Evaluating the Effectiveness of Gamification on Physical Activity: Systematic Review and Meta-analysis of Randomized Controlled Trials.
Gamification refers to the use of game elements in nongame contexts. The use of gamification to change behaviors and promote physical activity (PA) is a promising avenue for tackling the global physical inactivity pandemic and the current prevalence of chronic diseases. However, there is no evidence of the effectiveness of gamified interventions with the existence of mixed results in the literature. The aim of this systematic review and meta-analysis is to evaluate the effectiveness of gamified interventions and their health care potential by testing the generalizability and sustainability of their influence on PA and sedentary behavior. A total of 5 electronic databases (PubMed, Embase, Scopus, Web of Science, and the Cochrane Central Register of Controlled Trials) were searched for randomized controlled trials published in English from 2010 to 2020. Eligibility criteria were based on the components of the participants, interventions, comparators, and outcomes framework. Studies were included when they used gamified interventions in daily life with an active or inactive control group and when they assessed a PA or sedentary behavior outcome. We conducted meta-analyses using a random-effects model approach. Sensitivity analyses, influence analyses, and publication bias analyses were performed to examine the robustness of our results. The main meta-analysis performed on 16 studies and 2407 participants revealed a small to medium summary effect of gamified interventions on PA behavior (Hedges g=0.42, 95% CI 0.14-0.69). No statistical difference among different subgroups (adults vs adolescents and healthy participants vs adults with chronic diseases) and no interaction effects with moderators such as age, gender, or BMI were found, suggesting good generalizability of gamified interventions to different user populations. The effect was statistically significant when gamified interventions were compared with inactive control groups, such as waiting lists (Hedges g=0.58, 95% CI 0.08-1.07), and active control groups that included a nongamified PA intervention (Hedges g=0.23, 95% CI 0.05-0.41). This suggests that gamified interventions are not only efficient in changing behavior but also more effective compared with other behavioral interventions. The long-term effect (measured with follow-up averaging 14 weeks after the end of the intervention) was weaker, with a very small to small effect (Hedges g=0.15, 95% CI 0.07-0.23). This meta-analysis confirms that gamified interventions are promising for promoting PA in various populations. Additional analyses revealed that this effect persists after the follow-up period, suggesting that it is not just a novelty effect caused by the playful nature of gamification, and that gamified products appear effective compared with equivalent nongamified PA interventions. Future rigorous trials are required to confirm these findings.


Effect of chronic morphine treatment on thyrotropin and prolactin levels and acute hormone responses in the rat.
The effects of three chronic morphine regimens on basal and cold-stimulated thyrotropin (TSH) and on prolactin levels were studied in male rats with and without acute morphine challenge. All the chronic regimens decreased basal and cold-stimulated TSH levels, but only one regimen (10 mg/kg b.i.d.) significantly enhanced the prolactin levels. The altered cold-stimulated TSH and prolactin levels were recovered within about 96 and 48 hr, respectively, after the last morphine injection on the 7-day pretreatment period. After the chronic administration (14 days), acute morphine challenges were performed either when cold-stimulated TSH secretion was suppressed (12- and 24-hr lag time) or when the response to cold was normalized (96-hr lag time). When the TSH levels were still low, the small challenge doses of morphine (10 and 15 mg/kg) no longer suppressed the TSH secretion. This was due neither to the real tolerance nor to the persistent effect of chronic morphine but to the withdrawal stress, which was also reflected as elevated corticosterone levels. However, after a 96-hr lag time, the challenge doses of morphine decreased TSH secretion after each morphine regimen as effectively as in the naive rats. Thus, the regimens did not induce the development of tolerance to the effect of morphine on cold-stimulated TSH secretion. Most regimens seemed to cause some tolerance to the stimulatory effect of morphine on prolactin secretion irrespective of the duration of the lag time. Even after the mildest regimen (10 mg/kg once a day), a tolerance developed to the antinociceptive effect of morphine, and it lasted well up to 96 hr.(ABSTRACT TRUNCATED AT 250 WORDS)


sar: a genetic mouse model for human sarcosinemia generated by ethylnitrosourea mutagenesis.
A mouse mutant with sarcosinemia was found by screening the progeny of ethylnitrosourea-mutagenized mice for aminoacidurias. Paper chromatography, column chromatography, and gas chromatography-mass spectrometry identified high levels of sarcosine in the urine of the mutant mice. While sarcosine cannot be detected in the urine of plasma of normal mice, the urinary sarcosine level of 102 +/- 58 mmol per g of creatinine in the mutant mice was at the upper range of the urinary levels (1.5-4.5 mmol of sarcosine per g of creatinine) observed in humans with sarcosinemia. Similarly, the plasma sarcosine level of 785 +/- 153 mumol/liter in the sarcosinemic mice was at the upper range of the plasma sarcosine levels (53-760 mumol/liter) observed in affected humans. Sarcosine dehydrogenase [sarcosine:(acceptor) oxidoreductase (demethylating), EC 188.8.131.52] activity was deficient in sarcosinemic mice. The sarcosinuria phenotype in these mice was inherited as an autosomal recessive trait. This mouse mutant provides a useful genetic model for human sarcosinemia and for development of therapeutic approaches for genetic disease.


The color of death: race, observed skin tone, and all-cause mortality in the United States.
Objective: This paper examines how mortality covaries with observed skin tone among blacks and in relation to whites. Additionally, the study analyzes the extent to which social factors such as socioeconomic status affect this relationship. Design: This study uses data from the 1982 General Social Survey (N = 1,689) data linked to the National Death Index until 2008. We use this data to examine the links between race, observed skin tone among blacks, and all-cause mortality. Piecewise exponential hazard modeling was used to estimate disparities in skin tone mortality among blacks, and relative to whites. The multivariate models control for age, education, gender, region, metropolitan statistical area, marital status, labor force status, and household income. Results: Observed skin tone is a significant determinant of mortality among blacks and in relation to whites. Light skinned blacks had the lowest mortality hazards among blacks, while respondents with medium and dark brown skin experienced significantly higher mortality. The observed skin tone mortality disparities covaried with education; there are significant mortality disparities across observed skin tone groups among black respondents with high school or more education, and nonsignificant disparities among those with less education. Conclusion: It is crucial to identify the social processes driving racial disparities in health and mortality. The findings reveal that the nuanced social experiences of blacks with different observed skin tones markedly change the experience of racial inequality. Research on the nuanced social processes and biological mechanisms that connect differences in observed skin tone to mortality outcomes promises to better illuminate the experience of racial inequality and policy mechanisms we can use to undermine it.


The renin-angiotensin system and body function.
Angiotensin II, the biologically active component of the renin-angiotensin system, acts throughout the body to produce an impressive number of cardiovascular, endocrine, metabolic, and behavioral effects. Major actions include elevation of arterial pressure, stimulation of aldosterone secretion, and a variety of effects on the kidneys, brain, and pituitary. Investigation of the role of the renin-angiotensin system in physiological regulation has been greatly facilitated by the availability of specific inhibitors of the formation or actions of angiotensin II, most notably converting-enzyme inhibitors and angiotensin receptor antagonists. Studies with these agents have clearly shown that the renin-angiotensin system plays an important role in the defense of body balance and blood pressure in hypovolemic state, including sodium deficiency and hemorrhage. The inhibitors also lower blood pressure in some forms of hypertension, and converting-enzyme inhibitors are proving to be effective antihypertensive agents.


Casodex treatment induces hypoxia-related gene expression in the LNCaP prostate cancer progression model.
The changes in gene expression profile as prostate cancer progresses from an androgen-dependent disease to an androgen-independent disease are still largely unknown. We examined the gene expression profile in the LNCaP prostate cancer progression model during chronic treatment with Casodex using cDNA microarrays consisting of 2305 randomly chosen genes. Our studies revealed a representative collection of genes whose expression was differentially regulated in LNCaP cells upon treatment with Casodex. A set of 15 genes were shown to be highly expressed in Casodex-treated LNCaP cells compared to the reference sample. This set of highly expressed genes represents a signature collection unique to prostate cancer since their expression was significantly greater than that of the collective pool of ten cancer cell lines of the reference sample. The highly expressed signature collection included the hypoxia-related genes membrane metallo-endopeptidase (MME), cyclin G2, and Bcl2/adenovirus E1B 19 kDa (BNIP3). Given the roles of these genes in angiogenesis, cell cycle regulation, and apoptosis, we further analyzed their expression and concluded that these genes may be involved in the molecular changes that lead to androgen-independence in prostate cancer. Our data indicate that one of the mechanisms of Casodex action in prostate cancer cells is induction of hypoxic gene expression.


Biological understanding of the cardiovascular risk associated with major depression and panic disorder is important.
Panic disorder (PD) and major depressive disorders (MDD) are serious mental disorders but the mechanisms underlying the pathophysiology are poorly understood. Nitric oxide (NO) is a gas considered to play an important role in mediating anxiety and stress response and is synthesised from nitric oxide synthase (NOS). The endothelial isoform (eNOS) has been found also in platelets. Homocysteine (Hcy) is an amino acid which naturally occurs in the human body. Elevated levels are linked to increased risk of cardiovascular, neurological and psychiatric diseases. In this study we aimed to evaluate NO, platelet aggregation and Hcy levels in PD and MDD patients. Nineteen PD and 18 MDD patients participated in this study. NO levels were measured spectrophotometrically, platelet aggregation levels were measured in an aggregometer and Hcy levels were measured by HPLC. NO levels were significantly lower in patients with MDD and PD than in control subjects (P < 0.05). Hcy and platelet aggregation levels were significantly higher in patients with MDD and PD than in control subjects (P < 0.05). Further more detailed studies are needed to find out the effects of drugs on these parameters or to disclose the exact mechanism underlying the alteration of these parameters.


Comparing pinning in situ and capital realignment procedures for severe, stable slipped capital femoral epiphysis: a systematic review.
In severe, stable slipped capital femoral epiphysis, it is unclear whether pinning in situ (PIS) or capital realignment procedures (CRPs) are superior. Our primary aim was to compare patient-reported outcome measures (PROMs) following each strategy. Secondary aims were to compare rates of femoral head avascular necrosis (AVN) and complications. MEDLINE, Embase and Cochrane databases were searched according to an agreed strategy. Narrative review articles, case reports, letters to the editor and articles not written in English were excluded. The risk of bias was assessed using the Newcastle-Ottawa Scale. Of the 132 citations identified, 127 were excluded following de-duplication and application of the exclusion criteria. Three observational studies comparing PIS with CRP and two case series considering CRP alone were identified. One article was considered fair quality, and four articles were considered poor. In total, 198 hips from five studies were included (66 PIS, 132 CRP). PIS was associated with moderate-good functional outcomes, and CRP with good-high outcomes. Two comparative studies reported significantly better PROMs following CRP. AVN was reported in 1.5% following PIS and 10.6% following CRP. Regarding other complications, chondrolysis occurred in 3.0% following PIS and 2.4% following CRP. Femoroacetabular impingement rates were markedly higher following PIS (60.6% versus 2.3%). Reoperation rates were also greater following PIS (34.5% versus 13.3%). PIS tends to be associated with favourable AVN rates, but CRP with favourable PROMs and complication rates. However, comparisons were drawn from heterogeneous studies lacking long-term follow-up. Further high-quality research is required.


Predictors of poor outcomes after significant chest trauma in multiply injured patients: a retrospective analysis from the German Trauma Registry (Trauma Register DGU®).
Blunt thoracic trauma is one of the critical injury mechanisms in multiply injured trauma victims. Although these patients present a plethora of potential structural damages to vital organs, it remains debated which injuries actually influence outcome and thereby should be addressed initially. Hence, the aim of this study was to identify the influence of critical structural damages on mortality. All patients in the database of the TraumaRegister DGU® (TR-DGU) from 2002-2011 with AIS Chest ≥ 2, blunt trauma, age of 16 or older and an ISS ≥ 16 were analyzed. Outcome parameters were in-hospital mortality as well as ventilation time in patients surviving the initial 14 days after trauma. 22613 Patients were included (mean ISS 30.5 ± 12.6; 74.7% male; Mean Age 46.1 ± 197 years; mortality 17.5%; mean duration of ventilation 7.3 ± 11.5; mean ICU stay 11.7 ± 14.1 days). Only a limited number of specific injuries had a significant impact on survival. Major thoracic vessel injuries (AIS ≥5), bilateral lung contusion, bilateral flail chest, structural heart injury (AIS ≥3) significantly influence mortality in study patients. Several extrathoracic factors (age, blood transfusion, systolic blood pressure and extrathoracic severe injuries) were also predictive of increased mortality. Most injuries of the thoracic wall had no or only a moderate effect on the duration of ventilation. Injuries to the lung (laceration, contusion or pneumothoraces) had a moderate prolonging effect. Cardiac injuries and severe injuries to the thoracic vessels induced a substantially prolonged ventilation interval. We demonstrate quantitatively the influence of specific structural damages of the chest on critical outcome parameters. While most injuries of the chest wall have no or only limited impact in the study collective, injuries to the lung overall show adverse outcome. Injuries to the heart or thoracic vessels have a devastating prognosis following blunt chest trauma.


[Comparison of the field formation in the frontal area during prenatal period in Macaca (Macacus rhesus, S. Macaca mulatta) and man].
A developmental comparison of the fields 8, 9, 10, 11, 12, 47 during prenatal life in macacus rhesus and man was performed. Cytomyeloarchitectonic and morphometric methods of investigation were applied: the areas of the fields in question, the transverse thickness of the cortex, separate cortical layers, neuronal dimentions were measured. Alongside, general regularities in the development of these fields in both species (the frontal cortex is laid earlier than other parts of the cortex, the terms of the field differentiation coincide, uneven development of the fields with different phylogenic origin) some important differences are demonstrated. They are seen a) in the rate of the frontal field maturation--in man it takes longer time, b) in a more complex organization of the frontal part in man--during prenatal life new structural features of the fields, 44, 45, 46 and subfields in the field 47 appear. During prenatal life of monkey and man in the development of the frontal area the phenomena of heterochrony (common for both species) and anabolia (peculiar for man only) are observed (the appearance of new more complex structures). These phenomena display themselves in the shift of the laying of the frontal area towards earlier stages of embryogenesis.


Oral mucosal lesions and their association with sociodemographic, behavioral, and health status factors.
The aim of this study was to evaluate the frequency of oral mucosal lesions and their associations with sociodemographic, health, and behavioral factors in a southern Brazilian population. Information was collected from participants (n = 801) using a structured questionnaire during an oral cancer screening campaign held at an agribusiness show in southern Brazil in 2009. Data were described using frequency distributions or means and standard deviations. Associations between independent variables and outcomes were assessed using the Chi-squared test. A total of 465 lesions were detected (actinic cheilitis: n = 204, 25.5%; candidiasis: n = 50, 6.2%; fibrous inflammatory hyperplasia: n = 42, 5.2%; ulceration, n = 33, 4.1%; hemangioma: n = 14, 1.7%; leukoplakia: n = 11, 1.4%). Candidiasis, actinic cheilitis, and fibrous inflammatory hyperplasia were associated significantly with literacy. Actinic cheilitis was also associated significantly with sun exposure and hat use, and leukoplakia was associated with smoking. The high frequency of oral mucosal lesions observed highlights the importance of education about risk factors. Additionally, training of health professionals, mainly those from public health services, in the use of preventive and community education strategies is needed.


The mineralogy and structure of use-wear polish on chert.
Polished edges of archaeological stone tools are commonly investigated to obtain information on the tools' uses in prehistory. Yet to this day, it remains unclear what exactly such polishes are and how they form. Answering these questions should allow the elaboration of new interpretative methods based on objective measurements. Two major competing hypotheses of polish formation have been proposed: abrasion and the formation of a thin amorphous film on the chert or flint surface. We employ reflectance infrared spectroscopy, a technique particularly sensitive to thin amorphous films, to investigate these two hypotheses. We found no added amorphous layer that would have formed upon friction against bone, antler, ivory or wood. Our observations suggest polish formation by abrasion, notwithstanding previous claims of added amorphous surface structures. This has implications for our understanding of the physical processes taking place during friction of chert and flint against different materials. Our results also open the possibility to propose new pathways for identifying different use-wear processes, based on the degree of abrasion.


Long lasting control of viral rebound with a new drug ABX464 targeting Rev - mediated viral RNA biogenesis.
Current therapies have succeeded in controlling AIDS pandemic. However, there is a continuing need for new drugs, in particular those acting through new and as yet unexplored mechanisms of action to achieve HIV infection cure. We took advantage of the unique feature of proviral genome to require both activation and inhibition of splicing of viral transcripts to develop molecules capable of achieving long lasting effect on viral replication in humanized mouse models through inhibition of Rev-mediated viral RNA biogenesis. Current HIV therapies reduce viral load during treatment but titers rebound after treatment is discontinued. We devised a new drug that has a long lasting effect after viral load reduction. We demonstrate here that ABX464 compromises HIV replication of clinical isolates of different subtypes without selecting for drug resistance in PBMCs or macrophages. ABX464 alone, also efficiently compromised viral proliferation in two humanized mouse models infected with HIV that require a combination of 3TC, Raltegravir and Tenofovir (HAART) to achieve viral inhibition in current protocols. Crucially, while viral load increased dramatically just one week after stopping HAART treatment, only slight rebound was observed following treatment cessation with ABX464 and the magnitude of the rebound was maintained below to that of HAART for two months after stopping the treatment. Using a system to visualize single HIV RNA molecules in living cells, we show that ABX464 inhibits viral replication by preventing Rev-mediated export of unspliced HIV-1 transcripts to the cytoplasm and by interacting with the Cap Binding Complex (CBC). Deep sequencing of viral RNA from treated cells established that retained viral RNA is massively spliced but importantly, normal cellular splicing is unaffected by the drug. Consistently ABX464 is non-toxic in humans and therefore represents a promising complement to current HIV therapies. ABX464 represents a novel class of anti-HIV molecules with unique properties. ABX464 has a long lasting effect in humanized mice and neutralizes the expression of HIV-1 proviral genome of infected immune cells including reservoirs and it is therefore a promising drug toward a functional cure of HIV.


Ultrasonography Reliability in the Detection of Inflammatory and Structural Abnormalities: An Exercise in Multiple Joints.
Ultrasonography is an image technique that allows rheumatologists to visualize structural and inflammatory changes within a joint. The objective of this study was to assess the interobserver and intraobserver reliability of musculoskeletal ultrasound (US) in the detection of inflammatory and destructive joint changes in patients with polyarthritis. A Delphi exercise was undertaken to standardize and adapt the EULAR-OMERACT elementary US definitions of inflammatory lesions (effusion, synovial hypertrophy, power Doppler, bone erosions, and synovitis) for each joint. Fifteen patients were analyzed, and video clips of 600 joints were collected. Each joint was scored for the presence of each elementary component, on 2 separate occasions, by 6 examiners. Interobserver and intraobserver agreement analysis was assessed through Fleiss κ coefficient (κ). Considering all patients and all joints, the interobserver values were highest for erosions and lowest for effusion (κ = 0.7314 and κ = 0.6044, respectively). When analyzing different regions, the highest interobserver agreement was for tibiotalar joint (κ = 0.8043) and the lowest for wrist (κ = 0.6767). Intraobserver reliability was excellent for each and all elementary components and anatomical region. The present study showed either a good or excellent US interobserver and intraobserver reliability in elementary elements and anatomical region. This kind of US reliability exercises are important for standardization of exploration in everyday practice by reducing the variability associated with this imaging technique, and ensuring a greater degree of homogeneity and future comparability in the assessment of disease activity in polyarthritis patients.


Paracrystalline arrays of membrane-to-membrane cross bridges associated with the inner surface of plasma membrane.
In cultured cells of the rat kangaroo PtK2 line, veils of the cell surface were observed which consisted of only plasma membrane and paracrystalline arrays of membrane-associated particles sandwiched in between. These membrane-to-membrane cross-bridging 9-to 11-nm wide particles were somewhat coumellar-shaped and were arranged on a hexagonal lattice with an interparticle distance of 16nm. At higher magnification, they revealed an unstained core, thus suggesting a ringlike substructure. Similar arrays of paracrystal-containing veils, which were rather variable in size and frequency, were also observed in other cultured cells. It is hypothesized that these paracrystals represent protein macromolecular complexes associated with the inner plasma membrane surface which crystallize when plasma membranes come into close intracellular contact and other components of the subsurface network are removed.


Isolation, identification and characterization of a glyphosate-degrading bacterium, Bacillus cereus CB4, from soil.
A bacterial strain named CB4, with highly effective glyphosate degradation capability, was isolated from soil after enrichment. On the basis of the Biolog omniLog identification system (Biolog) and 16S ribosomal RNA (rRNA) gene sequencing methods, strain CB4 was identified as Bacillus cereus. Further experiments were carried out to optimize the growth of strain CB4 and the glyphosate degradation activity by high performance liquid chromatography (HPLC). The optimal conditions were found as follows: initial pH 6.0, incubation temperature 35°C, glyphosate concentration 6 g L(-1), inoculation amount 5% and incubation time 5 days. Under the optimal conditions, stain CB4 utilized 94.47% of glyphosate. This is the first report on B. cereus with a capacity to utilize herbicide glyphosate, and it can degrade glyphosate concentrations up to 12 g L(-1). Metabolization of glyphosate by strain B. cereus CB4 was studied. Results indicated that two concurrent pathways were capable of degrading glyphosate to AMPA, glyoxylate, sarcosine, glycine and formaldehyde as products. Glyphosate breakdown in B. cereus CB4 was achieved by the C-P lyase activity and the glyphosate oxidoreductase activity.


Repeated exposure to sevoflurane in neonatal rats impairs cognition in adulthood via the PKA-CREB-BDNF signaling pathway.
Sevoflurane (Sev) anesthesia is widely used in pediatrics due to its low blood-gas partition coefficient and lack of pungency. However, Sev treatment may lead to cognitive dysfunction in later life. The current study administered Sev to neonatal rats to investigate the effects of Sev treatment on cognitive performance in adulthood. In total, 6-day-old rats received 3% Sev for 2 h daily for 3 consecutive days. The cognitive function of rats in adulthood was evaluated in 56-day-old rats by Morris water maze test. The hippocampal neuron morphology was observed by Nissl staining. Hippocampal brain-derived neurotrophic factor (BDNF) levels were measured by ELISA. The protein expression of protein kinase A (PKA), cAMP response element binding protein (CREB), phosphorylated-CREB (p-CREB) and BDNF in hippocampus were assessed by western blotting. The water maze results demonstrated that neonatal treatment with Sev resulted in a significant impairment of cognition in 56-day-old adult rats. Behavioral analysis revealed that Sev treatment increased latency to first pass the platform and decreased residence in target quadrants and across platform frequency compared with the control group in Morris water maze tests. Furthermore, compared with the control group, neonatal exposure to Sev reduced the number of neurons and the concentration of BDNF in the hippocampus, a brain region important for learning and memory. Additionally, Sev significantly decreased the expression of PKA, p-CREB, BDNF and the p-CREB/CREB ratio. Treatment with bucladesine, a selective PKA agonist, partially reversed the deleterious effects of Sev. In summary, the results indicated that PKA-CREB-BDNF signaling served an important role in the cognitive decline caused by neonatal exposure to Sev.


Travelling along a road with obstacles: experiences of managing life to feel well while living with migraine.
Living a life with migraine can impair one's sense of feeling well, and migraine is a disorder that is associated with substantial disability. Earlier research on how people manage their migraine has given important insight into these people's preventive actions and how they handle their attacks, but there is still a lack of knowledge of how persons with migraine manage their lives to feel well from a more holistic viewpoint. Therefore, the aim of this study was to explore lived experiences of managing life to feel well while living with migraine. Nineteen persons with migraine were interviewed. A hermeneutic-phenomenological approach was used to explore their lived experiences. The findings reveal that persons with migraine not only used preventive strategies to abort and ease the consequences of migraine but also tried to amplify the good in life through increasing their energy and joy and through reaching peace with being afflicted with migraine. The findings of this study can encourage healthcare providers, as well as persons with migraine, to consider channeling their efforts into strategies aiming to amplify the good in life, including reaching peace of mind despite being afflicted.


[Yogurt as source of lactose autodigestion].
Microorganisms in yogurt have the capacity of quantitatively digest in vivo in the small intestine the lactose from yogurt. The process of autodigestion of lactose in yogurt reduces both lactose maldigestion and lactose intolerance in lactase deficient individuals. Enzyme activity in yogurt depends upon the buffer capacity of the yogurt, microbial cells resistance to acid and enzymatic activity and to the effect of bile in the microbial cell that release beta-galactosidase activity. Lactose autodigestion capacity of yogurt is significantly reduced in pasteurized yogurt and it is also affected by the type and amount of microorganism that is added to milk and by the presence of fat. Yogurt intake represents an important food alternative for lactase deficient individuals.


The genetics of hereditary colon cancer.
The genetic basis of sporadic colorectal cancer has illuminated our knowledge of human cancer genetics. This has been facilitated and catalyzed by an appreciation and deep understanding of the forms of colorectal cancer that harbor an inherited predisposition, including familial adenomatous polyposis (FAP), hereditary nonpolyposis colorectal cancer (HNPCC) or Lynch syndrome, the hamartomatous polyposis syndromes, and certain other rare syndromes. Identification of germline mutations in pivotal genes underlying the inherited forms of colorectal cancer has yielded many dividends, including functional dissection of critical molecular pathways that have been revealed to be important in development, cellular homeostasis, and cancer; new approaches in chemoprevention, molecular diagnostics and genetic testing, and therapy; and underscoring genotypic-phenotypic relationships.


Melatonin protects blood-brain barrier integrity and permeability by inhibiting matrix metalloproteinase-9 via the NOTCH3/NF-κB pathway.
The pathophysiological mechanism of white matter hyperintensities of cerebral small vessel disease (CSVD) includes an impaired blood-brain barrier (BBB) with increased permeability. Neuroinflammation likely contributes to the disruption of the BBB in CSVD. Therefore, understanding the molecular mechanism of how neuroinflammation causes BBB damage is essential to preventing BBB disruption in CSVD. Matrix metalloproteinase 9 (MMP-9) contributes to BBB damage in neuroinflammatory diseases. In this study, we observed that interleukin-1β (IL-1β)-induced MMP-9 secretion in pericytes increased BBB permeability to sodium fluorescein (Na-F) by damaging the disruption of VE-cadherin, occludin, claudin-5, and zonula occludin-1 (ZO-1). Melatonin reduced BBB permeability to Na-F and inhibited the disruption of the adherens and tight junction proteins. Melatonin also downregulated MMP-9 and upregulated tissue inhibitor of metalloproteinases 1 (TIMP-1) gene expression, which decreased the MMP-9/TIMP-1 ratio. In addition, nuclear translocation of NF-κB/p65 induced by IL-1β in pericytes upregulated MMP-9 expression, which was inhibited by the NF-κB inhibitor PDTC. However, the NOTCH3 inhibitor DAPT significantly inhibited NF-κB/p65 translocation to the nucleus, while melatonin in combination with DAPT significantly prevented NF-κB/p65 translocation than DAPT alone. Our results suggest that melatonin reduced MMP-9-induced permeability of the BBB. Melatonin reduced MMP-9 expression and activity, which was induced by IL-1β through the regulation of the NOTCH3/NF-κB signaling pathway in pericytes, suggesting that pericytes regulate BBB integrity and function.


Healthy Cities indicators--a suitable instrument to measure health?
The evidence-base for a health strategy should include information on the determinants of health and how they link together if it is to influence the health of the population. The WHO European Healthy Cities Network developed a set of 53 healthy city indicators (HCIs), to describe the health of its citizens and capture a range of local initiatives addressing the wider dimensions of health. This was the first systematic effort to collect and analyze a range of data from European cities. The analysis provided important insights into the interpretation, availability, and feasibility of collecting data, resulting in the development of a revised set of 32 indicators with improved definitions. An analysis of the revised indicators showed that this data was more complete and feasible to collect. It provided useful information to cities contributing to developing a description of health and thus helping to identify health problems. It also highlighted issues about the importance of collecting qualitative as well as quantitative data, the number of indicators and the appropriateness of using the indicators to compare different cities. HCIs facilitated the collection of routinely available health data in a systematic manner. The introduction of HCIs has encouraged cities to adopt a structured process of collecting information on the health of their citizens and build on this information by collecting appropriate local data for developing a city health profile to underpin a city health plan that would set out strategies and interventions to improve health and provide the evidence-base for health plans.


Opening a Pandora's Flask on a Prototype Catalytic Direct Arylation Reaction of Pentafluorobenzene: The Ag2CO3/Pd(OAc)2/PPh3 System.
Direct C-H functionalization reactions have opened new avenues in catalysis, removing the need for prefunctionalization of at least one of the substrates. Although C-H functionalization catalyzed by palladium complexes in the presence of a base is generally considered to proceed by the CMD/AMLA-6 mechanism, recent research has shown that silver(I) salts, frequently used as bases, can function as C-H bond activators instead of (or in addition to) palladium(II). In this study, we examine the coupling of pentafluorobenzene 1 to 4-iodotoluene 2a (and its analogues) to form 4-(pentafluorophenyl)toluene 3a catalyzed by palladium(II) acetate with the commonplace PPh3 ligand, silver carbonate as base, and DMF as solvent. By studying the reaction of 1 with Ag2CO3/PPh3 and with isolated silver (triphenylphosphine) carbonate complexes, we show the formation of C-H activation products containing the Ag(C6F5)(PPh3)n unit. However, analysis is complicated by the lability of the Ag-PPh3 bond and the presence of multiple species in the solution. The speciation of palladium(II) is investigated by high-resolution-MAS NMR (chosen for its suitability for suspensions) with a substoichiometric catalyst, demonstrating the formation of an equilibrium mixture of Pd(Ar)(κ1-OAc)(PPh3)2 and [Pd(Ar)(μ-OAc)(PPh3)]2 as resting states (Ar = Ph, 4-tolyl). These two complexes react stoichiometrically with 1 to form coupling products. The catalytic reaction kinetics is investigated by in situ IR spectroscopy revealing a two-term rate law and dependence on [Pdtot/nPPh3]0.5 consistent with the dissociation of an off-cycle palladium dimer. The first term is independent of [1], whereas the second term is first order in [1]. The observed rates are very similar with Pd(PPh3)4, Pd(Ph)(κ1-OAc)(PPh3)2, and [Pd(Ph)(μ-OAc)(PPh3)]2 catalysts. The kinetic isotope effect varied significantly according to conditions. The multiple speciation of both AgI and PdII acts as a warning against specifying the catalytic cycles in detail. Moreover, the rapid dynamic interconversion of AgI species creates a level of complexity that has not been appreciated previously.


Trocheliolide A, a Hydroperoxycembranoidal Diterpene from the Octocoral Sarcophyton trocheliophorum.
A new hydroperoxycembranoidal diterpene, trocheliolide A (1), was isolated from the octocoral Sarcophyton trocheliophorum. The structure of 1 was elucidated on the basis of spectroscopic and mass spectrometric methods.


Patterns and Predictors of Change in Moderate-to-Vigorous Physical Activity Over Time in Multiple Sclerosis.
Physical inactivity is common in persons with multiple sclerosis (MS), but there is very little known about the pattern and predictors of changes in physical activity over time. This study examined changes in moderate-to-vigorous physical activity (MVPA) over a 30-month time period and the demographic and clinical predictors of such changes in relapsing-remitting MS (RRMS). 269 persons with MS wore an accelerometer for a 7-day period and completed a demographic/clinical scale every 6 months over a 30-month period. Data were analyzed using latent class growth modeling (LCGM). LCGM identified a two-class model for changes in levels of MVPA over time. Class 1 involved higher initial levels of MVPA and linear decreases in MVPA over time, whereas Class 2 involved lower initial levels of MVPA and linear increases in MVPA over time. LCGM further indicated that males were more likely (OR = 5.8, P < .05) and those with higher disability status were less likely (OR = 0.51, P < .05) to belong to Class 1 than Class 2. Levels of MVPA change over time in persons with RRMS and the pattern of change suggests that behavioral physical activity interventions for persons with MS might target men and those with lower disability.


Eco-evolutionary perspectives on emergence, dispersion and dissolution of historical Dutch commons.
Historical commons represent self-governed governance regimes that regulate the use and management of natural and man-made shared resources. Despite growing scientific interests, analyses of commons evolution and temporal dynamics are rare and drivers of change (birth, adaptation, dissolution) remain obscure. We apply an interdisciplinary approach and address these issues from an eco-evolutionary perspective. Analyses of > 400 Dutch commons over more than a millennium (between the 9th and the 20th century) uncovered that most commons originated between 1200 and 1700, and that there was a particularly high rate of evolution during 1300-1550, a pattern intermediate to gradualism and punctuated equilibrium in biological evolution. Dissolutions of commons were rare prior to 1800 and peaked around 1850, comparable to a mass extinction in biology. Temporal trends in number, spatial distribution, density, and dispersion of historical commons were distinctive and resembled developments seen at the levels of species and individuals in the growth of biological communities and populations, in that they showed signs of saturation determined by the abundance and distribution of resources. The spatiotemporal dynamics of commons also pointed to important roles of social, economic and political factors, such as new reclamations of resources and pressure on resources due to population growth. Despite internal and external pressures, the self-governing commons studied here were very successful, in the sense that they persisted for on average >350 years. There was a weak positive relationship between the use of multiple resources and the lifespan of commons, resembling associations between diversity and persistence seen in biological systems. It is argued that eco-evolutionary perspectives can further the understanding of the long-term dynamics of commons as institutions for collective action, vitalize future research, improve management of shared goods, and advise about sustainable utilization of finite resources.


[Nocturnal stridor in multiple system atrophy].
Multiple system atrophy is a neurodegenerative disorder with parkinsonism, cerebellar ataxia and autonomic dysfunction. The occurrence of nocturnal stridor in patients with multiple system atrophy is associated with a decreased life expectancy. This is what makes adequate treatment so important. Nevertheless, stridor goes often unrecognized and there is frequent uncertainty about the therapeutic options. We present a 58-year-old woman with multiple system atrophy and nocturnal stridor. Direct laryngoscopy showed bilateral paresis of the vocal cords and glottic stenosis. Since the outcome of treatment with continuous positive airway pressure (CPAP) was unsatisfactory, a tracheotomy was performed after which the stridor disappeared. Polysomnography and laryngoscopy are the important diagnostic tools for this condition, after the clinical interview. Tracheotomy used to be the only therapeutic option for stridor. However, recent studies show a comparable effectiveness of non-invasive treatment with CPAP, provided there is a correct indication. The choice of therapy is a multidisciplinary process in which the neurologist, the sleep medicine specialist, the otolaryngologist, and the patient are involved.


Neural substrates of sound-touch synesthesia after a thalamic lesion.
Neural plasticity induced by stroke can mediate positive outcomes, such as recovery of function, but can also result in the formation of abnormal connections with negative consequences for perception and cognition. In three experiments using blood-oxygen level dependent (BOLD) functional magnetic resonance imaging, we examined the neural substrates of acquired auditory-tactile synesthesia, in which certain sounds can produce an intense somatosensory tingling sensation in a patient with a thalamic lesion. Compared with nine normal controls, the first experiment showed that the patient had a threefold greater BOLD response to sounds in the parietal operculum, the location of secondary somatosensory cortex. We hypothesized that this abnormal opercular activity might be the neural substrate of the patient's synesthesia. Supporting this hypothesis, the second experiment demonstrated that sounds that produced no somatosensation did not evoke a BOLD response in the operculum, while sounds that produced strong somatosensations evoked large BOLD responses. These abnormal responses may have resulted from plasticity induced by the loss of somatosensory inputs. Consistent with this idea, in the third experiment, BOLD responses to somatosensory stimulation were significantly weaker in the patient's operculum than in normal controls. These experiments demonstrate a double dissociation in the patient's secondary somatosensory cortex (increased responses to auditory stimulation and decreased responses to somatosensory stimulation), and suggest both that stroke-induced plasticity can result in abnormal connections between sensory modalities that are normally separate, and that synesthesia can be caused by inappropriate connections between nearby cortical territories.


Global public-private partnerships against neglected diseases: building governance structures for effective outcomes.
Focusing on the problem of pharmaceutical R&D for drugs and vaccines against neglected diseases in developing countries, this article argues that the effectiveness of global health partnerships potentially lies in their capacity to address the problem of dual market failures: on a first level they may tackle the poverty induced lack of effective demand for health products which impedes the creation of market-financed innovative products. On a second level, they may help overcoming hold-up problems and underinvestment induced by the complexity of neglected diseases R&D. Yet, organizing transactions within a partnership is not a panacea against these problems: a crucial determinant of success is proper ownership structures. They need to respond to (i) the degree to which the respective parties value the partnership outcome, (ii) the relative importance of the investment of the parties, and (iii) the nature of the partnership outcome. The argument developed in the analysis is built on an integrated framework combining insights from incomplete contracting theory and public goods economics. It is supported by a preliminary statistical analysis of 17 GHPs.


DAG lipase involvement in depolarization-induced suppression of inhibition: does endocannabinoid biosynthesis always meet the demand?
Hippocampal depolarization-induced suppression of inhibition (DSI) is a robust form of short-term synaptic plasticity. DSI is mediated by endocannabinoid signaling. Since this discovery, pinning down the endogenous cannabinoid receptor ligand that mediates DSI has been problematic. Blocking degradation of the endocannabinoid 2-arachidonoyl glycerol (2-AG) lengthens DSI, which seems to indicate that 2-AG mediates DSI. In contrast, pharmacological inhibition of the 2-AG-synthesizing enzyme diacylglycerol lipase (DAGL) has yielded conflicting results: DAGL inhibitors often fail to block hippocampal DSI. Recently, 2 studies seem to have cornered this problem using DAGL knockout mice. Hippocampal DSI is absent in DAGL-α knockout mice, pointing to a key role for 2-AG in DSI. However, these studies do not reconcile the discrepancy with pharmacological experiments. Here, we argue that the seeming contradiction between results from pharmacological and genetic approaches may be explained in several ways. We suggest that the contradiction may be resolved by taking a different perspective on endocannabinoid signaling: in some forms of endocannabinoid-mediated signaling endocannabinoids might not be necessarily produced "on demand" but presynthesized and stored until needed.


Disordered eating-related cognition and psychological flexibility as predictors of psychological health among college students.
The present cross-sectional study investigated the relation among disordered eating-related cognition, psychological flexibility, and poor psychological outcomes among a nonclinical college sample. As predicted, conviction of disordered eating-related cognitions was positively associated with general psychological ill-health and emotional distress in interpersonal contexts. Disordered eating-related cognition was also inversely related to psychological flexibility, which was inversely related to poor psychological health and emotional distress in interpersonal contexts. The combination of disordered eating-related cognition and psychological flexibility accounted for the proportion of variance of these poor psychological outcomes greater than disordered eating-related cognition alone. Finally, psychological flexibility accounted for the proportion of variance of these negative psychological variables greater than did disordered eating-related cognition.


Doxycycline, an Inhibitor of Mitochondrial Biogenesis, Effectively Reduces Cancer Stem Cells (CSCs) in Early Breast Cancer Patients: A Clinical Pilot Study.
Background and objectives: Cancer stem cells (CSCs) have been implicated in tumor initiation, recurrence, metastatic spread and poor survival in multiple tumor types, breast cancers included. CSCs selectively overexpress key mitochondrial-related proteins and inhibition of mitochondrial function may represent a new potential approach for the eradication of CSCs. Because mitochondria evolved from bacteria, many classes of FDA-approved antibiotics, including doxycycline, actually target mitochondria. Our clinical pilot study aimed to determine whether short-term pre-operative treatment with oral doxycycline results in reduction of CSCs in early breast cancer patients. Methods: Doxycycline was administered orally for 14 days before surgery for a daily dose of 200 mg. Immuno-histochemical analysis of formalin-fixed paraffin-embedded (FFPE) samples from 15 patients, of which 9 were treated with doxycycline and 6 were controls (no treatment), was performed with known biomarkers of "stemness" (CD44, ALDH1), mitochondria (TOMM20), cell proliferation (Ki67, p27), apoptosis (cleaved caspase-3), and neo-angiogenesis (CD31). For each patient, the analysis was performed both on pre-operative specimens (core-biopsies) and surgical specimens. Changes from baseline to post-treatment were assessed with MedCalc 12 (unpaired t-test) and ANOVA. Results: Post-doxycycline tumor samples demonstrated a statistically significant decrease in the stemness marker CD44 (p-value < 0.005), when compared to pre-doxycycline tumor samples. More specifically, CD44 levels were reduced between 17.65 and 66.67%, in 8 out of 9 patients treated with doxycycline. In contrast, only one patient showed a rise in CD44, by 15%. Overall, this represents a positive response rate of nearly 90%. Similar results were also obtained with ALDH1, another marker of stemness. In contrast, markers of mitochondria, proliferation, apoptosis, and neo-angiogenesis, were all similar between the two groups. Conclusions: Quantitative decreases in CD44 and ALDH1 expression are consistent with pre-clinical experiments and suggest that doxycycline can selectively eradicate CSCs in breast cancer patients in vivo. Future studies (with larger numbers of patients) will be conducted to validate these promising pilot studies.


Objective assessment of intraoperative skills for robot-assisted partial nephrectomy (RAPN).
RAPN training usually takes place in-vivo and methods vary across countries/institutions. No common system exists to objectively assess trainee capacity to perform RAPN at predetermined performance levels prior to in-vivo practice. The identification of objective performance metrics for RAPN training is a crucial starting point to improve training and surgical outcomes. The authors sought to examine the reliability, construct and discriminative validity of objective intraoperative performance metrics which best characterize the optimal and suboptimal performance of a reference approach for training novice RAPN surgeons. Seven Novice and 9 Experienced RAPN surgeons video recorded one or two independently performed RAPN procedures in the human. The videos were anonymized and two experienced urology surgeons were trained to reliably score RAPN performance, using previously developed metrics. The assessors were blinded to the performing surgeon, hospital and surgeon group. They independently scored surgeon RAPN performance. Novice and Experienced group performance scores were compared for procedure steps completed and errors made. Each group was divided at the median for Total Errors score, and subgroup scores (i.e., Novice HiErrs and LoErrs, Experienced HiErrs and LoErrs) were compared. The mean inter-rater reliability (IRR) for scoring was 0.95 (range 0.84-1). Compared with Novices, Experienced RAPN surgeons made 69% fewer procedural Total Errors. This difference was accentuated when the LoErr Expert RAPN surgeon's performance was compared with the HiErrs Novice RAPN surgeon's performance with an observed 170% fewer Total Errors. GEARS showed poor reliability (Mean IRR = 0.44; range 0.0-0.8), for scoring RAPN surgical performance. The RAPN procedure metrics reliably distinguish Novice and Experienced surgeon performances. They further differentiated performance levels within a group with similar experiences. Reliable and valid metrics will underpin quality-assured novice RAPN surgical training.


Arbitrary digital pulse sequence generator with delay-loop timing.
We propose an idea of an electronic multi-channel arbitrary digital sequence generator with temporal granularity equal to two clock cycles. We implement the generator with 32 channels using a low-cost ARM microcontroller and demonstrate its capability to produce temporal delays ranging from tens of nanoseconds to hundreds of seconds, with 24 ns timing granularity and linear scaling of delay with respect to the number of delay loop iterations. The generator is optionally synchronized with an external clock source to provide 100 ps jitter and overall sequence repeatability within the whole temporal range. The generator is fully programmable and able to produce digital sequences of high complexity. The concept of the generator can be implemented using different microcontrollers and applied for controlling of various optical, atomic, and nuclear physics measurement setups.


Comparison of pyrometry and thermography for thermal analysis of thermite reactions.
This study examines the thermal behavior of a laser ignited thermite composed of aluminum and bismuth trioxide. Temperature data were collected during the reaction using a four-color pyrometer and a high-speed color camera modified for thermography. The two diagnostics were arranged to collect data simultaneously, with similar fields of view and with similar data acquisition rates, so that the two techniques could be directly compared. Results show that at initial and final stages of the reaction, a lower signal-to-noise ratio affects the accuracy of the measured temperatures. Both diagnostics captured the same trends in transient thermal behavior, but the average temperatures measured with thermography were about 750 K higher than those from the pyrometer. This difference was attributed to the lower dynamic range of the thermography camera's image sensor, which was unable to resolve cooler temperatures in the field of view as well as the photomultiplier tube sensors in the pyrometer. Overall, while the camera could not accurately capture the average temperature of a scene, its ability to capture peak temperatures and spatial data make it the preferred method for tracking thermal behavior in thermite reactions.


Application of Mixed Methods in Health Services Management Research: A Systematic Review.
Mixed methods research (MMR) is versatile, pragmatic, and adaptable to constraints and opportunities during a research process. Although MMR has gain popularity in health services management research, little is known about how the research approach has been used and the quality of research. We conducted a systematic review of 198 MMR articles published in selected U.S.-based and international health services management journals from 2000 through 2018 to examine the extent of MMR application and scientific rigor. Results showed limited, yet increasing, use of MMR and a high degree of correspondence between MMR designs and study purposes. However, most articles did not clearly justify using MMR designs and the reporting of method details and research integration were inadequate in a significant portion of publications. We propose a checklist to assist the preparation and review of MMR manuscripts. Additional implications and recommendations to improve transparency, rigor, and quality in MMR are discussed.


Clinically relevant outcomes based on analysis of pooled data from 2 trials of duloxetine in patients with knee osteoarthritis.
To determine response with duloxetine versus placebo in patients with osteoarthritis (OA) of the knee using the Outcome Measures in Rheumatoid Arthritis Clinical Trials-Osteoarthritis Research Society International (OMERACT-OARSI) responder index and other clinically relevant outcomes including minimal clinically important improvement (MCII) and patient acceptable symptom state (PASS) for pain and function. Data were pooled from two 13-week, randomized, double-blind, placebo-controlled trials comparing duloxetine 60 to 120 mg/day with placebo in patients with symptomatic OA of the knee. Treatment response was determined according to the OMERACT-OARSI responder index, ≥ 30% pain reduction, ≥ 50% pain reduction, and MCII and PASS for pain and function. (ClinicalTrials.gov identifiers NCT00433290 and NCT00408421) Duloxetine-treated patients were 33% more likely to experience an OMERACT-OARSI response than placebo-treated patients [p < 0.001, number needed to treat (NNT) = 6]. A significantly greater percentage of duloxetine-treated patients, compared with placebo-treated patients, reported ≥ 30% improvement in pain from baseline to endpoint (p < 0.001, NNT = 5) and ≥ 50% improvement in pain relative to baseline (p < 0.001, NNT = 7). The duloxetine-treated patients were also more likely to fulfill MCII criteria for pain (p < 0.001, NNT = 6) and function (p < 0.001, NNT = 7), and to achieve PASS for pain (p < 0.001, NNT = 6) and function (p = 0.009, NNT = 9). More duloxetine-treated patients compared with placebo-treated patients experienced ≥ 1 treatment-emergent adverse event (p = 0.003, number needed to harm = 8). Significantly more patients receiving duloxetine than placebo achieved an OMERACT-OARSI response, improvements in pain and function exceeding the level accepted as MCII, and reached PASS. Results support the clinical relevance of outcomes of prior duloxetine studies in symptomatic OA of the knee.


Muscle patterns underlying voluntary modulation of co-contraction.
Manipulative actions involving unstable interactions with the environment require controlling mechanical impedance through muscle co-contraction. While much research has focused on how the central nervous system (CNS) selects the muscle patterns underlying a desired movement or end-point force, the coordination strategies used to achieve a desired end-point impedance have received considerably less attention. We recorded isometric forces at the hand and electromyographic (EMG) signals in subjects performing a reaching task with an external disturbance. In a virtual environment, subjects displaced a cursor by applying isometric forces and were instructed to reach targets in 20 spatial locations. The motion of the cursor was then perturbed by disturbances whose effects could be attenuated by increasing co-contraction. All subjects could voluntarily modulate co-contraction when disturbances of different magnitudes were applied. For most muscles, activation was modulated by target direction according to a cosine tuning function with an offset and an amplitude increasing with disturbance magnitude. Co-contraction was characterized by projecting the muscle activation vector onto the null space of the EMG-to-force mapping. Even in the baseline the magnitude of the null space projection was larger than the minimum magnitude required for non-negative muscle activations. Moreover, the increase in co-contraction was not obtained by scaling the baseline null space projection, scaling the difference between the null space projections in any block and the projection of the non-negative minimum-norm muscle vector, or scaling the difference between the null space projections in the perturbed blocks and the baseline null space projection. However, the null space projections in the perturbed blocks were obtained by linear combination of the baseline null space projection and the muscle activation used to increase co-contraction without generating any force. The failure of scaling rules in explaining voluntary modulation of arm co-contraction suggests that muscle pattern generation may be constrained by muscle synergies.


Dynamics of zebrafish fin regeneration using a pulsed SILAC approach.
The zebrafish owns remarkable regenerative capacities allowing regeneration of several tissues, including the heart, liver, and brain. To identify protein dynamics during fin regeneration we used a pulsed SILAC approach that enabled us to detect the incorporation of (13) C6 -lysine (Lys6) into newly synthesized proteins. Samples were taken at four different time points from noninjured and regrowing fins and incorporation rates were monitored using a combination of single-shot 4-h gradients and high-resolution tandem MS. We identified more than 5000 labeled proteins during the first 3 weeks of fin regeneration and were able to monitor proteins that are responsible for initializing and restoring the shape of these appendages. The comparison of Lys6 incorporation rates between noninjured and regrowing fins enabled us to identify proteins that are directly involved in regeneration. For example, we observed increased incorporation rates of two actinodin family members at the actinotrichia, which is a hairlike fiber structure at the tip of regrowing fins. Moreover, we used quantitative real-time RNA measurements of several candidate genes, including osteoglycin, si:ch211-288h17.3, and prostaglandin reductase 1 to correlate the mRNA expression to Lys6 incorporation data. This novel pulsed SILAC methodology in fish can be used as a versatile tool to monitor newly synthesized proteins and will help to characterize protein dynamics during regenerative processes in zebrafish beyond fin regeneration.


Comparison of XTT and Alamar blue assays in the assessment of the viability of various human cancer cell lines by AT-101 (-/- gossypol).
This study compared the two different commercially available in vitro viability assays: XTT and Alamar blue (AB), to detect anti-proliferative effects of AT-101, a cotton plant extract, on six different human carcinoma cell lines including: prostate (PC-3 and DU-145), breast (MCF-7 and MDA-MB-231), and ovary (OVCAR-3 and MDAH 2774) in a time- and dose-dependent manner. Cells were exposed to AT-101 in the concentration range of 2.5-40 µM for 24, 48, and 72 h. The AB assay was slightly more sensitive than the XTT assay in the evaluation of AT-101 at 24 h, suggesting that the AB assay might be used for detecting early changes in cell viability as compared to the XTT assay. Moreover, the AB assay showed less intra-assay variability as compared to the XTT. The non-toxic, non-radioactive AB metabolism assay allows rapid assessment of large numbers of samples, with simple equipment and at reduced cost for continuous monitoring of cancer cell viability, and, thus, should be accepted as a suitable alternative viability method.


Circulating Myeloperoxidase (MPO)-DNA complexes as marker for Neutrophil Extracellular Traps (NETs) levels and the association with cardiovascular risk factors in the general population.
Neutrophil extracellular traps (NETs) are DNA scaffolds enriched with antimicrobial proteins. NETs have been implicated in the development of various diseases, such as cardiovascular disease. Here, we investigate the association of demographic and cardiovascular (CVD) risk factors with NETs in the general population. Citrated plasma was collected from 6449 participants, aged ≥55 years, as part of the prospective population-based Rotterdam Study. NETs were quantified by measuring MPO-DNA complex using an ELISA. We used linear regression to determine the associations between MPO-DNA complex and age, sex, cardio-metabolic risk factors, and plasma markers of inflammation and coagulation. MPO-DNA complex levels were weakly associated with age (log difference per 10 year increase: -0.04 mAU/mL, 95% confidence interval [CI] -0.06;-0.02), a history of coronary heart disease (yes versus no: -0.10 mAU/mL, 95% CI -0.17;-0.03), the use of lipid-lowering drugs (yes versus no: -0.06 mAU/mL, 95% CI -0.12;-0.01), and HDL-cholesterol (per mmol/l increase: -0.07 mAU/mL/, 95% CI -0.12;-0.03). Older age, a history of coronary heart disease, the use of lipid-lowering drugs and higher HDL-cholesterol are weakly correlated with lower plasma levels of NETs. These findings show that the effect of CVD risk factors on NETs levels in a general population is only small and may not be of clinical relevance. This supports that NETs may play a more important role in an acute phase of disease than in a steady state situation.


[Smoke-free areas: persuasion and enforcement].
This paper presents the actions taken by the public health services in the city of Barcelona (Catalonia, Spain) to improve compliance with the requirements of the new 28/2005 tobacco control law. These were essentially informative at first, with a second phase where authority enforcement mechanisms were activated. In workplaces, educational settings and transport the law was incorporated without incidents nor relevant complaints, except for isolated incidents in some university or mass transport settings. In food establishments the process has been more complex. Estimating the frequency of related events, there are 17.5 formal citizen complaints for 100,000 person-years. Inspections generated by citizen complaints resulted in 3.3 administrative proceedings for 100,000 person-years, mostly for incurring in serious violations. Effectively enforcing the law required active information and communication policies, as well as the real enforcement by health authority. This caused an important workload to the public health services.


Nephrogenic adenoma: a form of adenomatous metaplasia of the bladder. A clinical and electron microscopical study.
3 additional cases of "nephrogenic adenoma" of the bladder are added to the 20 previously reported cases. Light and electron microscopic studies of the first 2 cases support the hypothesis of urothelial metaplasia. The name "adenomatous metaplasia" should replace "nephrogenic adenoma". Malignant change occurred in the third case, suggesting that this ought to be regarded as a potentially dangerous from of metaplasia. In the absence of malignant change, ileocaecocystoplasty with subtotal cystectomy afforded symptomatic cure in 2 cases: radical cystectomy was performed in the third because of malignant adenocarcinoma.


Compensatory induction of Fads1 gene expression in heterozygous Fads2-null mice and by diet with a high n-6/n-3 PUFA ratio.
In mammals, because they share a single synthetic pathway, n-6/n-3 ratios of dietary PUFAs impact tissue arachidonic acid (ARA) and DHA content. Likewise, SNPs in the human fatty acid desaturase (FADS) gene cluster impact tissue ARA and DHA. Here we tested the feasibility of using heterozygous Fads2-null-mice (HET) as an animal model of human FADS polymorphisms. WT and HET mice were fed diets with linoleate/α-linolenate ratios of 1:1, 7:1, and 44:1 at 7% of diet. In WT liver, ARA and DHA in phospholipids varied >2× among dietary groups, reflecting precursor ratios. Unexpectedly, ARA content was only <10% lower in HET than in WT livers, when fed the 44:1 diet, likely due to increased Fads1 mRNA in response to reduced Fads2 mRNA in HET. Consistent with the RNA data, C20:3n-6, which is elevated in minor FADS haplotypes in humans, was lower in HET than WT. Diet and genotype had little effect on brain PUFAs even though brain Fads2 mRNA was low in HET. No differences in cytokine mRNA were found among groups under unstimulated conditions. In conclusion, differential PUFA profiles between HET mice and human FADS SNPs suggest low expression of both FADS1 and 2 genes in human minor haplotypes.


Bacterial colonization immediately after installation on oral titanium implants.
Information on bacterial colonization immediately after dental implant insertion is limited. (1) To assess the early colonization on titanium implants immediately after placement and throughout the first 12 post-surgical weeks, (2) to compare the microbiota at interproximal subgingival implant and adjacent tooth sites. Subgingival plaque samples from implant and neighbouring teeth were studied by checkerboard DNA-DNA hybridization before surgery, 30 min after implant placement, and 1, 2, 4, 8, and 12 weeks after surgery. Comparing bacterial loads at implant sites between 30 min after placement with 1-week data showed that only the levels of Veillonella parvula (P<0.05) differed with higher loads at week 1 post-surgically. Week 12 data demonstrated significantly higher bacterial loads for 15/40 species at tooth sites compared with pre-surgery (P-values varying between 0.05 and 0.01). Between the period immediately after surgery and 12 weeks at implant sites, 29/40 species was more commonly found at 12 weeks. Included among these bacteria at implant sites were Porphyromonas gingivalis (P<0.05), Tannerella forsythia, (P<0.01), and Treponema denticola (P<0.001). Immediately post-surgery 5.9% of implants, and 26.2% of teeth, and at week 12, 15% of implants, and 39.1% of teeth harbored Staphylococcus aureus. Comparing tooth and implant sites, significantly higher bacterial loads were found at tooth sites for 27/40 species after 30 min following implant placement. This difference increased to 35/40 species at 12 weeks post-surgically. Bacterial colonization occurred within 30 min after implant placement. Early colonization patterns differed between implant and tooth surfaces.


[Postluminescence of the marine green alga Ulva fenestrata in millisecond and decisecond attenuation intervals].
Excitation and radiation spectra of the stationary level of postluminescence of marine green alga tallom were measured in millisecond and decisecond attenuation intervals. On the basis of these data and of temperature and light relationships of postluminescence the conclusion is drawn concerning the existence of three independent pigment systems within the range of Photosystem-2, which are responsible for luminescence generation within the given limits of attenuation.


Analysis of the apolipoprotein B gene and messenger ribonucleic acid in abetalipoproteinemia.
The apolipoprotein B-100 (apoB-100) gene in leukocytes and the apoB-100 messenger RNA (mRNA) and translated apolipoprotein in the livers from normal and abetalipoproteinemic individuals were evaluated. Four complementary DNA probes for apoB-100 covering the 5', middle, and 3' regions of the apoB-100 mRNA were utilized and Southern blot analysis indicated that the apoB-100 gene is present in abetalipoproteinemia without major insertions or deletions. Polyadenylated hepatic apoB-100 mRNA from two abetalipoproteinemic patients was normal in size, and the concentration of apoB-100 mRNA was increased sixfold compared with control hepatic apoB-100 mRNA levels. ApoB-100 was detected in hepatocytes of abetalipoproteinemic patients by immunohistochemical techniques. These results indicate that the biochemical defect in abetalipoproteinemic patients studied is most consistent with a posttranslational defect in apoB-100 processing or secretion with an up-regulation of the apoB-100 mRNA.


Efficient and Adaptive Methods for Computing Accurate Potential Surfaces for Quantum Nuclear Effects: Applications to Hydrogen-Transfer Reactions.
We present two sampling measures to gauge critical regions of potential energy surfaces. These sampling measures employ (a) the instantaneous quantum wavepacket density, an approximation to the (b) potential surface, its (c) gradients, and (d) a Shannon information theory based expression that estimates the local entropy associated with the quantum wavepacket. These four criteria together enable a directed sampling of potential surfaces that appears to correctly describe the local oscillation frequencies, or the local Nyquist frequency, of a potential surface. The sampling functions are then utilized to derive a tessellation scheme that discretizes the multidimensional space to enable efficient sampling of potential surfaces. The sampled potential surface is then combined with four different interpolation procedures, namely, (a) local Hermite curve interpolation, (b) low-pass filtered Lagrange interpolation, (c) the monomial symmetrization approximation (MSA) developed by Bowman and co-workers, and (d) a modified Shepard algorithm. The sampling procedure and the fitting schemes are used to compute (a) potential surfaces in highly anharmonic hydrogen-bonded systems and (b) study hydrogen-transfer reactions in biogenic volatile organic compounds (isoprene) where the transferring hydrogen atom is found to demonstrate critical quantum nuclear effects. In the case of isoprene, the algorithm discussed here is used to derive multidimensional potential surfaces along a hydrogen-transfer reaction path to gauge the effect of quantum-nuclear degrees of freedom on the hydrogen-transfer process. Based on the decreased computational effort, facilitated by the optimal sampling of the potential surfaces through the use of sampling functions discussed here, and the accuracy of the associated potential surfaces, we believe the method will find great utility in the study of quantum nuclear dynamics problems, of which application to hydrogen-transfer reactions and hydrogen-bonded systems is demonstrated here.


Management of asymptomatic severe aortic stenosis.
Patients with symptomatic severe aortic stenosis (AS) benefit from aortic valve replacement surgery, but the management of patients with asymptomatic severe AS is more controversial. While cholesterol and angiotensin have been linked to AS progression, we should await the results of ongoing randomized trials before medical therapy to lower cholesterol or inhibit angiotensin can be recommended to limit disease progression. Clinical factors, echocardiographic parameters, valve morphology, exercise stress testing results, and cardiac biomarkers may be useful in identifying patients who will have early development of symptoms during follow-up and require closer monitoring. The risks associated with aortic valve replacement outweigh the benefits in the majority of patients with asymptomatic severe AS.


Use of a spinal model to quantify the forces and motion that occur during therapists' tests of spinal motion.
Despite the widespread use of spinal mobilization, little is known about the forces used or the accuracy of therapists in estimating the forces they use in administering the technique. The purposes of this study were to quantify the forces used and to determine the accuracy of therapists in applying forces on a mechanical model. Ten physical therapists participated. A spinal model was used to measure applied force and displacement under different conditions of stiffness. The therapists applied oscillatory posteroanterior mobilizations to the model under three different conditions of stiffness. Mean peak forces across grades and stiffness levels ranged between 57.59 and 178.27 N. The forces were generally lower in the least stiff condition. Displacement varied with stiffness and mobilization grade. In the least stiff condition, the mean displacement varied between 2.25 and 3.45 mm for grades 1 to 4, respectively. Inter-therapist variability was high, and there was a systematic bias in underestimating the magnitude of applied force and in overestimating motion. The variability in force application and the general overestimation of motion detection may explain the poor reliability of measurements obtained with clinical tests based on motion palpation.


Back pain as the presenting symptom of generalised tetanus.
Back pain is a common presenting symptom in emergency departments and primary care across the UK. The extensive differential diagnosis includes mechanical, infective, thoracic, abdominal and vascular causes. This case report describes a patient who presented with lower back pain with a rare diagnosis, which is becoming more common in certain population groups, and emphasises the importance of clinical skills and insuring adequate tetanus prophylaxis.


Solving the Nonalignment of Methods and Approaches Used in Microplastic Research to Consistently Characterize Risk.
The lack of standard approaches in microplastic research limits progress in the abatement of plastic pollution. Here, we propose and test rescaling methods that are able to improve the alignment of methods used in microplastic research. We describe a method to correct for the differences in size ranges as used by studies reporting microplastic concentrations and demonstrate how this reduces the variation in aqueous-phase concentrations caused by method differences. We provide a method to interchange between number, volume, and mass concentrations using probability density functions that represent environmental microplastic. Finally, we use this method to correct for the incompatibility of data as used in current species sensitivity distributions (SSDs), caused by differences in the microplastic types used in effect studies and those in nature. We derived threshold effect concentrations from such a corrected SSD for freshwater species. Comparison of the rescaled exposure concentrations and threshold effect concentrations reveals that the latter would be exceeded for 1.5% of the known surface water exposure concentrations worldwide. Altogether, this toolset allows us to correct for the diversity of microplastic, to address it in a common language, and to assess its risks as one environmental material.


Strategies to Improve Information Transfer for Multitrauma Patients.
The aim of this multiphase mixed-method study was to improve access, flow, and consistency of information transfer for multitrauma patients leaving the Emergency Department. Methods included literature review, focus group interviews, chart audits, staff surveys, and a review of international trauma forms to inform an intervention developed with a researcher-led, clinician stakeholder group. Analysis included descriptive and inferential statistics. Baseline data revealed variability existed in patient-care documentation, showing little standardization. Improvement strategies implemented included a gold standard for information embedded in handover tools, raising staff awareness of complexities for information transfer. Improvement was seen in communication between wards coordinating transfer, improved documentation, decreased information duplication, improved legibility, and increased ease and efficiency in navigating to key information. Improvement in communication at patient transition is essential to continuity of safe, effective care, and is impacted by complex interactions between multiple factors. Difficulty increases for patients with high acuity.


The impact of anti-eosinophilic therapy on exercise capacity and inspiratory muscle strength in patients with severe asthma.
Exercise limitation is frequently described among asthmatic patients and could be related to different mechanisms of the pulmonary, cardiovascular and muscular systems. Despite this, cardiopulmonary exercise testing (CPET) does not have an established role in the management of severe asthma. The aim of our study was to investigate the role of CPET and inspiratory pressure measurement in exercise capacity and muscle strength in severe asthmatic patients treated with anti-IL-5 therapy. A monocentric observational study was conducted at Hanover Medical School, Germany, from April 2018 to June 2019. Patients affected by severe asthma treated with either mepolizumab or benralizumab were included. All patients underwent CPET before the initiation of antibody therapy and after 3 months, and follow-up visits were scheduled at 3, 6 and 12 months with plethysmography, inspiratory pressure measurement and blood gas analysis. 14 patients were enrolled: 10 (71.4%) females, median age 52 years (IQR 47-61). Seven patients were treated with benralizumab, seven with mepolizumab. Oxygen uptake (V'O2 peak) did not change significantly after 3 months of antibody treatment, while the mean value of the breathing reserve exhaustion reduced significantly from 78% to 60% (p=0.004). Whereas at baseline seven patients depleted the breathing reserve and two of them experienced oxygen desaturation during exercise, at 3 months no one presented any desaturation or breathing reserve exhaustion. The inspiratory pressure remained unchanged before and after the antibody therapy. CPET could show hints of alveolar recruitment and ventilatory efficiency in severe asthma patients treated with antibody therapy.


Investigation of relationship of the mitochondrial DNA 16189 T>C polymorphism with metabolic syndrome and its associated clinical parameters in Turkish patients.
Mitochondrial DNA (mtDNA) polymorphisms have been implicated in the pathophysiology of human diseases. Among them, a T>C nucleotide transition on the 16189 nucleotide position of mtDNA has been studied in several metabolic diseases including diabetes and obesity. In this study we aimed to investigate the association of this polymorphism among Turkish metabolic syndrome patients. A total of 220 cases (70 MetS patients and 150 healthy control subjects) were evaluated for their mtDNA 16189 variant by PCR-RFLP technique. In addition, clinical and biochemical variables, such as cholesterol levels, body fat percentage, insulin resistance and presence of type II diabetes, were also evaluated. Overall frequency of polymorphic C allele was determined as 0.19 without a significant association with type II diabetes and metabolic syndrome. This may be partly due to ethnical differences of populations studied and may also be related to other genetic and environmental factors. Moreover, there were no significant associations with biochemical variables among metabolic syndrome patients, except LDL and suppressed cortisol (sup-cortisol) levels. Low levels of LDL and sup-cortisol were significantly associated with the mtDNA 16189 variant, though the biochemical mechanism underlying this effect is not clear. This is the first study involving a Turkish population on the mtDNA 16189 T>C polymorphism. Further studies with larger cohorts will be needed to elucidate its relation with metabolic syndrome as well as lipid metabolism.


Nonlinear Growth Models as Measurement Models: A Second-Order Growth Curve Model for Measuring Potential.
Recent methodological work has highlighted the promise of nonlinear growth models for addressing substantive questions in the behavioral sciences. In this article, we outline a second-order nonlinear growth model in order to measure a critical notion in development and education: potential. Here, potential is conceptualized as having three components-ability, capacity, and availability-where ability is the amount of skill a student is estimated to have at a given timepoint, capacity is the maximum amount of ability a student is predicted to be able to develop asymptotically, and availability is the difference between capacity and ability at any particular timepoint. We argue that single timepoint measures are typically insufficient for discerning information about potential, and we therefore describe a general framework that incorporates a growth model into the measurement model to capture these three components. Then, we provide an illustrative example using the public-use Early Childhood Longitudinal Study-Kindergarten data set using a Michaelis-Menten growth function (reparameterized from its common application in biochemistry) to demonstrate our proposed model as applied to measuring potential within an educational context. The advantage of this approach compared to currently utilized methods is discussed as are future directions and limitations.


Association of collagenase and tissue inhibitor of metalloproteinases (TIMP) with hypertrophic cell enlargement in the growth plate.
In the transition from proliferating to hypertrophic cell zones in the growth plate, there is an increased in chondrocyte cell volume and a corresponding decrease in collagen content to allow for cell enlargement. To substantiate our hypothesis that collagenase is responsible for these changes, growth plates from rats treated with bisphosphonate (HEBP) were compared histologically and biochemically with growth plates from normal and vitamin D and phosphate deficient (-VDP) rats. HEBP-treated rats developed an expanded hypertrophic cell zone (HCZ) characterized by the presence of two distinct populations of hypertrophic cells. The proximal hypertrophic cells were only 2-fold enlarged compared to the proliferating cells, whereas 1/6 of the distal hypertrophic cells were enlarged almost 5-fold and appeared morphologically identical with hypertrophic cells from normal and -VDP rats. The HEBP growth plates were divided into cross-sectional thirds and analyzed for active and latent collagenase. The juxta-metaphyseal (lower 1/3) cartilage contained 100% of the fully enlarged hypertrophic cells and appeared identical to those found in normal and -VDP growth plates, along with 81% of the active and 77% of the total collagenase. Collagenase and tissue inhibitor of metalloproteinases (TIMP) were measured in extracts of similarly divided tissues. The presence of true collagenas was confirmed by using [3H]-telopeptide-free collagen. TIMP levels were inversely related to the presence of active collagenase and cellular hypertrophy. Substantial levels of latent collagenase were found in the extracellular fluid at sites of active collagenolysis, but not in the fluid phase surrounding the 2-fold enlarged hypertrophic cells. It is postulated that increased amounts of active collagenase and insufficient levels of TIMP may account for the reduced collagen content seen in the lower HCZ of both -VDP and HEBP rickets. Unlike active collagenase, which remains localized by binding to collagen, latent enzyme is probably restricted in its mobility throughout the extracellular space by diffusion, itself, or the interstices of the extracellular matrix.


Next-Generation Hypoxic Cell Radiosensitizers: Nitroimidazole Alkylsulfonamides.
Innovations in the field of radiotherapy such as stereotactic body radiotherapy, along with the advent of radio-immuno-oncology, herald new opportunities for classical oxygen-mimetic radiosensitizers. The role of hypoxic tumor cells in resistance to radiotherapy and in suppression of immune response continues to endorse tumor hypoxia as a bona fide, yet largely untapped, drug target. Only nimorazole is used clinically as a radiosensitizer, and there is a dearth of new radiosensitizers in development. Here we present a survey of novel nitroimidazole alkylsulfonamides and document their cytotoxicity and ability to radiosensitize anoxic tumor cells in vitro. We use a phosphate prodrug approach to increase aqueous solubility and to improve tumor drug delivery. A 2-nitroimidazole and a 5-nitroimidazole analogue demonstrated marked tumor radiosensitization in either ex vivo assays of surviving clonogens or tumor regrowth delay.


Paraptosis: a unique cell death mode for targeting cancer.
Programmed cell death (PCD) is the universal process that maintains cellular homeostasis and regulates all living systems' development, health and disease. Out of all, apoptosis is one of the major PCDs that was found to play a crucial role in many disease conditions, including cancer. The cancer cells acquire the ability to escape apoptotic cell death, thereby increasing their resistance towards current therapies. This issue has led to the need to search for alternate forms of programmed cell death mechanisms. Paraptosis is an alternative cell death pathway characterized by vacuolation and damage to the endoplasmic reticulum and mitochondria. Many natural compounds and metallic complexes have been reported to induce paraptosis in cancer cell lines. Since the morphological and biochemical features of paraptosis are much different from apoptosis and other alternate PCDs, it is crucial to understand the different modulators governing it. In this review, we have highlighted the factors that trigger paraptosis and the role of specific modulators in mediating this alternative cell death pathway. Recent findings include the role of paraptosis in inducing anti-tumour T-cell immunity and other immunogenic responses against cancer. A significant role played by paraptosis in cancer has also scaled its importance in knowing its mechanism. The study of paraptosis in xenograft mice, zebrafish model, 3D cultures, and novel paraptosis-based prognostic model for low-grade glioma patients have led to the broad aspect and its potential involvement in the field of cancer therapy. The co-occurrence of different modes of cell death with photodynamic therapy and other combinatorial treatments in the tumour microenvironment are also summarized here. Finally, the growth, challenges, and future perspectives of paraptosis research in cancer are discussed in this review. Understanding this unique PCD pathway would help to develop potential therapy and combat chemo-resistance in various cancer.


Classic vs. Novel Antibacterial Approaches for Eradicating Dental Biofilm as Adjunct to Periodontal Debridement: An Evidence-Based Overview.
Periodontitis is a multifactorial chronic inflammatory disease that affects tooth-supporting soft/hard tissues of the dentition. The dental plaque biofilm is considered as a primary etiological factor in susceptible patients; however, other factors contribute to progression, such as diabetes and smoking. Current management utilizes mechanical biofilm removal as the gold standard of treatment. Antibacterial agents might be indicated in certain conditions as an adjunct to this mechanical approach. However, in view of the growing concern about bacterial resistance, alternative approaches have been investigated. Currently, a range of antimicrobial agents and protocols have been used in clinical management, but these remain largely non-validated. This review aimed to evaluate the efficacy of adjunctive antibiotic use in periodontal management and to compare them to recently suggested alternatives. Evidence from in vitro, observational and clinical trial studies suggests efficacy in the use of adjunctive antimicrobials in patients with grade C periodontitis of young age or where the associated risk factors are inconsistent with the amount of bone loss present. Meanwhile, alternative approaches such as photodynamic therapy, bacteriophage therapy and probiotics showed limited supportive evidence, and more studies are warranted to validate their efficiency.


Altered Levels of mRNAs for Calcium-Binding/Associated Proteins, Annexin A1, S100A4, and TMEM64, in Peripheral Blood Mononuclear Cells Are Associated with Osteoporosis.
Osteoporosis is the most common metabolic bone disease in the world. Since osteoporosis is clinically symptomless until the first fracture occurs, early diagnosis is critical. Calcium, along with calcium-binding and calcium-associated proteins, plays an important role in homeostasis, maintaining healthy bone metabolism. This study is aimed at investigating the level of calcium-binding/associated proteins, annexin A1, S100A4, and TMEM64, in peripheral blood mononuclear cells associated with osteoporosis and its clinical significance. The levels of mRNAs of annexin A1, S100A4, and TMEM64 in human peripheral blood mononuclear cells were evaluated among 48 osteopenia and 23 osteoporosis patients compared to 17 nonosteoporotic controls. Total RNAs were isolated from clinical samples, and quantitation of mRNA levels was performed using real-time quantitative PCR. The levels of mRNAs for calcium-binding proteins, annexin A1 and S100A4, and calcium-associated protein, TMEM64, in human peripheral blood mononuclear cells were significantly reduced in osteopenia and osteoporosis patients compared with nonosteoporotic controls (one-way ANOVA, P < 0.0001, P = 0.039, and P = 0.0195, respectively). Annexin A1 and TMEM64 mRNAs were also significantly reduced in female osteoporosis patients over the age of 50 years compared to nonosteoporotic controls (one-way ANOVA, P = 0.004 and P = 0.0037, respectively). ROC analysis showed that the reduction in the level of mRNA for annexin A1, S100A4, or TMEM64 in the patients' peripheral blood mononuclear cells has a good diagnostic value for osteoporosis. The results show for the first time that calcium-binding/associated proteins, annexin A1 and TMEM64, could be future diagnostic biomarkers for osteoporosis.


Intermedin is overexpressed in hepatocellular carcinoma and regulates cell proliferation and survival.
Angiogenesis is one of the hallmarks of tumor growth and metastasis. Identification of tumor angiogenic factors has been a critical component in understanding cancer biology and treatment. Intermedin (IMD) has been reported to promote angiogenesis in a rat ischemic model and human umbilical vascular endothelial cells. Our study sought to determine the role of IMD in human hepatocellular carcinoma tumor progression. High IMD mRNA expression levels were observed in human hepatocellular carcinoma tumors, even in early stage disease, by real-time RT-PCR. Immunohistochemical analysis of hepatocellular carcinoma clinical samples demonstrated that the tumor regions were significantly more immunoreactive for IMD than adjacent benign liver. Inhibition of IMD expression using RNA interference reduced cell proliferation in SK-Hep-1 and SNU-398 cells. Blockage of IMD signaling using either an antagonist peptide or a neutralizing antibody inhibited growth in a dose-dependent manner with concomitant induction of apoptosis, causing cleavage of caspase-8 and downregulation of Gli1 and Bcl2. Conversely, addition of IMD active peptide increased the phosphorylation level of extracellular signal-regulated kinase. Thus, IMD might play an important role in cell proliferation and survival of hepatocellular carcinoma. Our data suggests that IMD is a potential biomarker and therapeutic target for hepatocellular carcinoma.


Catestatin, vasostatin, cortisol, and visual analog scale scoring for stress assessment in healthy dogs.
The neuroendocrine glycoprotein chromogranin A is a useful biomarker for stress in humans. Chromogranin A epitopes catestatin and vasostatin can be measured in dogs using radioimmunoassays. The objective of this study was to evaluate catestatin and vasostatin as canine stress biomarkers in a clinical setting. Blood and saliva were collected from 33 healthy dogs that were familiar with sampling procedures and the animal hospital environment (control group) and 30 healthy dogs that were unacquainted (stress group). During sampling, stress behavior was scored by the same observer using visual analog scale (VAS). Plasma was analyzed for catestatin and vasostatin, serum for cortisol, and saliva for catestatin. Differences between groups were analyzed using two-sample t-tests and P<0.05 was considered significant. Stress behavior VAS score in the control group was significantly lower than in the stress group during blood (P=0.002) and saliva (P=0.0009) sampling. Serum cortisol and saliva catestatin concentrations in the stress group were higher than the control group (P=0.003 and P<0.0001, respectively). Serum cortisol concentrations were correlated with those of saliva (r=0.34, P=0.04) and plasma catestatin (r=0.29, P=0.03). Plasma catestatin and vasostatin did not differ significantly between groups. In conclusion, concentrations of saliva catestatin, and serum cortisol, and stress behavior VAS scores were significantly higher in the stress group. The results indicate that saliva catestatin may be useful as a biomarker for acute psychological stress in dogs.


Comparison of 2 concentrations of levobupivacaine in postoperative patient-controlled epidural analgesia.
To evaluate the quality of analgesia and the incidence of side effects of 2 different concentrations of levobupivacaine given as an equal milligram-bolus dose (5 mg) via patient-controlled epidural analgesia after abdominal surgery. Prospective, randomized, blinded study. Postanesthesia care unit and surgical wards of a university hospital. Forty-nine patients (41 with complete file) undergoing major lower abdominal surgery. The patients were randomly assigned to 2 groups: 1.5 mg/mL (bolus 3.3 mL, lockout 20 minutes, n = 26) and 5 mg/mL (bolus 1 mL, lockout 20 minutes, n = 23). The epidural catheter was inserted in the low thoracic level (T9-T12) before induction of a standardized general anesthesia technique. Demography, upper sensory block, visual analog scale scores at rest and after coughing, levobupivacaine and rescue morphine consumption, motor blockade, hemodynamics, postoperative nausea and vomiting, sedation, and patient satisfaction were recorded within the first 48 hours. Both groups were similar with regard to demographics, upper level of sensory blockade (T8), and visual analog scale pain scores at rest and after coughing, as well as levobupivacaine and subcutaneous rescue morphine consumption. Motor blockade in the lower limbs was very low in both groups. Arterial blood pressure was slightly lower in the 5 mg/mL group during the first 24 hours (P = 0.052). Five patients in the 1.5 mg/mL and 7 in the 5 mg/mL group had postoperative nausea and vomiting (P = 0.43). No other side effects were recorded, and all of the patients were satisfied. Administering the same dose of levobupivacaine in either a low or high concentration via patient-controlled epidural analgesia mode provides an equal quality of analgesia with no difference in the incidence of side effects.


In vivo temperature rise and acute inflammatory response in anesthetized human pulp tissue of premolars having Class V preparations after exposure to Polywave® LED light curing units.
To evaluate the influence of light emitted from two Polywave®, LED light-curing units (LCU) on in vivo pulp temperature (PT) rise and signs of acute inflammatory response in pulps of human premolar having deep Class V preparations. Sixty intact, first premolars from 15 volunteers requiring extraction received infiltrative anesthesia. A sterile thermocouple probe was inserted within the pulp tissue through a minute occlusal pulp exposure in only 45 teeth (n = 9) to continuously monitor PT (°C). A deep buccal Class V preparation was created, and the surface was exposed to light from a commercial Polywave LCU (Bluephase 20i (20i), Ivoclar Vivadent) or from an experimental LCU (Exp) using the exposure modes (EM): 1s/Exp and 2s/Exp, 10s/20i, 20s/20i, and 60s/20i. Peak PT and PT rise values above baseline (ΔT) data were evaluated using a one-way ANOVA followed by Tukey's post-hoc test (α = 5%). Teeth used for histological and immunohistochemical analyses (n = 3) were extracted approximately 2 h after exposure to the LCU. No significant difference in peak PT and ΔT values was noted between 2s/Exp and 20s/20i groups, which both exhibited higher values than 1s/Exp and 10s/20i groups (p < 0.001). Dilated and congested blood vessels were seen after exposure to 1s/Exp, 2s/Exp, or 60s/20i EMs. The expression of IL-1β and TNF-α tended to be more intense when higher irradiance was delivered. Although higher irradiance delivered over a short exposure caused lower PT rise than 5.5 °C, such EMs should be used with caution, as they have more potential to harm the pulp tissue.