Identification and functional characterization of cardiac pacemaker cells in zebrafish.
In the mammalian heart a conduction system of nodes and conducting cells generates and transduces the electrical signals evoking myocardial contractions. Specialized pacemaker cells initiating and controlling cardiac contraction rhythmicity are localized in an anatomically identifiable structure of myocardial origin, the sinus node. We previously showed that in mammalian embryos sinus node cells originate from cardiac progenitors expressing the transcription factors T-box transcription factor 3 (Tbx3) and Islet-1 (Isl1). Although cardiac development and function are strikingly conserved amongst animal classes, in lower vertebrates neither structural nor molecular distinguishable components of a conduction system have been identified, questioning its evolutionary origin. Here we show that zebrafish embryos lacking the LIM/homeodomain-containing transcription factor Isl1 display heart rate defects related to pacemaker dysfunction. Moreover, 3D reconstructions of gene expression patterns in the embryonic and adult zebrafish heart led us to uncover a previously unidentified, Isl1-positive and Tbx2b-positive region in the myocardium at the junction of the sinus venosus and atrium. Through their long interconnecting cellular protrusions the identified Isl1-positive cells form a ring-shaped structure. In vivo labeling of the Isl1-positive cells by transgenic technology allowed their isolation and electrophysiological characterization, revealing their unique pacemaker activity. In conclusion we demonstrate that Isl1-expressing cells, organized as a ring-shaped structure around the venous pole, hold the pacemaker function in the adult zebrafish heart. We have thereby identified an evolutionary conserved, structural and molecular distinguishable component of the cardiac conduction system in a lower vertebrate.


COVID-19 Pulmonary Pathology: The Experience of European Pulmonary Pathologists throughout the First Two Waves of the Pandemic.
Autoptic studies of patients who died from COVID-19 constitute an important step forward in improving our knowledge in the pathophysiology of SARS-CoV-2 infection. Systematic analyses of lung tissue, the organ primarily targeted by the disease, were mostly performed during the first wave of the pandemic. Analyses of pathological lesions at different times offer a good opportunity to better understand the disease and how its evolution has been influenced mostly by new SARS-CoV-2 variants or the different therapeutic approaches. In this short report we summarize responses collected from a questionnaire survey that investigated important pathological data during the first two pandemic waves (spring-summer 2020; autumn-winter 2020-2021). The survey was submitted to expert lung pathologists from nine European countries involved in autoptic procedures in both pandemic waves. The frequency of each lung lesion was quite heterogeneous among the participants. However, a higher frequency of pulmonary superinfections, both bacterial and especially fungal, was observed in the second wave compared to the first. Obtaining a deeper knowledge of the pathological lesions at the basis of this complex and severe disease, which change over time, is crucial for correct patient management and treatment. Autoptic examination is a useful tool to achieve this goal.


Plantibacter auratus sp. nov., in the family Microbacteriaceae.
Strain NCIMB 9991(T) is a Gram-positive, short rod-shaped, yellow-pigmented bacterium, with a high DNA G+C content, and was originally deposited in 1967 as Arthrobacter sp. The bacterium is aerobic, non-motile, catalase-positive and oxidase-negative. Comparative 16S rRNA gene sequencing studies demonstrated that this strain was highly related genealogically to Plantibacter flavus DSM 14012(T). Strain IAM 14817(T) (=NCIMB 9991(T)) has the following characteristics: the predominant menaquinones are MK-9 and MK-10, the DNA G+C content is 68 mol%, the diamino acid in the cell wall is 2,4-l-diaminobutyric acid and the muramic acid in the peptidoglycan is of an acetyl type. The major fatty acid is 12-methyl tetradecanoic acid (anteiso-C(15 : 0)), followed by 14-methyl hexadecanoic acid (anteiso-C(17 : 0)), 14-methyl pentadecanoic acid (iso-C(16 : 0)) and hexadecanoic acid (C(16 : 0)). On the basis of morphological, physiological and chemotaxonomic characteristics, together with DNA-DNA hybridization and 16S rRNA gene sequence comparison, strain IAM 14817(T) represents a novel species within the genus Plantibacter, for which the name Plantibacter auratus sp. nov. is proposed, with the type strain IAM 14817(T) (=NCIMB 9991(T)=NBRC 15702(T)).


Sensitivity to posed and genuine displays of happiness and sadness: a fMRI study.
The ability to distinguish posed from genuine facial displays of emotion and to act accordingly is a fundamental social skill. To investigate the neural correlates underpinning this sensitivity, we compared changes in brain activity associated with judging posed and genuine facial displays of happiness and sadness using fMRI. Photographs of displays were presented to 7 right-handed females who judged whether the person was feeling the target emotion and made yes/no responses. Results showed activity increases during the observation of genuine compared to posed happy displays in the left medial superior frontal gyrus (BA 9) and the middle cingulate cortex bilaterally (BAs 24 and 31). The same comparison for sad displays showed increased activity in the left medial superior frontal gyrus (BA 8), and in the right middle and triangular inferior frontal gyri (both BA 46). Participants who exhibited higher sensitivity to sad displays showed larger activity difference in the left medial superior frontal gyrus (BA 8). The present study provides evidence of differential neural activity when judging posed versus genuine facial displays of emotions. Further research is required to elucidate how this might impact social affective neuroscience and in what ways genuine facial displays can enhance our understanding of emotion perception.


ABO incompatible renal transplant: Transfusion medicine perspective.
Our study presents an analysis of the trends of ABO antibody titers and the TPE (Therapeutic Plasma Exchange) procedures required pre and post ABO incompatible renal transplant. Twenty nine patients underwent ABO incompatible renal transplant during the study period. The ABO antibody titers were done using the tube technique and titer reported was the dilution at which 1+ reaction was observed. The baseline titers of anti-A and anti-B antibodies were determined. The titer targeted was ≤8. Patients were subjected to 1 plasma volume exchange with 5% albumin and 2 units of AB group FFP (Fresh Frozen Plasma) in each sitting. TPE procedures post-transplant were decided on the basis of rising antibody titer with/ without graft dysfunction. The average number of TPE procedures required was 4-5 procedures/patient in the pretransplant and 2-3/patient in the post-transplant period. An average titer reduction of 1 serial dilution/procedure was noted for Anti-A and 1.1/procedure for Anti-B. Number of procedures required to reach the target titer was not significantly different for Anti-A and Anti-B (P = 0.98). Outcome of the transplant did not differ significantly by reducing titers to a level less than 8 (P = 0.32). The difference in the Anti-A and Anti-B titers at 14th day post-transplant was found to be clinically significant (P = 0.042). With an average of 4-5 TPE procedures pretransplant and 2-3 TPE procedures post transplants, ABO incompatible renal transplantations can be successfully accomplished.


Changes in health conditions in the Huainan coal mine in the past three decades.
Health conditions in the Huainan Coal Mine have improved greatly since 1950. In particular, the work and living environments, the underground dust concentration, the water supply and waste disposal systems, and the nutritional value of miners' food have become much better. Thus the rates of occupational, infectious, and parasitic diseases and nutritional deficiency diseases have decreased considerably among miners. The decreases in the prevalence of anthracosilicosis and hookworm have been especially striking.


The performance frontier: innovating for a sustainable strategy.
A mishmash of sustainability tactics does not add up to a sustainable strategy. Too often, companies launch sustainability programs with the hope that they'll be financially rewarded for doing good, even when those programs aren't relevant to their strategy and operations. They fail to understand the trade-offs between financial performance and performance on environmental, social, and governance (ESG) issues. Improving one typically comes at a cost to the other. But it doesn't have to be this way. It's possible to simultaneously boost both financial and ESG performance-if you focus strategically on issues that are the most "material" to shareholder value, and you develop major innovations in products, processes, and business models that prioritize those concerns. Maps being developed by the Sustainability Accounting Standards Board, which rank the materiality of 43 issues for 88 industries, can provide valuable guidance. And broad initiatives undertaken by three companies-Natura, Dow Chemical, and CLP Group-demonstrate the kind of innovations that will push performance into new territory. Communicating the benefits to stakeholders is also critical, which is why integrated reports, which combine financial and ESG reporting, are now gaining in popularity.


Bleeding during gonioscopy after deep sclerectomy.
To show a new complication after deep sclerectomy (DS). We described two eyes of two patients with open-angle glaucoma and cataract who were operated on of an uneventful phacoemulsification and DS with SK-gel implantation. Bleeding during gonioscopic examination occurred in both eyes 7 and 8 months after combined surgery. The blood originated from the vessels around the Descemet window, and was probably due to manipulation or rocking of the goniolens. Pressure was immediately applied to the gonioscopic lens and the hyphema was interrupted. These cases show the presence of new vessels around the Descemet window after DS with SK-gel. Bleeding from the Descemet window vessels can occur during gonioscopy even months after DS. We recommend conducting a careful gonioscopic examination in patients who have undergone DS to avoid this complication.


Performance of High-Sensitivity Cardiac Troponin Assays to Reflect Comorbidity Burden and Improve Mortality Risk Stratification in Older Adults With Diabetes.
Incorporation of comorbidity burden to inform diabetes management in older adults remains challenging. High-sensitivity cardiac troponins are objective, quantifiable biomarkers that may improve risk monitoring in older adults. We assessed the associations of elevations in high-sensitivity cardiac troponin I (hs-cTnI) and T (hs-cTnT) with comorbidities and improvements in mortality risk stratification. We used logistic regression to examine associations of comorbidities with elevations in either troponin (≥85th percentile) among 1,835 participants in the Atherosclerosis Risk in Communities (ARIC) Study with diabetes (ages 67-89 years, 43% male, 31% black) at visit 5 (2011-2013). We used Cox models to compare associations of high cardiac troponins with mortality across comorbidity levels. Elevations in either troponin (≥9.4 ng/L for hs-cTnI, ≥25 ng/L for hs-cTnT) were associated with prevalent coronary heart disease, heart failure, chronic kidney disease, pulmonary disease, hypoglycemia, hypertension, dementia, and frailty. Over a median follow-up of 6.2 years (418 deaths), both high hs-cTnI and high hs-cTnT further stratified mortality risk beyond comorbidity levels; those with a high hs-cTnI or hs-cTnT and high comorbidity were at highest mortality risk. Even among those with low comorbidity, a high hs-cTnI (hazard ratio 3.0 [95% CI 1.7, 5.4]) or hs-cTnT (hazard ratio 3.3 [95% CI 1.8, 6.2]) was associated with elevated mortality. Many comorbidities were reflected by both hs-cTnI and hs-cTnT; elevations in either of the troponins were associated with higher mortality risk beyond comorbidity burden. High-sensitivity cardiac troponins may identify older adults at high mortality risk and be useful in guiding clinical care of older adults with diabetes.


Adenylyl cyclase expression and modulation of cAMP in rat taste cells.
cAMP is a second messenger implicated in sensory transduction for taste. The identity of adenylyl cyclase (AC) in taste cells has not been explored. We have employed RT-PCR to identify the AC isoforms present in taste cells and found that AC 4, 6, and 8 are expressed as mRNAs in taste tissue. These proteins are also expressed in a subset of taste cells as revealed by immunohistochemistry. Alterations of cAMP concentrations are associated with transduction of taste stimuli of several classes. The involvement of particular ACs in this modulation has not been investigated. We demonstrate that glutamate, which is a potent stimulus eliciting a taste quality termed umami, causes a decrease in cAMP in forskolin-treated taste cells. The potentiation of this response by inosine monophosphate, the lack of response to d-glutamate, and the lack of response to umami stimuli in nonsensory lingual epithelium all suggest that the cAMP modulation represents umami taste transduction. Because cAMP downregulation via ACs can be mediated through Galpha(i) proteins, we examined the colocalization of the detected ACs with Galpha(i) proteins and found that 66% of AC8 immunopositive taste cells are also positive for gustducin, a taste-specific Galpha(i) protein. Whether AC8 is directly involved in signal transduction of umami taste remains to be established.


Quantitative versus qualitative analysis of peak expiratory flow in occupational asthma.
Peak expiratory flow rates (PEF) are often used to confirm the diagnosis of occupational asthma. The records are usually analysed qualitatively, and this may lead to interobserver disagreement. In this study the diagnostic value of a qualitative assessment of change in PEF was compared with objective measures of change in PEF and the results of a specific inhalation challenge test with plicatic acid. Twenty five patients with possible red cedar asthma recorded PEF six times a day for three weeks at work and for two weeks away from work and underwent a challenge test with plicatic acid at the end of the recording period. Patients were considered to have cedar asthma if the FEV1 after inhalation of plicatic acid was 15% or more below that on the control day. PEF was plotted against time and assessed qualitatively by three physicians. The graph was considered positive for cedar asthma if two of the three physicians agreed that PEF was lower at work than away from work. The 95% confidence interval for variation in PEF between periods at work and away from work was also obtained from 15 asthmatic patients without occupational asthma. Differences in PEF between periods at work and away from work were considered positive for occupational asthma in the patients exposed to cedar when they were outside the 95% confidence interval for variations in PEF in the 15 patients whose asthma was nonoccupational. Of the 25 men studied, 15 had a positive response to plicatic acid. The qualitative PEF analysis had a sensitivity of 87% and a specificity of 90% in confirming red cedar asthma as diagnosed by the specific challenge test. Among the objective methods tested, only the difference in mean PEF between the maximum PEF at weekends and the minimum PEF on working days had a sensitivity (93%) greater than that of the qualitative method and a similar specificity. The qualitative assessment of PEF is a good diagnostic test for cedar asthma. Only one objective method of PEF analysis proved to be slightly more sensitive than the qualitative method and similar in specificity.


Effects of elevated ozone concentration on CH4 and N2O emission from paddy soil under fully open-air field conditions.
We investigated the effects of elevated ozone concentration (E-O3) on CH4 and N2O emission from paddies with two rice cultivars: an inbred Indica cultivar Yangdao 6 (YD6) and a hybrid one II-you 084 (IIY084), under fully open-air field conditions in China. A mean 26.7% enhancement of ozone concentration above the ambient level (A-O3) significantly reduced CH4 emission at tillering and flowering stages leading to a reduction of seasonal integral CH4 emission by 29.6% on average across the two cultivars. The reduced CH4 emission is associated with O3-induced reduction in the whole-plant biomass (-13.2%), root biomass (-34.7%), and maximum tiller number (-10.3%), all of which curbed the carbon supply for belowground CH4 production and its release from submerged soil to atmosphere. Although no significant difference was detected between the cultivars in the CH4 emission response to E-O3, a larger decrease in CH4 emission with IIY084 (-33.2%) than that with YD6 (-7.0%) was observed at tillering stage, which may be due to the larger reduction in tiller number in IIY084 by E-O3. Additionally, E-O3 reduced seasonal mean NOx flux by 5.7% and 11.8% with IIY084 and YD6, respectively, but the effects were not significant statistically. We found that the relative response of CH4 emission to E-O3 was not significantly different from those reported in open-top chamber experiments. This study has thus confirmed that increasing ozone concentration would mitigate the global warming potential of CH4 and suggested consideration of the feedback mechanism between ozone and its precursor emission into the projection of future ozone effects on terrestrial ecosystem.


Colostrinin decreases spontaneous and induced mutation frequencies at the hprt locus in Chinese hamster V79 cells.
Colostrinin (CLN), a uniform mixture of low-molecular weight, proline-rich polypeptides, induces neurite outgrowth of pheochromocytoma cells, extends the lifespan of diploid fibroblast cells, inhibits beta amyloid-induced apoptosis and resulted in improved cognitive function when administered to Alzheimer's patients. Here we investigated CLN's antimutagenic activity in cells stressed oxidatively or exposed to chemical or physical agents. Our data show that CLN did not alter cell cycle kinetics and cloning efficiency, while it inhibited the development of spontaneous mutations at the coding region of the hypoxanthine phosphoribosyl-transferase (hprt) gene in Chinese hamster V79 cells. In a dose-dependent manner, CLN lowered reactive oxygen species (ROS)-induced frequency of cells resistant to 6-thioguanine (6-TG) to nearly background level. Likewise, CLN decreased the frequency of methyl methanesulfonate- or mitomycin C-induced mutations in V79 cells. Notably, CLN (at 100, 250, and 500 ng per ml concentrations) decreased UVA-induced mutation frequency, while only the highest dose of CLN also decreased significantly the number of UVB-induced 6-TG-resistant mutant cells. Similar results were obtained using cell cultures of human origin. Overall, our data show that CLN significantly lowers the mutation frequency that develops spontaneously or is induced by ROS, chemical and physical agents. CLN itself has no mutagenic activity. Therefore, CLN may be used in human therapies systemically and/or locally for the prevention of diseases associated with sequence alterations in genomic and mitochondrial DNA.


Engineered polymer-media interfaces for the long-term self-renewal of human embryonic stem cells.
We have developed a synthetic polymer interface for the long-term self-renewal of human embryonic stem cells (hESCs) in defined media. We successfully cultured hESCs on hydrogel interfaces of aminopropylmethacrylamide (APMAAm) for over 20 passages in chemically-defined mTeSR™1 media and demonstrated pluripotency of multiple hESC lines with immunostaining and quantitative RT-PCR studies. Results for hESC proliferation and pluripotency markers were both qualitatively and quantitatively similar to cells cultured on Matrigel™-coated substrates. Mechanistically, it was resolved that bovine serum albumin (BSA) in the mTeSR™1 media was critical for cell adhesion on APMAAm hydrogel interfaces. This study uniquely identified a robust long-term culture surface for the self-renewal of hESCs without the use of biologic coatings (e.g., peptides, proteins, or Matrigel™) in completely chemically-defined media that employed practical culturing techniques amenable to clinical-scale cell expansion.


Supersaturated self-nanoemulsifying drug delivery systems (Super-SNEDDS) enhance the bioavailability of the poorly water-soluble drug simvastatin in dogs.
This study investigates the potential of supersaturated self-nanoemulsifying drug delivery systems (super-SNEDDS) to improve the bioavailability of poorly water-soluble drugs compared to conventional SNEDDS. Conventional SNEDDS contained simvastatin (SIM) at 75% of the equilibrium solubility (S (eq)). Super-SNEDDS containing SIM at 150 and 200% of S (eq) were produced by subjecting the SNEDDS preconcentrates to a heating and cooling cycle. The super-SNEDDS were physically stable over 10 months. During in vitro lipolysis of SNEDDS and super-SNEDDS the SIM concentration in the aqueous phase increased for the first 30 min almost proportional to the drug loads and amounts of preconcentrate employed. The 200% drug-loaded super-SNEDDS generated an amorphous SIM precipitate at the end of in vitro lipolysis. In vivo, the relative bioavailability of SIM from super-SEDDDS increased significantly to 180 ± 53.3% (p = 0.014) compared to the dosing of two capsules of (dose equivalent) 75% drug-loaded SNEDDS. A significant increase in the terminal half-life of elimination was observed for super-SNEDDS (2.3 ± 0.6 h) compared to conventional SNEDDS (1.4 ± 0.3 h) as well as a decreased area under the curve ratio of the SIM metabolite simvastatin acid to the parent compound (0.57 ± 0.20 and 0.90 ± 0.3), possibly due to a combination of saturation effects on presystemic metabolising enzymes and prolonged absorption along the small intestine. In summary, this study demonstrated that super-SNEDDS are a viable formulation option to enhance the bioavailability of poorly water-soluble drugs such as simvastatin while reducing the pill burden by an increased drug load of SNEDDS.


Structural insight into precursor tRNA processing by yeast ribonuclease P.
Ribonuclease P (RNase P) is a universal ribozyme responsible for processing the 5'-leader of pre-transfer RNA (pre-tRNA). Here, we report the 3.5-angstrom cryo-electron microscopy structures of Saccharomyces cerevisiae RNase P alone and in complex with pre-tRNAPhe The protein components form a hook-shaped architecture that wraps around the RNA and stabilizes RNase P into a "measuring device" with two fixed anchors that recognize the L-shaped pre-tRNA. A universally conserved uridine nucleobase and phosphate backbone in the catalytic center together with the scissile phosphate and the O3' leaving group of pre-tRNA jointly coordinate two catalytic magnesium ions. Binding of pre-tRNA induces a conformational change in the catalytic center that is required for catalysis. Moreover, simulation analysis suggests a two-metal-ion SN2 reaction pathway of pre-tRNA cleavage. These results not only reveal the architecture of yeast RNase P but also provide a molecular basis of how the 5'-leader of pre-tRNA is processed by eukaryotic RNase P.


Selection of a skin test antigen for American visceral leishmaniasis.
Studies were designed to examine skin test responses to leishmanial antigens in American visceral leishmaniasis (AVL) in Brazil. We found that after recovery from AVL, patients had positive delayed hypersensitivity reactions to Leishmania. Different amounts of a soluble extract obtained from Leishmania donovani chagasi promastigotes were compared with whole L. d. chagasi promastigotes in persons with past AVL. The most effective soluble preparations tested contained 25 and 50 micrograms leishmanial protein. These produced positive responses in 95%-100% of the individuals with past AVL. The 25 micrograms protein dose was used in further studies. This preparation produced no positive responses in either normal controls, tuberculosis patients, or schistosomiasis patients, and less than 5% positive responses in persons with Chagas' disease. The same amount of soluble extract prepared from L. mexicana amazonensis produced 82% positive skin test responses in persons with past AVL. When persons living in an area endemic for AVL were skin tested with the 25 micrograms preparation of L. d. chagasi extract, 34.1% yielded positive tests with a low number of positive responses in young children and 48% positive in adults. Only 3.1% of the population studied had a history of AVL. We have found that positive delayed hypersensitivity response to a soluble Leishmania extract is a sensitive and specific indicator of previous infection with AVL.


Angiogenesis inhibitor vasohibin-1 enhances stress resistance of endothelial cells via induction of SOD2 and SIRT1.
Vasohibin-1 (VASH1) is isolated as an endothelial cell (EC)-produced angiogenesis inhibitor. We questioned whether VASH1 plays any role besides angiogenesis inhibition, knocked-down or overexpressed VASH1 in ECs, and examined the changes of EC property. Knock-down of VASH1 induced premature senescence of ECs, and those ECs were easily killed by cellular stresses. In contrast, overexpression of VASH1 made ECs resistant to premature senescence and cell death caused by cellular stresses. The synthesis of VASH1 was regulated by HuR-mediated post-transcriptional regulation. We sought to define the underlying mechanism. VASH1 increased the expression of (superoxide dismutase 2) SOD2, an enzyme known to quench reactive oxygen species (ROS). Simultaneously, VASH1 augmented the synthesis of sirtuin 1 (SIRT1), an anti-aging protein, which improved stress tolerance. Paraquat generates ROS and causes organ damage when administered in vivo. More VASH1 (+/-) mice died due to acute lung injury caused by paraquat. Intratracheal administration of an adenovirus vector encoding human VASH1 augmented SOD2 and SIRT1 expression in the lungs and prevented acute lung injury caused by paraquat. Thus, VASH1 is a critical factor that improves the stress tolerance of ECs via the induction of SOD2 and SIRT1.


Enhanced specular peaks in diffuse light scattering from weakly rough metal surfaces.
We employ Monte Carlo techniques based on the reduced Rayleigh equations to study an enhanced specular peak that appears in the light scattered from weakly rough metal surfaces. This peak is not associated with the specular reflection but instead appears, with finite angular width, at the specular angle of the mean diffusely scattered intensity. As is the case with backscattering enhancement, the specular peak arises from the interference of contributions of multiple-scattering processes related to surface plasmon polariton excitation. We demonstrate that the specular peak is seen clearly for surface roughness that has a conventional Gaussian power spectrum. Further, we show that the peak appears more distinctly for roughness whose power spectrum has a new rectangular form, which is proposed here with the intent of better isolation of the scattering processes essential to the specular peak. Finally, for a pair of rough surfaces that have appropriately correlated surface roughness, it is found that the cross correlation of scattered amplitudes presents a well-isolated specular peak, which directly demonstrates the constructive interference that produces the effect.


Frequency and temporal resolution in elderly listeners with good and poor word recognition.
There is a subgroup of elderly listeners with hearing loss who can be characterized by exceptionally poor speech understanding. This study examined the hypothesis that the poor speech-understanding performance of some elderly listeners is associated with disproportionate deficits in temporal resolution and frequency resolution, especially for complex signals. Temporal resolution, as measured by gap detection, and frequency resolution, as measured by the critical ratio, were examined in older listeners with normal hearing, older listeners with hearing loss and good speech-recognition performance, and older listeners with hearing loss and poor speech-recognition performance. Listener performance was evaluated for simple and complex stimuli and for tasks of added complexity. In addition, syllable recognition was assessed in quiet and noise. The principal findings were that older listeners with hearing loss and poor word-recognition performance did not perform differently from older listeners with hearing loss and good word recognition on the temporal resolution measures nor on the spectral resolution measures for relatively simple stimuli. However, frequency resolution was compromised for listeners with poor word-recognition abilities when targets were presented in the context of complex signals. Group differences observed for syllable recognition in quiet were eliminated in the noise condition. Taken together, the findings support the hypothesis that unusual deficits in word-recognition performance among elderly listeners were associated with poor spectral resolution for complex signals.


Cost-effectiveness of Housing First Intervention With Intensive Case Management Compared With Treatment as Usual for Homeless Adults With Mental Illness: Secondary Analysis of a Randomized Clinical Trial.
In the At Home/Chez Soi trial for homeless individuals with mental illness, the scattered-site Housing First (HF) with Intensive Case Management (ICM) intervention proved more effective than treatment as usual (TAU). To evaluate the cost-effectiveness of the HF plus ICM intervention compared with TAU. This is an economic evaluation study of data from the At Home/Chez Soi randomized clinical trial. From October 2009 through July 2011, 1198 individuals were randomized to the intervention (n = 689) or TAU (n = 509) and followed up for as long as 24 months. Participants were recruited in the Canadian cities of Vancouver, Winnipeg, Toronto, and Montreal. Participants with a current mental disorder who were homeless and had a moderate level of need were included. Data were analyzed from 2013 through 2019, per protocol. Scattered-site HF (using rent supplements) with off-site ICM services was compared with usual housing and support services in each city. The analysis was performed from the perspective of society, with days of stable housing as the outcome. Service use was ascertained using questionnaires. Unit costs were estimated in 2016 Canadian dollars. Of 1198 randomized individuals, 795 (66.4%) were men and 696 (58.1%) were aged 30 to 49 years. Almost all (1160 participants, including 677 in the HF group and 483 in the TAU group) contributed data to the economic analysis. Days of stable housing were higher by 140.34 days (95% CI, 128.14-153.31 days) in the HF group. The intervention cost $14 496 per person per year; reductions in costs of other services brought the net cost down by 46% to $7868 (95% CI, $4409-$11 405). The incremental cost-effectiveness ratio was $56.08 (95% CI, $29.55-$84.78) per additional day of stable housing. In sensitivity analyses, adjusting for baseline differences using a regression-based method, without altering the discount rate, caused the largest change in the incremental cost-effectiveness ratio with an increase to $60.18 (95% CI, $35.27-$86.95). At $67 per day of stable housing, there was an 80% chance that HF was cost-effective compared with TAU. The cost-effectiveness of HF appeared to be similar for all participants, although possibly less for those with a higher number of previous psychiatric hospitalizations. In this study, the cost per additional day of stable housing was similar to that of many interventions for homeless individuals. Based on these results, expanding access to HF with ICM appears to be warranted from an economic standpoint. isrctn.org Identifier: ISRCTN42520374.


The analgesic and anticonvulsant effects of piperine in mice.
Piperine, is the major active principal of black pepper. In traditional medicine, black pepper has been used as an analgesic, anti-inflammatory agent and in the treatment of epilepsy. This study was conducted to evaluate the in vivo analgesic and anticonvulsant effects of piperine in mice. The analgesic and anticonvulsant effects of piperine were studied in mice using acetic acid-induced writhing, tail flick assay, pentylenetetrazole (PTZ)- and picrotoxin (PIC)-induced seizures models. The intraperitoneal (i.p.) administration of piperine (30, 50 and 70 mg/kg) significantly inhibited (P<0.01) the acetic acid-induced writhing in mice, similar to the effect of indomethacin (20 mg/kg i.p.). In the tail flick assay, piperine (30 and 50 mg/kg, i.p.) and morphine (5 mg/kg, i.p.) caused a significant increase (P<0.01) in the reaction time of mice. Pre-treatment of animals with naloxone (5 mg/kg i.p.), reversed the analgesic effects of both piperine and morphine in the tail flick assay. Piperine (30, 50 and 70 mg/kg, i.p.) and standard drugs, valproic acid (200 mg/kg, i.p.), carbamazepine (30 mg/kg, i.p.) and diazepam (1 mg/kg, i.p.) significantly (P<0.01) delayed the onset of PTZ-and PIC-induced seizures in mice. These findings indicate that piperine exhibits analgesic and anticonvulsant effects possibly mediated via opioid and GABA-ergic pathways respectively. Moreover, piperine being the main constituent of black pepper, may be contributing factor in the medicinal uses of black pepper in pain and epilepsy.


DOT1L Activity Promotes Proliferation and Protects Cortical Neural Stem Cells from Activation of ATF4-DDIT3-Mediated ER Stress In Vitro.
Growing evidence suggests that the lysine methyltransferase DOT1L/KMT4 has important roles in proliferation, survival, and differentiation of stem cells in development and in disease. We investigated the function of DOT1L in neural stem cells (NSCs) of the cerebral cortex. The pharmacological inhibition and shRNA-mediated knockdown of DOT1L impaired proliferation and survival of NSCs. DOT1L inhibition specifically induced genes that are activated during the unfolded protein response (UPR) in the endoplasmic reticulum (ER). Chromatin-immunoprecipitation analyses revealed that two genes encoding for central molecules involved in the ER stress response, Atf4 and Ddit3 (Chop), are marked with H3K79 methylation. Interference with DOT1L activity resulted in transcriptional activation of both genes accompanied by decreased levels of H3K79 dimethylation. Although downstream effectors of the UPR, such as Ppp1r15a/Gadd34, Atf3, and Tnfrsf10b/Dr5 were also transcriptionally activated, this most likely occurred in response to increased ATF4 expression rather than as a direct consequence of altered H3K79 methylation. While stem cells are particularly vulnerable to stress, the UPR and ER stress have not been extensively studied in these cells yet. Since activation of the ER stress program is also implicated in directing stem cells into differentiation or to maintain a proliferative status, the UPR must be tightly regulated. Our and published data suggest that histone modifications, including H3K4me3, H3K14ac, and H3K79me2, are implicated in the control of transcriptional activation of ER stress genes. In this context, the loss of H3K79me2 at the Atf4- and Ddit3-promoters appears to mark a point-of-no-return that activates the death program in NSCs.


Stereotactic radiosurgery of arteriovenous malformations: pathologic changes in resected tissue.
Both stereotactic radiosurgery and microsurgery are treatment modalities for arteriovenous malformations (AVM), and more recently, multimodality treatment using these approaches has been utilized. We surgically resected AVMs from 33 patients (ages 7-64 years old, mean age 30.4) 1-11 years after radiosurgery. AVM volumes were 0.8-117 cm3 (mean 21.6 cm3), and doses ranged from 4.6-45 GyE (mean 21.2 GyE). AVMs resected were submitted for pathologic review. Each AVM was evaluated for the following radiation changes, and the number of AVMs demonstrating these changes were noted: endothelial proliferation (27), hyaline (18) and calcium (10) in AVM vessel walls, partial (9) or complete (24) thrombosis of some AVM vessels, and necrosis of vessels (15) and adjacent brain tissue (11). A semiquantitative scale (mild, moderate, severe) incorporating the aforementioned changes present in each case classified the extent of radiation-induced change. There was a significant correlation (r = 0.624, p < 0.01) between extent of radiation change and dose of radiation received. There was no absolute radiation dose threshold below which radiation-induced changes were absent. However, all but one patient receiving greater than 20 GyE developed moderate to severe radiation vascular changes and the 3 patients treated with greater than 30 GyE all had severe radiation-induced changes. Radiation changes in AVMs following stereotactic radiosurgery appear to be dose-related. The correlation of dose to extent of radiation change may allow the determination of the optimal dose of radiation to treat AVMs.


Charcot arthropathy in spina bifida.
This multicenter study was undertaken to identify the prevalence of Charcot arthropathy in the spina bifida population; to evaluate the relationship of neurosegmental level, ambulatory level, and distribution of joint involvement; and to assess treatment results and make treatment recommendations. Sixteen patients were identified with Charcot arthropathy based on clinical and radiographic criteria ranging in age from 9 to 42 years. There were 15 ankles, seven knees, and four hips identified with Charcot arthropathy. Six patients underwent surgery and modification of orthoses, eight had a modification of orthoses only, one had no modification, and one was lost to follow-up. Mean follow-up was 4 years and 9 months (with four good, 17 fair, and five poor results). The best results were seen in 13 compliant patients with a brace modification, whereas poor results were seen in three patients with poor brace compliance. Based on our study, we have noted the prevalence of Charcot arthropathy in spina bifida to be one in 100 cases.


Genetic and environmental origins of gambling behaviors from ages 18 to 25: A longitudinal twin family study.
Gambling behaviors tend to increase in prevalence from late adolescence to young adulthood, and the underlying genetic and environmental influences during this period remain largely understudied. We examined the genetic and environmental influences on gambling behaviors contributing to stability and change from ages 18 to 25 in a longitudinal, behavioral genetic mixed-sex twin study design. Participants were enrolled in the Minnesota Twin Family Study. A range of gambling behaviors (maximum frequency, average frequency, money lost, and gambling problems) were assessed at ages 18 and 25. The results of our study support the following conclusions: (a) the genetic and environmental factors impacting a range of gambling behaviors are largely similar in men and women, (b) genetic factors increase in influence from 18 to 25 (21% at age 18 to 57% at age 25), (c) shared environmental factors are influential at age 18, but tend to decrease from ages 18 to 25 (55% at age 18 to 10% at age 25), and (d) nonshared environmental influences are similarly significant and are small to moderate in magnitude at both ages. The findings add to a small yet important research area regarding determinants of youth gambling behaviors and have the potential to inform prevention and intervention efforts. (PsycINFO Database Record


Young adults and higher education: barriers and breakthroughs to success.
Although access to higher education has increased substantially over the past forty years, student success in college--as measured by persistence and degree attainment--has not improved at all. Thomas Brock reviews systematic research findings on the effectiveness of various interventions designed to help at-risk students remain in college. Brock shows how changes in federal policy and public attitudes since the mid-1960s have opened up higher education to women, minorities, and nontraditional students and also shifted the "center of gravity" in higher education away from traditional four-year colleges toward nonselective community colleges. Students at two-year colleges, however, are far less likely than those at four-year institutions to complete a degree. Brock argues that the nation's higher education system must do much more to promote student success. Three areas, he says, are particularly ripe for reform: remedial education, student support services, and financial aid. In each of these three areas, Brock reviews programs and interventions that community colleges have undertaken in order to raise completion rates. Some colleges have, for example, experimented with remedial programs that build social cohesion between students and faculty and integrate content across courses. Other colleges have tested student support service programs that offer counseling and advising that are regular, intensive, and personalized. Still others have experimented with ways to simplify the financial aid application process and incentivize students to earn good grades and persist in school. Research shows that such programs and interventions can improve student outcomes, but Brock argues that more must be done to bring proven practices to scale and to test new ideas that might lead to better results. Institutions that most need help'are those that provide the greatest access to nontraditional and underprepared students in community colleges and less selective universities.


Sirtuin1 promotes osteogenic differentiation through downregulation of peroxisome proliferator-activated receptor γ in MC3T3-E1 cells.
Osteoporosis is a skeletal disorder characterized by bone loss, resulting in architectural deterioration of the skeleton, decreased bone strength and an increased risk of fragility fractures. Strengthening osteogenesis is an effective way to relieve osteoporosis. Sirtuin1 (Sirt1) is a nicotinamide adenine dinucleotide (NAD(+))-dependent deacetylase, which is reported to be involved in improving osteogenesis. Sirt1 targets peroxisome proliferator-activated receptor γ (PPARγ) in the regulation of adipose tissues; however, the molecular mechanism of Sirt1 in osteogenic differentiation is still unknown. PPARγ tends to induce more adipogenic differentiation rather than osteogenic differentiation. Hence, we hypothesized that Sirt1 facilitates osteogenic differentiation through downregulation of PPARγ signaling. Mouse pre-osteoblastic MC3T3-E1 cells were cultured under osteogenic medium. Sirt1 was overexpressed through plasmid transfection. The results showed that high expression of Sirt1 was associated with increased osteogenic differentiation, as indicated by quantitative PCR and Western blot analysis of osteogenic markers, and Von Kossa staining. Sirt1 overexpression also directly and negatively regulated the expression of PPARγ and its downstream molecules. Use of the PPARγ agonist Rosiglitazone, reversed the effects of Sirt1 on osteogenic differentiation. Using constructed luciferase plasmids, we demonstrated a role of Sirt1 in inhibiting PPARγ-induced activity and expression of adipocyte-specific genes, including acetyl-coenzyme A carboxylase (Acc) and fatty acid binding protein 4 (Fabp4). The interaction between Sirt1 and PPARγ was further confirmed using co-immunoprecipitation analysis. Together, these results reveal a novel mechanism for Sirt1 in osteogenic differentiation through downregulation of PPARγ activity. These findings suggest that the Sirt1-PPARγ pathway may represent a potential target for enhancement of osteogenesis and treatment of osteoporosis.


Evaluation and use of epidemiological evidence for environmental health risk assessment: WHO guideline document.
Environmental health risk assessment is increasingly being used in the development of environmental health policies, public health decision making, the establishment of environmental regulations, and research planning. The credibility of risk assessment depends, to a large extent, on the strength of the scientific evidence on which it is based. It is, therefore, imperative that the processes and methods used to evaluate the evidence and estimate health risks are clear, explicit, and based on valid epidemiological theory and practice. Epidemiological Evidence for Environmental Health Risk Assessment is a World Health Organization (WHO) guideline document. The primary target audiences of the guidelines are expert review groups that WHO (or other organizations) might convene in the future to evaluate epidemiological evidence on the health effects of environmental factors. These guidelines identify a set of processes and general approaches to assess available epidemiological information in a clear, consistent, and explicit manner. The guidelines should also help in the evaluation of epidemiological studies with respect to their ability to support risk assessment and, consequently, risk management. Conducting expert reviews according to such explicit guidelines would make health risk assessment and subsequent risk management and risk communication processes more readily understood and likely to be accepted by policymakers and the public. It would also make the conclusions reached by reviews more readily acceptable as a basis for future WHO guidelines and other recommendations, and would provide a more rational basis for setting priorities for future research.


Laparoscopic Ovarian Drilling Improves Endometrial Homeobox Gene Expression in PCOS.
The study was designed to investigate whether laparoscopic ovarian drilling (LOD) of ovaries alters the expression levels of HOXA-10 and HOXA-11 mRNA in the endometrium of infertile women with clomiphene-resistant PCOS. Expression of HOXA-10 and HOXA-11 mRNA in the endometrium obtained before and after LOD during the midsecretory phase was measured. Expression of each gene was evaluated using real-time reverse transcriptase polymerase chain reaction (RT-PCR). Expression levels of HOXA-10 and HOXA-11 mRNA were lower in endometrium of patients with PCOS before LOD compared with fertile controls. But the differences failed to show statistical significance. Compared with fertile subjects, LOD of PCOS ovaries up-regulated endometrial HOXA-10 and HOXA-11 mRNA expression. Fold changes of HOXA-10 and HOXA-11 mRNA after LOD were found to be 4.46 and 4.19, respectively. Fold change increase in HOXA-10 and HOXA-11 mRNA was found to be statistically significant (P < .01, P < .02). There is a receptivity defect in the endometrium of women with PCOS that affects fertility regardless of other causes of infertility. LOD increases endometrial HOXA-10 and HOXA-11 mRNA expressions and improves receptivity in patients with clomiphene-resistant PCOS.


Input-driven oscillations in networks with excitatory and inhibitory neurons with dynamic synapses.
Previous work has shown that networks of neurons with two coupled layers of excitatory and inhibitory neurons can reveal oscillatory activity. For example, Börgers and Kopell (2003) have shown that oscillations occur when the excitatory neurons receive a sufficiently large input. A constant drive to the excitatory neurons is sufficient for oscillatory activity. Other studies (Doiron, Chacron, Maler, Longtin, & Bastian, 2003; Doiron, Lindner, Longtin, Maler, & Bastian, 2004) have shown that networks of neurons with two coupled layers of excitatory and inhibitory neurons reveal oscillatory activity only if the excitatory neurons receive correlated input, regardless of the amount of excitatory input. In this study, we show that these apparently contradictory results can be explained by the behavior of a single model operating in different regimes of parameter space. Moreover, we show that adding dynamic synapses in the inhibitory feedback loop provides a robust network behavior over a broad range of stimulus intensities, contrary to that of previous models. A remarkable property of the introduction of dynamic synapses is that the activity of the network reveals synchronized oscillatory components in the case of correlated input, but also reflects the temporal behavior of the input signal to the excitatory neurons. This allows the network to encode both the temporal characteristics of the input and the presence of spatial correlations in the input simultaneously.


Rapid, Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components.
The recent Zika virus outbreak highlights the need for low-cost diagnostics that can be rapidly developed for distribution and use in pandemic regions. Here, we report a pipeline for the rapid design, assembly, and validation of cell-free, paper-based sensors for the detection of the Zika virus RNA genome. By linking isothermal RNA amplification to toehold switch RNA sensors, we detect clinically relevant concentrations of Zika virus sequences and demonstrate specificity against closely related Dengue virus sequences. When coupled with a novel CRISPR/Cas9-based module, our sensors can discriminate between viral strains with single-base resolution. We successfully demonstrate a simple, field-ready sample-processing workflow and detect Zika virus from the plasma of a viremic macaque. Our freeze-dried biomolecular platform resolves important practical limitations to the deployment of molecular diagnostics in the field and demonstrates how synthetic biology can be used to develop diagnostic tools for confronting global health crises. PAPERCLIP.


[Evaluation of mass vaccination programmes: the experience of Hepatitis A in Catalonia].
The hepatitis A + B vaccination programme of preadolescents was introduced in 1998 in Catalonia. During the following years, one of the main objectives has been to quantify the reduction in the incidence of hepatitis A caused by the vaccination programme. A retrospective analysis applying the space-time scan statistic to reported incidence rates of hepatitis A was carried out in the counties of Catalonia from 1992 to 2007. The relative risk (RR) was calculated and the spatial autocorrelation was estimated using Moran's I statistic. Six of the 7 space-time clusters identified by the scan statistic occurred in the pre-vaccine era (1992-1998) and only one in the post-vaccine era (1992-2007). In the first 10 four-weekly periods of the post-vaccine era (1999-2005) there was a significant reduction in the incidence of hepatitis A in Catalonia with respect to the pre-vaccine era (1992-1998) (p<0,01). Moran's I statistic showed no pattern of global spatial dependence and was useful in detecting local clusters. These results corroborate previous studies that attributed most of the reduction in the incidence of hepatitis A in Catalonia to the effect of vaccination.


The interplay of StyR and IHF regulates substrate-dependent induction and carbon catabolite repression of styrene catabolism genes in Pseudomonas fluorescens ST.
In Pseudomonas fluorescens ST, the promoter of the styrene catabolic operon, PstyA, is induced by styrene and is subject to catabolite repression. PstyA regulation relies on the StyS/StyR two-component system and on the IHF global regulator. The phosphorylated response regulator StyR (StyR-P) activates PstyA in inducing conditions when it binds to the high-affinity site STY2, located about -40 bp from the transcription start point. A cis-acting element upstream of STY2, named URE, contains a low-affinity StyR-P binding site (STY1), overlapping the IHF binding site. Deletion of the URE led to a decrease of promoter activity in inducing conditions and to a partial release of catabolite repression. This study was undertaken to assess the relative role played by IHF and StyR-P on the URE, and to clarify if PstyA catabolite repression could rely on the interplay of these regulators. StyR-P and IHF compete for binding to the URE region. PstyA full activity in inducing conditions is achieved when StyR-P and IHF bind to site STY2 and to the URE, respectively. Under catabolite repression conditions, StyR-P binds the STY1 site, replacing IHF at the URE region. StyR-P bound to both STY1 and STY2 sites oligomerizes, likely promoting the formation of a DNA loop that closes the promoter in a repressed conformation. We found that StyR and IHF protein levels did not change in catabolite repression conditions, implying that PstyA repression is achieved through an increase in the StyR-P/StyR ratio. We propose a model according to which the activity of the PstyA promoter is determined by conformational changes. An open conformation is operative in inducing conditions when StyR-P is bound to STY2 site and IHF to the URE. Under catabolite repression conditions StyR-P cellular levels would increase, displacing IHF from the URE and closing the promoter in a repressed conformation. The balance between the open and the closed promoter conformation would determine a fine modulation of the promoter activity. Since StyR and IHF protein levels do not vary in the different conditions, the key-factor regulating PstyA catabolite repression is likely the kinase activity of the StyR-cognate sensor protein StyS.


Genetic Evidence for Early Peritoneal Spreading in Pelvic High-Grade Serous Cancer.
Most pelvic high-grade serous (HGS) carcinomas have been proposed to arise from tubal primaries that progress rapidly to advanced disease. However, the temporal sequence of ovarian and peritoneal metastases is not well characterized. To establish the sequence of metastases, phylogenetic relationships among the ovarian and peritoneal carcinomas were determined from single-nucleotide variations (SNVs) in nine tumor regions from each patient with pelvic HGS carcinomas. Somatic SNVs from each tumor sample were used to reconstruct phylogenies of samples from each patient. Variant allele frequencies were used to reconstruct subclone phylogenies in each tumor sample. We show that pelvic HGS carcinomas are highly heterogeneous, only sharing less than 4% of somatic SNVs among all nine carcinoma implants in one patient. TP53 mutations are found in all nine carcinoma implants in each patient. The phylogenetic analyses reveal that peritoneal metastases arose from early branching events that preceded branching events for ovarian carcinomas in some patients. Finally, subclone phylogenies indicate the presence of multiple subclones at each tumor implant and early tumor clones in peritoneal implants. The genetic evidence that peritoneal implants arose before or concurrently with ovarian implants is consistent with the emerging concept of the extra-ovarian origin of pelvic HGS cancer. Our results challenge the concept of stepwise spatial progression from the fallopian primary to ovarian carcinomas to peritoneal dissemination and suggest an alternative progression model where peritoneal spreading of early clones occurs before or in parallel with ovarian metastases.


Virus-induced behavioral alteration of mice.
Neonatal mice were inoculated intracerebrally with lymphocytic choriomeningitis virus (LCM). These mice developed long-term persistent tolerant infections and when tested at 3.5 to 6.0 months of age they showed significant increases in behavioral latency when subjected to open-field tests, and significant decreases in the current level required to elicit to startle response and in locomotor activity in a running wheel. Comparable results were obtained with mice in which persistent infection was induced at 8 weeks of age and which were tested at 3.5 to 6.0 months of age. It was concluded that mice infected with LCM at birth or as adults exhibited long-lasting behavioral abnormalities.


Storage of hemopoietic stem cells.
Autologous, and in some cases allogeneic, hemopoietic stem cells (HSC) are stored for varying periods of time prior to infusion. For periods of greater than 48 h, storage requires cryopreservation. It is essential to optimize cell storage and ensure the quality of the product for subsequent reinfusion. A number of important variables may affect the subsequent quality of infused HSC and therapeutic cells (TC). This review discusses these and also reviews the regulatory framework that now aims to ensure the quality of stem cells and TC for transplantation. Important variables included cell concentration, temperature, interval from collection to cryopreservation, manipulations performed. They also included rate of freezing and whether controlled-rate freezing was employed. Parameters studied were type of cryoprotectant utilized [dimethyl sulphoxide (DMSO) is most commonly used, sometimes in combination with hydroxyethyl starch (HES)]; and storage conditions. It is also important to assess the quality of stored stem cells. Measurements employed included the total cell count (TNC), mononuclear cell count (MNC), CD34+ cells and colony-forming units - granulocyte macrophage (CFU-GM). Of these, TNC and CD34+ are the most useful. However, the best measure of the quality of stored stem cells is their subsequent engraftment. The quality systems used in stem cell laboratories are described in the guidance of the Joint Accreditation Committee of ISCT (Europe) and the EBMT (JACIE) and the EU Directive on Tissues and Cells plus its supporting commission directives. Inspections of facilities are carried out by the appropriate national agencies and JACIE. For high-quality storage of HSC and TC, processing facilities should use validated procedures that take into account critical variables. The quality of all products must be assessed before and after storage.


Homicidal sex offenders: psychological, phallometric, and diagnostic features.
Homicidal sex offenders represent an understudied population in the forensic literature. Forty-eight homicidal sex offenders assessed between 1982 and 1992 were studied in relation to a comparison group of incest offenders. Historical features, commonly used psychological inventories, criminal histories, phallometric assessments, and DSM diagnoses were collected on each group. The homicidal sex offenders, compared with the incest offenders, self-reported that they had more frequently been removed from their homes during childhood and had more violence and forensic psychiatric contact in their histories. On the self-report psychological inventories, the homicidal sex offenders portrayed themselves as functioning significantly better in the areas of sexuality (Derogatis Sexual Functioning Inventory) and aggression/hostility (Buss-Durkee Hostility Inventory). However, on the Psychopathy Checklist-Revised (PCL-R), researchers rated the homiciders significantly more psychopathic than the incest offenders on Factor 1 (personality traits) and Factor 2 (antisocial history). Police records revealed the homicidal subjects also had been charged or convicted of more violent and nonviolent nonsexual offenses. The phallometric assessments indicated that the homicidal sex offenders demonstrated higher levels of response to pedophilic stimuli and were significantly more aroused to stimuli depicting assaultive acts to children, relative to the incest offenders. Despite the homiciders' self-reports of fairly good psychological functioning, DSM-III diagnoses reliably discriminated between the groups. A large number of homicidal sex offenders were diagnosed as suffering from psychosis, antisocial personality disorder, paraphilias, sexual sadism, sexual sadism with pedophilia, and substance abuse. Seventy-five percent of the homicidal sex offenders had three or more diagnoses compared with six percent of the incest offenders. The article addresses the role of "hard" versus "soft" measures in the assessment and treatment of violent sex offenders. In addition, the usefulness of phallometric assessments and the PCL-R and its subscales are considered.


Modelling distribution in European stream macroinvertebrates under future climates.
Climate change is predicted to have profound effects on freshwater organisms due to rising temperatures and altered precipitation regimes. Using an ensemble of bioclimatic envelope models (BEMs), we modelled the climatic suitability of 191 stream macroinvertebrate species from 12 orders across Europe under two climate change scenarios for 2080 on a spatial resolution of 5 arc minutes. Analyses included assessments of relative changes in species' climatically suitable areas as well as their potential shifts in latitude and longitude with respect to species' thermal preferences. Climate-change effects were also analysed regarding species' ecological and biological groupings, namely (1) endemicity and (2) rarity within European ecoregions, (3) life cycle, (4) stream zonation preference and (5) current preference. The BEMs projected that suitable climate conditions would persist in Europe in the year 2080 for nearly 99% of the modelled species regardless of the climate scenario. Nevertheless, a decrease in the amount of climatically suitable areas was projected for 57-59% of the species. Depending on the scenario, losses could be of 38-44% on average. The suitable areas for species were projected to shift, on average, 4.7-6.6° north and 3.9-5.4° east. Cold-adapted species were projected to lose climatically suitable areas, while gains were expected for warm-adapted species. When projections were analysed for different species groupings, only endemics stood out as a particular group. That is, endemics were projected to lose significantly larger amounts of suitable climatic areas than nonendemic species. Despite the uncertainties involved in modelling exercises such as this, the extent of projected distributional changes reveals further the vulnerability of freshwater organisms to climate change and implies a need to understand the consequences for ecological function and biodiversity conservation.


Preferential accumulation of single-stranded regions in telomeres of human fibroblasts.
We have demonstrated recently that chronic hyperoxic treatment accelerates the rate of aging of fibroblasts and the rate of telomere shortening in parallel. It was hypothesized that accelerated telomere shortening is due to preferential accumulation of oxidative damage in telomeres. To test this hypothesis, we measured the accumulation of sites sensitive to S1 nuclease treatment in telomeres, in minisatellites, and in the bulk of the genome of fibroblasts under different models of oxidative stress as well as after treatment with the alkylating agent, N-methyl-N'-nitro-N-nitrosoguanidine. A comparison with qualitative data obtained by alkaline electrophoresis reveals that the sites transferred to double-strand breaks by treatment with low concentrations of S1 nuclease are, in fact, single-stranded regions in the DNA. These regions may resemble single-stranded overhangs, gaps, or conventional single-strand breaks. The frequency of single-stranded regions is significantly higher in telomeres than in minisatellites or in the bulk of the genome under all conditions tested. Those regions induced in minisatellites or in the overall genome by a bolus dose of hydrogen peroxide are completely repaired within 24 h. On the other hand, 50 +/- 12% of H2O2-induced single-stranded regions remain unrepaired for at least 19 days in telomeres of human fibroblasts, leading to a significant increase of the telomeric steady-state level of these lesions. This preferential accumulation might significantly contribute to telomere shortening.


Pliocene oceanic seaways and global climate.
Tectonically induced changes in oceanic seaways had profound effects on global and regional climate during the Late Neogene. The constriction of the Central American Seaway reached a critical threshold during the early Pliocene ~4.8-4 million years (Ma) ago. Model simulations indicate the strengthening of the Atlantic Meridional Overturning Circulation (AMOC) with a signature warming response in the Northern Hemisphere and cooling in the Southern Hemisphere. Subsequently, between ~4-3 Ma, the constriction of the Indonesian Seaway impacted regional climate and might have accelerated the Northern Hemisphere Glaciation. We here present Pliocene Atlantic interhemispheric sea surface temperature and salinity gradients (deduced from foraminiferal Mg/Ca and stable oxygen isotopes, δ18O) in combination with a recently published benthic stable carbon isotope (δ13C) record from the southernmost extent of North Atlantic Deep Water to reconstruct gateway-related changes in the AMOC mode. After an early reduction of the AMOC at ~5.3 Ma, we show in agreement with model simulations of the impacts of Central American Seaway closure a strengthened AMOC with a global climate signature. During ~3.8-3 Ma, we suggest a weakening of the AMOC in line with the global cooling trend, with possible contributions from the constriction of the Indonesian Seaway.


The genetic organisation of prokaryotic two-component system signalling pathways.
Two-component systems (TCSs) are modular and diverse signalling pathways, involving a stimulus-responsive transfer of phosphoryl groups from transmitter to partner receiver domains. TCS gene and domain organisation are both potentially informative regarding biological function, interaction partnerships and molecular mechanisms. However, there is currently little understanding of the relationships between domain architecture, gene organisation and TCS pathway structure. Here we classify the gene and domain organisation of TCS gene loci from 1405 prokaryotic replicons (>40,000 TCS proteins). We find that 200 bp is the most appropriate distance cut-off for defining whether two TCS genes are functionally linked. More than 90% of all TCS gene loci encode just one or two transmitter and/or receiver domains, however numerous other geometries exist, often with large numbers of encoded TCS domains. Such information provides insights into the distribution of TCS domains between genes, and within genes. As expected, the organisation of TCS genes and domains is affected by phylogeny, and plasmid-encoded TCS exhibit differences in organisation from their chromosomally-encoded counterparts. We provide here an overview of the genomic and genetic organisation of TCS domains, as a resource for further research. We also propose novel metrics that build upon TCS gene/domain organisation data and allow comparisons between genomic complements of TCSs. In particular, 'percentage orphaned TCS genes' (or 'Dissemination') and 'percentage of complex loci' (or 'Sophistication') appear to be useful discriminators, and to reflect mechanistic aspects of TCS organisation not captured by existing metrics.


Rosetting of human T lymphocytes with sheep and human erythrocytes. Comparison of human and sheep ligand binding using purified E receptor.
Previous studies have shown that the purified T lymphocyte glycoprotein, cluster differentiation 2 (CD2) (also known as T11, lymphocyte function-associated antigen (LFA)-2, and the erythrocyte (E) rosette receptor) interacts with the LFA-3 molecule on human E. We have examined the interaction of the purified CD2 molecule with the T11 target structure (T11TS) molecule on sheep E, and compared the two interactions. Purified, 125I-labeled CD2 bound to sheep E and the binding was inhibited by anti-T11TS monoclonal antibody (mAb). Reciprocally, the binding of T11TS mAb to sheep E was inhibited by pretreatment of sheep E with purified CD2. High concentrations of purified CD2 aggregated sheep E, possibly by inserting into the membrane, and the aggregation was inhibited by T11TS mAb. The affinity and number of binding sites for purified CD2 on sheep and human E was found to be similar, with Ka of 9 X 10(7)/M and 6 X 10(7)/M and 9800 and 8300 CD2 binding sites/E, respectively. Thus, the human T lymphocyte CD2 molecule is a receptor that cross-reacts between LFA-3 on human E and T11TS on sheep E, suggesting that LFA-3 and T11TS are functionally homologous ligands. As measured by saturation mAb binding, there are 8100 and 3900 ligand molecules/sheep and human E, respectively. Human and sheep E have surface areas of 145 and 54 micron 2, respectively. The 3.2- to 5.6-fold higher ligand density on sheep E appears to account for the ability of sheep but not human E to rosette with certain types of human T lymphocytes.


Long-term secretion of therapeutic proteins from genetically modified skeletal muscles.
Protein delivery from genetically modified skeletal muscle has been reported previously. However, a stable and prolonged secretion was obtained in immunocompromised or newborn animals only. To evaluate the clinical relevance of this approach, we have transduced myoblasts from an adult beta-glucuronidase-deficient (MPS VII) mouse with retroviral vectors carrying either the human beta-glucuronidase cDNA or the murine erythropoietin (Epo) cDNA. The cells were then grafted into the tibialis anterior muscle of adult immunocompetent MPS VII recipients. Protein expression was controlled either by ubiquitous or muscle-specific transcriptional regulatory elements. Animals were analyzed over an 8-month period. The in situ detection of beta-glucuronidase activity revealed up to 60% of genetically modified myofibers in the recipient muscles. The human desmin promoter and enhancer showed the highest in vivo expression. Secretion of beta-glucuronidase induced a disappearance of lysosomal storage lesions in the liver and spleen of recipient animals. Delivery of Epo led to a permanent increase of hematocrit values over 3 months. These results showed that the transplantation of genetically modified myoblasts allowed a sustained secretion of recombinant proteins at therapeutic levels in immunocompetent adult mice. They suggest that the approach may be considered for human applications.


Operant procedures for assessing behavioral flexibility in rats.
Executive functions consist of multiple high-level cognitive processes that drive rule generation and behavioral selection. An emergent property of these processes is the ability to adjust behavior in response to changes in one's environment (i.e., behavioral flexibility). These processes are essential to normal human behavior, and may be disrupted in diverse neuropsychiatric conditions, including schizophrenia, alcoholism, depression, stroke, and Alzheimer's disease. Understanding of the neurobiology of executive functions has been greatly advanced by the availability of animal tasks for assessing discrete components of behavioral flexibility, particularly strategy shifting and reversal learning. While several types of tasks have been developed, most are non-automated, labor intensive, and allow testing of only one animal at a time. The recent development of automated, operant-based tasks for assessing behavioral flexibility streamlines testing, standardizes stimulus presentation and data recording, and dramatically improves throughput. Here, we describe automated strategy shifting and reversal tasks, using operant chambers controlled by custom written software programs. Using these tasks, we have shown that the medial prefrontal cortex governs strategy shifting but not reversal learning in the rat, similar to the dissociation observed in humans. Moreover, animals with a neonatal hippocampal lesion, a neurodevelopmental model of schizophrenia, are selectively impaired on the strategy shifting task but not the reversal task. The strategy shifting task also allows the identification of separate types of performance errors, each of which is attributable to distinct neural substrates. The availability of these automated tasks, and the evidence supporting the dissociable contributions of separate prefrontal areas, makes them particularly well-suited assays for the investigation of basic neurobiological processes as well as drug discovery and screening in disease models.


Biliary stents: models and methods for endoscopic stenting.
Biliary stenting is widely used to palliate malignant obstruction or to treat benign biliary diseases. Recently, the most important changes have related to self-expandable metal stents (SEMSs), which are now available in a wide variety of designs, and to treatment techniques. This article is part of a combined publication that expresses the current view of the European Society of Gastrointestinal Endoscopy (ESGE) about endoscopic biliary stenting: a technology review describes the stent models and stenting techniques, and a separate clinical guideline states the evidence and recommendations regarding stenting.


Climate-driven trends in contemporary ocean productivity.
Contributing roughly half of the biosphere's net primary production (NPP), photosynthesis by oceanic phytoplankton is a vital link in the cycling of carbon between living and inorganic stocks. Each day, more than a hundred million tons of carbon in the form of CO2 are fixed into organic material by these ubiquitous, microscopic plants of the upper ocean, and each day a similar amount of organic carbon is transferred into marine ecosystems by sinking and grazing. The distribution of phytoplankton biomass and NPP is defined by the availability of light and nutrients (nitrogen, phosphate, iron). These growth-limiting factors are in turn regulated by physical processes of ocean circulation, mixed-layer dynamics, upwelling, atmospheric dust deposition, and the solar cycle. Satellite measurements of ocean colour provide a means of quantifying ocean productivity on a global scale and linking its variability to environmental factors. Here we describe global ocean NPP changes detected from space over the past decade. The period is dominated by an initial increase in NPP of 1,930 teragrams of carbon a year (Tg C yr(-1)), followed by a prolonged decrease averaging 190 Tg C yr(-1). These trends are driven by changes occurring in the expansive stratified low-latitude oceans and are tightly coupled to coincident climate variability. This link between the physical environment and ocean biology functions through changes in upper-ocean temperature and stratification, which influence the availability of nutrients for phytoplankton growth. The observed reductions in ocean productivity during the recent post-1999 warming period provide insight on how future climate change can alter marine food webs.


A 17-year review of foodborne outbreaks: describing the continuing decline in England and Wales (1992-2008).
Systematic national surveillance of foodborne disease outbreaks effectively serves the development of public health policy on food safety. The Health Protection Agency has maintained a collaborative surveillance system for foodborne outbreaks in England and Wales since 1992. Up to 2008, 2429 foodborne outbreaks were identified, described and analysed for changes over time. Salmonella spp. accounted for half of the outbreaks, although the proportion of these decreased over the surveillance period. Similarly, the proportion of outbreaks caused by Clostridium perfringens decreased, while those attributed to Campylobacter spp. and Vero cytotoxin-producing Escherichia coli O157 increased. Although poultry meat was the most frequently implicated food vehicle in outbreaks followed by miscellaneous foods and red meats, the proportion of outbreaks attributed to meats in fact decreased over time but those linked to miscellaneous foods did not. Over the surveillance period, the proportion of outbreaks linked to eggs and S. Enteritidis non-phage-type 4, particularly in food service establishments, increased, highlighting the importance of this organism/setting/vehicle association. Contributory factors in most outbreaks were cross-contamination, inadequate heat treatment, and inappropriate food storage. This study describes the overall decline in foodborne outbreaks, providing evidence that the introduction and adherence to effective control measures provide the best means of minimizing the risk of foodborne infection.


Bioluminescence in mesopelagic squid: diel color change during counterillumination.
Two species of mesopelagic squid greatly altered the color of their bio-luminescence during counterillumination. The color change was triggered by changes in water temperature corresponding to those normally encountered by these vertically migrating animals. These squid can probably conceal themselves under the different colors of downwelling light that they encounter in their day and night habitats.


Restrictive compared with liberal red cell transfusion strategies in cardiac surgery: a meta-analysis.
To determine whether a restrictive strategy of red blood cell (RBC) transfusion at lower haemoglobin concentrations is inferior to a liberal strategy of RBC transfusion at higher haemoglobin concentrations in patients undergoing cardiac surgery. We conducted a systematic review, meta-analysis, and trial sequential analysis of randomized controlled trials of the effect of restrictive and liberal RBC transfusion strategies on mortality within 30 days of surgery as the primary outcome. Secondary outcomes were those potentially resulting from anaemia-induced tissue hypoxia and transfusion outcomes. We searched the electronic databases MEDLINE, EMBASE, and the Cochrane Library until 17 November 2017. Thirteen trials were included. The risk ratio (RR) of mortality derived from 4545 patients assigned to a restrictive strategy when compared with 4547 transfused according to a liberal strategy was 0.96 [95% confidence interval (CI) 0.76-1.21, I2 = 0]. A restrictive strategy did not have a statistically significant effect on the risk of myocardial infarction (RR 1.01, 95% CI 0.81-1.26; I2=0), stroke (RR 0.93, 95% CI 0.68-1.27, I2 = 0), renal failure (RR 0.96, 95% CI 0.76-1.20, I2 = 0), or infection (RR 1.12, 95% CI 0.98-1.29, I2 = 0). Subgroup analysis of adult and paediatric trials did not show a significant interaction. At approximately 70% of the critical information size, the meta-analysis of mortality crossed the futility boundary for inferiority of the restrictive strategy. The current evidence does not support the notion that restrictive RBC transfusion strategies are inferior to liberal RBC strategies in patients undergoing cardiac surgery.


Adaptive plasticity in mate preference linked to differences in reproductive effort.
There is abundant evidence for the existence of marked mate preferences in natural populations, but the occurrence of within-population variation in mate preferences has received little attention and is often regarded as nonadaptive deviation from the optimal norm. Here we show experimentally that the preference of female collared flycatchers Ficedula albicollis for male forehead patch size, a sexually selected trait, varies with the time of breeding, an environmental factor with strong effects on reproductive success. Contrary to expectations based on time-constrained choice models, only late-breeding females prefer males with a large patch size. The variation in mate preference matches a seasonal change in female reproductive success: long-term data reveal a positive relationship between female reproductive success and male patch size exclusively in late breeders. In addition, female reproductive effort, as assessed by clutch size, appears to be adjusted relative to both timing of breeding and male phenotype. We conclude that not only can mate preferences display adaptive plasticity within populations, but this plasticity can also be linked to differences in reproductive investment.


Constraints and opportunities for implementing nutrition-specific, agricultural and market-based approaches to improve nutrient intake adequacy among infants and young children in two regions of rural Kenya.
Several types of interventions can be used to improve nutrient intake adequacy in infant and young child (IYC) diets, including fortified foods, home fortification, nutrition education and behaviour change communication (BCC) in addition to agricultural and market-based strategies. However, the appropriate selection of interventions depends on the social, cultural, physical and economic context of the population. Derived from two rural Kenyan populations, this analysis combined information from: (1) a quantitative analysis to derive a set of food-based recommendations (FBRs) to fill nutrient intake gaps in IYC diets and identify 'problem nutrients' for which intake gaps require solutions beyond currently available foods and dietary patterns, and (2) an ethnographic qualitative analysis to identify contextual factors posing opportunities or constraints to implementing the FBRs, including perceptions of cost, convenience, accessibility and appropriateness of the recommended foods for IYC diets and other social or physical factors that determine accessibility of those foods. Opportunities identified included BCC to increase the acceptability and utilisation of green leafy vegetables (GLV) and small fish and agronomic interventions to increase the productivity of GLV and millet. Value chains for millet, beans, GLV, milk and small fish should be studied for opportunities to increase their accessibility in local markets. Processor-level interventions, such as partially cooked fortified dry porridge mixes or unfortified cereal mixes incorporating millet and beans, may increase the accessibility of foods that provide increased amounts of the problem nutrients. Multi-sectoral actors and community stakeholders should be engaged to assess the feasibility of implementing these locally appropriate strategies.


Strength in numbers? Cognitive reappraisal tendencies and psychological functioning among Latinos in the context of oppression.
The typically positive relationship between cognitive reappraisal and psychological functioning may be nullified for Latinos embedded within multiple contexts of oppression (Perez & Soto, 2011). Multiply oppressive contexts are characterized by exposure to oppression at a societal level (distal oppression), in the immediate environment (proximal oppression), and at an individual level (personal oppression). We replicated and extended Perez and Soto's (2011) findings by examining whether the reappraisal-psychological functioning association was moderated by (a) relative numerical representation of Latinos within the environment (proximal oppression) and (b) personal perceptions of discrimination (personal oppression) among 425 Latino college students throughout the United States. For Latinos in high-Latino counties, greater use of reappraisal was associated with better psychological functioning, regardless of perceived discrimination; this relationship was absent for Latinos in low-Latino counties who perceived greater discrimination. Findings highlight the importance of considering how contextual factors can alter the adaptive functions of emotion regulation strategies.


Implications of the Fracture Risk Assessment Algorithm for the assessment and improvement of bone health in patients with prostate cancer: A comprehensive review.
Maintaining the optimum bone health is one of the important concerns in patients with prostate cancer, but it usually remains neglected. Failure to screen these patients is detrimental to both the length and the quality of life. The estimation of bone mineral density (BMD) and more recently the World Health Organization's fracture risk assessment (FRAX) algorithm in appropriate patients is recommended by several specialty organizations/associations at the time of instituting androgen deprivation therapy (ADT) for metastatic and high-risk individuals. It provides a 10-year risk evaluation of hip and major osteoporotic fractures (MOF). Using this web-based new investigating tool, candidates at high risk of fractures can be predicted more accurately according to clinical risk factors (CRF) alone or in combination with the femoral neck BMD. The FRAX application for senile osteoporosis has been studied and reviewed extensively, but no systematic review has ever been conducted for assessing the implication of FRAX in prostate cancer. This review article will give insight about the validity, role, and utility of this investigating tool in clinical practice for fracture risk assessment in these individuals. This systematic review was carried out as per the Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines and Cochrane review principles. We searched the PubMed, Cochrane database of systematic reviews, and the EMBASE electronic database until December 2018 using the medical subject heading terms prostate cancer and FRAX. A total of nine studies meet the inclusion criteria and were included in the review. These studies enrolled a total of 3704 patients (sample size range, 78-1220) of localized, metastatic, castration resistant prostate cancer with or without ADT and/or on photon or radiotherapy. The factors that influenced FRAX included age, ethnicity, baseline BMD, duration of ADT, presence of CRF, and measurement methods (CRF, with/without BMD, computed tomography based). An advanced age and duration of ADT were the most robust risk factors. A 10-year MOF and hip fracture risk estimation was higher when the femoral neck BMD was not incorporated in the FRAX measurement. Despite several well-known strengths of using FRAX in the fracture risk assessment of suitable candidates with prostate cancer, several risk factors such as the mode/duration of ADT, mode of radiotherapy, Vitamin D levels, bone remodeling markers, and recent/recurrent fractures need to be incorporated in the FRAX calculator for improving the predictive ability. In contrast to senile osteoporosis with a longer life expectancy, the fracture risk in patients with prostate cancer need to be measured more frequently and for a shorter time. Therefore, models like Garvan calculator with both 5- and 10-year risk estimates have to be developed for these patients. Additionally, its utilization is of limited value in the presence of recurrent fractures or falls. The FRAX algorithm is beneficial in identifying patients who require early intervention or bone-directed therapy as an early step to decrease skeletal-related events and other morbidity. Several risk factors need to be added for improving the FRAX predictive value. This model is still underutilized in the clinical practice and increasing the awareness among treating physicians will help in optimizing the bone health and the quality of life of this important population subgroup.


Temporary suture tarsorrhaphy.
To describe a technique for performing a bolsterless temporary tarsorrhaphy. Retrospective analysis and surgical technique description. Temporary suture tarsorrhaphy (TST), which consists of a suture through the upper and lower eyelid posterior lamella, was performed after eyelid or socket surgery. Over 15 years, >1000 patients in the practice of one of the authors (R.L.A.) had TST that successfully maintained corneal coverage without complications in all but four eyes. In two patients, replacement was required because of tissue erosion; in two patients, the suture was placed too posteriorly and caused corneal irritation that required replacement. The TST is functionally equivalent to, or superior to, traditional bolster temporary tarsorrhaphy. The TST is faster and simpler, requires fewer materials, and avoids the risks of bolsters, which include eyelid margin necrosis, irregularities, and lash loss from vascular compromise.


Fullerene and the origin of life.
The role of carbon in the development of life and as the structural backbone of all organisms is universally accepted and an essential part of evolution. However, the molecular basis is largely unknown and the interactions of carbon with nitrogen and oxygen in space are enigmatic. In 1985, the previously unknown form of carbon, coined fullerene, was discovered. We hypothesize that by virtue of the unique properties of fullerene, this hollow, ultra-robust, large, purely carbon molecule was the earliest progenitor of life. It acted as a stable universal biologic template on which small molecules spontaneously assembled and then formed, by further assembly, a surface mantle (here termed rosasome) of larger molecules. We submit that this process, by its inherent flexibility, initiated evolution, allowing the emergence of parallel diverse rosasome lines responding selectively to varying spatial environments. For example, rosasomal lines mantled with nucleotide and peptide layers are conceived as primordial forerunners of the ubiquitous ribosome. Moreover, the parallel independent and interdependent evolution of rosasome lines would be more rapid than sequential development, refute precedence of either DNA or RNA, and explain the evolution of integration of two subunits with different structures and functions in ribosomes and of the triplet nature of the codon. Based on recent astronomical data, this hypothesis supports the concept that life is not a singularity. This concept also suggests a potential vehicle for therapeutics, biotechnology and genetic engineering.


A novel DNA-binding protein modulating methicillin resistance in Staphylococcus aureus.
Methicillin resistance in Staphylococcus aureus is conferred by the mecA-encoded penicillin-binding protein PBP2a. Additional genomic factors are also known to influence resistance levels in strain specific ways, although little is known about their contribution to resistance phenotypes in clinical isolates. Here we searched for novel proteins binding to the mec operator, in an attempt to identify new factor(s) controlling methicillin resistance phenotypes. Analysis of proteins binding to a DNA fragment containing the mec operator region identified a novel, putative helix-turn-helix DNA-binding protein, SA1665. Nonpolar deletion of SA1665, in heterogeneously methicillin resistant S. aureus (MRSA) of different genetic backgrounds, increased methicillin resistance levels in a strain dependent manner. This phenotype could be fully complemented by reintroducing SA1665 in trans. Northern and Western blot analyses, however, revealed that SA1665 had no visible influence on mecA transcription or amounts of PBP2a produced. SA1665 is a new chromosomal factor which influences methicillin resistance in MRSA. Although SA1665 bound to the mecA promoter region, it had no apparent influence on mecA transcription or translation, suggesting that this predicted DNA-binding protein modulates resistance indirectly, most likely through the control of other genomic factors which contribute to resistance.


Clarification of the characteristics of needle-tip movement during vacuum venipuncture to improve safety.
Complications resulting from venipuncture include vein and nerve damage, hematoma, and neuropathic pain. Although the basic procedures are understood, few analyses of actual data exist. It is important to improve the safety standards of this technique during venipuncture. This study aimed to obtain data on actual needle movement during vacuum venipuncture in order to develop appropriate educational procedures. Six experienced nurses were recruited to collect blood samples from 64 subjects. These procedures were recorded using a digital camera. Software was then used to track and analyze motion without the use of a marker in order to maintain the sterility of the needle. Movement along the X- and Y-axes during blood sampling was examined. Approximately 2.5 cm of the needle was inserted into the body, of which 6 mm resulted from advancing or moving the needle following puncture. The mean calculated puncture angle was 15.2°. Given the hazards posed by attaching and removing the blood collection tube, as well as by manipulating the needle to fix its position, the needle became unstable whether it was fixed or not fixed. This study examined venipuncture procedures and showed that the method was influenced by increased needle movement. Focusing on skills for puncturing the skin, inserting the needle into the vein, and changing hands while being conscious of needle-tip stability may be essential for improving the safety of venipuncture.


Biofilm formation and changes in bacterial cell surface hydrophobicity during growth in a CAPD model system.
Peritonitis is a frequent complication of continuous ambulatory peritoneal dialysis (CAPD), with patients suffering recurrent attacks. The microorganisms most frequently implicated in the infection are the skin microflora, in particular, the coagulase-negative staphylococci such as Staphylococcus epidermidis. These microorganisms gain access to the peritoneal cavity via the in-dwelling silicone rubber catheter in the abdominal wall and often persist as biofilms on the surface of the catheter. The surface characteristics of S. epidermidis were monitored during growth in a CAPD in-vitro model together with their ability to adhere to silicone rubber substrata. Fresh dialysis fluid exerted an injurious effect on the cells leading to a decrease in cell numbers but during the simulated dialysis period the cells adapted to the applied stresses. Over a 96-h period in the model both a clinical isolate and a skin isolate of S. epidermidis adopted a more hydrophobic phenotype. The data presented here show that the bacteria grown in this in-vivo reflective CAPD model continually adapt to their environment and become more tolerant to the stresses imposed. The adapted cells were seen to colonise silicone rubber substrata.


Developmental changes in the balance of glycolytic ATP production and oxidative phosphorylation in ventricular cells: A simulation study.
The developmental program of the heart requires accurate regulation to ensure continuous circulation and simultaneous cardiac morphogenesis, because any functional abnormalities may progress to congenital heart malformation. Notably, energy metabolism in fetal ventricular cells is regulated in a manner that differs from adult ventricular cells: fetal cardiomyocytes generally have immature mitochondria and fetal ventricular cells show greater dependence on glycolytic ATP production. However, although various characteristics of energy metabolism in fetal ventricular cells have been reported, to our knowledge, a quantitative description of the contributions of these factors to fetal ventricular cell functions has not yet been established. Here, we constructed a mathematical model to integrate various characteristics of fetal ventricular cells and predicted the contribution of each characteristic to the maintenance of intracellular ATP concentration and sarcomere contraction under anoxic conditions. Our simulation results demonstrated that higher glycogen content, higher hexokinase activity, and lower creatine concentration helped prolong the time for which ventricular cell contraction was maintained under anoxic conditions. The integrated model also enabled us to quantitatively assess the contributions of factors related to energy metabolism in ventricular cells. Because fetal cardiomyocytes exhibit similar energy metabolic profiles to stem cell-derived cardiomyocytes and those in the failing heart, an improved understanding of these fetal ventricular cells will contribute to a better comprehension of the processes in stem cell-derived cardiomyocytes or under pathological conditions.


Phase II trial of yttrium-90-DOTA-biotin pretargeted by NR-LU-10 antibody/streptavidin in patients with metastatic colon cancer.
A Phase II study of yttrium-90-tetra-azacyclododecanetetra-acetic acid-biotin (90Y-DOTA-biotin) pretargeted by NR-LU-10 antibody/streptavidin (SA) was performed. The primary objectives of the study were to evaluate the efficacy and safety of this therapy in patients with metastatic colon cancer. Twenty-five patients were treated with a single dose of 110 mCi/m2 (mean administered dose, 106.5 +/- 10.3 mCi/m2) of 90Y-DOTA-biotin. There were three components of the therapy. Patients first received NR-LU-10/SA on day 1. A clearing agent (biotin-galactose-human serum albumin) was administered approximately 48 h after the NR-LU-10/SA to remove residual circulating unbound NR-LU-10/SA. Lastly, 24 h after administration of clearing agent, patients received biotin-DOTA-labeled with 110 mCi/m2 90Y. All three components of the therapy were administered i.v. Both hematological and nonhematological toxicities were observed. Diarrhea was the most frequent grade 4 nonhematological toxicity (16%; with 16% grade 3 diarrhea). Hematological toxicity was less severe with 8% grade 3 and 8% grade 4 neutropenia and 8% grade 3 and 16% grade 4 thrombocytopenia. The overall response rate was 8%. Two partial responders had freedom from progression of 16 weeks. Four patients (16%) had stable disease with freedom from progression of 10-20 weeks. Despite the relatively disappointing results of this study in terms of therapeutic efficacy and toxicity, proof of principle was obtained for the pretargeting approach. In addition, valuable new information was obtained about normal tissue tolerance to low-dose-rate irradiation that will help to provide useful guidelines for future study designs.


Modular total syntheses of mycolactone A/B and its [2H]-isotopologue.
A modular total synthesis of mycolactone A/B, the exotoxin produced by Mycobacterium ulcerans, has been achieved through the orchestration of several Pd-catalyzed key steps. While this route leads to a mixture of the natural product and its C12 epimer (4 : 1 ratio), this was inconsequential from the biological activity standpoint. Compared to the previously reported routes, this synthetic blueprint allows the late-stage modification of the toxin, as exemplified by the preparation of [22,22,22-2H3]-mycolactone A/B.


Cyclooxygenase-1 is overexpressed and promotes angiogenic growth factor production in ovarian cancer.
Inhibition of cyclooygenase-2 (COX-2) catalytic activity has proven successful in restricting the growth of epithelial-derived cancers in vivo. Whether COX-2 inhibitor therapy would be beneficial in the prevention and/or treatment of ovarian cancer, the most lethal gynecological malignancy worldwide, is not known. Most patients with ovarian cancer undergo cytoreductive therapy. Because many of the cytotoxic drugs used to treat ovarian cancer induce COX-2 expression, samples from patients that had not undergone cytoreductive therapy were specifically chosen for COX isoform expression analysis. A majority of specimens exhibited elevated levels of COX-1, not COX-2, mRNA, and protein compared with normal ovarian tissue. Focal regions within the tumor expressing high COX-1 also had elevated levels of pro-angiogenic proteins. Selective inhibition of COX-1, not COX-2, inhibited arachidonic acid-stimulated vascular endothelial growth factor production, which could be reversed by cotreatment with prostaglandin E(2). Thus, COX-1 may contribute to carcinoma development in the ovary through stimulation of neovascularization. Clinical studies testing the efficacy of COX inhibition as adjuvant therapy for ovarian cancer may see more beneficial effects with adjuvant therapy with either a COX-1 selective or nonselective cyclooxygenase inhibitor as compared with a COX-2 selective drug.