Biomarkers for screening of lung cancer and pre-neoplastic lesions in a high risk Chilean population.
The mortality of lung cancer (LC), increases each year in the world, in spite of any advances, in development of new drugs to advance stages of LC. The high incidence of LC has been associated with smoking habit, genetic diversity and environmental pollution. Antofagasta region has been reported to have the highest LC mortality rate in Chile and its inhabitants were exposed to arsenic in their drinking water in concentrations as high as 870 μg/L. Non-invasive techniques such as biomarkers (Automatic Quantitative Cytometry: AQC and DR70) and Auto Fluorescence Bronchoscopy (AFB) might be potentially useful as a supplementary diagnostic approach and early detection. Early detection is one of the most important factors to intervene and prevent cancer progression in LC. This is a work of an ongoing prospective bimodality cancer surveillance study in high risk LC volunteers. Enrolment was done in subjects from Antofagasta and Metropolitan regions. In addition, we enrolled subjects who were suspected of having lung cancer. AQC, DR70 and AFB were used as tools in the detection of pre-neoplastic (PNL) and neoplastic lesions (NL). Half of the samples, classified as suspicious by AFB, were confirmed as metaplasia or dysplasia by histopathology. For LC, DR70 showed a higher sensitivity (95.8%) and specificity (91.9%) than AQC. However, for PNL AQC showed a higher sensitivity (91.9%) than DR70 (27.3%), although both with low PPV values. As a pre screener, both biomarkers might be employed as complementary tools to detect LC, especially as serially combined tests, with a sensitivity of 60% and a PPV of 65.2%. Additionally, the use of parallel combined tests might support the detection of PNL (sensitivity 91.2%; PPV 49.1%). This work adds information on cellular and molecular biomarkers to complement imaging techniques for early detection of LC in Latin America that might contribute to formulate policies concerning screening of LC. Supported by INNOVA-CORFO, Chile.


Gene expression patterns in a rodent model for depression.
Disturbances in sleep are encountered in the majority of patients with depression. To elucidate the possible molecular mechanisms behind this relationship we examined gene expression changes in a rodent model for depression and disturbed sleep. Animals were treated with daily injections of clomipramine in their early infancy, after which gene expression in basal forebrain was examined using Affymetrix Rat 230.2 chips. We tested the levels of both single transcripts and involved pathways, and searched for common nominators (i.e. transcription factors) that could explain these changes. We identified 72 differentially expressed gene transcripts, many of which are involved in epigenetic regulation, such as DNMT2. Analysis of functional pathways revealed statistically significant changes of the biological process of synaptic transmission, the cellular compartment of the synapse and the molecular function of GABA signalling, showing that transcripts with altered expression are functionally related. Finally, promoter analysis of the differentially expressed genes showed a clear enrichment of binding sites for the transcription factor CREB1, a molecule also involved in epigenetic regulation (cAMP response element-binding protein induces histone modifications). These results indicate that CREB1 may constitute one of the major links between disturbed sleep and mood. The results also highlight the molecular mechanisms in the murine clomipramine model, previously shown to be a valid model for depression.


Chemical and physical guidance of fish spermatozoa into the egg through the micropyle†,‡.
Eggs of teleost fish, unlike those of many other animals, allow sperm entry only at a single site, a narrow canal in the egg's chorion called the micropyle. In some fish (e.g., flounder, herring, and Alaska pollock), the micropyle is a narrow channel in the chorion, with or without a shallow depression around the outer opening of micropyle. In some other fish (e.g., salmon, pufferfish, cod, and medaka), the micropyle is like a funnel with a conical opening. Eggs of all the above fish have a glycoprotein tightly bound to the chorion surface around the micropyle. This glycoprotein directs spermatozoa into the micropylar canal in a Ca2+-dependent manner. This substance, called the micropylar sperm attractant or MISA, increases fertilization efficiency and is essential in herring. In flounder, salmon, and perhaps medaka, fertilization is possible without MISA, but its absence makes fertilization inefficient because most spermatozoa swim over the micropyle without entering it. The mechanism underlying sperm-MISA interactions is yet to be determined, but at least in herring the involvement of Ca2+ and K+ channel proteins, as well as CatSper and adenylyl cyclase, is very likely. In some other fish (e.g., zebrafish, loach, and goldfish), the chorion around the micropyle is deeply indented (e.g., zebrafish and loach) or it has radially or spirally arranged grooves around the outer opening of the micropyle (e.g., goldfish). MISA is absent from the eggs of these fish and sperm entry into micropylar canal seems to be purely physical.


Reversible bond formation in a gold-atom-organic-molecule complex as a molecular switch.
We report on the formation of a metal-molecule complex that can be used as a molecular switch. Using a cryogenic scanning tunneling microscope, a covalent bond was formed reversibly between a gold atom and a perylene-3,4,9,10-tetracarboxylic dianhydride molecule supported by a thin insulating film. The bonded and the nonbonded state of the complex were found to be associated with different charge states, and the switching between the two states was accompanied by a considerable change in the tunneling current. Atomic force microscopy molecular imaging was employed to determine precisely the atomic structure of the complex, and the experimental results were corroborated by density functional theory calculations.


Associated anomalies in asymmetric crying facies and 22q11 deletion.
Congenital asymmetric crying facies, a minor congenital anomaly due to unilateral absence or hypoplasia of the depressor anguli oris muscle, is associated at times with major congenital anomalies. A large number of asymmetric crying facies cases with chromosome 22q11 microdeletions have presently been reported. Fluorescence in situ hybridization (FISH) analysis for 22q11 deletion was performed on 8 infants with asymmetric crying facies. Five of our patients had at least one associated systemic anomaly. Two of 5 patients had conotruncal heart disease (Cayler cardiofacial syndrome). In three of the affected infants, we failed to reveal additional congenital malformation. The 22q11 deletion was present in only one patient. This baby had congenital hypoparathyroidism, severe neonatal hypocalcaemia and tetralogy of Fallot. We suggest, a 22q11 deletion should be excluded not in all cases but in cases with Cayler cardiofacial syndrome and in ACF associated with additional congenital anomalies.


Transient bile duct obstruction. Response of serum bilirubin and alkaline phosphatase levels in the rat.
The immediate evolution of changes in serum bilirubin and alkaline phosphatase levels during and after brief temporary occlusion (12 hours or less) of the main bile duct was studied in the rat. Transient obstruction of the main bile duct of more than one hour was characterized by a simultaneous increase in serum levels of both bilirubin and alkaline phosphatase. The time required for return to normal values was less for bilirubin than the duration of a brief obstruction and was more prolonged for alkaline phosphatase than the duration of obstruction. In obstruction of four or more hours, it was rare to find an isolated elevation of serum bilirubin; this occurred when the control value for alkaline phosphatase levels was low and their relative increase was obscured. Thus, an isolated increase in the serum bilirubin level is infrequently compatible with obstruction of the bile duct, even when transient and of brief duration. Conversely, isolated elevation of serum alkaline phosphatase levels in an acute abdominal syndrome may indicate relieved biliary obstruction.


Estimating Emissions of Toxic Hydrocarbons from Natural Gas Production Sites in the Barnett Shale Region of Northern Texas.
Oil and natural gas operations have continued to expand and move closer to densely populated areas, contributing to growing public concerns regarding exposure to hazardous air pollutants. During the Barnett Shale Coordinated Campaign in October, 2013, ground-based whole air samples collected downwind of oil and gas sites revealed enhancements in several potentially toxic volatile organic compounds (VOCs) when compared to background values. Molar emissions ratios relative to methane were determined for hexane, benzene, toluene, ethylbenzene, and xylene (BTEX compounds). Using methane leak rates measured from the Picarro mobile flux plane (MFP) system and a Barnett Shale regional methane emissions inventory, the rates of emission of these toxic gases were calculated. Benzene emissions ranged between 51 ± 4 and 60 ± 4 kg h-1. Hexane, the most abundantly emitted pollutant, ranged from 642 ± 45 to 1070 ± 340 kg h-1. While observed hydrocarbon enhancements fall below federal workplace standards, results may indicate a link between emissions from oil and natural gas operations and concerns about exposure to hazardous air pollutants. The larger public health risks associated with the production and distribution of natural gas are of particular importance and warrant further investigation, particularly as the use of natural gas increases in the United States and internationally.


Transcriptional control of the heme oxygenase gene in mouse M1 cells during their TPA-induced differentiation into macrophages.
It has long been known that heme oxygenase (HO) is a key enzyme in heme catabolism and recently it was also found to acts as an oxidative stress protein to produce carbon monoxide (CO), which has similar actions to those of nitrogen monoxide (NO). Therefore, we examined transcriptional control of the HO gene in mouse M1 (myeloleukemia) cells during their differentiation into macrophages. Since the promoter region of this gene is known to have a TPA-responsive element (TRE), its expression might be regulated by a C-kinase signal transduction pathway. Then we investigated the activation of the HO gene after treatment of M1 cells with TPA and inhibitors of C-kinase. When M1 cells were treated with TPA, they differentiated into macrophage-like cells. Upon treatment with TPA, H2O2 was produced first, the nuclear proto-oncogenes fos and jun were activated, and then the HO gene was activated. The extent of transcriptional activation of the fos, jun, and HO genes in M1 cells treated with TPA was reduced by a specific inhibitor of C-kinase and a scavenger of oxygen radicals. When M1 cells were treated with H2O2, essentially the same level of transcription of the HO gene was observed, but the extent of transcriptional activation of the fos and jun genes was about half of the treatment with TPA. Super-shift assays using the TRE of the HO gene revealed that the Fos and Jun proteins from nuclei of M1 cells treated with TPA bound to the TRE, and same assays using DNA with the NF-kappa B motif also revealed that the active NF-kappa B protein from M1 cells treated with H2O2 or TPA also bound to the corresponding motif. These results strongly suggest that the HO gene in M1 cells is activated by TPA through a production of H2O2, an oxidative activation pathway of NF-kappa B, and a signal-transduction pathway that involves C-kinase during the differentiation of macrophages that occurs upon treatment with TPA.


Factors affecting the region of most isometric femoral attachments. Part I: The posterior cruciate ligament.
We measured how the distance between selected tibial and femoral attachments of the PCL changes with knee flexion in six intact cadaver knee. The femoral location was the primary determinant of whether the distance increased, decreased, or remained nearly constant. The proximal-distal location of a fiber's femoral attachment had a stronger effect than had the anterior-posterior location. The tibial location had only a small statistically significant effect. These results suggest that the function of fibers within the PCL is determined primarily by their femoral attachment location. We determined all femoral attachments whose tibio-femoral distance changed 0, 2, 4, 6, 8, and 10 mm during flexion from 0 degrees to 90 degrees. No absolutely isometric point existed. Attachments whose separation distance changed less than 2 mm formed a bullet-shaped region whose base was against the roof of the intercondylar notch and whose nose pointed posteriorly and slightly distally. The axis of the "bullet" was near the proximal edge of the femoral insertion of the PCL. Along the axis, anterior attachments, located near the roof of the intercondylar notch, were more isometric than were posterior attachments, located near the cartilage. Attachments located distal to the axis moved away from the tibial insertion of the PCL when the knee was flexed. The more distal the femoral attachment, the larger the increase in tibiofemoral distance that occurred with flexion. The opposite was true of attachments proximal to the 2 mm region.


[Antibacterial therapy for acute cystitis in the age of growing pathogen resistance].
Acute cystitis refers to uncomplicated lower urinary tract infections, with the recurrence rates after the first cystitis episode being 50%. The basic treatment for the above diseases is antibacterial therapy, whose efficiency depends entirely on the right choice of a drug during initial empiric therapy. The paper gives the European Association of Urology guidelines and Russian guidelines, which are based on the results of both international (ARESC) and Russian (DARMIS) studies of urinary tract infection pathogens and their susceptibility to antibacterial drugs. Phosphomycin trometamol and furasidine potassium are the drugs of choice to treat acute cystitis in Russia now.


[High-level sports and anorexia nervosa].
Due to intensive physical training, and the pressure exerted by coaches, parents and the athlete him/herself during pre-competitive and competitive periods, a large number of high-level sporting activities can trigger or participate in anorexia nervosa. That said, some of these activities are more likely than others to promote this eating disorder. It is worth highlighting the sports most at risk for the onset or development of anorexia nervosa, and then addressing the means of prevention and regulation adapted to these circumstances.


A Qualitative Study of NIMBYism for Waste in Smaller Urban Areas of a Low-Income Country, Mzuzu, Malawi.
When waste management infrastructure is built, there can be resistance from the local affected populations, often termed the Not in My Backyard (NIMBY) phenomenon. This study aims to understand the forms of resistance that may develop in such contexts, focusing on 2 solid waste and 1 liquid waste management site within Mzuzu City, Malawi. At the newest solid waste site, community resistance had grown to the extent that the site was reportedly destroyed by the local community. Interviews and observations of the sites are complemented by examining historic and recent satellite images. It was found that, at the new solid waste site, community engagement had not been conducted effectively prior to construction and as part of ongoing site operations. This was compounded by poor site management and the non-delivery of the promised benefits to the community. In contrast, at the liquid waste site, the community could access untreated sludge for use as fertilizer and were happier to live within its vicinity. While NIMBYism is a frustrating phenomenon for city planners, it is understandable that communities want to protect their health and well-being when there is a history of mismanagement of waste sites which is sadly common in low-income settings. It is difficult for government agencies to deliver these services and broader waste management. In this study, an unsuccessful attempt to do something better with a legitimate goal is not necessarily a failure, but part of a natural learning process for getting things right.


Retarded eruption of maxillary second premolars associated with late development of the germs.
This study focused on five unusual cases with anomalous eruptions of the maxillary second premolars that related to late development of the germs. In four cases, the affected premolars erupted between the ages of 12y3m and 14y6m. In one case, which was suspected to include bilateral anomalies, the eruption of the premolar was excessively late, continuing up to 17y8m old. It was surmised that the degree of tooth formation in the contralateral side might help to predict approximately at what age the late premolars will erupt.


MALDI-TOF MS analysis of isoflavones in soy products.
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a new technique having a number of advantages for food analysis. This study is the first to demonstrate the use of MALDI-TOF MS to identify isoflavones in soy samples. 2',4',6'-Trihydroxyacetophenone (THAP) and 2,5-dihydroxybenzoic acid (DHB) were both good matrices for isoflavones, but DHB was chosen as the best because it worked well for sample extracts, with good spot-to-spot repeatability. Isoflavones were predominantly ionized in a protonated form with a very small amount of sodium or potassium adduct ions. Fragmentation occurred only through loss of glycosidic residues. Daidzin showed more than twice the response of genistin using MALDI-TOF MS. A simple solid phase extraction of isoflavones from soy samples was developed for MALDI-TOF MS analysis. MALDI-TOF MS can provide an isoflavone profile in 2 min and serves as a powerful tool to identify and study processing changes of isoflavones in soy products.


Postdecision Evidence Integration and Depressive Symptoms.
Background: Metacognition, or the ability to reflect on one's own thoughts, may be important in the development of depressive symptoms. Recent work has reported that depressive symptoms were associated with lower metacognitive bias (overall confidence) during perceptual decision making and a trend toward a positive association with metacognitive sensitivity (the ability to discriminate correct and incorrect decisions). Here, we extended this work, investigating whether confidence judgments are more malleable in individuals experiencing depressive symptoms. We hypothesized that depressive symptoms would be associated with greater adjustment of confidence in light of new evidence presented after a perceptual decision had been made. Methods: Participants (N = 416) were recruited via Amazon Mechanical Turk. Metacognitive confidence was assessed through two perceptual decision-making tasks. In both tasks, participants made a decision about which of two squares contained more dots. In the first task, participants rated their confidence immediately following the decision, whereas in the second task, participants observed new evidence (always in the same direction as initial evidence) before rating their confidence. Participants also completed questionnaires measuring depressive symptoms and self-esteem. Analysis: Metacognitive bias was calculated as overall mean confidence, whereas metacognitive sensitivity was calculated using meta-d' (a response-bias free measure of how closely confidence tracks task performance) in the first task. Postdecision evidence integration (PDEI) was defined as the change in confidence following postdecision evidence on the second task. Results: Participants with more depressive symptoms made greater confidence adjustments (i.e., greater PDEI) in light of new evidence (β = 0.119, p = 0.045), confirming our main hypothesis. We also observed that lower overall confidence was associated with greater depressive symptoms, although this narrowly missed statistical significance (β = -0.099, p = 0.056), and we did not find an association between metacognitive sensitivity (meta-d') and depressive symptoms. Notably, self-esteem was robustly associated with overall confidence (β = 0.203, p < 0.001), which remained significant when controlling for depressive symptoms. Conclusions: We found that individuals with depressive symptoms were more influenced by postdecisional evidence, adjusting their confidence more in light of new evidence. Individuals with low self-esteem were less confident about their initial decisions. This study should be replicated in a clinically depressed sample.


Challenges of Tobacco Control Program in Iran.
This study assessed the status of cigarette marketing in Iran as well as the attitude of smokers toward smoking and the policies and tobacco control programs in the country. This cross-sectional study was conducted on 3480 volunteer smokers in six provinces, using a stratified cluster random sampling method. The study population consisted of smokers who used at least one cigarette per day. The data collection tool was an anonymous self-administered questionnaire including basic information and 20 five-choice questions related to participants' attitude toward smoking habits (Cronbach's alpha; 79.73%.). The majority (66.9%) of participants started smoking at the age of 10 to 19 years and 61.1% used foreign cigarettes. Of 160 marketed brands, 38 (23.8%) were domestic and 122 (76.2%) were foreign, including 63 (39.3%) imported and 59 (36.9%) smuggled brands. Being tempted (25.0%), getting nervous (24.1%), and seeking euphoria (24.1%) were the most common reasons for restarting cigarette smoking after cessation. The majority of participants believed that smoking in public places was a violation of the rights of others and smoking should be avoided in such places. Smoking prevention programs should focus on adolescents as the most vulnerable age group. Raising the retail price of tobacco products through increasing taxes can reduce consumption, particularly among first starters and youth. However, increasing taxes and prices of tobacco products may be effective when simultaneous effective measures are implemented to eliminate all kinds of illicit trade in all forms of tobacco products.


Influence of maternal depressive symptoms on ratings of childhood behavior.
This study uses information collected on two occasions from a probability sample of families with 5- to 12-year-old children (N = 1151) participating in a general population study in 1983 and follow-up in 1987. It evaluated the importance of maternal bias in the assessment of child behavior by comparing the relative strengths of association between maternal depression and childhood behavior and between maternal depression and mother reporting errors. Conduct problems and hyperactivity were measured as latent criterion variables constructed from mother, teacher, and youth (aged 12 to 16 years) ratings and their associations with maternal depression were modeled using covariance structure analysis. The analyses revealed that maternal depression was associated significantly with conduct problems (phi = .35) and hyperactivity (phi = .38) among 5- to 7-years-olds in 1983 but not 4 years later in 1987. None of the associations between maternal depression and mother reporting errors were significant. Among 8- to 12-year-olds in 1983, maternal depression was associated significantly not only with conduct problems (phi = .17) and hyperactivity (phi = .15) but also with mother rating errors of these behaviors (psi = .13 and .17, respectively). Four years later in 1987 when this cohort was 12 to 16 years old, the only significant association was between maternal depression and conduct disorder (phi = .25). Although evidence exists for associations between maternal depressed mood and mother rating errors, there also appears to be a substantive association between maternal depression and childhood behavior.


Constitutive modeling of human femoropopliteal artery biaxial stiffening due to aging and diabetes.
Atherosclerotic obstructive disease of the femoropopliteal artery (Peripheral Arterial Disease, PAD) is notorious for high treatment failure rates. Older age and diabetes mellitus (DM) are among the major risk factors for PAD, and both are associated with increased arterial stiffness. Our goal was to develop a constitutive model describing multiaxial arterial stiffening, and use it to portray aging of normal and diabetic human femoropopliteal arteries (FPA). Fresh human FPAs (n=744) were obtained from 13-82-year-old donors. Arteries were tested using planar biaxial extension, and their behavior was modeled with a constitutive relation that included stiffening functions of age. FPA diameter, wall thickness, circumferential, and longitudinal opening angles increased with age, while longitudinal pre-stretch decreased. Diameter and circumferential opening angle did not change with age in subjects with DM. Younger FPAs were more compliant longitudinally but became more isotropic with age. Arteries with DM stiffened significantly faster in the circumferential direction than arteries without DM. Constitutive model accurately portrayed orthotropic stiffening with age of both normal and diabetic arteries. Constitutive description of FPA aging contributes to understanding of arterial pathophysiology and can help improve fidelity of computational models investigating device-artery interaction in PAD repair by providing more personalized arterial properties. We have analyzed n=744 human femoropopliteal artery (FPA) specimens using biaxial tensile testing to derive constitutive description of FPA aging in diabetic and non-diabetic subjects. The proposed model allows determination of FPA mechanical properties for subjects of any given age in the range of 13-82years. These results contribute to understanding of FPA pathophysiology and can help improve fidelity of computational models investigating device-artery interaction in peripheral arterial disease repair by providing more personalized arterial properties. In addition, they can guide the development of new materials tunable to diabetic and non-diabetic arteries.


The use of electrically stimulated skeletal muscle to pump blood.
Electrically stimulated skeletal muscle can be used in many ways to pump blood. The important physiological characteristics of skeletal muscle for use in a cardiac assist role are presented. The force developed by a twitch and tetanic contraction are quantitated and the considerations in the choice of the stimulus applied to the innervating motor nerve are discussed. The importance of stimulus frequency and duration of the stimulus train are presented in terms of the stroke volume produced by skeletal muscle contracting around a pouch. The effect that preload has on pouch stroke volume and muscle blood flow are demonstrated. Finally an example is given in which cardiac output is augmented by an untrained, electrically stimulated, canine rectus abdominis muscle wrapped around a pouch connected between the apex of the left ventricle and the aorta. In this study, the augmentation in cardiac output was measured when the skeletal muscle was contracted after each second, third, and fourth ventricular contraction. The augmentation was 46% +/- 4, 31% +/- 7, and 25% +/- 4, respectively, for the three contraction regimens. Although electrically stimulated skeletal muscle can be used as a power source to assist the failing heart, the optimum application is yet to be discovered. Important considerations in selecting the optimal application are preload, stimulus train duration, train rate, and muscle blood flow.


Clopidogrel treatment before coronary artery bypass graft surgery increases postoperative morbidity and blood product requirements.
An increasing number of patients are referred for coronary artery bypass graft surgery while treated with clopidogrel. This agent inhibits the platelet P2Y12 adenosine-5'-diphosphate (ADP) receptor, which results in an inhibition of platelet aggregation. The aim of this study was to determine the effect of preoperative clopidogrel treatment on postoperative bleeding, mortality, and morbidity in patients after coronary artery bypass graft surgery. Retrospective cohort study. University hospital (single institution). One hundred forty-four patients who underwent isolated coronary artery bypass graft surgery. Seventy-two patients who received clopidogrel during the preoperative period formed the study group. Seventy-two patients (matched based on age, sex, and preoperative risk profile) served as the control group. Clopidogrel-treated patients received significantly more platelet (4.4 +/- 5.7 v 1.3 +/- 3.2 U, p < 0.001) and red blood cell (5.1 +/- 4.2 v 2.6 +/- 2.6 U, p < 0.001) transfusions compared with the control group. All-cause mortality and morbidity were significantly higher in clopidogrel-treated patients (n = 7, 9% v n = 1, 1%; p = 0.031). In addition, the lengths of stay in the intensive care unit and the hospital were significantly longer in these patients (2.5 +/- 2.7 v 1.4 +/- 0.9 days, p = 0.002; 9.9 +/- 11 v 6 +/- 2.5 days, p = 0.003). Despite an increased morbidity in the clopidogrel group, the midterm survival was similar between the 2 groups (1-year and 5-year survival 97% +/- 2% and 95.7% +/- 3% v 100% +/- 0% and 87% +/- 10%, respectively; p = 0.885). Preoperative clopidogrel is associated with increased transfusion requirement after coronary artery bypass graft surgery. The present data suggest that all-cause mortality and major morbidity may also increase in these patients. In clopidogrel-treated patients, coronary artery bypass graft surgery should be delayed in the absence of specific medical indications as recommended by recent American Heart Association guidelines.


The Maturation of the Superior Collicular Map of Auditory Space in the Guinea Pig is Disrupted by Developmental Visual Deprivation.
In the normal guinea pig a map of auditory space appears, in the deeper layers of the superior colliculus, at 32 days after birth (DAB). The animal is unable to construct this collicular map of auditory space in the absence of developmental visual experience. Auditory receptive fields of animals dark-reared from birth are typically large, occupying most of the contralateral hemifield. There is no topographic relationship between the collicular location of the recording electrode and the spatial position from which auditory stimuli elicit a maximal response. The fields of dark-reared animals resemble, in their tuning parameters, the spatially undifferentiated fields typical of young postnatal normal guinea pigs. To investigate the time-course during which visual experience is required for map emergence, animals received normal visual experience until either 18 or 26 DAB and were then dark-reared until the terminal mapping experiment. Maps developed in neither group. Animals provided with a normal visual environment until 30 DAB, and then placed in the dark did, however, construct topographically organized spatial maps with discrete spatial receptive fields. Maps also failed to emerge in animals receiving normal visual experience both before and after a 4-day period of visual deprivation between 26 and 30 DAB. We conclude that this 4-day period, or part of it, constitutes a 'crucial' period during which visual experience is required for the normal elaboration of the collicular map of auditory space.


Skeletal myogenic progenitors originating from embryonic dorsal aorta coexpress endothelial and myogenic markers and contribute to postnatal muscle growth and regeneration.
Skeletal muscle in vertebrates is derived from somites, epithelial structures of the paraxial mesoderm, yet many unrelated reports describe the occasional appearance of myogenic cells from tissues of nonsomite origin, suggesting either transdifferentiation or the persistence of a multipotent progenitor. Here, we show that clonable skeletal myogenic cells are present in the embryonic dorsal aorta of mouse embryos. This finding is based on a detailed clonal analysis of different tissue anlagen at various developmental stages. In vitro, these myogenic cells show the same morphology as satellite cells derived from adult skeletal muscle, and express a number of myogenic and endothelial markers. Surprisingly, the latter are also expressed by adult satellite cells. Furthermore, it is possible to clone myogenic cells from limbs of mutant c-Met-/- embryos, which lack appendicular muscles, but have a normal vascular system. Upon transplantation, aorta-derived myogenic cells participate in postnatal muscle growth and regeneration, and fuse with resident satellite cells.The potential of the vascular system to generate skeletal muscle cells may explain observations of nonsomite skeletal myogenesis and raises the possibility that a subset of satellite cells may derive from the vascular system.


A multilevel analysis of effect of neighbourhood and individual wealth status on sexual behaviour among women: evidence from Nigeria 2003 Demographic and Health Survey.
Nigeria is home to more people living with HIV than any other country in the world, except South Africa and India - where an estimated 2.9 million [1.7 million - 4.2 million] people were living with the virus in 2005. Women bear the greatest burden of frequent high-risk pregnancies, raising large families, and increasingly, the AIDS epidemic. Thus, there is a need for better understanding of the determinants of high risk sexual behaviour among women. In this study, we examined factors associated with extra-marital sex among women in Nigeria and investigated how much variation in reported extra-marital sex can be attributed to individual-, and community-level factors. We analyzed data from 6362 sexually active women aged 15 - 49 years who participated in the Nigeria 2003 Demographic and Health Survey using multilevel logistic regression models. Results are presented as odds ratio with 95% confidence interval. Independent of other factors, compared to women aged 15-24 years, those 25 - 34 years (odds ratio [OR] 0.59; 95% CI: 0.44 - 0.79) and 35 years or older (OR 0.36; 95% CI: 0.24 - 0.54) were less likely to have reported multiple concurrent sex partners in the last 12 years. As expected, women currently or formerly married were less likely to have reported multiple concurrent sex partners than women never married. Women who drank alcohol in the last three months were more likely to have reported multiple concurrent sex partners. Compared to women from richest household, women from poorest and middle household were 83% and 51% more likely to multiple concurrent sex partners in the last 12 month respectively. After individual compositional and contextual factors, community wealth status was statistically significant with sexual behaviour. The study has demonstrated that individual and community wealth status are independent predictors of women's sexual behaviour, and that there is significant neighbourhood variation in odds of multiple concurrent sex partners, even after controlling for effects of both individual- and community-level characteristics. Scholars trying to understand variation individual high risk sexual behaviour should pay attention to the characteristics of both individuals and places of residence.


Effects of dronedarone started rapidly after amiodarone discontinuation.
Multiple studies have shown that amiodarone is effective in treating atrial fibrillation (AF), but is associated with a relatively high incidence of side effects; however, due to amiodarone's long elimination half-life (20-100 days), physicians may hesitate to start other drugs until it has fully cleared. A rapid switch from amiodarone to dronedarone is feasible. EURIDIS and ADONIS were double-blind, multinational, parallel-group trials comparing the efficacy and safety of dronedarone with placebo over 12 months. This retrospective subanalysis of EURIDIS/ADONIS compared the effects of dronedarone in patients discontinuing amiodarone within 2 days before randomization ("rapid switch") with results in patients who had received no amiodarone during the 2 months preceding randomization. In total, 1237 patients were enrolled ("rapid switch", n = 154; "no amiodarone", n = 1014). In both the "rapid switch" and the "no amiodarone" groups, dronedarone users had significantly lower AF recurrence than patients receiving placebo (HR = 0.64, 95% CI, 0.44-0.95; P = 0.0224 and HR = 0.79, 95% CI, 0.67-0.92; P = 0.0027, respectively). Dronedarone users had a higher incidence of bradyarrhythmic events than placebo-treated patients. A "rapid switch" from amiodarone to dronedarone was associated with a higher incidence of serious heart failure events and heart failure hospitalizations versus all other groups. Overall event rates were low and there was no significant difference in total adverse event rates or deaths between groups. In this patient population, a switch from amiodarone to dronedarone within a 2-day time frame might be feasible in certain patient categories, but further investigation is warranted.


Malignant glioma-induced neuronal cell death in an organotypic glioma invasion model. Technical note.
Rapid growth and diffuse brain infiltration are hallmarks of malignant gliomas. The underlying molecular pathomechanisms of these tumors, however, remain to be determined. The authors present a novel glioma invasion model that allows researchers to monitor consecutively tumor cell proliferation and migration in an organotypic brain environment. Enhanced green fluorescent protein-labeled F98 rat glioma cells were implanted into slice cultures obtained from a rat hippocampus, and tumor growth was microscopically documented up to 20 days in vitro. Invasion along radially oriented migratory streams could be observed 5 days after implantation of rat F98, human U87MG, and mouse GL261 glioma cells, whereas human Be(2)c neuroblastoma cells and mouse HT22 hippocampal neurons failed to invade the brain parenchyma. Following implantation of F98 glioma cells into the entorhinal cortex, cell death was observed within the infiltrated brain parenchyma as well as in the neuroanatomically connected dentate gyrus. Application of the N-methyl-D-aspartate receptor antagonist MK801 to the culture medium significantly reduced neuronal degeneration in the dentate gyrus, whereas the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor antagonist GYKI 52466 inhibited peritumoral cytotoxicity. This new model allows researchers to address in a systematic manner the molecular pathways of brain invasion as well as specific tumor-host interactions such as necrosis.


LRRK2 G2019S mutations are associated with an increased cancer risk in Parkinson disease.
Leucine rich repeat kinase (LRRK2) G2019S mutations are presumed to cause PD through a toxic gain of function of the protein kinase. Small molecule kinase inhibitors have been developed for the treatment of certain cancers, and some antioncogenic agents such as sunitinib, may nonspecifically inhibit LRRK2. Few studies, however, have assessed cancer risk in LRRK2 mutation carriers. To explore this risk, we evaluated records of Ashkenazi Jewish (AJ) PD patients participating in genetic research. Charts were reviewed for 163 unrelated AJ PD patients, 31 of whom harbored the G2019S mutation. History of cancer was queried at baseline intake using a form reviewing medical conditions, and charts were reviewed for all follow-up visits. 9/31 LRRK2 G2019S mutation carriers had nonskin cancers, whereas 15/132 without mutations had nonskin cancers, representing an almost threefold increased risk in this group (HR 2.9, 95% CI 1.3-6.6). Age at first nonskin cancer was younger in the LRRK2 carriers (56.0 years) than the noncarriers (62.0 years), but was not significant. 67% of the LRRK2 carriers had their cancer before the onset of PD, whereas only 40% of noncarriers developed their first nonskin cancer before onset of PD. While further evaluation is warranted, our findings indicate an increased risk of nonskin cancers in LRRK2 G2019S mutation carriers, which may be related to toxic gain of function of mutated LRRK2.


Polycomb target genes are silenced in multiple myeloma.
Multiple myeloma (MM) is a genetically heterogeneous disease, which to date remains fatal. Finding a common mechanism for initiation and progression of MM continues to be challenging. By means of integrative genomics, we identified an underexpressed gene signature in MM patient cells compared to normal counterpart plasma cells. This profile was enriched for previously defined H3K27-tri-methylated genes, targets of the Polycomb group (PcG) proteins in human embryonic fibroblasts. Additionally, the silenced gene signature was more pronounced in ISS stage III MM compared to stage I and II. Using chromatin immunoprecipitation (ChIP) assay on purified CD138+ cells from four MM patients and on two MM cell lines, we found enrichment of H3K27me3 at genes selected from the profile. As the data implied that the Polycomb-targeted gene profile would be highly relevant for pharmacological treatment of MM, we used two compounds to chemically revert the H3K27-tri-methylation mediated gene silencing. The S-adenosylhomocysteine hydrolase inhibitor 3-Deazaneplanocin (DZNep) and the histone deacetylase inhibitor LBH589 (Panobinostat), reactivated the expression of genes repressed by H3K27me3, depleted cells from the PRC2 component EZH2 and induced apoptosis in human MM cell lines. In the immunocompetent 5T33MM in vivo model for MM, treatment with LBH589 resulted in gene upregulation, reduced tumor load and increased overall survival. Taken together, our results reveal a common gene signature in MM, mediated by gene silencing via the Polycomb repressor complex. The importance of the underexpressed gene profile in MM tumor initiation and progression should be subjected to further studies.


From Gut to Heart: Havoc in a Young Patient with Typhoid-associated Cardiomyopathy.
Typhoid is an endemic hassle, especially in third-world countries like Pakistan. It is an enteric fever characterized by systemic manifestations that include high temperature and abdominal pain. If not properly treated, at times, it can transgress into complications predominantly involving the gut, where the site of pathology lies. Sometimes, however, it can involve other organ systems and pose diagnostic challenges owing to unfamiliar modes of presentation. Here in, we present a case of a 14-year-old male, previously afflicted and treated for typhoid who presented to the medical consult service with abdominal pain, high-grade fever, and mild chest discomfort. His hemodynamic parameters deteriorated within weeks as he developed pulmonary edema and hypoxemia. He was later diagnosed with echocardiography which earlier on, showed signs of acute myocarditis and eventually dilated cardiomyopathy. The patient was treated with antibiotics coupled with supportive and intensive care which yielded relief in his symptoms. He was later followed up with serial echocardiograms and showed improvement in the cardiac parameters.


The Human G Protein-Coupled ATP Receptor P2Y11 Is Associated With IL-10 Driven Macrophage Differentiation.
The G protein-coupled P2Y11 receptor is known to sense extracellular ATP during inflammatory and immune responses. The dinucleotide NAD+ has also been proposed to be a P2Y11 receptor ligand but its role is less clear. Here, we have examined for the first time human P2Y11 receptor protein levels and show that the receptor was upregulated during polarization of M2 macrophages. IL-10 reinforced P2Y11 receptor expression during differentiation of M2c macrophages expressing CD163, CD16, and CD274 (PD-L1). Nutlin-3a mediated p53 stabilization further increased P2Y11 receptor, CD16, and PD-L1 expression. AMP-activated kinase (AMPK), which mediates anti-inflammatory effects of IL-10, and nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme of the NAD+ salvage pathway, which is under the control of AMPK, were also required for P2Y11 receptor expression. The P2Y11 receptor agonist ATPγS and NAD+ could independently stimulate the production of IL-8 in M2 macrophages, however, only the ATPγS-induced response was mediated by P2Y11 receptor. Both in a recombinant system and in macrophages, P2Y11 receptor-driven IL-8 production predominantly depended on IkB kinase (IKK), and extracellular signal-regulated kinase (ERK). In conclusion, our data indicate that an AMPK-NAMPT-NAD+ signaling axis promotes P2Y11 receptor expression during M2 polarization of human macrophages in response to IL-10. PD-L1 expressing M2c macrophages that secrete the cancer-promoting chemokine IL-8 in response to P2Y11 receptor stimulation may represent an important target in checkpoint blockade immunotherapy.


Active smoking may negatively affect response rate, progression-free survival, and overall survival of patients with metastatic renal cell carcinoma treated with sunitinib.
Obesity, smoking, hypertension, and diabetes are risk factors for renal cell carcinoma development. Their presence has been associated with a worse outcome in various cancers. We sought to determine their association with outcome of sunitinib treatment in metastatic renal cell carcinoma (mRCC). An international multicenter retrospective study of sunitinib-treated mRCC patients was performed. Multivariate analyses were performed to determine the association between outcome and the pretreatment status of smoking, body mass index, hypertension, diabetes, and other known prognostic factors. Between 2004 and 2013, 278 mRCC patients were treated with sunitinib: 59 were active smokers, 67 were obese, 73 were diabetic, and 165 had pretreatment hypertension. Median progression-free survival (PFS) was 9 months, and overall survival (OS) was 22 months. Factors associated with PFS were smoking status (past and active smokers: hazard ratio [HR]: 1.17, p = .39; never smokers: HR: 2.94, p < .0001), non-clear cell histology (HR: 1.62, p = .011), pretreatment neutrophil-to-lymphocyte ratio >3 (HR: 3.51, p < .0001), use of angiotensin system inhibitors (HR: 0.63, p = .01), sunitinib dose reduction or treatment interruption (HR: 0.72, p = .045), and Heng risk (good and intermediate risk: HR: 1.07, p = .77; poor risk: HR: 1.87, p = .046). Factors associated with OS were smoking status (past and active smokers: HR: 1.25, p = .29; never smokers: HR: 2.7, p < .0001), pretreatment neutrophil-to-lymphocyte ratio >3 (HR: 2.95, p < .0001), and sunitinib-induced hypertension (HR: 0.57, p = .002). Active smoking may negatively affect the PFS and OS of sunitinib-treated mRCC. Clinicians should consider advising patients to quit smoking at initiation of sunitinib treatment for mRCC.


Multiple functions of Osterix are required for bone growth and homeostasis in postnatal mice.
The transcription factor Osterix (Osx) is required for osteoblast differentiation and bone formation during embryonic development, but it is not known whether Osx has an essential function in postnatal bone growth and in bone homeostasis. Conditional deletion of Osx at several time points postnatally revealed that Osx was essential for osteoblast differentiation and new bone formation in growing and adult bones. Additionally, inactivation of Osx in bones severely disrupted the maturation, morphology, and function of osteocytes. These findings identify Osx as having an essential role in the cell-specific genetic program of osteocytes. Interestingly, Osx inactivation also led to the massive accumulation of unresorbed calcified cartilage in a large area below the growth plate of endochondral bones. This specific area was also marked by an unanticipated almost complete lack of bone marrow cells and a marked decrease in the density and size of osteoclasts. This diminished density of osteoclasts could contribute to the lack of resorption of mineralized cartilage. In addition, we speculate that the abnormally accumulated, mainly naked cartilage represents an unfavorable substrate for osteoclasts. Our study identifies Osx as an essential multifunctional player in postnatal bone growth and homeostasis.


Detection of mutant K-ras DNA in plasma or serum of patients with colorectal cancer.
Increased understanding of the molecular basis of colorectal cancer and recognition that extracellular DNA circulates in the plasma and serum of cancer patients enables new approaches to detection and monitoring. We used a polymerase chain reaction (PCR) assay to demonstrate mutant K-ras DNA in the plasma or serum of patients with colorectal cancer. Plasma or serum was fractionated from the blood of 31 patients with metastatic or unresected colorectal cancer and from 28 normal volunteers. DNA was extracted using either a sodium chloride or a gelatin precipitation method and then amplified in a two-stage PCR assay using selective restriction enzyme digestion to enrich for mutant K-ras DNA. Mutant K-ras DNA was detected in the plasma or serum of 12 (39%) patients, all confirmed by sequencing, but was not detected in any of the normal volunteers. K-ras mutations were detected in plasma or serum regardless of sex, primary tumour location, principal site of metastasis or proximity of chemotherapy and surgery to blood sampling. Tumour specimens available for 19 of the patients were additionally assayed for ras mutations and compared with blood specimens. Our results indicate mutant K-ras DNA is readily detectable by PCR in the plasma or serum of patients with advanced colorectal cancer. Thus, plasma- or serum-based nucleic acid amplification assays may provide a valuable method of monitoring and potentially detecting colorectal cancer.


Adapting and enhancing PAX Good Behavior Game for First Nations communities: a mixed-methods study protocol developed with Swampy Cree Tribal Council communities in Manitoba.
High rates of mental health problems, such as suicidal behaviours, among First Nations youth in Canada are a major public health concern. The Good Behavior Game (GBG) is a school-based intervention that provides a nurturing environment for children and has been shown to promote positive outcomes. PAX Good Behavior Game (PAX GBG) is an adaptation and enhancement of the GBG. While PAX GBG has been implemented in Indigenous communities, little research exists examining the cultural and contextual appropriateness and effectiveness of the intervention in First Nations communities. The present paper describes a protocol of the mixed-methods approach guided by an Indigenous ethical engagement model adopted to implement, adapt and evaluate PAX GBG in First Nations communities in Manitoba, Canada. First, implementation outcomes (eg, acceptability, adoption) of PAX GBG will be evaluated using qualitative interviews with teachers, principals and community members from Swampy Cree Tribal Council (SCTC) communities. Second, by linking administrative databases to programme data from schools in 38 First Nations communities, we will compare PAX GBG and control groups to evaluate whether PAX GBG is associated with improved mental health and academic outcomes. Third, the qualitative results will help inform a cultural and contextual adaptation of PAX GBG called First Nations PAX (FN PAX). Fourth, FN PAX will be implemented in a few SCTC communities and evaluated using surveys and qualitative interviews followed by the remaining communities the subsequent year. Ethical approval was obtained from the University of Manitoba Health Research Ethics Board and will be obtained from the Health Information Privacy Committee and respective data providers for the administrative database linkages. Dissemination and knowledge translation will include community and stakeholder engagement throughout the research process, reports and presentations for policymakers and community members, presentations at scientific conferences and journal publications.


Fingerprint Database Reconstruction Based on Robust PCA for Indoor Localization.
The indoor localization method based on the Received Signal Strength (RSS) fingerprint is widely used for its high positioning accuracy and low cost. However, the propagation behavior of radio signals in an indoor environment is complicated and always leads to the existence of outliers and noises that deviate from a normal RSS value in the database. The fingerprint database containing outliers and noises will severely degrade the performance of an indoor localization system. In this paper, an approach to reconstruct the fingerprint database is proposed with the purpose of mitigating the influences of outliers. More specifically, by exploiting the spatial and temporal correlations of RSS data, the database can be transformed into a low-rank matrix. Therefore, the RPCA (Robust Principle Component Analysis) technique can be applied to recover the low-rank matrix from a noisy matrix. In addition, we propose an improved RPCA model which takes advantage of the prior knowledge of a singular value and could remove outliers and structured noise simultaneously. The experimental results show that the proposed method can eliminate outliers and structured noise efficiently.


[Speech perception learning in patients with cochlear implants].
A model of speech apprehensibility of postlingual patients after cochlear implantation has been developed. The study of filtrated speech comprehension by persons with normal hearing has used speech material in three 50 Hz spectral bands. The central frequencies were selected by tonotopics of the basilar cochlear membrane. Characteristics of speech comprehension and apprehensibility by recipients of cochlear implants are demonstrated. The model reveals problems of acoustic comprehension facing the implanted patients and shows importance of learning and personality features of the implanted patients for early rehabilitation.


Salt-inducible kinase 3, SIK3, is a new gene associated with hearing.
Hearing function is known to be heritable, but few significant and reproducible associations of genetic variants have been identified to date in the adult population. In this study, genome-wide association results of hearing function from the G-EAR consortium and TwinsUK were used for meta-analysis. Hearing ability in eight population samples of Northern and Southern European ancestry (n = 4591) and the Silk Road (n = 348) was measured using pure-tone audiometry and summarized using principal component (PC) analysis. Genome-wide association analyses for PC1-3 were conducted separately in each sample assuming an additive model adjusted for age, sex and relatedness of subjects. Meta-analysis was performed using 2.3 million single-nucleotide polymorphisms (SNPs) tested against each of the three PCs of hearing ability in 4939 individuals. A single SNP lying in intron 6 of the salt-inducible kinase 3 (SIK3) gene was found to be associated with hearing PC2 (P = 3.7×10(-8)) and further supported by whole-genome sequence in a subset. To determine the relevance of this gene in the ear, expression of the Sik3 protein was studied in mouse cochlea of different ages. Sik3 was expressed in murine hair cells during early development and in cells of the spiral ganglion during early development and adulthood. Our results suggest a developmental role of Sik3 in hearing and may be required for the maintenance of adult auditory function.


Differentially expressed platelet activation-related genes in dogs with stage B2 myxomatous mitral valve disease.
Peripheral blood carries a reservoir of mRNAs that regulate cardiac structure and function potential. Although it is well recognized that the typical symptoms of Myxomatous Mitral Valve Disease (MMVD) stage B2 are long-standing hemodynamic disorder and cardiac structure remodeling caused by mitral regurgitation, the transcriptomic alterations in blood from such dogs are not understood. In the present study, comparative high-throughput transcriptomic profiling of blood was performed from normal control (NC) and naturally-occurring MMVD stage B2 (MMVD) dogs. Using Weighted Gene Co-expression Network Analyses (WGCNA), Gene Ontology (GO), and Kyoto Encyclopedia of Gene and Genomes (KEGG), we identified that the turquoise module was the most highly correlated with echocardiographic features and found 64 differentially expressed genes (DEGs) that were significantly enriched in platelet activation related pathways. Therefore, from the turquoise module, we selected five DEGs (MDM2, ROCK1, RIPK1, SNAP23, and ARHGAP35) that, according to real-time qPCR, exhibited significant enrichment in platelet activation related pathways for validation. The results showed that the blood transcriptional abundance of MDM2, ROCK1, RIPK1, and SNAP23 differed significantly (P < 0.01) between NC and MMVD dogs. On the other hand, Correlation Analysis revealed that MDM2, ROCK1, RIPK1, and SNAP23 genes negatively regulated the heart structure parameters, and followed the same trend as observed in WGCNA. We screened four platelet activation related genes, MDM2, ROCK1, RIPK1, and SNAP23, which may be considered as the candidate biomarkers for the diagnosis of MMVD stage B2. These findings provided new insights into MMVD pathogenesis.


Computerized Adaptive Tests: Efficient and Precise Assessment of the Patient-Centered Impact of Diabetic Retinopathy.
Evaluate efficiency, precision, and validity of RetCAT, which comprises ten diabetic retinopathy (DR) quality of life (QoL) computerized adaptive tests (CATs). In this cross-sectional clinical study, 183 English and/or Mandarin-speaking participants with DR (mean age ± standard deviation [SD] 56.4 ± 11.9 years; 38% proliferative DR [worse eye]) were recruited from retinal clinics in Singapore. Participants answered the RetCAT tests (Symptoms, Activity Limitation, Mobility, Emotional, Health Concerns, Social, Convenience, Economic, Driving, and Lighting), which were capped at seven items each, and other questionnaires, and underwent eye tests. Our primary evaluation focused on RetCAT efficiency (i.e. standard error of measurement [SEM] ± SD achieved and time needed to complete each CAT). Secondary evaluations included an assessment of RetCAT's test precision and validity. Mean SEM across all RetCAT tests was 0.351, ranging from 0.272 ± 0.130 for Economic to 0.484 ± 0.130 for Emotional. Four tests (Mobility, Social, Convenience, and Driving) had a high level of measurement error. The median time to take each RetCAT test was 1.79 minutes, ranging from 1.12 (IQR [interquartile range] 1.63) for Driving to 3.28 (IQR 2.52) for Activity Limitation. Test precision was highest for participants at the most impaired end of the spectrum. Most RetCAT tests displayed expected correlations with other scales (convergent/divergent validity) and were sensitive to DR and/or vision impairment severity levels (criterion validity). RetCAT can provide efficient, precise, and valid measurement of DR-related QoL impact. Future application of RetCAT will employ a stopping rule based on SE rather than number of items to ensure that all tests can detect meaningful differences in person abilities. Responsiveness of RetCAT to treatment interventions must also be determined. RetCAT may be useful for measuring the patient-centered impact of DR severity and disease progression and evaluating the effectiveness of new therapies.


Targeting Mitochondria for Treatment of Chemoresistant Ovarian Cancer.
Ovarian cancer is the leading cause of death from gynecologic malignancy in the Western world. This is due, in part, to the fact that despite standard treatment of surgery and platinum/paclitaxel most patients recur with ultimately chemoresistant disease. Ovarian cancer is a unique form of solid tumor that develops, metastasizes and recurs in the same space, the abdominal cavity, which becomes a unique microenvironment characterized by ascites, hypoxia and low glucose levels. It is under these conditions that cancer cells adapt and switch to mitochondrial respiration, which becomes crucial to their survival, and therefore an ideal metabolic target for chemoresistant ovarian cancer. Importantly, independent of microenvironmental factors, mitochondria spatial redistribution has been associated to both tumor metastasis and chemoresistance in ovarian cancer while specific sets of genetic mutations have been shown to cause aberrant dependence on mitochondrial pathways in the most aggressive ovarian cancer subtypes. In this review we summarize on targeting mitochondria for treatment of chemoresistant ovarian cancer and current state of understanding of the role of mitochondria respiration in ovarian cancer. We feel this is an important and timely topic given that ovarian cancer remains the deadliest of the gynecological diseases, and that the mitochondrial pathway has recently emerged as critical in sustaining solid tumor progression.


Oligosaccharides and glycoconjugates in bovine milk and colostrum.
Oligosaccharides and glycoconjugates are some of the most important bioactive components in milk. A great deal of information is available on the biological function of the components from human milk. Their primary role seems to be in providing protection against pathogens by acting as competitive inhibitors for the binding sites on the epithelial surfaces of the intestine. Evidence is also available to support the role of some of these components as growth promoters for genera of beneficial microflora in the colon. Compared with human milk, levels of oligosaccharides in bovine milk are very low. Nevertheless, a number of neutral and acidic oligosaccharides have been isolated from bovine milk and characterised. The highest concentration of these molecules is found in early postparturition milk (colostrum). The chemical structure of the oligosaccharides and many of the glycoconjugates from bovine milk are similar to those in human milk. It is likely that bovine oligosaccharides and glycoconjugates can be used in milk products as bioactive components in human nutrition.


Pes anserinus transfer for the unstable knee. A 7-year follow-up of 43 cases of ruptured anterior cruciate ligament.
On average 7 years postoperatively, we evaluated 43 consecutive patients who had had pesoplasty for chronic injury of the anterior cruciate ligament. Fourteen patients had undergone further surgery during the follow-up period; ten of these had extra- or intra-articular stabilizing procedures, and eight meniscectomies were performed after the index operation. At follow-up of the 33 knees that had not had additional stabilizing surgery, only one-third of the knees had normal radiograms, and the majority had anterolateral rotational instability (pivot shift) which, however, did not exclude satisfactory knee function. We conclude that improvement after transfer of the pes anserinus tendons was due to the simultaneous meniscectomy; we do not believe that the pesoplasty changed the natural course of the chronic rupture of the anterior cruciate ligament.


WNT16B is a new marker of cellular senescence that regulates p53 activity and the phosphoinositide 3-kinase/AKT pathway.
Senescence is a tumor suppression mechanism that is induced by several stimuli, including oncogenic signaling and telomere shortening, and controlled by the p53/p21(WAF1) signaling pathway. Recently, a critical role for secreted factors has emerged, suggesting that extracellular signals are necessary for the onset and maintenance of senescence. Conversely, factors secreted by senescent cells may promote tumor growth. By using expression profiling techniques, we searched for secreted factors that were overexpressed in fibroblasts undergoing replicative senescence. We identified WNT16B, a member of the WNT family of secreted proteins. We found that WNT16B is overexpressed in cells undergoing stress-induced premature senescence and oncogene-induced senescence in both MRC5 cell line and the in vivo murine model of K-Ras(V12)-induced senescence. By small interfering RNA experiments, we observed that both p53 and WNT16B are necessary for the onset of replicative senescence. WNT16B expression is required for the full transcriptional activation of p21(WAF1). Moreover, WNT16B regulates activation of the phosphoinositide 3-kinase (PI3K)/AKT pathway. Overall, we identified WNT16B as a new marker of senescence that regulates p53 activity and the PI3K/AKT pathway and is necessary for the onset of replicative senescence.


Laparoscopic sleeve gastrectomy for morbid obesity.
The incidence of obesity is steadily rising, and it has been estimated that 40% of the US population will be obese by the year 2025 if the current trend continues. In recent years there has been renewed interest in the surgical treatment of morbid obesity in concomitance with the epidemic of obesity. Bariatric surgery proved effective in providing weight loss of large magnitude, correction of comorbidities and excellent short-term and long-term outcomes, decreasing overall mortality and providing a marked survival advantage. The Laparoscopic Sleeve Gastrectomy (LSG) has increased in popularity and is currently very "trendy" among laparoscopic surgeons involved in bariatric surgery. As LSG proved to be effective in achieving considerable weight loss in the short-term, it has been proposed by some as a sole bariatric procedure. This editorial focuses on the particular advantages of LSG in the treatment of morbid obesity.


TGF-β1 increases sialidase 3 expression in human lung epithelial cells by decreasing its degradation and upregulating its translation.
Purpose: We previously found extensive desialylation of glycoconjugates and upregulation of the sialidase enzyme NEU3 in fibrotic lesions in human and mouse lungs. However, studies using microarray analysis of whole lung tissue mRNA and single cell RNA-seq found no significant difference in levels of NEU3 mRNA between IPF patients and controls. This study aimed to elucidate how NEU3 was upregulated in fibrotic lungs.Materials and methods: Transforming growth factor-β1 (TGF-β1), a key driver of fibrosis, was added to A549 human alveolar basal epithelial adenocarcinoma cells and human small airway epithelial cells (HSAEpC). NEU3 expression in A549 cells and HSAEpC was detected by immunofluorescence staining. NEU3 translation and degradation were assessed by polysome profiling (polysomes efficiently translate mRNAs; monosomes poorly translate mRNAs) and cycloheximide chase after treating cells with or without TGF-β1 for 48 h.Results: TGF-β1 increased NEU3 expression and secretion in A549 cells and HSAEpC but did not change total (nuclear + cytosolic) NEU3 mRNA levels. TGF-β1 decreased the degradation rate of NEU3 in A549 cells. TGF-β1 decreased NEU3 mRNA levels in monosomes and increased NEU3 mRNA level in polysomes.Conclusion: TGF-β1 upregulates levels of NEU3 in epithelial cells by both decreasing NEU3 degradation and by increasing the translation of NEU3 mRNA, explaining the apparent paradox of high levels of NEU3 protein in pulmonary fibrosis without a concomitant increase in the expression of NEU3 mRNA.


Predictors of Number of Healthcare Professionals Consulted by Individuals with Mental Disorders or High Psychological Distress.
This study assesses the contribution of predisposing, enabling, and needs factors and related variables that predicted the number of healthcare professionals consulted for mental health reasons among 746 individuals with mental disorders and high psychological distress. The data were drawn from the third (T3) and fourth data collection periods (T4) of a longitudinal study conducted in a Quebec/Canada epidemiological catchment area. Hierarchical linear regression was performed on the number of types of healthcare professionals consulted in the 12 months prior to T4. Predictors were identified at T3, classified as predisposing, enabling, and needs factors (i.e., clinical and related variables) according to the Andersen Behavioral Model. Three needs factors were associated with the number of types of healthcare professionals consulted: Post-traumatic stress disorder, stressful events, and marginally suicide ideation. Three enabling factors: Having a family physician, previous use of mental health services, and employment status were also related to the dependent variable. Poor self-perception of mental health status was the only predisposing factor retained. While needs factors were the main predictors of the number of types of healthcare professionals consulted, enabling factors may reduce the influence of needs factors, by the deployment of various strategies that facilitate continuous and appropriate care.


Physical and technical demands of Australian football: an analysis of maximum ball in play periods.
This study compares ball in play (BiP) analyses and both whole game (WG) and quarter averaged data for physical and technical demands of sub-elite Australian football (AF) players competing in the West Australian Football League across playing positions. Microsensor data were collected from 33 male AF players in one club over 19 games of the 2019 season. BiP time periods and technical performance data (e.g., kicks) were acquired from the Champion Data timeline of statistics, and time matched to the microsensor data. Linear mixed modelling was utilised to establish differences between maximum BiP periods and averaged data. The analyses indicated significant differences (p < 0.0001) between maximum BiP and WG data for all metrics and all playing position (half-line, key position, and midfielders). The percentage difference was greatest for very high-speed running (171-178%), accelerations (136-142%), high-intensity efforts (128-139%), and high-speed running (134-147%) compared to PlayerLoad™ (50-56%) and total running distance (56-59%). No significant (p > 0.05) differences were evident for maximum BiP periods when they were compared between playing positions (i.e., half line vs key position vs midfield). Significant (p < 0.0001) differences were also noted between maximum BiP phases and averaged data across all 4 quarters, for each microsensor metric, and all playing positions. Technical actions (e.g., kicks and handballs) were observed in 21-48% of maximum BiP phases, depending on playing positions and microsensor metric assessed, with kicks and handballs constituting > 50% of all actions performed. These results show the BiP analysis method provides a more accurate assessment of the physical demands and technical actions performed by AF players, which are underestimated when using averaged data. The data presented in this study may be used to inform the design and monitoring of representative practice, ensuring that athletes are prepared for both the physical and technical demands of the most demanding passages of play.


A putative ATP/GTP binding protein affects Leishmania mexicana growth in insect vectors and vertebrate hosts.
Leishmania virulence factors responsible for the complicated epidemiology of the various leishmaniases remain mainly unidentified. This study is a characterization of a gene previously identified as upregulated in two of three overlapping datasets containing putative factors important for Leishmania's ability to establish mammalian intracellular infection and to colonize the gut of an insect vector. The investigated gene encodes ATP/GTP binding motif-containing protein related to Leishmania development 1 (ALD1), a cytosolic protein that contains a cryptic ATP/GTP binding P-loop. We compared differentiation, growth rates, and infective abilities of wild-type and ALD1 null mutant cell lines of L. mexicana. Loss of ALD1 results in retarded growth kinetics but not defects in differentiation in axenic culture. Similarly, when mice and the sand fly vector were infected with the ALD1 null mutant, the primary difference in infection and colonization phenotype relative to wild type was an inability to achieve maximal host pathogenicity. While ability of the ALD1 null mutant cells to infect macrophages in vitro was not affected, replication within macrophages was clearly curtailed. L. mexicana ALD1, encoding a protein with no assigned functional domains or motifs, was identified utilizing multiple comparative analyses with the related and often experimentally overlooked monoxenous flagellates. We found that it plays a role in Leishmania infection and colonization in vitro and in vivo. Results suggest that ALD1 functions in L. mexicana's general metabolic network, rather than function in specific aspect of virulence as anticipated from the compared datasets. This result validates our comparative genomics approach for finding relevant factors, yet highlights the importance of quality laboratory-based analysis of genes tagged by these methods.


Potential Use of Senolytics for Pharmacological Targeting of Precancerous Lesions.
Senescence is a cell state that contributes to several homeostatic and pathologic processes. In addition to being induced in somatic cells in response to replicative exhaustion (replicative senescence) as part of organismal aging, senescence can also be triggered prematurely by oncogene hyperactivation or tumor suppressor dysfunction [oncogene-induced senescence (OIS)]. Consequently, senescent cells comprise a major component of precancerous lesions of skin, oral mucosa, nasopharynx, prostate, gut, and lung. Unfortunately, invasive (or minimally invasive) interventions are currently the only available approach employed to eradicate premalignant lesions that carry the potential for cancer progression. Senolytics are a newly emerging drug class capable of selectively eliminating senescent cells. Although senolytics have been successfully demonstrated to mitigate a myriad of aging-related pathologies and to cull senescent cancer cells, there is a paucity of evidence for the potential use of senolytics as a novel approach to eliminate oncogene-induced senescent cells. This Emerging Concepts commentary will 1) summarize evidence in established models of OIS including B-Raf-induced nevi, transgenic lung cancer, and pancreatic adenocarcinoma models, as well as evidence from clinical precancerous lesions; 2) suggest that OIS is targetable; and 3) propose the utilization of senolytic agents as a revolutionary means to interfere with the ability of senescent premalignant cells to progress to cancer in vitro and in vivo If proven to be effective, senolytics will represent an emerging tool to pharmacologically treat precancerous lesions. SIGNIFICANCE STATEMENT: The treatment of premalignant lesions is largely based on the utilization of invasive (or minimally invasive) measures. Oncogene-induced senescence (OIS) is one form of senescence that occurs in response to oncogene overexpression in somatic cells and is present in precancerous lesions. Although the contribution of OIS to disease progression is undetermined, recent evidence suggests that senescent cells are permissive for malignant transformation. Accordingly, the pharmacological targeting of oncogene-induced senescent cells could potentially provide a novel, less invasive, means for the treatment of premalignant disease.


Beyond NMDA Receptors: Homeostasis at the Glutamate Tripartite Synapse and Its Contributions to Cognitive Dysfunction in Schizophrenia.
Cognitive deficits are core symptoms of schizophrenia but remain poorly addressed by dopamine-based antipsychotic medications. Glutamate abnormalities are implicated in schizophrenia-related cognitive deficits. While the role of the NMDA receptor has been extensively studied, less attention was given to other components that control glutamate homeostasis. Glutamate dynamics at the tripartite synapse include presynaptic and postsynaptic components and are tightly regulated by neuron-astrocyte crosstalk. Here, we delineate the role of glutamate homeostasis at the tripartite synapse in schizophrenia-related cognitive dysfunction. We focus on cognitive domains that can be readily measured in humans and rodents, i.e., working memory, recognition memory, cognitive flexibility, and response inhibition. We describe tasks used to measure cognitive function in these domains in humans and rodents, and the relevance of glutamate alterations in these domains. Next, we delve into glutamate tripartite synaptic components and summarize findings that implicate the relevance of these components to specific cognitive domains. These collective findings indicate that neuron-astrocyte crosstalk at the tripartite synapse is essential for cognition, and that pre- and postsynaptic components play a critical role in maintaining glutamate homeostasis and cognitive well-being. The contribution of these components to cognitive function should be considered in order to better understand the role played by glutamate signaling in cognition and develop efficient pharmacological treatment avenues for schizophrenia treatment-resistant symptoms.


Combined postmenopausal hormone therapy and cardiovascular disease: toward resolving the discrepancy between observational studies and the Women's Health Initiative clinical trial.
Observational research on postmenopausal hormone therapy suggests a 40-50% reduction in coronary heart disease incidence among women using these preparations. In contrast, the Women's Health Initiative clinical trial of estrogen plus progestin found an elevated incidence over a 5.6-year intervention period through July 7, 2002. Toward explaining this discrepancy, the authors analyzed data from this trial, which included 16,608 postmenopausal women aged 50-79 years, and corresponding data from 53,054 women in the Women's Health Initiative observational study, 33% of whom were estrogen-plus-progestin users at baseline. Estrogen-plus-progestin hazard ratio estimates for coronary heart disease, stroke, and venous thromboembolism in the observational study were 39-48% lower than those in the clinical trial following age adjustment. However, hazard ratios tended to decrease with increasing time from initiation of estrogen-plus-progestin use, and observational study hazard ratio estimates are heavily weighted by longer-term use while clinical trial hazard ratio estimates reflect shorter-term use. Following control for time from estrogen-plus-progestin initiation and confounding, hazard ratio estimates were rather similar for the two cohorts, although there was evidence of some remaining difference for stroke. These analyses have implications for both the design and the analysis of observational studies.


Femoral fracture fixation in developing countries: an evaluation of the Surgical Implant Generation Network (SIGN) intramedullary nail.
The Surgical Implant Generation Network (SIGN) intramedullary nailing system was designed to treat femoral fractures in developing countries where real-time imaging, power equipment, and fracture tables are often not available. We performed a retrospective analysis of prospectively collected data on femoral shaft fractures treated with the SIGN intramedullary nailing system. Seventy consecutive patients with a closed diaphyseal femoral fracture were treated with the SIGN intramedullary nail at Mulago National Hospital in Uganda between February 2007 and March 2008, and fifty of these patients (the study cohort) were followed for at least six months or until fracture-healing. The mean time to surgery was 13.2 days (range, zero to thirty-three days). All fractures healed, although two required dynamization for treatment of delayed union. No hardware failures occurred. An interlocking screw missed the nail in two patients, but both fractures healed without complications. One superficial and one deep infection developed; the latter required nail removal after fracture union. Including these patients, complications requiring further treatment occurred in 14% (seven) of the fifty patients. The SIGN intramedullary nailing system promotes predictable healing of femoral fractures in settings with limited resources including lack of real-time imaging, lack of power reaming, and delayed presentation to the operating room.


An immunocytochemical and ultrastructural study of the larval anterior intestine of the frog Rana temporaria, with especial reference to endocrine cells.
Endocrine cells of the larval intestine of Rana temporaria tadpoles have been identified by argyrophilic, immunocytochemical and electron-microscopical techniques. Scarce endocrine cells have been found in both the short non-absorptive zone immediately following the stomach, and in the rest of the anterior intestine. Endocrine cells are frequently seen to extend a cytoplasmic process towards the lumen. Immunoreactivity for serotonin, somatostatin, bombesin and cholecystokinin-8 has been detected. According to the ultrastructural traits of the endocrine granules, three larval intestinal endocrine populations have been differentiated.


Territorial expansion and primary state formation.
A major research problem in anthropology is the origin of the state and its bureaucratic form of governance. Of particular importance for evaluating theories of state origins are cases of primary state formation, whereby a first-generation state evolves without contact with any preexisting states. A general model of this process, the territorial-expansion model, is presented and assessed with archaeological data from six areas where primary states emerged in antiquity: Mesoamerica, Peru, Egypt, Mesopotamia, the Indus Valley, and China. In each case, the evidence shows a close correspondence in time between the first appearance of state institutions and the earliest expansion of the state's political-economic control to regions lying more than a day's round-trip from the capital. Although additional research will add detail and clarity to the empirical record, the results to date are consistent with the territorial-expansion model, which argues that the success of such long-distance expansion not only demanded the bureaucratization of central authority but also helped provide the resources necessary to underwrite this administrative transformation.


Phylogeography and postglacial recolonization of Europe by Rhinolophus hipposideros: evidence from multiple genetic markers.
The demographic history of Rhinolophus hipposideros (lesser horseshoe bat) was reconstructed across its European, North African and Middle-Eastern distribution prior to, during and following the most recent glaciations by generating and analysing a multimarker data set. This data set consisted of an X-linked nuclear intron (Bgn; 543 bp), mitochondrial DNA (cytb-tRNA-control region; 1630 bp) and eight variable microsatellite loci for up to 373 individuals from 86 localities. Using this data set of diverse markers, it was possible to determine the species' demography at three temporal stages. Nuclear intron data revealed early colonization into Europe from the east, which pre-dates the Quaternary glaciations. The mtDNA data supported multiple glacial refugia across the Mediterranean, the largest of which were found in the Ibero-Maghreb region and an eastern location (Anatolia/Middle East)-that were used by R. hipposideros during the most recent glacial cycles. Finally, microsatellites provided the most recent information on these species' movements since the Last Glacial Maximum and suggested that lineages that had diverged into glacial refugia, such as in the Ibero-Maghreb region, have remained isolated. These findings should be used to inform future conservation management strategies for R. hipposideros and show the power of using a multimarker data set for phylogeographic studies.


Therapeutic proposals for the treatment of idiopathic constipation.
The objectives to be achieved by a medical therapy of constipation are: 1) to protect the patient from excessive use of dangerous drugs (laxatives), 2) to help the patient to understand what is a "normal" intestinal function, 3) to reduce or eliminate pain and 4) to avoid complications. The first step consists in general measures (reduced stress, regular meals and physical exercise) and some modifications in diet habits (greater than 1.5 1 of water a day, vegetables, fruits, whole wheat bread). The pharmacological therapy is based on drugs which act in different ways: a) some contain unabsorbable substances (i.e. cellulose, emicellulose) that increase the volume of the stools: b) unabsorbable sugars (i.e. lactulose, lactose) or salts (Mg-sulphate, citrate and Na-sulphate) that provoke an osmotic effect and stimulate the colonic motility; c) suppositories that stimulate the defecation reflex; d) drugs able to stimulate colonic secretion and propulsive motility (i.e. anthraquinones, oral bisacodyl, phenolphthalein, castor oil, prokinetics). There are many conditions in which medical therapy fails its objective: in these cases it is important to exclude other causes of constipation (i.e. drug-related constipation, endocrine disorders, metabolic diseases, systemic illnesses or lesion of the enteric plexus) in order to obtain an improvement of this symptom.


The effect of alpha-bungarotoxin on acetylcholine receptors.
Fluctuations of membrane potential ('membrane noise') during acetylcholineinduced depolarization have been studied by intracellular recording from endplates of frog muscle fibres. When acetylcholine sensitivity had been greatly reduced by an irreversible inhibitor (alpha-bungarotoxin), analysis of the membrane noise showed that neither the amplitude nor the time course of the 'elementary' potential change (due to the molecular depolarizing action of acetylcholine) were altered.


Modulation of self-appraisal of illness, medication adherence, life quality and autonomic functioning by transcranial direct current stimulation in schizophrenia patients.
Little is known about the impact of fronto-temporal transcranial direct current stimulation (tDCS) on attitudes toward mental illness, psychosocial and autonomic functioning, life quality, and medication adherence among schizophrenia patients. Sixty schizophrenia patients were randomly allocated to receive 10 sessions of active (2 mA, 20 min, 2 sessions/day for five weekdays) or sham fronto-temporal tDCS. Self-Appraisal of Illness Questionnaire (SAIQ), Medication Adherence Rating Scale (MARS), World Health Organization Quality of Life-BREF (WHOQOL-BREF) and indices of heart rate variability (HRV) were measured at baseline, immediately after tDCS and at one-month follow-up visit. There were significant group-by-time interactions for scores of SAIQ presence/outcome subscale, total MARS and its subscale of subjective response to taking medication, WHOQOL-BREF psychological domain. Relative to sham, tDCS significantly improved self-awareness of presence/outcome of schizophrenia (Cohen's d = 0.465, p = 0.0011), subjective response to taking medication (Cohen's d = 0.639, p < 0.001) and psychological domain of life quality (Cohen's d = 0.459, p = 0.00114). These effects lasted for less than one month. The group-by-time interactions were non-significant for clinician-rated psychosocial functioning, mean RR intervals, and all HRV indices. Fronto-temporal tDCS briefly optimizes self-reported insight levels, beliefs about treatment adherence, and psychological domain of life quality in patients with schizophrenia. Further studies are required to confirm whether patients treated with 5-day, 10-session tDCS in combination with multisession "maintenance" stimulation every month would attain favourable outcomes. We provide novel evidence for the potential utility of tDCS in schizophrenia.


Measuring oxygen reduction/evolution reactions on the nanoscale.
The efficiency of fuel cells and metal-air batteries is significantly limited by the activation of oxygen reduction and evolution reactions. Despite the well-recognized role of oxygen reaction kinetics on the viability of energy technologies, the governing mechanisms remain elusive and until now have been addressable only by macroscopic studies. This lack of nanoscale understanding precludes optimization of material architecture. Here, we report direct measurements of oxygen reduction/evolution reactions and oxygen vacancy diffusion on oxygen-ion conductive solid surfaces with sub-10 nm resolution. In electrochemical strain microscopy, the biased scanning probe microscopy tip acts as a moving, electrocatalytically active probe exploring local electrochemical activity. The probe concentrates an electric field in a nanometre-scale volume of material, and bias-induced, picometre-level surface displacements provide information on local electrochemical processes. Systematic mapping of oxygen activity on bare and platinum-functionalized yttria-stabilized zirconia surfaces is demonstrated. This approach allows direct visualization of the oxygen reduction/evolution reaction activation process at the triple-phase boundary, and can be extended to a broad spectrum of oxygen-conductive and electrocatalytic materials.


Pulmonary ground-glass nodules: increase in mass as an early indicator of growth.
To compare manual measurements of diameter, volume, and mass of pulmonary ground-glass nodules (GGNs) to establish which method is best for identifying malignant GGNs by determining change across time. In this ethics committee-approved retrospective study, baseline and follow-up CT examinations of 52 GGNs detected in a lung cancer screening trial were included, resulting in 127 GGN data sets for evaluation. Two observers measured GGN diameter with electronic calipers, manually outlined GGNs to obtain volume and mass, and scored whether a solid component was present. Observer 1 repeated all measurements after 2 months. Coefficients of variation and limits of agreement were calculated by using Bland-Altman methods. In a subgroup of GGNs containing all resected malignant lesions, the ratio between intraobserver variability and growth (growth-to-variability ratio) was calculated for each measurement technique. In this subgroup, the mean time for growth to exceed the upper limit of agreement of each measurement technique was determined. The kappa values for intra- and interobserver agreement for identifying a solid component were 0.55 and 0.38, respectively. Intra- and interobserver coefficients of variation were smallest for GGN mass (P < .001). Thirteen malignant GGNs were resected. Mean growth-to-variability ratios were 11, 28, and 35 for diameter, volume, and mass, respectively (P = .03); mean times required for growth to exceed the upper limit of agreement were 715, 673, and 425 days, respectively (P = .02). Mass measurements can enable detection of growth of GGNs earlier and are subject to less variability than are volume or diameter measurements.


Ankle joint mechanics and foot proportions differ between human sprinters and non-sprinters.
Recent studies of sprinters and distance runners have suggested that variations in human foot proportions and plantarflexor muscle moment arm correspond to the level of sprint performance or running economy. Less clear, however, is whether differences in muscle moment arm are mediated by altered tendon paths or by variation in the centre of ankle joint rotation. Previous measurements of these differences have relied upon assumed joint centres and measurements of bone geometry made externally, such that they would be affected by the thickness of the overlying soft tissue. Using magnetic resonance imaging, we found that trained sprinters have shorter plantarflexor moment arms (p = 0.011) and longer forefoot bones (p = 0.019) than non-sprinters. The shorter moment arms of sprinters are attributable to differences in the location of the centre of rotation (p < 0.001) rather than to differences in the path of the Achilles tendon. A simple computer model suggests that increasing the ratio of forefoot to rearfoot length permits more plantarflexor muscle work during plantarflexion that occurs at rates expected during the acceleration phase following the sprint start.


De novo birth of functional microproteins in the human lineage.
Small open reading frames (sORFs) can encode functional "microproteins" that perform crucial biological tasks. However, their size makes them less amenable to genomic analysis, and their origins and conservation are poorly understood. Given their short length, it is plausible that some of these functional microproteins have recently originated entirely de novo from noncoding sequences. Here we sought to identify such cases in the human lineage by reconstructing the evolutionary origins of human microproteins previously found to have measurable, statistically significant fitness effects. By tracing the formation of each ORF and its transcriptional activation, we show that novel microproteins with significant phenotypic effects have emerged de novo throughout animal evolution, including two after the human-chimpanzee split. Notably, traditional methods for assessing coding potential would miss most of these cases. This evidence demonstrates that the functional potential intrinsic to sORFs can be relatively rapidly and frequently realized through de novo gene emergence.


Once-weekly intravenous paricalcitol in the treatment of secondary hyperparathyroidism in hemodialysis patients.
Paricalcitol, a vitamin D analog, is commonly administered three times weekly to control secondary hyperparathyroidism in hemodialysis patients. Less frequent dosing would be more convenient, require less nursing time, and be an option in other dialysis modalities. No studies have examined the efficacy of once-weekly dosing of paricalcitol. Chronic hemodialysis patients receiving a stable dose of paricalcitol three times weekly with intact PTH (iPTH) 100-500 ng/l were monitored during a two-week baseline, then were converted to a single mid-week paricalcitol dose equal to the previous cumulative weekly dose. Serum calcium and phosphorus were monitored weekly and iPTH levels determined during study Weeks 4 and 8. A single paricalcitol dose adjustment was made during study Week 5 based on iPTH to achieve a target value of 150-300 ng/l. Phosphate binders and calcium dialysate bath were kept constant during the study. In the 25 patients, mean iPTH was 295 +/- 107 ng/l at baseline, and not significantly different at Week 4 (307 +/- 111 ng/l) or Week 8 (285 +/- 98 ng/l). Paricalcitol dose increases mid-study were almost exclusively in patients with iPTH > 300 ng/l. Calcium, phosphorus, and calcium x phosphorus product were not significantly different on weekly therapy. (Only one patient developed a calcium > 2.55 mmol/l during the study.) Once-weekly dosing of paricalcitol is an effective option in treatment of secondary hyperparathyroidism. Less frequent dosing may better allocate nursing time and potentially benefit other patient populations with CKD and ESRD.


Identification of macrophage-specific infectivity loci (mil) of Legionella pneumophila that are not required for infectivity of protozoa.
We have recently shown that many mutants of Legionella pneumophila exhibit similar defective phenotypes within both U937 human-derived macrophages and the protozoan host Acanthamoeba (L.-Y. Gao, O. S. Harb, and Y. Abu Kwaik, Infect. Immun. 65:4738-4746, 1997). These observations have suggested that many of the mechanisms utilized by L. pneumophila to parasitize mammalian and protozoan cells are similar, but our data have not excluded the possibility that there are unique mechanisms utilized by L. pneumophila to survive and replicate within macrophages but not protozoa. To examine this possibility, we screened a bank of 5,280 miniTn10::kan transposon insertion mutants of L. pneumophila for potential mutants that exhibited defective phenotypes of cytopathogenicity and intracellular replication within macrophage-like U937 cells but not within Acanthamoeba polyphaga. We identified 32 mutants with various degrees of defects in cytopathogenicity, intracellular survival, and replication within human macrophages, and most of the mutants exhibited wild-type phenotypes within protozoa. Six of the mutants exhibited mild defects in protozoa. The defective loci were designated mil (for macrophage-specific infectivity loci). Based on their intracellular growth defects within macrophages, the mil mutants were grouped into five phenotypic groups. Groups I to III included the mutants that were severely defective in macrophages, while members of the other two groups exhibited a modestly defective phenotype within macrophages. The growth kinetics of many mutants belonging to groups I to III were also examined, and these were shown to have a similar defective phenotype in peripheral blood monocytes and a wild-type phenotype within another protozoan host, Hartmannella vermiformis. Transmission electron microscopy of A. polyphaga infected by three of the mil mutants belonging to groups I and II showed that they were similar to the parent strain in their capacity to recruit the rough endoplasmic reticulum (RER) around the phagosome. In contrast, infection of macrophages showed that the three mutants failed to recruit the RER around the phagosome during early stages of the infection. None of the mil mutants was resistant to NaCl, and the dot or icm NaCl(r) mutants are severely defective within mammalian and protozoan cells. Our data indicated that in addition to differences in mechanisms of uptake of L. pneumophila by macrophages and protozoa, there were also genetic loci required for L. pneumophila to parasitize mammalian but not protozoan cells. We hypothesize that L. pneumophila has evolved as a protozoan parasite in the environment but has acquired loci specific for intracellular replication within macrophages. Alternatively, ecological coevolution with protozoa has allowed L. pneumophila to possess multiple redundant mechanisms to parasitize protozoa and that some of these mechanisms do not function within macrophages.