Deptor knockdown enhances mTOR Activity and protein synthesis in myocytes and ameliorates disuse muscle atrophy.
Deptor is an mTOR binding protein that affects cell metabolism. We hypothesized that knockdown (KD) of Deptor in C2C12 myocytes will increase protein synthesis via stimulating mTOR-S6K1 signaling. Deptor KD was achieved using lentiviral particles containing short hairpin (sh)RNA targeting the mouse Deptor mRNA sequence, and control cells were transfected with a scrambled control shRNA. KD reduced Deptor mRNA and protein content by 90%, which increased phosphorylation of mTOR kinase substrates, 4E-BP1 and S6K1, and concomitantly increased protein synthesis. Deptor KD myoblasts were both larger in diameter and exhibited an increased mean cell volume. Deptor KD increased the percentage of cells in the S phase, coincident with an increased phosphorylation (S807/S811) of retinoblastoma protein (pRb) that is critical for the G(1) to S phase transition. Deptor KD did not appear to alter basal apoptosis or autophagy, as evidenced by the lack of change for cleaved caspase-3 and light chain (LC)3B, respectively. Deptor KD increased proliferation rate and enhanced myotube formation. Finally, in vivo Deptor KD (~50% reduction) by electroporation into gastrocnemius of C57/BL6 mice did not alter weight or protein synthesis in control muscle. However, Deptor KD prevented atrophy produced by 3 d of hindlimb immobilization, at least in part by increasing protein synthesis. Thus, our data support the hypothesis that Deptor is an important regulator of protein metabolism in myocytes and demonstrate that decreasing Deptor expression in vivo is sufficient to ameliorate muscle atrophy.


Bone turnover markers and prediction of fracture: a prospective follow-up study of 1040 elderly women for a mean of 9 years.
Osteoporosis is characterized by compromised bone mass and strength, predisposing to an increased risk of fracture. Increased bone metabolism has been suggested to be a risk factor for fracture. The aim of this study was to evaluate whether baseline bone turnover markers are associated with long-term incidence of fracture in a population-based sample of 1040 women who were 75 years old (Malmö OPRA study). Seven bone markers (S-TRACP5b, S-CTX-I, S-OC[1-49], S-TotalOC, S-cOC, S-boneALP, and urinary osteocalcin) were measured at baseline and 1-year follow-up visit. During the mean follow-up of 9.0 years (range 7.4-10.9), 363 women sustained at least one fracture of any type, including 116 hip fractures and 103 clinical vertebral fractures. High S-TRACP5b and S-CTX-I levels were associated with increased risk of any fracture with hazard ratios [HRs (95% confidence interval)] of 1.16 (1.04-1.29) and 1.13 (1.01-1.27) per SD increase, respectively. They also were associated with increased risk of clinical vertebral fracture with HRs of 1.22 (1.01-1.48) and 1.32 (1.05-1.67), respectively. Markers were not associated with risk for hip fracture. Results were similar when we used resorption markers, including urinary osteocalcin, measured at the 1-year visit or an average of the two measurements. The HRs were highest for any fracture in the beginning of the follow-up period, 2.5 years from baseline. For vertebral fractures, the association was more pronounced and lasted for a longer period of time, at least for 5 years. In conclusion, elevated levels of S-TRACP5b, S-CTX-I, and urinary osteocalcin are associated with increased fracture risk for up to a decade in elderly women.


Predicting recovery from head injury in young children: a prospective analysis.
It has been argued that young children's brains are "plastic," and may sustain substantial brain insult with little loss of function. Recent research suggests that this notion may not apply for generalized cerebral pathology. The present study aimed to evaluate this proposition using a sample of 73 young children, divided into 3 groups: severe head injury (HI; N = 17); mild-moderate HI (N = 32); and noninjured controls (N = 24). Preinjury screening established equivalence across groups for age, sex, preinjury ability, behavioral adjustment, socioeconomic status, and family functioning. Children were evaluated as soon as possible postinjury, and again 12 months postinjury, in three domains: intellectual ability, language, and memory. Results indicated that severe HI was associated with substantial, persisting difficulties in all areas. In contrast, children with mild-moderate HI experienced fewer difficulties, and often performed similarly to controls, both acutely and 12 months postinjury. There was no evidence of differential recovery of function associated with injury severity, with performance increments consistent across groups and probably due to either age-appropriate developmental gains, or test-related practice effects. Poorer outcome at 12 months postinjury was predicted by injury severity primarily, with earlier age at injury, and premorbid ability associated with outcome in specific domains.


The effectiveness of interventions during and after residence in women's shelters: a meta-analysis.
Due to intimate partner violence (IPV), a proportion of some women seek safety from their abuser in shelters. Little is known yet about whether the received shelter interventions, offered during and after women's stay, are effective. Therefore, a meta-analysis of studies was performed, testing the effectiveness of interventions administered to female IPV victims during and/or after their residence in shelters, in terms of mental health, re-abuse and social outcomes. From January 1985 through July 2011, five databases were searched for English-language articles. Original research articles evaluating the effects of interventions provided to IPV victims during and after shelter residence were identified. Hedges' g effect sizes and 95% confidence intervals (CIs) were calculated and pooled if three or more studies including one outcome type were available. Ten original studies, including a total number of 726 female IPV victims, were analysed. We found that shelter interventions were effective in improving mental health outcomes (g = -0.39; 95% CI: 0.24-0.54), in decreasing abuse (g = 0.32; 95% CI: 0.08-0.55) and in improving social outcomes (g = 0.71; 95% CI: 0.54-0.88) in shelter-based abused women. This analysis suggests that interventions provided during and after stay in a shelter are effective in improving mental health, abuse and social outcomes, but further research has to confirm this. Moreover, future studies should compare different type of interventions and should evaluate cost-effectiveness.


Dog response to inactivated canine parvovirus and feline panleukopenia virus vaccines.
Inactivated canine parvovirus (CPV) and inactivated feline panleukopenia virus (FPV) vaccines were evaluated in dogs. Maximal serologic response occurred within 1-2 weeks after vaccination. Antibody titers then declined rapidly to low levels that persisted at least 20 weeks. Immunity to CPV, defined as complete resistance to infection, was correlated with serum antibody titer and did not persist longer than 6 weeks after vaccination with inactivated virus. However, protection against generalized infection was demonstrated 20 weeks after vaccination. In unvaccinated dogs, viremia and generalized infection occurred after oronasal challenge with virulent CPV. In contrast, viral replication was restricted to the intestinal tract and gut-associated lymphoid tissue of vaccinated dogs. Canine parvovirus was inactivated by formalin, beta-propiolactone (BPL), and binary ethylenimine (BEI) in serum-free media; inactivation kinetics were determined. Formalin resulted in a greater loss of viral HA than either BEI of BPL, and antigenicity was correspondingly reduced.


[The Activity Theory as a conceptual model for nursing].
The search for theoretical models in order to provide inputs and to reconsider the nursing practice, as well as to found research and teaching, has been a concern among contemporaneous nursing researchers. This article proposes a glance at the pedagogical assumption that supports the application of the Activity Theory, i.e., qualifying subjects to learn how to learn, to build and to search for knowledge. This theory also assumes actions in dynamic contexts of action-operation, a feature that must be emphasized in nursing education, as well as in the development of the abilities needed for group work.


Subnanometer vacancy defects introduced on graphene by oxygen gas.
The basal plane of graphene has been known to be less reactive than the edges, but some studies observed vacancies in the basal plane after reaction with oxygen gas. Observation of these vacancies has typically been limited to nanometer-scale resolution using microscopic techniques. This work demonstrates the introduction and observation of subnanometer vacancies in the basal plane of graphene by heat treatment in a flow of oxygen gas at low temperature such as 533 K or lower. High-resolution transmission electron microscopy was used to directly observe vacancy structures, which were compared with image simulations. These proposed structures contain C═O, pyran-like ether, and lactone-like groups.


Music training relates to the development of neural mechanisms of selective auditory attention.
Selective attention decreases trial-to-trial variability in cortical auditory-evoked activity. This effect increases over the course of maturation, potentially reflecting the gradual development of selective attention and inhibitory control. Work in adults indicates that music training may alter the development of this neural response characteristic, especially over brain regions associated with executive control: in adult musicians, attention decreases variability in auditory-evoked responses recorded over prefrontal cortex to a greater extent than in nonmusicians. We aimed to determine whether this musician-associated effect emerges during childhood, when selective attention and inhibitory control are under development. We compared cortical auditory-evoked variability to attended and ignored speech streams in musicians and nonmusicians across three age groups: preschoolers, school-aged children and young adults. Results reveal that childhood music training is associated with reduced auditory-evoked response variability recorded over prefrontal cortex during selective auditory attention in school-aged child and adult musicians. Preschoolers, on the other hand, demonstrate no impact of selective attention on cortical response variability and no musician distinctions. This finding is consistent with the gradual emergence of attention during this period and may suggest no pre-existing differences in this attention-related cortical metric between children who undergo music training and those who do not.


Plasma and Brain Concentrations of Doxycycline after Single and Repeated Doses in Wild-Type and APP23 Mice.
Repurposing doxycycline for the treatment of amyloidosis has recently been put forward because of the antiaggregating and anti-inflammatory properties of the drug. Most of the investigations of the therapeutic potential of doxycycline for neurodegenerative amyloidosis, e.g., prion and Alzheimer disease (AD), have been carried out in mouse models, but surprisingly no data are available regarding the concentrations reached in the brain after systemic administration. We filled this gap by analyzing the pharmacokinetic profile of doxycycline in plasma and brain after single and repeated intraperitoneal injections of 10 and 100 mg/kg, in wild-type mice and the APP23 mouse model of AD. The main outcomes of our study are: 1) Peak plasma concentrations ranged from 2 to10 μg/ml, superimposable to those in humans; 2) brain-to-plasma ratio was ∼0.2, comparable to the cerebrospinal fluid/serum ratios in humans; 3) brain Cmax 4-6 hours after a single dose was ∼0.5 (10 mg/kg) and ∼5 μM (100 mg/kg). Notably, these concentrations are lower than those required for the drug's antiaggregating properties as observed in cell-free studies, suggesting that other features underlie the positive cognitive effects in AD mice; 4) elimination half-life was shorter than in humans (3-6 vs. 15-30 hours), therefore no significant accumulation was observed in mouse brain following repeated treatments; and 5) there were no differences between doxycycline concentrations in brain areas of age-matched wild-type and APP23 mice. These data are useful for planning preclinical studies with translational validity, and to identify more reliably the mechanism(s) of action underlying the central in vivo effects of doxycycline.


[Genetic study of Escherichia coli K-12 mutants resistant to 2,6-diaminopurine].
Mutants, resistant to the inhibitory effect of 2,6-diaminopurine and incapable of utilizing adenine as a purine source, are obtained from purinenucleoside phosphorylase-defective purine-dependent Escherichia coli K-12 strains. The mutations obtained (apt) disturb the uptake of adenosine and inosine only in the presence of a mutation for purinenucleoside phosphorylase (pup gene) in the genome of purine-dependent bacteria. The introduction of pup+ allele into the genome of mutants obtained (genotype purDpup apt) results in the restoration of the ability to uptake adenine and purine ribosides. Strains of purDpup+apt genotype are characterized by more short generation time under the growth in the presence of adenine as compared with purDpup apt+ strains which indicates the existance of an efficient pathway of adenine utilization in E. coli with the cooperation of purinenucleoside phosphorylase. Mutations apt have revealed a combined transfer with purE marker under the transduction frequency of approximately 5%. The gene order on E. coli K-12 chromosome is apt-purE-gal, as estimated from the data on conjugation crosses.


Fibrosing interstitial pneumonia predicts survival in patients with rheumatoid arthritis-associated interstitial lung disease (RA-ILD).
Rheumatoid arthritis (RA) is a systemic autoimmune disorder with a variety of extra-articular manifestations. The lung is a common target and diffuse parenchymal lung disease can appear as any of the patterns found with idiopathic interstitial pneumonia. Controversy exists as to the prognostic significance of these patterns among patients with RA-ILD. We retrospectively identified 48 patients with a diagnosis of RA-ILD confirmed by surgical lung biopsy. The pathology was reviewed by four expert pulmonary pathologists. We examined survival after stratifying on the presence or absence of fibrotic ILD, and contrasted it with a matched idiopathic pulmonary fibrosis (IPF) population. The Cox proportional hazards model was used to identify independent predictors of survival. The majority of subjects were male smokers with physiologic restriction. A usual interstitial pneumonia (UIP)-pattern was identified in 31% of subjects. Median survival time for the entire cohort was 1360 days. Subjects with fibrotic ILD had worse survival than subjects with non-fibrotic ILD (log rank p = 0.02). There was no difference in survival between UIP-pattern RA-ILD subjects and IPF controls (log rank p = 0.94). Multivariable analysis revealed that age (hazard ratio [HR] = 1.04, p = 0.01) and fibrosis (HR = 2.1, p = 0.02) were independent predictors of mortality. Both cellular and fibrosing ILD patterns are common among RA-ILD patients who undergo surgical lung biopsy. These patients have a shortened survival when compared to the general population and all-comers with RA. Age and the presence of a fibrosing interstitial pneumonia predict shortened survival in these patients. Survival in UIP is similar to matched IPF patients.


Extraction and separation of volatile and fixed oils from seeds of Myristica fragrans by supercritical CO₂: chemical composition and cytotoxic activity on Caco-2 cancer cells.
Isolation of volatile and fixed oils from nutmeg have been obtained by supercritical fractioned extraction with carbon dioxide. Extraction experiments were carried out at pressures of 90 and 250 bar and temperature of 40 °C. The extraction step performed at 90 bar produced a volatile fraction mainly formed by myristicin (32.8%), sabinene (16.1%), α-pinene (9.8%), β-pinene (9.4%), β-phellandrene (4.9%), safrole (4.1%) and terpinen-4-ol (3.6%). The oil yield relative to this step of the process was 1.4% by weight of the charge. The last extraction step at 250 bar produced a butter-like material (nutmeg butter). The yield of this step was 14.4% by weight. The most represented fatty acids of fixed oil from nutmeg were 14:0 (79.2%), 18:1 n-9 (7.4%) and 16:0 (6.1%), and in particular the unsaturated fatty acids 18:1 n-9 averaged 32.96 μg/mg of oil. The level of myristicin in the nutmeg essential and fixed oils was also directly quantified by reversed HPLC-DAD. Moreover, the essential oil obtained from nutmeg, as well as myristicin, showed a significant in vitro inhibitory effect on the growth of a colon cancer cell line (undifferentiated Caco-2 cells). In this study, the chemical characterization and the anticancer activity of nutmeg oils obtained by supercritical extraction with carbon dioxide were investigated. This is important for their potential application in food and pharmaceutical industries.


Theory of mind, empathy and eye gaze strategies during observation of a work of art: neurodegenerative pathologies.
Theory of mind (ToM) and empathy are severely impaired in the behavioural variant of frontotemporal dementia (bvFTD) and more mildly in Alzheimer's (AD) and Parkinson's diseases (PD). Such impairments are associated with behavioural disorders. Modification of visual scanning strategies of complex visual scenes is also found in these pathologies. We hypothesised that these patients applied atypical gaze strategies when observing social events, which does not allow them to properly process social cues and results in the production of erroneous inferences and lack of empathy towards others. Fifty-five participants were divided into four groups: five bvFTD subjects, 19 AD subjects, 17 PD subjects and 14 matched control subjects. ToM and empathy were assessed by eye movement recording (eye-tracking) and by a questionnaire during a painting observation. Scores obtained were compared between each group and to social cognition reference tests and correlated to the NeuroPsychiatric Inventory. Our paradigm was suitable for assessing cognitive ToM while it lacked sensitivity for empathy assessment. Severe ToM impairment was highlighted in bvFTD, while milder difficulties were observed in AD and for PD. Subjects with bvFTD and AD produced erroneous inferences from cognitive mental states. ToM performances were linked to visual exploration strategies of the painting. Atypical visual observation was highlighted in the bvFTD and AD groups causing a time delay in taking the perspective of the other person. Finally, we highlighted that social cognition performances, gaze strategies and behavioural disorders were correlated. Observing a painting in association with eye-tracking technology can be a good support for social cognition assessment. We highlighted a link between atypical visual scanning strategies, ToM impairment and behavioural disorders in these pathologies. ToM skills could be improved by training in the search for visual social cues. Therefore, this kind of remediation could have positive effects on behavioural disorders.


Heterologous passive cutaneous anaphylaxis with bovine immunoglobulin E.
Immunoglobulin (Ig) E was detected in sera of cows previously inoculated with ovalbumin in alum or in Freund's incomplete adjuvant. Titers greater than 64 were produced by homologous passive cutaneous anaphylaxis (PCA) reactions on calf skin, using sera from these cows. This bovine IgE was heat labile. Bovine sera containing IgE were used to examine cross-species reactivity by heterologous PCA. Rats, rabbits, guinea pigs, and dogs failed to develop PCA reactions when sensitized by intradermal injections of the bovine IgE and challenge exposed at intervals from 2 to 72 hours later with IV ovalbumin and Evan's blue dye. In contrast, sheep and goats became passively sensitized by the bovine IgE. Considerable variation in latent period and titer was apparent after comparison of results in heterologous species with each other and with the homologous recipient. Nonetheless, the ability of this homocytotropic antibody to cross species is important to the phylogenetic relationship of IgE in different species.


Hepatocyte expression of the senescence marker p21 is linked to fibrosis and an adverse liver-related outcome in alcohol-related liver disease.
Alcohol-related liver disease (ALD) remains a leading cause of liver-related morbidity and mortality. Age, fibrosis stage, MELD score and continued alcohol consumption predict outcome in everyday clinical practice. In previous studies increased hepatocyte nuclear area and hepatocyte expression of p21, both markers of senescence, were associated with increased fibrosis stage and a poor outcome in non-alcohol-related fatty liver disease, while increased hepatocyte nuclear area was related to liver dysfunction in ALD cirrhosis. This study, therefore, investigated the pattern of hepatocyte cell cycle phase distribution and hepatocyte p21 expression in relation to outcome in ALD. Liver sections from two cohorts were studied. The first comprised 42 patients across the full spectrum of ALD. The second cohort comprised 77 patients with ALD cirrhosis. Immunohistochemistry assessed hepatocyte expression of cell cycle phase markers and p21. Regenerating liver (n=12) and "normal" liver sections (n=5) served as positive and negative controls, respectively. In the first cohort there was little cell cycle progression beyond G1/S phase and increased hepatocyte p21 expression (p<0.0001), which correlated independently with fibrosis stage (p=0.005) and an adverse liver-related outcome (p=0.03). In the second cohort, both hepatocyte p21 expression (p<0.001) and MELD score (p=0.006) were associated independently with an adverse liver-related outcome; this association was stronger with hepatocyte p21 expression (AUROC 0.74; p=0.0002) than with MELD score (AUROC 0.59; p=0.13). Further, hepatocyte p21 expression co-localised with increased hepatic stellate cell activation. The findings are consistent with impaired cell cycle progression beyond the G1/S phase in ALD. The striking independent associations between increased hepatocyte p21 expression and both fibrosis stage and an adverse liver-related outcome in both cohorts suggests hepatocyte senescence plays an important role in ALD. Measuring hepatocyte p21 expression is simple and cheap and in this series was a useful measure of long-term prognosis in ALD.


An atypical E3 ligase zinc finger protein 91 stabilizes and activates NF-kappaB-inducing kinase via Lys63-linked ubiquitination.
The NF-κB transcription factors control many physiological processes, including inflammation, immunity, and apoptosis. Its activity contributes to the development of various cell malignancies. NF-κB-inducing kinase (NIK) plays a pivotal role in NF-κB activation. However, the molecular mechanism to stabilize and activate NIK remains elusive, although it is known that cIAP1/2 (cellular inhibitor of apoptosis 1 and 2) ubiquitinate NIK for degradation. Here, we report a novel NF-κB-related zinc finger protein 91 (ZFP91) that stabilizes and activates NIK in a ubiquitination-dependent manner. We show that ZFP91 interacts with and promotes the Lys(63)-linked ubiquitination of NIK and subsequent processing of p100 to p52. The results of in vitro biochemical assays indicate that ZFP91 functions as an E3 ligase directly to NIK. Remarkably, the ubiquitination of NIK coincides with its Thr(559) phosphorylation. Furthermore, knockdown of ZFP91 expression by RNA interference inhibits the CD40 ligation-induced activation of NIK and p100 processing as well as the expression of noncanonical NF-κB target genes. These data clearly indicate that ZFP91 is an important regulator of the noncanonical NF-κB pathway.


Thermally Induced Encapsulation of Food Nutrients into Phytoferritin through the Flexible Channels without Additives.
The cavity of phytoferritin provides a nanospace to encapsulate and deliver food nutrient molecules. However, tranditional methods to prepare the ferritin-nutrient complexes must undergo acid/alkaline conditions or apply additives. In this work, we provide a novel guideline that thermal treatment at 60 °C can expand ferritin channels by uncoiling the surrounding α-helix. Upon reduction of the temperature to 20 °C, food nutrient rutin can be encapsulated in apo-soybean seed ferritin (apoSSF) at pH 7.0 through channels without disassembly of the protein cage and with no addition of additives. Results indicated that one apoSSF could encapsulate about 10.5 molecules of rutin, with an encapsulation ratio of 8.08% (w/w). In addition, the resulting rutin-loaded SSF complexes were monodispersed in a size of 12 nm in aqueous solution. This work provides a novel pathway for the encapsulation of food nutrient molecules into the nanocavity of ferritin under a neutral pH condition induced by thermal treatment.


Fine elasticity imaging utilizing the iterative RF-echo phase matching method.
To non-invasively quantify elasticity of soft tissue, we previously developed the iterative two-dimensional (2-D) rf-echo phase matching method for accurately measuring a 2-D displacement vector field generated in vivo in soft tissue during acquisition of two successive rf-echo data frames. We also developed a stable method for uniquely reconstructing a shear modulus distribution using strains derived from the measurement data. However, as in our measurement method a displacement is determined by using the phase characteristics of the finite local echo data as the index to iteratively search for the corresponding local data, change of the local phase characteristics due to tissue deformation deteriorates the accuracy of the determination. Thus, we improve the previous method such that, in principle, the displacement can be determined using an infinitesimal phase characteristics. That is, we incorporate an effective mechanism into the previous iterative phase matching scheme: the local size is made suitably smaller during the iterative phase matching. The demonstrated ability of measurement and reconstruction in simulation, and experiments on in vitro in pork rib and in vivo in breast tissue, shows this refinement allows not only better spatial resolution of the shear modulus image but also improved accuracy, and indicates that the improved method has a high potential to be applied for various soft tissues.


Effects of maternal identity and incubation temperature on snapping turtle (Chelydra serpentina) metabolism.
Individual variation in physiological traits may have important consequences for offspring survivorship and adult fitness. Variance in offspring phenotypes is due to interindividual differences in genotype, environment, and/or maternal effects. This study examined the contributions of incubation environment, maternal effects, and clutch identity to individual variation in metabolic rates in the common snapping turtle, Chelydra serpentina. We measured standard metabolic rate, as determined by oxygen consumption, for 246 individuals representing 24 clutches at 15 degrees and 25 degrees C, and we measured standard metabolic rates additionally for 34 individuals at 20 degrees and 30 degrees C. Standard metabolic rate for 34 snapping turtles measured at 15 degrees, 20 degrees, 25 degrees, and 30 degrees C increased with increasing temperature. Mean standard metabolic rate for 246 individuals was 0.247 microL O(2) min(-1) g(-1) at 15 degrees C and 0.919 microL O(2) min(-1) g(-1) at 25 degrees C. At 15 degrees C, mass at hatching, individual mass, and egg mass had no significant effects on metabolic rate, but at 25 degrees C, mass at hatching, individual mass, and egg mass did have significant effects on metabolic rate. Incubation temperature had no significant effect on metabolic rate at 15 degrees, but it did have a significant effect at 25 degrees C. Clutch identity had a significant effect on metabolic rate at both 15 degrees and 25 degrees C. Interindividual variation in standard metabolic rate due to incubation temperature, and especially clutch identity, could have large effects on energy budgets. Results suggest that there were both environmental and genetic effects on standard metabolic rate.


Knowledge, group-based medical mistrust, future expectations, and perceived disadvantages of medical genetic testing: perspectives of Black African immigrants/refugees.
Reasons for low participation of ethnic minorities in genetic studies are multifactorial and often poorly understood. Based on published literature, participation in genetic testing is low among Black African immigrants/refugees although they are purported to bear disproportionate disease burden. Thus, research involving Black African immigrant/refugee populations that examine their perspectives on participating in genetic studies is needed. This report examines and describes the knowledge of medical genetics, group-based medical mistrust, and future expectations of genetic research and the influence of these measures on the perceived disadvantages of genetic testing among Black African immigrants/refugees. Using a cross-sectional survey design, a nonprobability sample (n = 212) of Black African immigrants/refugees was administered a questionnaire. Participants ranged in age from 18 to 61 years (mean = 38.91, SD = 9.78). The questionnaire consisted of 5 instruments: (a) sociodemographic characteristics, (b) Knowledge of Medical Genetics scale, (c) Group-Based Medical Mistrust Scale, (d) Future Expectations/Anticipated Consequences of Genetics Research scale, and (e) Perceived Disadvantages of Genetic Testing scale. Participants were concerned that genetic research may result in scientists 'playing God,' interfering with the natural order of life. In multivariate analyses, the perceived disadvantages of genetic testing increased as medical mistrust and anticipated negative impacts of genetic testing increased. Increase in genetic knowledge contributed to a decrease in perceived disadvantages. Our findings suggest that recruitment of Black African immigrants/refugees in genetic studies should address potential low knowledge of genetics, concerns about medical mistrust, the expectations/anticipated consequences of genetic research, and the perceived disadvantages of genetic testing.


Nutritional implications of D-xylose in pigs.
Hemicellulose consists primarily of pentose sugars, joined together in a polysaccharide chain with D-xylose as the most abundant component. Ileal digestibility and urinary excretion of D-xylose and associated effects of this pentose sugar on ileal and faecal digestibility of dry matter (DM), organic matter (OM), gross energy (GE) and nitrogen were studied in pigs. Castrated pigs were prepared with a post-valvular T-caecum cannula to measure ileal digestibility. Faecal digestibility was measured in non-cannulated pigs. D-Xylose was given at dietary inclusion levels of 100 and 200 g/kg, and the control sugar, D-glucose, at a rate of 200 g/kg diet. Ileal digestibility of D-xylose as well as that of D-glucose was found to be close to 100%. The presence of D-xylose in the diet decreased ileal digesta pH and increased ileal flow of volatile fatty acids, suggesting the occurrence of microbial degradation of D-xylose in the pig small intestine. In pigs fed on the 100 g D-xylose/kg diet, 44.5% of the D-xylose intake appeared in the urine. This percentage increased significantly to 52.6 when pigs were fed on the 200 g D-xylose/kg diet. Ileal and faecal digestibility of DM, OM, GE and N, as well as N retention, decreased significantly in pigs fed on the 200 g D-xylose/kg diet.


Firefly "femmes fatales" acquire defensive steroids (lucibufagins) from their firefly prey.
Female fireflies of the genus Photuris, the so-called firefly "femmes fatales," prey on male fireflies of the genus Photinus. The females are able to entrap the males by faking the flash signal characteristics of the Photinus female. We found that by feeding on Photinus males, Photuris females gain more than nutrients. They also acquire defensive steroidal pyrones called lucibufagins, which are contained in Photinus but which Photuris fireflies are unable to produce on their own. Photuris females that eat Photinus males or lucibufagin are rejected by Phidippus jumping spiders. Lucibufagin itself proved to be a deterrent to such spiders. Field-collected Photuris females contain lucibufagin in varying amounts. The more lucibufagin they contain the more unacceptable they are to Phidippus.


The spatial and temporal pattern of fatty acid amide hydrolase expression in rat hippocampus during postnatal development.
GABAergic synaptic transmission is efficiently controlled by endogenous cannabinoids in cortical structures. Fatty acid amide hydrolase (FAAH) is one of the metabolizing enzymes of endocannabinoids in the brain. In this study we investigated the cellular and subcellular distribution of FAAH at various timepoints during the first postnatal weeks, when GABA is still depolarizing, and plays a crucial role in network events. FAAH immunoreactivity is strong in the CA3 region already at postnatal day 0 (P0), but in CA1 only after P8. During this period, FAAH levels in hilar mossy cells decrease and in granule cells slowly increase. Pyramidal cells express FAAH first in the soma and proximal dendrites, and gradually in more distal segments, reaching adult levels in the most distal dendrites only at P22. Transient expression of FAAH was found in a small number of stratum radiatum cells that may be interneurons and in ependymal cells at the border of the alveus and corpus callosum between P2 and P8. At the ultrastructural level, FAAH distribution at all ages was very similar to the adult pattern, i.e. it was largely associated with the membrane of cytoplasmic vesicles, mitochondria and endoplasmic reticulum. During postnatal development of the hippocampus, the spatio-temporal expression of FAAH correlates well with the general pattern of neuronal maturation, but not with the arrival of afferent pathways, which suggests that FAAH - and its major endocannabinoid substrate, anandamide - is unlikely to be involved in the presynaptic control of neurotransmission. Instead, FAAH may subserve general roles as the inactivating enzyme for many fatty acid amides, in addition to anandamide.


Proline biosynthesis in Streptococcus thermophilus: characterization of the proBA operon and its products.
The presence of proline in the medium was not essential for growth of Streptococcus thermophilus, indicating that there is a proline biosynthetic pathway in this organism. Genetic and biochemical analysis identified and characterized this pathway. Two genes, designated proB and proA, were cloned, sequenced and characterized. Biochemical analysis of the proB- and proA-encoded enzymes showed that the proline biosynthetic pathway of S. thermophilus is similar to the one previously described in Escherichia coli. The deduced amino acid sequence of a 2-408 kb DNA region containing the genes revealed the similarity of the S. thermophilus gene products to ProB and ProA of E. coli and Serratia marcescens, and to the corresponding N- and C-terminal domains of the bifunctional plant enzyme delta 1-pyrroline-5-carboxylate synthetase of Vigna aconitifolia. Northern blot analysis showed that the two genes in S. thermophilus are organized in a single operon with proB proximal and proA distal to the promoter; primer extension analysis indicated that proBA transcription is not under repressive control by exogenously supplied proline.


Fluorescent silica nanoparticles improve optical imaging of stem cells allowing direct discrimination between live and early-stage apoptotic cells.
Highly bright and photostable cyanine dye-doped silica nanoparticles, IRIS Dots, are developed, which can efficiently label human mesenchymal stem cells (hMSCs). The application procedure used to label hMSCs is fast (2 h), the concentration of IRIS Dots for efficient labeling is low (20 μg mL(-1) ), and the labeled cells can be visualized by flow cytometry, confocal microscopy, and transmission electron microscopy. Labeled hMSCs are unaffected in their viability and proliferation, as well as stemness surface marker expression and differentiation capability into osteocytes. Moreover, this is the first report that shows nonfunctionalized IRIS Dots can discriminate between live and early-stage apoptotic stem cells (both mesenchymal and embryonic) through a distinct external cell surface distribution. On the basis of biocompatibility, efficient labeling, and apoptotic discrimination potential, it is suggested that IRIS Dots can serve as a promising stem cell tracking agent.


Alteration of brain viscoelasticity after shunt treatment in normal pressure hydrocephalus.
Normal pressure hydrocephalus (NPH) represents a chronic neurological disorder with increasing incidence. The symptoms of NPH may be relieved by surgically implanting a ventriculoperitoneal shunt to drain excess cerebrospinal fluid. However, the pathogenesis of NPH is not yet fully elucidated, and the clinical response of shunt treatment is hard to predict. According to current theories of NPH, altered mechanical properties of brain tissue seem to play an important role. Magnetic resonance elastography (MRE) is a unique method for measuring in vivo brain mechanics. In this study cerebral MRE was applied to test the viscoelastic properties of the brain in 20 patients with primary (N = 14) and secondary (N = 6) NPH prior and after (91 ± 16 days) shunt placement. Viscoelastic parameters were derived from the complex modulus according to the rheological springpot model. This model provided two independent parameters μ and α, related to the inherent rigidity and topology of the mechanical network of brain tissue. The viscoelastic parameters μ and α were found to be decreased with -25% and -10%, respectively, compared to age-matched controls (P < 0.001). Interestingly, α increased after shunt placement (P < 0.001) to almost normal values whereas μ remained symptomatically low. The results indicate the fundamental role of altered viscoelastic properties of brain tissue during disease progression and tissue repair in NPH. Clinical improvement in NPH is associated with an increasing complexity of the mechanical network whose inherent strength, however, remains degraded.


Optimal management of nausea and vomiting of pregnancy.
Nausea and vomiting of pregnancy (NVP) is a common medical condition in pregnancy with significant physical and psychological morbidity. Up to 90% of women will suffer from NVP symptoms in the first trimester of pregnancy with up to 2% developing hyperemesis gravidarum which is NVP at its worst, leading to hospitalization and even death in extreme cases. Optimal management of NVP begins with nonpharmacological approaches, use of ginger, acupressure, vitamin B6, and dietary adjustments. The positive impact of these noninvasive, inexpensive and safe methods has been demonstrated. Pharmacological treatments are available with varying effectiveness; however, the only drug marketed specifically for the treatment of NVP in pregnancy is Diclectin(®) (vitamin B6 and doxylamine). In addition, the Motherisk algorithm provides a guideline for use of safe and effective drugs for the treatment of NVP. Optimal medical management of symptoms will ensure the mental and physical wellbeing of expecting mothers and their developing babies during this often stressful and difficult time period. Dismissing NVP as an inconsequential part of pregnancy can have serious ramifications for both mother and baby.


Epigallocatechin-3-Gallate Reduces Hepatic Oxidative Stress and Lowers CYP-Mediated Bioactivation and Toxicity of Acetaminophen in Rats.
Epigallocatechin-3-gallate (EGCG) is the most abundant polyphenol in green tea. To investigate the effects of dietary EGCG on oxidative stress and the metabolism and toxicity of acetaminophen in the liver, rats were fed diets with (0.54%) or without EGCG supplementation for four weeks and were then injected intraperitoneally with acetaminophen (1 g/kg). The results showed that EGCG lowered hepatic oxidative stress and cytochrome P450 (CYP) 1A2, 2E1, and 3A, and UDP-glucurosyltransferase activities prior to acetaminophen injection. After acetaminophen challenge, the elevations in plasma alanine aminotransferase activity and histological changes in the liver were ameliorated by EGCG treatment. EGCG reduced acetaminophen-induced apoptosis by lowering the Bax/Bcl2 ratio in the liver. EGCG mildly increased autophagy by increasing the LC3B II/I ratio. Lower hepatic acetaminophen-glutathione and acetaminophen-protein adducts contents were observed after EGCG treatment. EGCG increased glutathione peroxidase and NAD(P)H quinone 1 oxidoreductase activities and reduced organic anion-transporting polypeptides 1a1 expression in the liver after acetaminophen treatment. Our results indicate that EGCG may reduce oxidative stress and lower the metabolism and toxicity of acetaminophen. The reductions in CYP-mediated acetaminophen bioactivation and uptake transporter, as well as enhanced antioxidant enzyme activity, may limit the accumulation of toxic products in the liver and thus lower hepatotoxicity.


Investigation of potential antibacterial properties of methanol extracts from fungus Ganoderma applanatum.
In this study we searched for antibacterial compounds in methanol extracts of three layers (tube layer, context and cutis) of Ganoderma applanatum fruit body against Bacillus subtilis ATCC 6051 as a gram-positive bacterium and Escherichia coli ATCC 25922 and Pseudomonas syringae pv. syringae DPIC 219 as gram-negative bacteria. We found antibacterial properties in all three layers only against gram-negative bacteria. The fractions were isolated and purified by thin layer chromatography (TLC) and high-performance liquid chromatography. After purification, four major compounds - A, B, C and D - were tested for antibacterial properties, and B and D were found to possess antibacterial properties. The results of TLC and spectral data, including mass spectra and (1)H NMR of compounds B and C, showed that they were saturated fatty acids and compound D was palmitic acid (hexadecanoic acid) (16:0).


The limitations imposed on reading by low vision aids.
The changes that occur in the reading eye movements of normally sighted readers were measured as they used hand magnifiers to identify how these devices contribute to the slow reading of visually-impaired patients. Subjects inexperienced in magnifier use read texts containing two sizes of print, using hand magnifiers of two different powers, held at two different eye-to-magnifier distances. The effect of magnification (up to 13.5x) and field-of-view (FOV) (2-45 characters) could be assessed independently. Reading speed decreased with increasing magnification because the size of the saccades did not increase in proportion to the magnification: for a given level of magnification, decreasing the FOV and decreasing the viewing distance both reduce the size of the saccades even further. The overall reading speed is only slowed significantly when the FOV restriction is extreme (two characters' width). Two mechanisms seem to be used spontaneously by normally sighted readers to mitigate the limitation of reading speed created by the shortened saccades: head movement in the direction of reading and retinal image slip during fixation.


Characteristics of multidimensional holographic associative memory in retrieval with dynamically localizable attention.
This paper presents the performance analysis (capacity and retrieval accuracy) of multidimensional holographic associative memory (MHAC). MHAC has the unique ability to retrieve pattern-associations with changeable attention. In attention actuated retrieval the user can dynamically select any subset of the elements in the example query pattern and expect the memory to confine its associative match only within the specified field of attention. Existing artificial associative memories lack this ability. Also most of these models need at least 50% of bits in the input pattern to be correct for successful retrieval. MHAC, with the unique ability of localizable attention, can retrieve information correctly even with cues as small as 10% of the query frame. This paper investigates the performance of MHAC in attention actuated retrieval both analytically and experimentally. Besides confirmation, the experiments also identify an operational range space (ORS) for this memory within which various attention based applications can be built with a performance guarantee.


Neuronal protein tyrosine phosphatase 1B deficiency results in inhibition of hypothalamic AMPK and isoform-specific activation of AMPK in peripheral tissues.
PTP1B(-/-) mice are resistant to diet-induced obesity due to leptin hypersensitivity and consequent increased energy expenditure. We aimed to determine the cellular mechanisms underlying this metabolic state. AMPK is an important mediator of leptin's metabolic effects. We find that alpha1 and alpha2 AMPK activity are elevated and acetyl-coenzyme A carboxylase activity is decreased in the muscle and brown adipose tissue (BAT) of PTP1B(-/-) mice. The effects of PTP1B deficiency on alpha2, but not alpha1, AMPK activity in BAT and muscle are neuronally mediated, as they are present in neuron- but not muscle-specific PTP1B(-/-) mice. In addition, AMPK activity is decreased in the hypothalamic nuclei of neuronal and whole-body PTP1B(-/-) mice, accompanied by alterations in neuropeptide expression that are indicative of enhanced leptin sensitivity. Furthermore, AMPK target genes regulating mitochondrial biogenesis, fatty acid oxidation, and energy expenditure are induced with PTP1B inhibition, resulting in increased mitochondrial content in BAT and conversion to a more oxidative muscle fiber type. Thus, neuronal PTP1B inhibition results in decreased hypothalamic AMPK activity, isoform-specific AMPK activation in peripheral tissues, and downstream gene expression changes that promote leanness and increased energy expenditure. Therefore, the mechanism by which PTP1B regulates adiposity and leptin sensitivity likely involves the coordinated regulation of AMPK in hypothalamus and peripheral tissues.


Rapid formation of inositol 1,3,4,5-tetrakisphosphate following muscarinic receptor stimulation of rat cerebral cortical slices.
Carbachol stimulation of muscarinic receptors in rat cortical slices prelabelled with myo-[2-3H]inositol caused the rapid formation of a novel inositol polyphosphate. Evidence derived from its chromatographic behaviour, and from the structure of the products formed in partial dephosphorylation experiments, suggests that it is probably D-myo-inositol 1,3,4,5-tetrakisphosphate. An enzyme in human red cell membranes specifically removes the 5-phosphate from it to form inositol 1,3,4-trisphosphate. It is suggested that inositol 1,3,4,5-tetrakisphosphate is likely to be a second messenger, and that it is the precursor of inositol 1,3,4-trisphosphate and possibly of inositol 1,4,5-trisphosphate.


Facilitated nucleocytoplasmic shuttling of the Ran binding protein RanBP1.
The Ran binding protein RanBP1 is localized to the cytosol of interphase cells. A leucine-rich nuclear export signal (NES) near the C terminus of RanBP1 is essential to maintain this distribution. We now show that RanBP1 accumulates in nuclei of cells treated with the export inhibitor, leptomycin B, and collapse of the nucleocytoplasmic Ran:GTP gradient leads to equilibration of RanBP1 across the nuclear envelope. Low temperature prevents nuclear accumulation of RanBP1, suggesting that import does not occur via simple diffusion. Glutathione S-transferase (GST)-RanBP1(1-161), which lacks the NES, accumulates in the nucleus after cytoplasmic microinjection. In permeabilized cells, nuclear accumulation of GST-RanBP1(1-161) requires nuclear Ran:GTP but is not inhibited by a dominant interfering G19V mutant of Ran. Nuclear accumulation is enhanced by addition of exogenous karyopherins/importins or RCC1, both of which also enhance nuclear Ran accumulation. Import correlates with Ran concentration. Remarkably, an E37K mutant of RanBP1 does not import into the nuclei under any conditions tested despite the fact that it can form a ternary complex with Ran and importin beta. These data indicate that RanBP1 translocates through the pores by an active, nonclassical mechanism and requires Ran:GTP for nuclear accumulation. Shuttling of RanBP1 may function to clear nuclear pores of Ran:GTP, to prevent premature release of import cargo from transport receptors.


Measuring movements in adolescents with psychosis using the Microsoft Kinect sensor: a pilot study exploring a new tool for assessing aspects of antipsychotic-induced parkinsonism.
The assessment of motor disturbances in antipsychotic-treated adolescent patients is often limited to the use of observer-based rating scales with interobserver variability. The objectives of this pilot study were to measure movement patterns associated with antipsychotic-induced parkinsonism in young patients with psychosis and initiating/treated with antipsychotics, using a computer application connected with the Microsoft Kinect sensor (Motorgame). All participants were assessed by neurological examination, clinical side effect rating scales (Udvalg for Kliniske Undersøgelser Side Effect Rating Scale, Barnes Akathisia Rating Scale, Simpson Angus Scale (SAS), and Abnormal Involuntary Movement Scale), and the Motorgame. Furthermore, speed of information processing and motor speed with subtests from the Brief Assessment of Cognition in Schizophrenia test battery was assessed. We included 21 adolescents with first-episode psychosis (62% treated with antipsychotics; males 38%; mean age 16 ± 1.4 years) and 69 healthy controls (males 36%; mean age 16 ± 1.5 years). Prolonged time of motor performance (TOMP) in the Motorgame was associated with higher SAS scores for arm dropping (p = .009). A consistent practice effect was detected (p < .001). We found no significant associations between TOMP and age, height, body weight, sex, antipsychotic dosage, or information processing speed. We found an uncorrected significant association between prolonged TOMP and shoulder bradykinesia. The Motorgame was found useful in assessing parkinsonian symptoms in early-onset psychosis and accepted by participants. Future studies of larger cohorts, including patients with high scores in clinical motor side effect scales, are required to establish solid validity of the novel test.


The Role of Human Papillomavirus Genotyping in Cervical Cancer Screening: A Large-Scale Evaluation of the cobas HPV Test.
The cobas HPV Test ("cobas"; Roche Molecular Systems) detects HPV16 and HPV18 individually, and a pool of 12 other high-risk (HR) HPV types. The test is approved for (i) atypical squamous cells of undetermined significance (ASC-US) triage to determine need for colposcopy, (ii) combined screening with cytology ("cotesting"), and (iii) primary HPV screening. To assess the possible value of HPV16/18 typing, >17,000 specimens from a longitudinal cohort study of initially HPV-positive women (HC2, Qiagen) were retested with cobas. To study accuracy, cobas genotyping results were compared with those of an established method, the Linear Array HPV Genotyping Test (LA, Roche Molecular Systems). Clinical value of the typing strategy was evaluated by linking the cobas results (supplemented by other available typing results) to 3-year cumulative risks of CIN3+. Grouped hierarchically (HPV16, else HPV18, else other HR types, else negative), the κ statistic for agreement between cobas and LA was 0.86 [95% confidence interval (CI), 0.86-0.87]. In all three scenarios, HPV16-positive women were at much higher 3-year risk of CIN3+ than HPV16-negative women: women ages 21 and older with ASC-US (14.5%; 95% CI, 13.5%-15.5% vs. 3.5%; 95% CI, 3.3-3.6); women ages 30 years and older that were HPV-positive cytology-negative (10.3%; 95% CI, 9.6-11.1 vs. 2.3%; 95% CI, 2.2-2.4); and all women 25 years and older that were HPV-positive (18.5%; 95% CI, 17.8-19.2 vs. 4.3%; 95% CI, 4.2-4.4). The cobas and LA results show excellent agreement. The data support HPV16 typing. HPV16 typing is useful in the management of HPV-positive/cytology-negative women in cotesting, of all HPV-positive women in primary HPV testing, and perhaps in the management of HPV-positive women with ASC-US. Cancer Epidemiol Biomarkers Prev; 24(9); 1304-10.


Influence of cryoprecipitate, Factor XIII, and fibrinogen concentrate on hyperfibrinolysis.
Hyperfibrinolysis is a potentially life-threatening condition associated with poor clot integrity and excessive bleeding. Although antifibrinolytics are an effective treatment, more liberal use of these drugs may lead to a prothrombotic risk, and an earlier and potentially safer treatment option would be desirable. Hyperfibrinolysis has been shown to be attenuated by in vitro supplementation of purified human Factor (F)XIII concentrate. Cryoprecipitate represents an alternative source of FXIII and the only approved source of concentrated FXIII in some countries. The aim of this study was to investigate whether cryoprecipitate, FXIII, and fibrinogen concentrate mitigate hyperfibrinolysis. Ten citrate blood specimens from healthy subjects were spiked with tissue plasminogen activator (t-PA) and subsequently supplemented with cryoprecipitate, FXIII, fibrinogen concentrate, and ɛ-aminocaproic acid (EACA). Thromboelastometry tests were performed at baseline, after t-PA, and after supplementation. Hyperfibrinolysis was assessed using the clot lysis index at 60 minutes (LI60; reciprocal of maximum lysis). The LI60 was significantly improved (fibrinolysis attenuated) after cryoprecipitate supplementation compared to t-PA alone and compared to FXIII and fibrinogen concentrate. Hyperfibrinolysis was only fully reversed using EACA. In addition, cryoprecipitate demonstrated the least variability in the attenuation of hyperfibrinolysis among 10 healthy subjects, compared to FXIII and fibrinogen concentrate. This is the first study to show that cryoprecipitate is able to significantly mitigate hyperfibrinolysis in an in vitro model. Further investigations are warranted to determine whether cryoprecipitate may have a previously unrecognized benefit and should be administered earlier in resuscitation protocols.


Bronchoalveolar lavage in the assessment of pulmonary Hodgkin's disease.
Abnormal chest radiographs in patients with Hodgkin's disease are occasionally due to pulmonary Hodgkin's disease. The fluids recovered from bronchoalveolar lavages (BALs) from 50 patients prior to autologous bone marrow transplantation for advanced Hodgkin's disease were examined. Abnormal chest roentgenograms were present in 24 patients (48%); 4 (17%) of these had Reed-Sternberg cells or their mononucleated variants in the lavage fluid and an alveolar lymphocytosis averaging 31.4% (normal: 11.5%). The lymphocytes were small and monotonous. Of the 20 patients with abnormal chest roentgenograms but no Reed-Sternberg cells in the lavage fluid, the lymphocyte count was 10.88%, with only 3 patients exceeding 17%. Two patients with normal chest roentgenograms had Reed-Sternberg-like cells in their lavage fluids and averaged 23% lymphocytes in their lavage differential count. Eosinophils averaged 1% or less of the lavage differential and were not predictive of pulmonary Hodgkin's disease. This experience suggests that pulmonary Hodgkin's disease can be diagnosed by BAL. Reed-Sternberg cells and their mononucleated variants can be recognized by their characteristic cytomorphologic features, although care must be taken not to misinterpret reactive binucleated macrophages as neoplastic cells. In patients with Hodgkin's disease, Reed-Sternberg cells should be sought when an alveolar lymphocytosis is present.


Systemic administration of doxycycline versus metronidazole plus amoxicillin in the treatment of localized aggressive periodontitis: a clinical and microbiologic study.
The use of antibiotics as an adjunctive therapy in the management of periodontal diseases has always been of interest for dental practitioners. Although studies have demonstrated the importance of different antibiotics as adjunct in the treatment of periodontitis, few studies have reported comparative results of antibiotics in localized aggressive periodontitis (LAP). The aim of the present study was to evaluate the comparative effects of systemic tetracycline derivative doxycycline versus a combination of metronidazole and amoxicillin in the elimination/suppression of Actinobacillus actinomycetemcomitans and their clinical effects during 90 days of follow-up in patients with LAP. Thirty patients with untreated LAP participated in the study. Patients were randomly divided into 2 groups and given doxycycline or metronidazole plus amoxicillin, and periodontal clinical parameters were achieved at baseline and 10, 30, 60, and 90 days after microbiologic sampling. Patients were also given mechanical debridement after measurement at baseline. Both groups of patients demonstrated improvement in clinical parameters, and microbiologic samples did not show any A actinomycetemcomitans at days 30, 60, and 90. Systemic use of doxycycline or metronidazole plus amoxicillin as an adjunct to conventional therapy may be used for the treatment of patients with LAP.


CD27 distinguishes two phases in bone marrow infiltration of splenic marginal zone lymphoma.
To investigate CD27 expression in splenic marginal zone lymphoma (SMZL), an indolent low-grade B-cell lymphoma with constant involvement of the bone marrow, especially with an intrasinusoidal pattern. It is not clear if the neoplastic clone is composed of virgin or somatically mutated B cells. CD27 is reported to be a hallmark of memory B cells. We evaluated 64 bone marrow biopsy specimens (BMBs) from 36 patients with SMZL for the expression of CD27. For comparison, splenectomy specimens of patients with traumatic splenic rupture or with SMZL were used. All BMBs showed lymphomatous infiltration. When located in the marrow sinusoids, neoplastic cells were CD27- in all cases and therefore corresponded to naive B cells. In nodular/interstitial infiltration, the cells were CD27+ and therefore corresponded to memory B cells. No difference in immunohistochemical expression of B and T antibodies was found between intrasinusoidal and interstitial/nodular infiltration. CD27 was constantly expressed in the splenic marginal zone of normal spleen, surgically removed for trauma, and in seven out of 10 spleens with SMZL. We propose the existence of two different phases of neoplastic progression with, first, expansion of a virgin B clone in the bone marrow and, following exposure to antigen, a re-colonization of the bone marrow.


Fmoc-modified amino acids and short peptides: simple bio-inspired building blocks for the fabrication of functional materials.
Amino acids and short peptides modified with the 9-fluorenylmethyloxycarbonyl (Fmoc) group possess eminent self-assembly features and show distinct potential for applications due to the inherent hydrophobicity and aromaticity of the Fmoc moiety which can promote the association of building blocks. Given the extensive study and numerous publications in this field, it is necessary to summarize the recent progress concerning these important bio-inspired building blocks. Therefore, in this review, we explore the self-organization of this class of functional molecules from three aspects, i.e., Fmoc-modified individual amino acids, Fmoc-modified di- and tripeptides, and Fmoc-modified tetra- and pentapeptides. The relevant properties and applications related to cell cultivation, bio-templating, optical, drug delivery, catalytic, therapeutic and antibiotic properties are subsequently summarized. Finally, some existing questions impeding the development of Fmoc-modified simple biomolecules are discussed, and corresponding strategies and outlooks are suggested.


Transgenic or plant expression vector-mediated recombination of Plum Pox Virus.
Different mutants of an infectious full-length clone (p35PPV-NAT) of Plum pox virus (PPV) were constructed: three mutants with mutations of the assembly motifs RQ and DF in the coat protein gene (CP) and two CP chimeras with exchanges in the CP core region of Zucchini yellow mosaic virus and Potato virus Y. The assembly mutants were restricted to single infected cells, whereas the PPV chimeras were able to produce systemic infections in Nicotiana benthamiana plants. After passages in different transgenic N. benthamiana plants expressing the PPV CP gene with a complete (plant line 4.30.45.) or partially deleted 3'-nontranslated region (3'-NTR) (plant line 17.27. 4.), characterization of the viral progeny of all mutants revealed restoration of wild-type virus by recombination with the transgenic CP RNA only in the presence of the complete 3'-NTR (4.30.45.). Reconstitution of wild-type virus was also observed following cobombardment of different assembly-defective p35PPV-NAT together with a movement-defective plant expression vector of Potato virus X expressing the intact PPV-NAT CP gene transiently in nontransgenic N. benthamiana plants. Finally, a chimeric recombinant virus was detected after cobombardment of defective p35PPV-NAT with a plant expression vector-derived CP gene from the sour cherry isolate of PPV (PPV-SoC). This chimeric virus has been established by a double recombination event between the CP-defective PPV mutant and the intact PPV-SoC CP gene. These results demonstrate that viral sequences can be tested for recombination events without the necessity for producing transgenic plants.


Structural changes and biological responsiveness of an injectable and mouldable monetite bone graft generated by a facile synthetic method.
Brushite (dicalcium phosphate dihydrate) and monetite (dicalcium phosphate anhydrous) are of considerable interest in bone augmentation owing to their metastable nature in physiological fluids. The anhydrous form of brushite, namely monetite, has a finer microstructure with higher surface area, strength and bioresorbability, which does not transform to the poorly resorbable hydroxyapatite, thus making it a viable alternative for use as a scaffold for engineering of bone tissue. We recently reported the formation of monetite cements by a simple processing route without the need of hydrothermal treatment by using a high concentration of sodium chloride in the reaction mix of β-tricalcium phosphate and monocalcium phosphate monohydrate. In this paper, we report the biological responsiveness of monetite formed by this method. The in vitro behaviour of monetite after interaction and ageing both in an acellular and cellular environment showed that the crystalline phase of monetite was retained over three weeks as evidenced from X-ray diffraction measurements. The crystal size and morphology also remained unaltered after ageing in different media. Human osteoblast cells seeded on monetite showed the ability of the cells to proliferate and express genes associated with osteoblast maturation and mineralization. Furthermore, the results showed that monetite could stimulate osteoblasts to undergo osteogenesis and accelerate osteoblast maturation earlier than cells cultured on hydroxyapatite scaffolds of similar porosity. Osteoblasts cultured on monetite cement also showed higher expression of osteocalcin, which is an indicator of the maturation stages of osteoblastogenesis and is associated with matrix mineralization and bone forming activity of osteoblasts. Thus, this new method of fabricating porous monetite can be safely used for generating three-dimensional bone graft constructs.


Pathology of the peripheral neuropathy induced by p-bromophenylacetylurea.
p-Bromophenylacetylurea (BPAU) is a neurotoxin known to cause intraaxonal accumulations of tubulovesicular materials and distal axonal degeneration. This study focused on two aspects of the pathology in the peripheral nerves in rats: (1) the comparative pathology of acute and chronic intoxication, and (2) the ability of intoxicated neurons to regenerate. The rats were divided into acute and chronic intoxication groups. Rats acutely intoxicated received one intraperitoneal injection (400 mg. per kg.) and developed paraparesis after 2 weeks. Rats chronically intoxicated were given daily intraperitoneal injections (10 mg. per kg.) for 2 to 7 months; paraparesis appeared after 8 weeks. Both groups showed degeneration of distal nerves. By 3 weeks, rats in the acute intoxication group showed an 18 to 50 per cent loss of axons in the distal phrenic nerve. Axons contained membranous trabeculae (tubules and vesicles); at later times, the tubulovesicular profiles increased and, subsequently, axons degenerated. In these animals, regenerating sprouts were first seen 3 weeks after stopping BPAU; these sprouts also contained tubulomembranous structures which persisted for 18 months. After 2 to 3 months, the chronic intoxication group showed a reduction in the number of axons in the distal phrenic nerve. Surviving axons showed tubulovesicular profiles similar to those observed in acutely intoxicated rats. In this group of rats, axonal regeneration was also characterized by the presence of membranous profiles in the axonal sprouts. Our observations indicate that the pathologic changes caused by BPAU are independent of the rate of administration of the drug. The long-standing abnormalities in the regenerating sprouts suggest a persistent toxic effect on axons. Since membranes are delivered and retrieved through fast anterograde and retrogade transport, the accumulation of membranes may be related to abnormalities in these systems. BPAU appears to be a useful agent to study the pathogenetic mechanisms causing tubulovesicular neuropathies.


Prenatal diagnosis of Smith-Lemli-Opitz syndrome in a pregnancy with low maternal serum oestriol and a sex-reversed fetus.
A cytogenetically normal male fetus was subsequently found to have female external genitalia, a cardiac malformation and mid-trimester intra-uterine growth retardation by ultrasound examination. The maternal serum oestriol level was low. The combination of low oestriol and sonographic findings suggested Smith Lemli Opitz syndrome (SLO), which was confirmed by a markedly increased amniotic fluid level of 7-dehydrocholesterol. We review the differential diagnosis of apparent sex reversal in a fetus and low maternal serum oestriol level. To further examine the specificity of low maternal oestriol level as a marker for SLO a follow-up study of 12141 pregnancies screened for Down syndrome using three biochemical markers: alpha-fetoprotein, beta-human chorionic gonadotrophin and oestriol was performed. 26 pregnancies had an oestriol level that was 0.25 MoM or less. SLO was not diagnosed clinically in any of the liveborn children ascertained through a low maternal oestriol level. Nine of the pregnancies ended in spontaneous miscarriage. Although the frequency of SLO in pregnancies with low maternal oestriol levels or sex-reversed fetuses is unknown, the diagnosis of SLO should, nevertheless, be considered in both clinical settings.


[Polyamines and collagenes interaction in different processes of extracellular matrix production and functioning in norm and with pathology].
A great file of experimental data on tight interrelations between polyamines and main components of extracellular matrix (collagenes, laminin, fibronectin, etc.) metabolisms has been accumulated in the recent twenty years. Consequences of such tight interaction of collagenes, polyamines, enzymes of their metabolisms during formation and functioning of extracellular matrix are realised not only at a cellular level (proliferation, differentiation, adhesion, apoptosis), but also at the level of the whole organism (immunity, connective tissues and skeleton formation, tumors). Search for the ways of cells behavior modulation via influence on the levels of polyamines and contents of extracellular matrix at present is new direction in pharmacology of antitumor preparations that enables regulation of organism-tumor relationships, inhibition of angiogenesis, tumor growth and metastasing.


Chromogranin-A: a multifaceted cardiovascular role in health and disease.
Chromogranin A (CgA), a major component of the chromaffin granules, is co-stored and co-released with catecholamines. It is also expressed in extra-adrenal sites, including the heart. In the rat, CgA localizes in atrial myoendocrine cells, associated with Atrial Natriuretic Peptide (ANP), and in the conduction system. In the human heart it is present in the ventricular myocardium, co-localized with B-type NP (BNP). CgA is the precursor of several biologically active peptides generated by proteolytic processing also in the heart. Two of them, vasostatin-1 (VS-1) and catestatin (Cst), inhibit cardiac contraction and relaxation, counter-regulate beta-adrenergic and endothelinergic stimulation, and protect the heart against ischemia/reperfusion damages. Recently, clinical studies have suggested CgA to be involved also in cardiovascular pathologies. High plasma CgA levels were found in hypertension, chronic and acute heart failure, myocardial infarction, decompensated and hypertrophic heart, and acute coronary syndromes. These alterations correlate with those of conventional cardiovascular biomarkers, such as NP and endothelin-1 (ET-1), and have prognostic relevance, being indicative of both severity of the disease and mortality. Accordingly, the current knowledge indicates CgA as a multifaceted peptide in cardiovascular homeostasis. Whether the influence elicited by the protein on both normal and failing heart is beneficial and/or detrimental, as well as its implication in the cardiac neuroendocrine scenario is under intense investigation. This review will focus on: i) the involvement of CgA and its derived peptides in the mechanisms which sustain cardiac function and compensation, ii) CgA clinical relevance, and iii) its putative value as a clinical biomarker.


[Pre-operative period of the heart surgical patient: nursing approach making a difference].
This is a study that was developed in a Cardiology Unit of a University Hospital in the country area of Rio Grande do Sul. Its aim was to identify the perception of the cardiac surgical patient about the pre-surgical orientations given by the nursing staff. The population was of male and female adult patients who were submitted to any cardiac surgical process. Semi-structured interviews were conducted to grasp the perception of the patient about the given information. The following themes emerged: tranquilizing awakening; stressful environment; nursing: being with and humanization; privileged feeling; technology in orientation; and orientation: meaning of clarifying, tranquility and courage.


Loss of TREM2 rescues hyperactivation of microglia, but not lysosomal deficits and neurotoxicity in models of progranulin deficiency.
Haploinsufficiency of the progranulin (PGRN)-encoding gene (GRN) causes frontotemporal lobar degeneration (GRN-FTLD) and results in microglial hyperactivation, TREM2 activation, lysosomal dysfunction, and TDP-43 deposition. To understand the contribution of microglial hyperactivation to pathology, we used genetic and pharmacological approaches to suppress TREM2-dependent transition of microglia from a homeostatic to a disease-associated state. Trem2 deficiency in Grn KO mice reduced microglia hyperactivation. To explore antibody-mediated pharmacological modulation of TREM2-dependent microglial states, we identified antagonistic TREM2 antibodies. Treatment of macrophages from GRN-FTLD patients with these antibodies led to reduced TREM2 signaling due to its enhanced shedding. Furthermore, TREM2 antibody-treated PGRN-deficient microglia derived from human-induced pluripotent stem cells showed reduced microglial hyperactivation, TREM2 signaling, and phagocytic activity, but lysosomal dysfunction was not rescued. Similarly, lysosomal dysfunction, lipid dysregulation, and glucose hypometabolism of Grn KO mice were not rescued by TREM2 ablation. Synaptic loss and neurofilament light-chain (NfL) levels, a biomarker for neurodegeneration, were further elevated in the Grn/Trem2 KO cerebrospinal fluid (CSF). These findings suggest that TREM2-dependent microglia hyperactivation in models of GRN deficiency does not promote neurotoxicity, but rather neuroprotection.


CBP Activity Mediates Effects of the Histone Deacetylase Inhibitor Butyrate on WNT Activity and Apoptosis in Colon Cancer Cells.
Mutations in the WNT/beta-catenin pathway are responsible for initiating the majority of colorectal cancers (CRCs). We have previously shown that hyperactivation of this signaling by histone deacetylase inhibitors (HDACis) such as butyrate, a fermentation product of dietary fiber, promotes CRC cell apoptosis. The extent of association between beta-catenin and the transcriptional coactivator CREB-binding protein (CBP) influences WNT/catenin signaling and, therefore, colonic cell physiology. CBP functions as a histone acetylase (HAT); therefore, we hypothesized that the modulation of WNT/catenin activity by CBP modifies the ability of the HDACi butyrate to hyperinduce WNT signaling and apoptosis in CRC cells. Our findings indicate that CBP affects the hyperinduction of WNT activity by butyrate. ICG-001, which specifically blocks association between CBP and beta-catenin, abrogates the butyrate-triggered increase in the number of CRC cells with high levels of WNT/catenin signaling. Combination treatment of CRC cells with ICG-001 and butyrate results in cell type-specific effects on apoptosis. Further, both butyrate and ICG-001 repress CRC cell proliferation, with additive effects in suppressing cell growth. Our study strongly suggests that ICG-001-like agents would be effective against butyrate/HDACi-resistant CRC cells. Therefore, ICG-001-like agents may represent an important therapeutic option for CRCs that exhibit low-fold hyperactivation of WNT activity and apoptosis in the presence of HDACis. The findings generated from this study may lead to approaches that utilize modulation of CBP activity to facilitate CRC therapeutic or chemopreventive strategies.


Recommended joint and meta-analysis strategies for case-control association testing of single low-count variants.
In genome-wide association studies of binary traits, investigators typically use logistic regression to test common variants for disease association within studies, and combine association results across studies using meta-analysis. For common variants, logistic regression tests are well calibrated, and meta-analysis of study-specific association results is only slightly less powerful than joint analysis of the combined individual-level data. In recent sequencing and dense chip based association studies, investigators increasingly test low-frequency variants for disease association. In this paper, we seek to (1) identify the association test with maximal power among tests with well controlled type I error rate and (2) compare the relative power of joint and meta-analysis tests. We use analytic calculation and simulation to compare the empirical type I error rate and power of four logistic regression based tests: Wald, score, likelihood ratio, and Firth bias-corrected. We demonstrate for low-count variants (roughly minor allele count [MAC] < 400) that: (1) for joint analysis, the Firth test has the best combination of type I error and power; (2) for meta-analysis of balanced studies (equal numbers of cases and controls), the score test is best, but is less powerful than Firth test based joint analysis; and (3) for meta-analysis of sufficiently unbalanced studies, all four tests can be anti-conservative, particularly the score test. We also establish MAC as the key parameter determining test calibration for joint and meta-analysis.


Enhanced growth of ginger plants by an eco- friendly nitrogen-fixing Pseudomonas protegens inoculant in glasshouse fields.
Excessive nitrogen (N) fertilization in glasshouse fields greatly increases N loss and fossil-fuel energy consumption resulting in serious environmental risks. Microbial inoculants are strongly emerging as potential alternatives to agrochemicals and offer an eco-friendly fertilization strategy to reduce our dependence on synthetic chemical fertilizers. Effects of a N-fixing strain Pseudomonas protegens CHA0-ΔretS-nif on ginger plant growth, yield, and nutrient uptake, and on earthworm biomass and the microbial community were investigated in glasshouse fields in Shandong Province, northern China. Application of CHA0-ΔretS-nif could promote ginger plant development, and significantly increased rhizome yields, by 12.93% and 7.09%, respectively, when compared to uninoculated plants and plants treated with the wild-type bacterial strain. Inoculation of CHA0-ΔretS-nif had little impact on plant phosphorus (P) acquisition, whereas it was associated with enhanced N and potassium (K) acquisition by ginger plants. Moreover, inoculation of CHA0-ΔretS-nif had positive effects on the bacteria population size and the number of earthworms in the rhizosphere. Similar enhanced performances were also found in CHA0-ΔretS-nif-inoculated ginger plants even when the N-fertilizer application rate was reduced by 15%. A chemical N input of 573.8 kg ha-1 with a ginger rhizome yield of 1.31 × 105 kg ha-1 was feasible. The combined application of CHA0-ΔretS-nif and a reduced level of N-fertilizers can be employed in glasshouse ginger production for the purpose of achieving high yields while at the same time reducing the inorganic-N pollution from traditional farming practices. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


From the community to the classroom: the Aboriginal health curriculum at the Northern Ontario School of Medicine.
More undergraduate medical education programs are including curricula concerning the health, culture and history of Aboriginal people. This is in response to growing international recognition of the large divide in health status between Aboriginal and non-Aboriginal people, and the role medical education may play in achieving health equity. In this paper, we describe the development and delivery of the Aboriginal health curriculum at the Northern Ontario School of Medicine (NOSM). We describe a process for curriculum development and delivery, which includes ongoing engagement with Aboriginal communities as well as faculty expertise. Aboriginal health is delivered as a core curriculum, and learning is evaluated in summative assessments. Aboriginal health objectives are present in 4 of 5 required courses, primarily in years 1 and 2. Students attend a required 4-week Aboriginal cultural immersion placement at the end of year 1. Resources of Aboriginal knowledge are integrated into learning. In this paper, we reflect on the key challenges encountered in the development and delivery of the Aboriginal health curriculum. These include differences in Aboriginal and non-Aboriginal knowledge; risk of reinforcing stereotypes in case presentations; negotiation of curricular time; and faculty readiness and development. An organizational commitment to social accountability and the resulting community engagement model have been instrumental in creating a robust, sustainable program in Aboriginal health at NOSM.


Improving outcomes of first-episode psychosis: an overview.
Outcomes of psychotic disorders are associated with high personal, familiar, societal and clinical burden. There is thus an urgent clinical and societal need for improving those outcomes. Recent advances in research knowledge have opened new opportunities for ameliorating outcomes of psychosis during its early clinical stages. This paper critically reviews these opportunities, summarizing the state-of-the-art knowledge and focusing on recent discoveries and future avenues for first episode research and clinical interventions. Candidate targets for primary universal prevention of psychosis at the population level are discussed. Potentials offered by primary selective prevention in asymptomatic subgroups (stage 0) are presented. Achievements of primary selected prevention in individuals at clinical high risk for psychosis (stage 1) are summarized, along with challenges and limitations of its implementation in clinical practice. Early intervention and secondary prevention strategies at the time of a first episode of psychosis (stage 2) are critically discussed, with a particular focus on minimizing the duration of untreated psychosis, improving treatment response, increasing patients' satisfaction with treatment, reducing illicit substance abuse and preventing relapses. Early intervention and tertiary prevention strategies at the time of an incomplete recovery (stage 3) are further discussed, in particular with respect to addressing treatment resistance, improving well-being and social skills with reduction of burden on the family, treatment of comorbid substance use, and prevention of multiple relapses and disease progression. In conclusion, to improve outcomes of a complex, heterogeneous syndrome such as psychosis, it is necessary to globally adopt complex models integrating a clinical staging framework and coordinated specialty care programmes that offer pre-emptive interventions to high-risk groups identified across the early stages of the disorder. Only a systematic implementation of these models of care in the national health care systems will render these strategies accessible to the 23 million people worldwide suffering from the most severe psychiatric disorders.


Acceleration of scrapie disease in mice by an adenovirus.
Coinfected mice were examined for a possible interaction between the scrapie agent and an adenovirus. A low titer (10(2) TCD50) of mouse adenovirus (MAdV) caused a significant acceleration of clinical signs of scrapie in mice infected 128 days previously with scrapie. In this experiment, the coinfected mice died 19 days earlier than mice infected with scrapie alone. When a higher titer of MAdV (10(4)-10(5) PFU) was used, a more drastic acceleration of scrapie disease was seen in mice infected 85 and 110 days previously with scrapie. At 85 days, coinfection caused mice to die 37 days earlier than mice infected with scrapie alone, whereas at 110 days, coinfection caused mice to die 52 days earlier than mice infected with scrapie alone. MAdV alone caused no clinical disease in normal mice. The brains of coinfected mice and mice that had been infected with scrapie alone showed a histopathology consistent with scrapie. A possible explanation for these findings is that the replication of the scrapie agent is accelerated by adenovirus. Defective parvoviruses are known to be helped by adenoviruses. Spleens from coinfected mice but not from mice infected with MAdV alone yielded, in cultures of BALB 3T3 cells, infectious MAdV and one or two smaller agents with the dimension and shape of a parvovirus.


Sulfate reduction controlled by organic matter availability in deep sediment cores from the saline, alkaline Lake Van (Eastern Anatolia, Turkey).
As part of the International Continental Drilling Program deep lake drilling project PaleoVan, we investigated sulfate reduction (SR) in deep sediment cores of the saline, alkaline (salinity 21.4‰, alkalinity 155 m mEq(-1), pH 9.81) Lake Van, Turkey. The cores were retrieved in the Northern Basin (NB) and at Ahlat Ridge (AR) and reached a maximum depth of 220 m. Additionally, 65-75 cm long gravity cores were taken at both sites. SR rates (SRR) were low (≤22 nmol cm(-3) day(-1)) compared to lakes with higher salinity and alkalinity, indicating that salinity and alkalinity are not limiting SR in Lake Van. Both sites differ significantly in rates and depth distribution of SR. In NB, SRR are up to 10 times higher than at AR. SR could be detected down to 19 mblf (meters below lake floor) at NB and down to 13 mblf at AR. Although SRR were lower at AR than at NB, organic matter (OM) concentrations were higher. In contrast, dissolved OM in the pore water at AR contained more macromolecular OM and less low molecular weight OM. We thus suggest, that OM content alone cannot be used to infer microbial activity at Lake Van but that quality of OM has an important impact as well. These differences suggest that biogeochemical processes in lacustrine sediments are reacting very sensitively to small variations in geological, physical, or chemical parameters over relatively short distances.


Functional characteristics of Peyer's patch cells. III. Carrier priming of T cells by antigen feeding.
Peyer's patch T cells may serve an important role in the interaction of the host with intraluminal gut antigens. Studies presented in this paper demonstrate that T cells in murine Peyer's patches can be carrier primed for helper function in the induction of an antihapten response by feeding antigen. Carrier priming was assessed by measuring the ability of Peyer's patch cells from mice fed heterologous erythrocytes to enhance an antitrinitrophenyl (TNP) response in vitro when added to normal Peyer's patch cells cultured with TNP coupled to the erythrocyte used for feeding. Priming of T helper cells in Peyer's patches was antigen specific and occurred when erythrocytes were administered orally but not when erythrocytes were injected intravenously or intraperitoneally. Murine Peyer's patches are naturally deficient in a cooperating accessory adherent cell type(s) required for B-cell induction to humoral antibody synthesis in vitro and antigen feeding does not result in significant induction of Peyer's patch B cells to humoral antibody synthesis in vivo. Since Peyer's patch T cells can be carrier-antigen primed for helper function in the absence of B-cell induction to humoral antibody synthesis, these studies may indicate that T-cell priming is less dependent than B-cell induction on cooperating accessory adherent cells.


Insect pathogenic properties of Serratia marcescens: phage-resistant mutants with a decreased resistance to Cecropia immunity and a decreased virulence to Drosophila.
A non-pigmented strain of Serratia marcescens (Db10) was isolated from moribund Drosophila flies. From this strain were isolated spontaneous mutants resistant to streptomycin (Db11) and nalidixic acid (Db12). Mutant Db11 was used for the isolation of two phages, phi J and phi K, which grew on Db10, Db11 and Db12, but not on three reference strains of S. marcescens. Mutant Db11 was demonstrated to fulfil koch's postulates. Strain Db10 and its antibiotic-resistant derivatives were lethal to Drosophila whether given in the food or by injection. Evidence for toxin(s) was found only in sterile supernatants from 7 d cultures. Such extracts contained proteolytic activity and inactivated the antibacterial activity in immune haemolymph from Cecropia. Phages phi J and phi K were used to isolate phage-resistant mutants of Db11. Three such mutants and their parental strain were investigated for their susceptibility to immune haemolymph from Cecropia. The parental strain was resistant to incubation with 90% haemolymph for 2 h at 37 degrees C; all phage-resistant mutants were susceptible to the immune haemolymph with "killing times" (i.e. the time required to kill 90% of the viable cells) ranging from 15 to 55 min. When the same strains were compared for their virulence to Drosophila, the phage-resistant mutants had significantly reduced virulence. It is concluded that resistance to insect immunity plays an important role in the overall pathogenicity of S. marcescens.


Laparoscopically assisted colorectal resection in Hirschsprung's disease.
Minimally invasive surgery (MIS) has been adapted to virtually every kind of abdominal operation. Colorectal resections of any extent are possible. The reduction in local and systemic surgical trauma in MIS suggests this method could be valuable for rectal resection for Hirschsprung's disease. - Diagnostic work-up is similar to that in open surgery. Three trocars are sufficient, and a fourth may be helpful. Dissection encompasses the entire aganglionic segment and is extended orally to normal bowel.. After complete dissection down to the pelvic floor the bowel is everted transanally, resected in due length and the coloanal anastomosis sutured from outside. Since 1996 four children out of nine (age 11 weeks - 18 years) have been operated laparoscopically at our institution. - There were no intraoperative complications. Recovery time is impressively rapid. Oral intake may be begun on the first postoperative day and physical activity is rapidly restored. There are however problems typical to this method: anastomotic leakage, stricture formation, and the necessity of continuing dilation. - The feasibility of laparoscopic colorectal resections of any extent is unquestionable. Benefits for the patient seem evident. Prospective randomized studies to provide a higher level of evidence for the benefit of laparoscopy as compared to open technique are difficult to perform due to the small number of patients.


[White Paper of the European Commission--strategy for a new chemicals policy].
The current chemicals regulation in EU member states provides sufficient data on new chemicals but almost non on the about 100.000 existing chemicals. To improve the situation the EU Commission is proposing in a recent White Paper on a strategy for a new chemicals policy to regulate new and existing chemicals in an identical manner. The Commission suggests to use in vitro methods to provide information rapidly on hazardous health and environmental properties of chemicals. The fundamental change in the testing strategy, which is relying on in vitro methods rather than testing in animals is most welcome not only from the scientific and economic point of view but also from the perspective of the protection of animals and the environment. A calculation of the costs and time for testing all existing chemicals in animal experiments shows that this task is unrealistic from a financial point of view and also when taking into account the time required for testing. Taking into account the proposals provided by several working groups of the EU validation centre ECVAM during the past year, a concept is described that will allow the testing of all existing chemicals applying only non-animal tests. Since the new testing strategy will be cheaper and faster it will be more acceptable from both the financial and the ethical point of view. Finally, establishing the proposed new in vitro tests in the safety testing strategy will require considerable funding of validation studies by the EU Commission.


Exposure to toluene in the printing industry is associated with subfecundity in women but not in men.
To examine the possible influence of exposure to toluene on human fertility. In a cross sectional study, a sample of 150 male and 90 female printing industry workers were interviewed retrospectively on reproductive experience with a modified version of the European study of infertility and subfecundity questionnaire. Exposure categories comprised job descriptions and information on exposure measurements obtained by industrial hygienists. The fecundability ratio (FR) was estimated on the basis of time to pregnancy (TTP) or periods of unprotected intercourse not leading to pregnancy (PUNP) by means of survival analysis with proportional hazard models. Confounders such as age, ethnicity, smoking, parity, pelvic inflammatory diseases, and frequency of sexual intercourse were controlled for in the analyses. 256 Periods of TTP or PUNP were reported by men and 174 by women. After exclusion of induced abortions, birth control failures, and periods without employment for female workers we were able to analyse 169 periods in men and 100 periods in women. Male workers who had been exposed to different concentrations of toluene and their partners did not show a reduction in fecundity. In women (39 periods occurred during exposure) fecundity was reduced (FR 0.47, 95% confidence interval (95% CI) 0.29 to 0.77). Neither, restriction to only the first period nor exclusion of PUNPs changed the results (FR 0.48, 95% CI 0.24 to 0.97). After considering possible biases, low daily exposure to toluene in women seems to be associated with reduced fecundity. This result is in accordance with other findings for organic solvents and supports both the hypotheses that (a) organic solvents could affect hormonal regulation, and that (b) organic solvents increase early fetal losses which in turn contributes to longer times of unprotected intercourse.


Neferine hinders choriocarcinoma cell proliferation, migration and invasion through repression of long noncoding RNA-CHRF.
The comprehensive pathological peculiarities of Neferine (NEF) have been testified in disparate diseases. But, the functions of NEF in choriocarcinoma progression remain unexplored. The research endeavoured to uncover the anti-tumour action of NEF in choriocarcinoma cells. NEF at diverse doses was employed to dispose JEG-3 and HTR-8 cells, and cell viability assessment adopted CCK-8 assay. After 60 μg/mL NEF management, BrdU-positive cells, apoptosis, migration, invasion and correlative factors were assessed. CHRF expression in choriocarcinoma tumour and choriocarcinoma cell lines was estimated via RT-qPCR. Then, the functions of overexpressed CHRF in NEF-disposed cells were determined. At last, impacts of NEF on PI3K/AKT/mTOR and ERK1/2 pathways were evaluated. Results showed that NEF restrained cell proliferation, triggered apoptosis and repressed migration and invasion in JEG-3 and HTR-8 cells. CHRF was ascended in choriocarcinoma tissues and NEF repressed CHRF expression in choriocarcinoma cell lines. Additionally, overexpressed CHRF abolished the above functions of NEF in choriocarcinoma cells proliferation, apoptosis, migration and invasion. Further, NEF impeded PI3K/AKT/mTOR and ERK1/2 pathways via repressing CHRF. The explorations testified that NEF exhibited the anti-tumour action in JEG-3 and HTR-8 cells via hindering PI3K/AKT/mTOR and ERK1/2 pathways by mediating CHRF.


Feedlot performance and tissue residues of cattle consuming diets containing aflatoxins.
A comparative slaughter feeding trial was conducted using crossbred Hereford-Angus steers. Cottonseed meal known to be contaminated with aflatoxins was fortified with long-grain rice cultures of Aspergillus flavus to produce complete rations that contained 0, 60, 300 and 600 ppb aflatoxin B1 (AFB1). Forty steers, weighing approximately 250 kg were randomly assigned to four equal treatment groups. Animals were housed in individual pens and fed the assigned contaminated ration ad libitum for 155 d. On d 156 all animals were placed on the control ration for 14 d. Animals were weighed and blood samples collected every month for clinical chemistry analysis. Liver, muscle and fat biopsies were collected at 6-wk intervals. These samples were stored in liquid N2 prior to analysis for aflatoxin M1 and B1 content. Separate liver samples were taken for histopathological examination. Aflatoxin was withdrawn from the diet 2 wk prior to slaughter. After 1 wk of withdrawal liver, muscle and fat samples were collected and assayed for AFB1 and aflatoxin M1 (AFM1). Liver samples also were taken for histopathological examination. Growth rates and feed intakes in the 60 and 300 ppb aflatoxin treatments were not significantly different from controls. The 600-ppb treatment group had a decrease in feed intake and rate of gain. Rations containing 60 and 300 ppb of aflatoxin had no significant influence on blood components and enzyme patterns. The 600-ppb treatment caused a slight, but consistent, increase in serum glutamic oxaloacetic transaminase and alkaline phosphatase.(ABSTRACT TRUNCATED AT 250 WORDS)


The acute side effects of bright light therapy: a placebo-controlled investigation.
Despite the emergence of numerous clinical and non-clinical applications of bright light therapy (LT) in recent decades, the prevalence and severity of LT side effects have not yet been fully explicated. A few adverse LT effects-headache, eye strain, irritability, and nausea-have been consistently reported among depressed individuals and other psychiatric cohorts, but there exists little published evidence regarding LT side effects in non-clinical populations, who often undergo LT treatment of considerably briefer duration. Accordingly, in the present study we examined, in a randomized sample of healthy young adults, the acute side effects of exposure to a single 30-minute session of bright white light (10,000 lux) versus dim red light (< 500 lux). Across a broad range of potential side effects, repeated-measures analyses of variance revealed no significant group-by-time (Pre, Post) interactions. In other words, bright light exposure was not associated with a significantly higher incidence of any reported side effect than was the placebo control condition. Nevertheless, small but statistically significant increases in both eye strain and blurred vision were observed among both the LT and control groups. Overall, these results suggest that the relatively common occurrence of adverse side effects observed in the extant LT literature may not fully extend to non-clinical populations, especially for healthy young adults undergoing LT for a brief duration.