolic diseases, and no chronic or acute medication. Subjects served as their own controls. After 7 days on a self-selected standardized diet, a 24-hour baseline urine sample was collected. A second 24-hour urine sample was collected after 5 days on the prescribed dietary or supplemental challenge. These were analyzed for biochemical and physicochemical risk factors. Additionally, 24-hour dietary recall questionnaires were recorded at baseline and after the 5-day test period, and were analyzed using a food analysis program. Statistical analysis of variance was performed on all of the data. Urine composition, relative supersaturation of urinary salts, calcium oxalate metastable limit, and Tiselius risk index. None of the protocols altered any of the urinary biochemical or physicochemical risk factors in black subjects. In white subjects, the calcium diet significantly increased urinary potassium (P =.0001) and decreased the relative supersaturation of brushite (P =.035); the calcium supplement significantly decreased the Tiselius risk index (P =.014); vitamin B6 supplement significantly decreased urinary calcium (P =.016), urinary phosphate (P =.027), and the relative supersaturation of brushite (P =.004); L-glutamine supplement significantly decreased relative supersaturation of calcium oxalate (P =.01); L-cystine supplement significantly decreased urinary calcium (P =.031) and the Tiselius risk index (P =.013). Because none of the challenges had an effect on the urinary risk factors in black subjects, it is speculated that a renal or gastrointestinal homeostatic adjustment occurs in this group, thereby keeping urinary concentration of substances in balance.


Eggplant Domestication: Pervasive Gene Flow, Feralization, and Transcriptomic Divergence.
In the context of food security, examining the genomics of domestication will help identify genes underlying adaptive and economically important phenotypes, for example, larger fruit, improved taste, and loss of agronomically inferior phenotypes. Examination of genome-scale single nucleotide polymorphisms demonstrates the relationships between wild ancestors of eggplant (Solanum melongena L.), confirming that Solanum insanum L. is the wild progenitor. This species is split roughly into an Eastern (Malaysian, Thai, and Vietnamese) and Western (Indian, Madagascan, and Sri Lankan) group, with domesticates derived from the former. Additional "wild" accessions from India appear to be feral escapes, derived multiple times from domesticated varieties through admixture. Accessions with small egg-shaped fruit are generally found intermixed with East Asian Solanum insanum confirming they are primitive relative to the large-fruited domesticates. Comparative transcriptomics was used to track the loci under selection. Sequence analysis revealed a genetic bottleneck reducing variation by almost 50% in the primitive accessions relative to the wild species and a further 10% in the landraces. We also show evidence for selection on genes with a role in response to wounding and apoptosis. Genes showing a significant difference in expression between wild and primitive or between primitive and landrace genepools were mostly (>75%) downregulated in the derived populations and enriched for gene ontologies related to defense, flowering, signaling, and response to biotic and abiotic stimuli. This work reveals genomic changes involved in crop domestication and improvement, and the population genetics work explains why defining the eggplant domestication trajectory has been so challenging.


Antibiotic prescriptions for outpatient acute rhinosinusitis in Canada, 2007-2013.
Acute rhinosinusitis (ARS) is a respiratory disease commonly caused by viral infections. Physicians regularly prescribe antibiotics despite bacterial etiologies being uncommon. This is of concern, as this use adds to the selection pressure for resistance. Here we present the descriptive epidemiology of acute rhinosinusitis and corresponding antibiotic prescribing practices by Canadian outpatient physicians from 2007-2013. Diagnosis and antibiotic prescription data for ARS were extracted from the Canadian Disease and Therapeutic Index for 2007 to 2013, and population data were acquired from Statistics Canada. ARS diagnosis and antibiotic prescription rates and frequencies of antibiotic classes were calculated. Eighty-eight percent of patients diagnosed with ARS in 2013 were adults, with a greater rate of antibiotic prescriptions observed among the adults relative to the pediatric patients (1632.9 and 468.6 antibiotic prescriptions per 10,000 inhabitants). Between 2007 and 2013, the ARS diagnosis rate decreased from 596 to 464 diagnoses per 10,000 inhabitants, while the percentage of diagnoses with antibiotic prescriptions at the national level remained stable (87% to 84%). From 2007 to 2013, prescription rates for macrolides decreased from 203.5 to 105.4 prescriptions per 10,000 inhabitants. In 2013, penicillins with extended spectrum were more commonly prescribed compared to macrolides among adult patients (153.5 and 105.4 prescriptions per 10,000 inhabitants, respectively). This study is the first to describe physician antibiotic prescribing practices for treatment of ARS in Canada. Results show that antibiotic treatment for ARS represents an area for implementing antimicrobial stewardship, and through it, managing antibiotic resistance. Further work is required to better understand diagnosing practices and treatment criteria for ARS, and use this information to further assist physicians to limit unnecessary antibiotic prescribing practices.


"One-stop-shop" staging: should we prefer FDG-PET/CT or MRI for the detection of bone metastases?
The aim of this study was to compare the diagnostic accuracy of fully diagnostic, contrast-enhanced whole-body FDG-PET/CT and whole-body MRI for detection of bone metastases in patients suffering from newly diagnosed non-small cell lung cancer and malignant melanoma. 109 consecutive non-small cell lung cancer (n=54) and malignant melanoma (n=55) patients underwent whole-body FDG-PET/CT and whole-body MRI for initial tumor staging. All images were evaluated by four experienced physicians (three radiologists, one nuclear medicine physician). The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy for detection of bone metastases were determined for both modalities. Statistically significant differences between FDG-PET/CT and MRI were calculated with Fisher's Exact test (p<0.05). Clinical and imaging follow-up data with a mean follow-up time of 434 days served as the reference standard. According to the reference standard 11 patients (10%) suffered from bone metastases. The sensitivity, specificity, PPV, NPV, and accuracy for the detection of osseous metastases was 45%, 99%, 83%, 94%, and 94% with whole-body FDG-PET/CT and 64%, 94%, 54%, 96%, and 91% with whole-body MRI. The difference was not statistically significant (p=0.6147). FDG-PET/CT and MRI seem to be equally suitable for the detection of skeletal metastases in patients suffering from newly diagnosed non-small cell lung cancer and malignant melanoma. Both modalities go along with a substantial rate of false-negative findings requiring a close follow-up of patients who are staged free of bone metastases at initial staging.


A novel plasmid (pEMCJH03) isolated from moraxella catarrhalis possibly useful as a cloning and expression vector within this species.
A preliminary screening study of six Moraxella catarrhalis isolates from primary school children in the Netherlands identified a small 3.5 kb plasmid (pEMCJH03), containing four open reading frames, which encoded three mobilizing and one replicase protein. Insertion of a kanamycin containing transposon (yielding pEMCJH04) allowed selection and isolation of the plasmid in Escherichia coli. Natural transformation of pEMCJH04 into M. catarrhalis was successful for 25% (3/12) of non-isogenic isolates, with no link between (lack of) transformability and genetic lineage or (lack of) transformability and complement phenotype, though the transformation efficiency was found to be rather low at approximately 615CFU/microg (range=60-1040CFU/microg ). This is only the second publication detailing a plasmid isolated from this important respiratory pathogen, and the ability to clone and express foreign proteins in M. catarrhalis using pEMCJH04 could help in the development of a vaccine expression vector, as well as providing a useful tool for studying promoter activity and in complementation studies of gene knockout mutants.


Reactivity of complex communities can be more important than stability.
Understanding stability-whether a community will eventually return to its original state after a perturbation-is a major focus in the study of various complex systems, particularly complex ecosystems. Here, we challenge this focus, showing that short-term dynamics can be a better predictor of outcomes for complex ecosystems. Using random matrix theory, we study how complex ecosystems behave immediately after small perturbations. Our analyses show that many communities are expected to be 'reactive', whereby some perturbations will be amplified initially and generate a response that is directly opposite to that predicted by typical stability measures. In particular, we find reactivity is prevalent for complex communities of mixed interactions and for structured communities, which are both expected to be common in nature. Finally, we show that reactivity can be a better predictor of extinction risk than stability, particularly when communities face frequent perturbations, as is increasingly common. Our results suggest that, alongside stability, reactivity is a fundamental measure for assessing ecosystem health.


Central vs. peripheral metabolic control of estrous cycles in Syrian hamsters. II. Glucoprivation.
Estrous cycles in Syrian hamsters are inhibited by food deprivation or treatment with pharmacological inhibitors of intracellular glucose utilization (glucoprivic treatments). These same metabolic challenges increase neural stimulation in areas of the caudal brain stem thought to be involved in detection of metabolic signals. Experiment 1 was designed to examine whether vagally transmitted signals are important for glucoprivic effects on estrous cycles and on neural stimulation in the caudal brain stem. Vagotomized or sham-operated hamsters were treated with 2-deoxy-D-glucose (2-DG) at a dose known to decrease cellular glucose utilization and inhibit estrous cycles (1,750 mg/kg). Vagotomized and sham-operated hamsters did not differ significantly in incidence of 2-DG-induced anestrus or in neural stimulation in the caudal brain stem, but the effects of 2-DG on estrous cycles and neural stimulation appeared to have been attenuated in vagotomized hamsters. In experiment 2, hamsters were injected intracerebroventricularly with 2-DG or glucose at doses that did not induce anestrus when injected systemically (125 and 250 mg/kg). Groups treated with intracerebroventricular injections of 2-DG showed a significantly higher incidence of anestrus than those treated with glucose. In experiment 3, effects of systemic injections of 2-DG were prevented by prior injection of glucose or fructose at the same concentration, indicating that 2-DG acts via effects on glucose metabolism, rather than via a nonspecific pharmacological effect or generalized stress response. Results of these experiments and those reported elsewhere (J. E. Schneider, A. J. Hall, and G. N. Wade. Am. J. Physiol. 272 (Regulatory Integrative Comp. Physiol, 41) R400-R405, 1997] are consistent with the notion that central glucoprivation is sufficient, whereas peripheral lipoprivation is not critical, for metabolic effects on estrous cycles.


Current management of pediatric acute otitis media.
Acute otitis media (AOM) is the most common childhood bacterial infection for which antibiotics are prescribed worldwide. The most common pathogens causing AOM in children are Streptococcus pneumoniae, nontypeable Haemophilus influenzae, Moraxella catarrhalis and Group A streptococcus. Antibiotic resistance is increasing among the bacterial pathogens causing AOM, with percentages of penicillin- and macrolide-resistant S. pneumoniae strains estimated to be between 30 and 70%, and of beta-lactamase-producing H. influenzae ranging between 20 and 40%. The introduction of the seven-valent pneumococcal conjugated vaccine had a major role in decreasing the number of vaccine-related S. pneumoniae AOM episodes, recurrent AOM cases and cases requiring the insertion of ventilation tubes. In parallel, it caused a rapid shift in the microbiology of AOM, characterized by an increase in the number of non-vaccine S. pneumoniae serotypes and H. influenzae isolates. The management of AOM in childhood has evolved considerably during recent years as a result of the new insights provided by the publication of the American Academy of Pediatrics and American Academy of Family Physicians guidelines for the treatment of AOM. The new treatment guidelines establish a clear hierarchy among various antibacterials used in the treatment of AOM and also the use of an age-stratified approach to AOM by recommending an observation strategy ('watchful waiting') without the use of antibacterials for some groups of AOM patients. Adherence to such a policy in patients with uncertain/questionable AOM diagnosis and/or mild-to-moderate symptoms, in addition to its implementation in patients over 2 years of age, could substantially reduce the use of antibacterials for the treatment of AOM and play a major role in the strategy of decreasing antibacterial resistance.


Ion migration assisted inscription of high refractive index contrast waveguides by femtosecond laser pulses in phosphate glass.
In this Letter, we report on the successful fabrication of low loss, high refractive index contrast waveguides via ion migration upon femtosecond laser writing in phosphate glass. Waveguides were produced in two different phosphate glass compositions with high and low La(2)O(3) content. In the La-rich glass, a large refractive index increase in the guiding region was observed due to the incoming migration of La accompanied by the out-diffusion of K. The much smaller refractive index change in the La-less glass is caused by rearrangements of the glass structure. These results confirm the feasibility of adapting the glass composition for enabling the laser writing of high refractive index contrast structures via spatially selective modification of the glass composition.


Structured review of post-cardiotomy extracorporeal membrane oxygenation: part 1-Adult patients.
Cardiogenic shock, cardiac arrest, acute respiratory failure, or a combination of such events, are all potential complications after cardiac surgery which lead to high mortality. Use of extracorporeal temporary cardio-circulatory and respiratory support for progressive clinical deterioration can facilitate bridging the patient to recovery or to more durable support. Over the last decade, extracorporeal membrane oxygenation (ECMO) has emerged as the preferred temporary artificial support system in such circumstances. Many factors have contributed to widespread ECMO use, including the relative ease of implantation, effectiveness, versatility, low cost relative to alternative devices, and potential for full, not just partial circulatory support. While there have been numerous publications detailing the short and midterm outcomes of ECMO support, specific reports about post-cardiotomy ECMO (PC-ECMO), are limited, single-center experiences. Etiology of cardiorespiratory failure leading to ECMO implantation, associated ECMO complications, and overall patient outcomes may be unique to the PC-ECMO population. Despite the rise in PC-ECMO use over the past decade, short-term survival has not improved. This report, therefore, aims to present a comprehensive overview of the literature with respect to the prevalence of ECMO use, patient characteristics, ECMO management, and in-hospital and early post-discharge patient outcomes for those treated for post-cardiotomy heart, lung, or heart-lung failure.


Immunogenicity to the bivalent HPV-16/18 vaccine among adolescent African students exposed to helminths and malaria.
Efficacious vaccines that prevent human papillomavirus (HPV) infection, the recognized cause of cervical cancer, are now available. However, in sub-Saharan Africa, immune-modulating infections such as helminths and malaria may affect immunogenicity to the HPV vaccine. This study aimed to evaluate the effect of helminth infections and exposure to malaria on the immune response to the bivalent HPV-16/18 vaccine. AS04-adjuvanted HPV-16/18 vaccinated students between 10 and 16 years of age from western Uganda, at 18 months-post vaccination were followed up for six months. After consent was obtained, demographic data, blood, and stool samples were collected. Multiplex HPV serology technology was used to determine HPV-16/18 antibody levels expressed as median fluorescent intensity (MFI). The malaria antibody immunoassay test was used to detect antibodies to malaria parasites. The Kato-Katz method was used to detect the presence of helminths. HPV-16/18 antibody levels among students exposed to malaria or helminths were compared with those who were not exposed using the Student's t-test. A total of 211 students participated in the study. There was no difference between MFI levels to HPV-16/18 antibodies at 18- and 24-month follow-ups among students who were positive and negative to malaria or helminth exposure. There was an increase in HPV-18 MFI antibody levels at month 24 among the students who were positive for malaria at enrolment (p = 0.05). Immune-modulating parasites (malaria/helminths) were not associated with reduced immune response to the bivalent HPV-16/18 vaccine. The data may support the use of this vaccine in sub-Saharan Africa.


Added sugar and dietary fiber consumption are associated with creativity in preadolescent children.
Objectives: Creativity requires the ability to combine existing mental representations in new ways and depends, in part, on the hippocampus. Hippocampal function is, in turn, affected by a number of health factors, including aerobic fitness, excess adiposity, and diet. Specifically, in rodent studies, diets high in saturated fatty acids and sugar - hallmarks of a western diet- have been shown to negatively impact hippocampal function and thereby impair performance on cognitive tasks that require the hippocampus. Yet relatively few studies have examined the relationship between diet and hippocampal-dependent cognition in children. Methods: The current study therefore sought to explore the relationship of several diet quality markers including dietary lipids (saturated fatty acids and omega-3 fatty acids), simple carbohydrates (added sugars), and dietary fiber with creativity in preadolescent children. Participants (N = 57; mean age = 9.1 years) completed the Verbal Form of the Torrance Test of Creative Thinking (TTCT), a standardized test of creativity known to require the hippocampus. Additionally, participants completed a 3-day food intake record with the assistance of a parent, underwent dual energy x-ray absorptiometry (DXA) to assess central adiposity, and VO2max testing to assess aerobic fitness. Results: Added sugar intake was negatively associated, and dietary fiber was positively associated with overall TTCT performance. These relationships were sustained even after controlling for key covariates. Discussion: These findings are among the first to report an association between added sugar consumption and hippocampal-dependent cognition during childhood and, given the key role of the hippocampus in learning and memory, as well as creative thinking, have potential educational and public health implications.


Can serology diagnose upper genital tract Chlamydia trachomatis infections? Studies on women with pelvic pain, with or without chlamydial plasmid DNA in endometrial biopsy tissue.
Upper genital tract chlamydial infections in women are on the increase, and serology might be a convenient tool for diagnosis. Evaluations of this approach are needed in women with or without microbiologic evidence of organisms in the upper genital tract. To compare the results of antibody assays with cervical culture and upper genital tract histopathology in women with pelvic pain and chlamydial plasmid DNA in endometrial biopsies. Chlamydia trachomatis plasmid DNA was detected by polymerase chain reaction (PCR) on extracted deparaffinized endometrial biopsy tissue. Five antichlamydial antibody assays were performed measuring total antibodies or immunoglobulin G (IgG), IgM, and IgA classes on sera from 14 women with plasmid DNA as well as 31 without plasmid DNA. Accepting the presence of plasmid DNA as the gold standard, no single test had total diagnostic accuracy. The best sensitivity and specificity occurred with the following assays: whole inclusion fluorescence (WIF) (100% and 80.6%); microimmunofluorescence IgM (MIF IgM) (78.6% and 93.6%); and heatshock protein-60 enzyme immunoassay (42.9% and 100%). Although recombinant anti-lipopolysaccharide enzyme-linked immunosorbent assays measured anti-chlamydial antibodies in a large proportion of these women, specificity was low. The sensitivity and specificity of cervical culture was 28.6% and 100% and of endometrial histopathology was 71.4% and 48.4%. Analysis of patient serological profiles suggested that and 6 women without plasmid DNA may have been cases that were missed by PCR. Evaluations of assays to diagnosis Chlamydia trachomatis upper genital tract infections could use the presence of organisms or their markers in the upper genital tract as a standard of comparison. Some of these serological assays, such as WIF or MIF IgM, may be helpful in diagnosis, but more studies are needed.


The value of cholangiography through Jackson-Pratt drains in the management of postoperative biliary injuries.
Iatrogenic biliary injury is the most significant complication after laparoscopic cholecystectomy. We present our experience with an alternative diagnostic approach using transcatheter cholangiography (TCC) through a Jackson-Pratt (JP) drain and discuss potential benefits and limitations of the technique. From March 2002 to February 2012, 40 patients with major postoperative biliary injury underwent biliary reconstruction at our institution. Mean age was 51.7 ± 18.1 years (range, 19 to 86 years) with 30 (75%) females. Seventeen (42.5%) injuries were detected intraoperatively and in 13 (32.5%) cases, JP drains were placed for biliary drainage. Lesions were classified according to Bismuth grade: I (10 patients [25%]), II (10 patients [25%]), III (six patients [15%]), IV (10 patients [25%]), and V (four patients [10%]). TCC was performed in seven patients with JP drains (53.8%). It fully defined the injury site in three cases of limited magnetic resonance cholangiopancreatography (MRCP) such as common hepatic duct and common bile duct leaks and in four cases (57.1%) that endoscopic retrograde cholangiopancreatography (ERCP) was limited as a result of clipping of the distal common bile duct. TCC showed promising results in cases of limited MRCP and ERCP such as fistulous orifices or leakage. It may represent an alternative adjunct in the diagnostic armamentarium of complex biliary injuries.


The cannabinoid CB1 receptor antagonist SR141716A attenuates the memory impairment produced by delta9-tetrahydrocannabinol or anandamide.
The administration of delta9-tetrahydrocannabinol (THC), the principle psychoactive ingredient in marijuana, or the endogenous cannabinoid anandamide, has been shown to impair recent memory. The purpose of the present investigation was to determine if the cannabinoid CB1 receptor antagonist SR141716A could attenuate THC- or anandamide-induced memory impairment, and to assess the effects on memory of SR141716A alone. Memory was assessed in rats well-trained in a two-component instrumental discrimination task, consisting of a conditional discrimination, and a non-match-to-position to assess recent or working memory. SR141716A (0.0-2.0 mg/kg) had no effect on either the conditional discrimination or the non-match-to-position. However, SR141716A (0.0-2.0 mg/kg) attenuated the memory impairment produced by THC (2.0 or 4.0 mg/kg) as indexed by an enhancement of performance in the non-match-to-position. When administered to rats pretreated with anandamide (2.0 mg/kg), SR141716A (0.0-2.5 mg/kg) impaired performance in the conditional discrimination at the highest dose. This was interpreted as a deficit in some capacity unrelated to memory (e.g., motor impairment). However, lower doses of SR141716A (0.1 and 0.5 mg/kg) attenuated the anandamide-induced impairment of performance in the non-match-to-position without affecting the conditional discrimination. This is the first report that the memory impairment produced by anandamide can be attenuated by a cannabinoid antagonist; results suggest that anandamide-induced memory disruption is mediated by CB receptors.


Thermal acclimation of tropical coral reef fishes to global heat waves.
As climate-driven heat waves become more frequent and intense, there is increasing urgency to understand how thermally sensitive species are responding. Acute heating events lasting days to months may elicit acclimation responses to improve performance and survival. However, the coordination of acclimation responses remains largely unknown for most stenothermal species. We documented the chronology of 18 metabolic and cardiorespiratory changes that occur in the gills, blood, spleen, and muscles when tropical coral reef fishes are thermally stressed (+3.0°C above ambient). Using representative coral reef fishes (Caesio cuning and Cheilodipterus quinquelineatus) separated by >100 million years of evolution and with stark differences in major life-history characteristics (i.e. lifespan, habitat use, mobility, etc.), we show that exposure duration illicited coordinated responses in 13 tissue and organ systems over 5 weeks. The onset and duration of biomarker responses differed between species, with C. cuning - an active, mobile species - initiating acclimation responses to unavoidable thermal stress within the first week of heat exposure; conversely, C. quinquelineatus - a sessile, territorial species - exhibited comparatively reduced acclimation responses that were delayed through time. Seven biomarkers, including red muscle citrate synthase and lactate dehydrogenase activities, blood glucose and hemoglobin concentrations, spleen somatic index, and gill lamellar perimeter and width, proved critical in evaluating acclimation progression and completion, as these provided consistent evaluation of thermal responses across species.


Induction of heme oxygenase-1 attenuates lipopolysaccharide-induced inflammasome activation in human gingival epithelial cells.
Interleukin-1β (IL-1β) is a pathogenic factor for the destruction of periodontal tissue in periodontitis. The processing of IL-1β is regulated by cytosolic machinery termed as the inflammasome, which recruits and activates caspase-1 and then cleaves pro-IL-1β to produce mature IL-1β. Porphyromonas gingivalis (P. gingivalis) infection and lipopolysaccharide (LPS) have been shown to activate the NLRP3 inflammasome and stimulate IL-1β production in human oral cells. Heme oxygenase-1 (HO-1) is an ubiquitous cytoprotective enzyme. The products of HO-1 exhibit protective biological activities, including antioxidant and anti-inflammatory effects. In the present study, we investigated the hypothesis that the induction of HO-1 inhibits the activation of the inflammasome and protects against LPS-induced inflammatory damage in cultured human gingival epithelial cells (GECs). Our results revealed that LPS induced the overexpression of the inflammasome components, NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and caspase-1, by western blot analysis. Co-immunoprecipitation analysis indicated that LPS increased the binding of NLRP3 and ASC, and confocal imaging revealed that LPS increased the immunostaining and co-localization of ASC and caspase-1, indicating that LPS enhanced the assembly/formation of the inflammasome components. Hemin, a potent HO-1 inducer, blocked the LPS-induced overexpression and the formation of the NLRP3 inflammasome. Furthermore, hemin also inhibited the LPS-induced increase in the production of IL-1β, as shown by enzyme-linked immunosorbent assay and blocked the nuclear translocation of pro-inflammatory nuclear factor-κB (NF-κB), as shown by confocal assays. As a result, hemin protected the cells from LPS-induced damage, which was demonstrated by the immunostaining pattern of the cell junction protein, E-cadherin. LPS produced a disturbed staining pattern of E-cadherin, suggesting the disruption of epithelial integrity, which was abolished in the hemin-treated cells. In conclusion, our data demonstrate that the induction of HO-1 by hemin attenuates LPS-induced inflammatory damage in human GECs through the inhibition of inflammasome activation.


Akt Regulates Axon Wrapping and Myelin Sheath Thickness in the PNS.
The signaling pathways that regulate myelination in the PNS remain poorly understood. Phosphatidylinositol-4,5-bisphosphate 3-kinase 1A, activated in Schwann cells by neuregulin and the extracellular matrix, has an essential role in the early events of myelination. Akt/PKB, a key effector of phosphatidylinositol-4,5-bisphosphate 3-kinase 1A, was previously implicated in CNS, but not PNS myelination. Here we demonstrate that Akt plays a crucial role in axon ensheathment and in the regulation of myelin sheath thickness in the PNS. Pharmacological inhibition of Akt in DRG neuron-Schwann cell cocultures dramatically decreased MBP and P0 levels and myelin sheath formation without affecting expression of Krox20/Egr2, a key transcriptional regulator of myelination. Conversely, expression of an activated form of Akt in purified Schwann cells increased expression of myelin proteins, but not Krox20/Egr2, and the levels of activated Rac1. Transgenic mice expressing a membrane-targeted, activated form of Akt under control of the 2',3'-cyclic nucleotide 3'-phosphodiesterase promoter, exhibited thicker PNS and CNS myelin sheaths, and PNS myelin abnormalities, such as tomacula and myelin infoldings/outfoldings, centered around the paranodes and Schmidt Lanterman incisures. These effects were corrected by rapamycin treatmentin vivo Importantly, Akt activity in the transgenic mice did not induce myelination of nonmyelinating Schwann cells in the sympathetic trunk or Remak fibers of the dorsal roots, although, in those structures, they wrapped membranes redundantly around axons. Together, our data indicate that Akt is crucial for PNS myelination driving axonal wrapping by unmyelinated and myelinated Schwann cells and enhancing myelin protein synthesis in myelinating Schwann cells. Although the role of the key serine/threonine kinase Akt in promoting CNS myelination has been demonstrated, its role in the PNS has not been established and remains uncertain. This work reveals that Akt controls several key steps of the PNS myelination. First, its activity promotes membrane production and axonal wrapping independent of a transcriptional effect. In myelinated axons, it also enhances myelin thickness through the mTOR pathway. Finally, sustained Akt activation in Schwann cells leads to hypermyelination/dysmyelination, mimicking some features present in neuropathies, such as hereditary neuropathy with liability to pressure palsies or demyelinating forms of Charcot-Marie-Tooth disease. Together, these data demonstrate the role of Akt in regulatory mechanisms underlying axonal wrapping and myelination in the PNS.


Effect of a medical student emergency ultrasound clerkship on number of emergency department ultrasounds.
To determine whether a medical student emergency ultrasound clerkship has an effect on the number of patients undergoing ultrasonography and the number of total scans in the emergency department. We conducted a prospective, single-blinded study of scanning by emergency medicine residents and attendings with and without medical students. Rotating ultrasound medical students were assigned to work equally on all days of the week. We collected the number of patients scanned and the number of scans, as well as participation of resident and faculty. In seven months 2,186 scans were done on the 109 days with students and 707 scans on the 72 days without them. Data on 22 days was not recorded. A median of 13 patients per day were scanned with medical students (CI 12-15) versus seven (CI 6-9) when not. In addition, the median number of scans was 18 per day with medical students (CI 16-20) versus eight (CI 6-10) without them. There were significantly more patients scanned and scans done when ultrasound medical students were present.


Long-term course of probable PTSD after the 9/11 attacks: a study in urban primary care.
Although the short- and midterm psychological effects of the attacks on September 11, 2001 (9/11) have been well described, less is known about the long-term effects. This study examines the course of probable posttraumatic stress disorder (PTSD), its predictors and clinical consequences in a cohort of 455 primary care patients in New York City, interviewed approximately 1 and 4 years after 9/11. The rate of PTSD decreased from 9.6% to 4.1%. Pre-9/11 major depressive disorder emerged as the strongest predictor of PTSD, particularly late-PTSD. At follow-up, late-PTSD was associated with major depressive and anxiety disorders, and PTSD regardless of timing was associated with impaired functioning. Findings highlight the importance of ongoing evaluation of mental health needs in primary care settings in the aftermath of disasters.


A study on the application of near infrared hyperspectral chemical imaging for monitoring moisture content and water activity in low moisture systems.
Moisture content and water activity are key parameters in predicting the stability of low moisture content products. However, conventional methods for moisture content and water activity determination (e.g., loss on drying method, Karl Fischer titration, dew point method) are time consuming, demand specialized equipment and are not amenable to online processing. For this reason they are typically applied at-line on a limited number of samples. Near infrared hyperspectral chemical imaging is an emerging technique for spatially characterising the spectral properties of samples. Due to the fast acquisition of chemical images, many samples can be evaluated simultaneously, thus providing the potential for online evaluation of samples during processing. In this study, the potential of NIR chemical imaging for predicting the moisture content and water activity of a selection of low moisture content food systems is evaluated.


Guidelines for reporting methods to estimate metabolic rates by aquatic intermittent-flow respirometry.
Interest in the measurement of metabolic rates is growing rapidly, because of the importance of metabolism in advancing our understanding of organismal physiology, behaviour, evolution and responses to environmental change. The study of metabolism in aquatic animals is undergoing an especially pronounced expansion, with more researchers utilising intermittent-flow respirometry as a research tool than ever before. Aquatic respirometry measures the rate of oxygen uptake as a proxy for metabolic rate, and the intermittent-flow technique has numerous strengths for use with aquatic animals, allowing metabolic rate to be repeatedly estimated on individual animals over several hours or days and during exposure to various conditions or stimuli. There are, however, no published guidelines for the reporting of methodological details when using this method. Here, we provide the first guidelines for reporting intermittent-flow respirometry methods, in the form of a checklist of criteria that we consider to be the minimum required for the interpretation, evaluation and replication of experiments using intermittent-flow respirometry. Furthermore, using a survey of the existing literature, we show that there has been incomplete and inconsistent reporting of methods for intermittent-flow respirometry over the past few decades. Use of the provided checklist of required criteria by researchers when publishing their work should increase consistency of the reporting of methods for studies that use intermittent-flow respirometry. With the steep increase in studies using intermittent-flow respirometry, now is the ideal time to standardise reporting of methods, so that - in the future - data can be properly assessed by other scientists and conservationists.


Viperin has species-specific roles in response to herpes simplex virus infection.
Viperin is a gene with a broad spectrum of antiviral functions and various mechanisms of action. The role of viperin in herpes simplex virus type 1 (HSV-1) infection is unclear, with conflicting data in the literature that is derived from a single human cell type. We have addressed this gap by investigating viperin during HSV-1 infection in several cell types, spanning species and including immortalized, non-immortalized and primary cells. We demonstrate that viperin upregulation by HSV-1 infection is cell-type-specific, with mouse cells typically showing greater increases compared with those of human origin. Further, overexpression and knockout of mouse, but not human viperin significantly impedes and increases HSV-1 replication, respectively. In primary mouse fibroblasts, viperin upregulation by infection requires viral gene transcription and occurs in a predominantly IFN-independent manner. Further we identify the N-terminal domain of viperin as being required for the anti-HSV-1 activity. Interestingly, this is the region of viperin that differs most between mouse and human, which may explain the apparent species-specific activity against HSV-1. Finally, we show that HSV-1 virion host shutoff (vhs) protein is a key viral factor that antagonises viperin in mouse cells. We conclude that viperin can be upregulated by HSV-1 in mouse and human cells, and that mouse viperin has anti-HSV-1 activity.


Cell culture-selected substitutions in influenza A(H3N2) neuraminidase affect drug susceptibility assessment.
Assessment of drug susceptibility has become an integral part of influenza virus surveillance. In this study, we describe the drug resistance profile of influenza A(H3N2) virus, A/Mississippi/05/2011, collected from a patient treated with oseltamivir and detected via surveillance. An MDCK cell-grown isolate of this virus exhibited highly reduced inhibition by the neuraminidase (NA) inhibitors (NAIs) oseltamivir (8,005-fold), zanamivir (813-fold), peramivir (116-fold), and laninamivir (257-fold) in the NA inhibition assay. Sequence analysis of its NA gene revealed a known oseltamivir-resistance marker, the glutamic acid-to-valine substitution at position 119 (E119V), and an additional change, threonine to isoleucine at position 148 (T148I). Unlike E119V, T148I was not detected in the clinical sample but acquired during viral propagation in MDCK cells. Using recombinant proteins, T148I by itself was shown to cause only a 6-fold increase in the zanamivir 50% inhibitory concentration (IC50) and had no effect on inhibition by other drugs. The T148I substitution reduced NA activity by 50%, most likely by affecting the positioning of the 150 loop at the NA catalytic site. Using pyrosequencing, changes at T148 were detected in 35 (23%) of 150 MDCK cell-grown A(H3N2) viruses tested, which was lower than the frequency of changes at D151 (85%), an NA residue previously implicated in cell selection. We demonstrate that culturing of the A(H3N2) viruses (n = 11) at a low multiplicity of infection delayed the emergence of the NA variants with changes at position 148 and/or 151, especially when conducted in MDCK-SIAT1 cells. Our findings highlight the current challenges in monitoring susceptibility of influenza A(H3N2) viruses to the NAI class of antiviral drugs.


Determining the genome-wide kinship coefficient seems unhelpful in distinguishing consanguineous couples with a high versus low risk for adverse reproductive outcome.
Offspring of consanguineous couples are at increased risk of congenital disorders. The risk increases as parents are more closely related. Individuals that have the same degree of relatedness according to their pedigree, show variable genomic kinship coefficients. To investigate whether we can differentiate between couples with high- and low risk for offspring with congenital disorders, we have compared the genomic kinship coefficient of consanguineous parents with a child affected with an autosomal recessive disorder with that of consanguineous parents with only healthy children, corrected for the degree of pedigree relatedness. 151 consanguineous couples (73 cases and 78 controls) from 10 different ethnic backgrounds were genotyped on the Affymetrix platform and passed quality control checks. After pruning SNPs in linkage disequilibrium, 57,358 SNPs remained. Kinship coefficients were calculated using three different toolsets: PLINK, King and IBDelphi, yielding five different estimates (IBDelphi, PLINK (all), PLINK (by population), King robust (all) and King homo (by population)). We performed a one-sided Mann Whitney test to investigate whether the median relative difference regarding observed and expected kinship coefficients is bigger for cases than for controls. Furthermore, we fitted a mixed effects linear model to correct for a possible population effect. Although the estimated degrees of genomic relatedness with the different toolsets show substantial variability, correlation measures between the different estimators demonstrated moderate to strong correlations. Controls have higher point estimates for genomic kinship coefficients. The one-sided Mann Whitney test did not show any evidence for a higher median relative difference for cases compared to controls. Neither did the regression analysis exhibit a positive association between case-control status and genomic kinship coefficient. In this case-control setting, in which we compared consanguineous couples corrected for degree of pedigree relatedness, a higher degree of genomic relatedness was not significantly associated with a higher likelihood of having an affected child. Further translational research should focus on which parts of the genome and which pathogenic mutations couples are sharing. Looking at relatedness coefficients by determining genome-wide SNPs does not seem to be an effective measure for prospective risk assessment in consanguineous parents.


Effects of mirror therapy combined with neuromuscular electrical stimulation on motor recovery of lower limbs and walking ability of patients with stroke: a randomized controlled study.
To investigate the effectiveness of mirror therapy combined with neuromuscular electrical stimulation in promoting motor recovery of the lower limbs and walking ability in patients suffering from foot drop after stroke. Randomized controlled study. Inpatient rehabilitation center of a teaching hospital. Sixty-nine patients with foot drop. Patients were randomly divided into three groups: control, mirror therapy, and mirror therapy + neuromuscular electrical stimulation. All groups received interventions for 0.5 hours/day and five days/week for four weeks. 10-Meter walk test, Brunnstrom stage of motor recovery of the lower limbs, Modified Ashworth Scale score of plantar flexor spasticity, and passive ankle joint dorsiflexion range of motion were assessed before and after the four-week period. After four weeks of intervention, Brunnstrom stage ( P = 0.04), 10-meter walk test ( P < 0.05), and passive range of motion ( P < 0.05) showed obvious improvements between patients in the mirror therapy and control groups. Patients in the mirror therapy + neuromuscular electrical stimulation group showed better results than those in the mirror therapy group in the 10-meter walk test ( P < 0.05). There was no significant difference in spasticity between patients in the two intervention groups. However, compared with patients in the control group, patients in the mirror therapy + neuromuscular electrical stimulation group showed a significant decrease in spasticity ( P < 0.001). Therapy combining mirror therapy and neuromuscular electrical stimulation may help improve walking ability and reduce spasticity in stroke patients with foot drop.


Holistic assessment of anger in adolescents - Development of a rating scale.
Anger related issues in adolescents are an important contributor to adolescent mortality and many negative developmental outcomes. Anger being the most primitive defence mechanism, determining its maladaptive nature is a complex phenomenon. Further, it lacks clarity in terms of its definition, demarcation from aggression and hostility, and also its assessment. Available anger rating scales do not represent anger adequately or comprehensively. The concept of anger is dealt with exhaustively in Indian classical texts. According to these texts, anger is an evil virtue that is born out of Rajasic nature and leads to momentary loss of cognitive functional abilities. Manifestation of anger occurs at behavioural (kayika), verbal (vachika) and mental (manasika) levels. Based on these principles, a psychometric scale is developed for assessment of anger in adolescents. The objective of this study was to develop a psychometric scale for assessment of anger in adolescents based on behavioural (kayika), verbal (vachika) and mental (manasika) levels. Item generation was done referring to the existing scales, texts on spirituality and psychology. It also included discussions with children, teachers and parents. Item reduction was carried out using expert opinion, focused group discussion and factor analysis. Testretest reliability was checked with 127 children (63 girls and 64 boys). The final scale was administered to 757 children (427 girls and 330 boys) to ascertain reliability co-efficient values. Co-efficient of Alpha value for final scale recorded 0.804. Test-retest reliability showed .835 correlation. Anger can be assessed holistically using Trikarana (kayika, vachika and manasika) concept as dealt in almost all Indian classical texts. A 23-item adolescent anger scale is a comprehensive tool to assess behavioural, verbal and mental anger in adolescents.


Yersinia pseudotuberculosis YopE prevents uptake by M cells and instigates M cell extrusion in human ileal enteroid-derived monolayers.
Many pathogens use M cells to access the underlying Peyer's patches and spread to systemic sites via the lymph as demonstrated by ligated loop murine intestinal models. However, the study of interactions between M cells and microbial pathogens has stalled due to the lack of cell culture systems. To overcome this obstacle, we use human ileal enteroid-derived monolayers containing five intestinal cell types including M cells to study the interactions between the enteric pathogen, Yersinia pseudotuberculosis (Yptb), and M cells. The Yptb type three secretion system (T3SS) effector Yops inhibit host defenses including phagocytosis and are critical for colonization of the intestine and Peyer's patches. Therefore, it is not understood how Yptb traverses through M cells to breach the epithelium. By growing Yptb under two physiological conditions that mimic the early infectious stage (low T3SS-expression) or host-adapted stage (high T3SS-expression), we found that large numbers of Yptb specifically associated with M cells, recapitulating murine studies. Transcytosis through M cells was significantly higher by Yptb expressing low levels of T3SS, because YopE and YopH prevented Yptb uptake. YopE also caused M cells to extrude from the epithelium without inducing cell-death or disrupting monolayer integrity. Sequential infection with early infectious stage Yptb reduced host-adapted Yptb association with M cells. These data underscore the strength of enteroids as a model by discovering that Yops impede M cell function, indicating that early infectious stage Yptb more effectively penetrates M cells while the host may defend against M cell penetration of host-adapted Yptb.


Electrically induced verbal perseveration: A striatal deafferentation model.
The present study aimed to elucidate the neural correlates of the deafferentation cognitive model of verbal perseveration (VP) by analyzing the connectomics of the sites where electric stimulation elicited VP during awake left glioma surgery. We retrospectively reviewed the anatomic sites that generated VP when electrically stimulated in a series of 21 patients operated on while awake for a left glioma. Each stimulation point was manually located on the postoperative MRI and then registered to the Montreal Neurological Institute template. Connectomics of these sites were further analyzed with Tractotron and disconnectome maps. VP stimulation sites were located within the white matter surrounding the posterosuperior head of the caudate nucleus, as well as within the white matter of the external capsule and the superolateral wall of the temporal horn of the ventricle. Furthermore, Tractotron and disconnectome maps revealed the connectome of these stimulation sites: the inferior fronto-occipital fasciculus, frontostriatal tract, and anterior thalamic radiations. On the basis of these results and other data, we propose the following anatomic implementation of the deafferentation cognitive model: the lexico-semantic system, comprising different areas linked together through direct cortico-cortical connections, sends information to the striatum; the striato-thalamic system acts as a tunable filter of this lexico-semantic input; and the thalamus projects back to the lexico-semantic system, amplifying the targeted response and inhibiting its competitors.


Liquid biopsy reveals collateral tissue damage in cancer.
Cancer inflicts damage to surrounding normal tissues, which can culminate in fatal organ failure. Here, we demonstrate that cell death in organs affected by cancer can be detected by tissue-specific methylation patterns of circulating cell-free DNA (cfDNA). We detected elevated levels of hepatocyte-derived cfDNA in the plasma of patients with liver metastases originating from different primary tumors, compared with cancer patients without liver metastases. In addition, patients with localized pancreatic or colon cancer showed elevated hepatocyte cfDNA, suggesting liver damage inflicted by micrometastatic disease, by primary pancreatic tumor pressing the bile duct, or by a systemic response to the primary tumor. We also identified elevated neuron-, oligodendrocyte-, and astrocyte-derived cfDNA in a subpopulation of patients with brain metastases compared with cancer patients without brain metastasis. Cell type-specific cfDNA methylation markers enabled the identification of collateral tissue damage in cancer, revealing the presence of metastases in specific locations and potentially assisting in early cancer detection.


When does acute pain become chronic?
The transition from acute to chronic pain appears to occur in discrete pathophysiological and histopathological steps. Stimuli initiating a nociceptive response vary, but receptors and endogenous defence mechanisms in the periphery interact in a similar manner regardless of the insult. Chemical, mechanical, and thermal receptors, along with leucocytes and macrophages, determine the intensity, location, and duration of noxious events. Noxious stimuli are transduced to the dorsal horn of the spinal cord, where amino acid and peptide transmitters activate second-order neurones. Spinal neurones then transmit signals to the brain. The resultant actions by the individual involve sensory-discriminative, motivational-affective, and modulatory processes in an attempt to limit or stop the painful process. Under normal conditions, noxious stimuli diminish as healing progresses and pain sensation lessens until minimal or no pain is detected. Persistent, intense pain, however, activates secondary mechanisms both at the periphery and within the central nervous system that cause allodynia, hyperalgesia, and hyperpathia that can diminish normal functioning. These changes begin in the periphery with upregulation of cyclo-oxygenase-2 and interleukin-1β-sensitizing first-order neurones, which eventually sensitize second-order spinal neurones by activating N-methyl-d-aspartic acid channels and signalling microglia to alter neuronal cytoarchitecture. Throughout these processes, prostaglandins, endocannabinoids, ion-specific channels, and scavenger cells all play a key role in the transformation of acute to chronic pain. A better understanding of the interplay among these substances will assist in the development of agents designed to ameliorate or reverse chronic pain.


Update on imaging of vulvar squamous cell carcinoma.
The purpose of this article is to describe the advantages and diagnostic and prognostic implications of imaging in the management of vulvar carcinoma. As the treatment of vulvar carcinoma evolves to tailored surgery and chemoradiation therapy to reduce morbidity, the importance of pretreatment assessment of vulvar carcinoma increases. Advances in imaging, such as pelvic MRI and PET, add to the benefits of clinical evaluation in pretreatment planning, assessment of response to chemoradiation therapy, and posttreatment surveillance.


Genetic control of human NK cell repertoire.
Through differential killer cell Ig-like receptor (KIR) and CD94:NKG2 gene expression, human NK cells generate diverse repertoires, each cell having an inhibitory receptor for autologous HLA class I. Using a new method for measuring repertoire difference that integrates multiple flow cytometry parameters, we found individual repertoire stability, but population variability. Correlating repertoire differences with KIR and HLA genotype for 85 sibling pairs reveals the dominant influence of KIR genotype; HLA genotype having a subtle, modulating effect on relative KIR expression frequencies. HLA and/or KIR genotype also influences CD94:NKG2A expression. After HLA-matched stem cell transplantation, KIR repertoires either recapitulated that of the donor or were generally depressed for KIR expression. Human NK cell repertoires are defined by combinations of variable KIR and HLA class I genes and conserved CD94:NKG2 genes.


Presenting features of meningococcal disease, public health messages and media publicity: are they consistent?
To investigate whether the presenting features of meningococcal disease as promoted in public health awareness material and in the print media accurately reflect the clinical features in patients admitted to Auckland hospitals with meningococcal disease January 1998 to June 1999. Hospital record, public health message and newspaper article review, with analysis by presenting feature, age group and disease complex. The most common presenting features were fever (95%), rash (65%), vomiting and nausea (64%), lethargy (62%), headache (44%), refusing food and drink (35%), irritability (33%), muscle ache and joint pains (27%) and stiff neck (26%). Public health messages gave appropriate emphasis to the key features, whereas newspaper articles under-emphasised these. The term 'meningitis' was used more frequently in newspapers (65%) than in public health messages (30%), despite meningitis alone presenting less frequently (38% of cases) than meningococcal septicaemia, and having a less serious prognosis. Presenting features currently noted in the Ministry of Health's health education resource material are appropriate. Public health specialists dealing with the media should ensure that appropriate messages are incorporated into media reports. A greater use of the term 'meningococcal disease' by both public health agencies and media would convey to the public the message that this disease has a spectrum of presenting features, with those of septicaemia more common, but also indicating an even greater need for urgency of action than with 'classical' meningitic features.


A Humorously Serious Take on Plight of Postgraduate Medical Entrance Examinees in India.
Following is a satire on the plight of postgraduate medical entrance examinees in India. It highlights the questions they have to face during their entrance examination preparations and the annoying explanations they have to read in the entrance preparation books. Though the system of examination is improving in India, few loopholes do exist and need to be rectified. This article, thus, attracts the attention of readers towards one such backdrop regarding controversial/wrong/misprinted questions which has come a long way but still has scope of improvement.