Patterns of coping in families of psychotic children.
The nature of family adaptation to severely and chronically disabling cognitive and behavioral conditions is considered. The relationship between atypical development and family response is examined to derive basic concepts--parents' perceptions of their child's unusual characteristics, effects on social relationships, etc.--and suggest strategies for intervention and research.


Acromegaly due to production of a growth hormone releasing factor by a bronchial carcinoid tumor.
We have studied growth hormone production in a patient with a bronchial carcinoid and acromegaly. The absence of growth hormone from the carcinoid tumour was demonstrated by extraction, cell culture and immunoperosidase techniques. Using a linked perfusion culture system, effluent from the bronchial carcinoid tumour culture stimulated a rapid release of growth hormone from a rat pituitary monolayer. This is the first time evidence of growth hormone releasing activity by a bronchial carcinoid has been demonstrated in a production.


Dietary intake of yogurt and cheese in children at age 1 year and sleep duration at age 1 and 3 years: the Japan Environment and Children's Study.
We previously reported that 1-year-old infants born to mothers who regularly consumed fermented food during pregnancy had a lower risk of sleep deprivation. However, it is not known if these positive effects are enhanced when infants themselves eat fermented foods or the long-term effects of such consumption. In this study, we examined the association between the frequency of fermented food intake during the child's weaning period and sleep deprivation at age 1 and 3 years. This birth cohort study used data from a nationwide, government-funded study called the Japan Environment and Children's Study (JECS), covering 65,210 mother-child pairs. We examined the association between infants' consumption of fermented foods at 1 year of age and sleep deprivation at 1 and 3 years of age. There was no association between yogurt or cheese intake and sleep duration at age 1; at age 3, there was no group difference, although a trend test showed that yogurt intake at age 1 was significantly associated with sleep duration at age 3. There was also no association between the frequency of cheese intake and inadequate sleep duration at age 3. Frequency of children's yogurt and cheese intake at age 1 was not associated with sleep duration at age 1 or 3. However, a trend test showed a significant association between the frequency of yogurt intake at age 1 and sleep duration at age 3.


Role of semi-volatile particulate matter in gas-particle partitioning leading to change in oxidative potential.
Atmospheric semi-volatile organic compounds (SVOCs) are complex in their chemical and toxicological characteristics with sources from both primary combustion emissions and secondary oxygenated aerosol formation processes. In this study, thermal desorption of PM2.5 in association with online measurement of reactive oxygen species (ROS) was carried out to study the role of SVOCs in its gas-particle partitioning. The mass concentrations of PM2.5, black carbon (BC) and p-PAHs downstream of a thermodenuder were measured online at different temperature settings (25, 50, 100, and 200 °C) to characterize PM physico-chemical properties. While the mass concentrations of PM2.5 and p-PAHs reduced to ∼34% at 200 °C compared to that in ambient temperature, BC mass concentration has decreased by 30% at the highest temperature. Furthermore, the submicron particle size distribution showed reduced particle number concentration in Aitken mode at 200 °C heating. The ROS, measured by Particle-into-Nitroxide-Quencher, also showed reduction and followed a similar trend with PM measurements, where the total ROS decreased by 12%, 31%, and 53% at 50 °C, 100 °C, and 200 °C, respectively, compared to the ambient sample. When a HEPA filter was included in the upstream of samples, 39% of gas phase ROS reduction was observed at 200 °C. This provided a good estimate of the contribution of SVOCs in ROS production in PM2.5, where decreased SVOCs concentration at 200 °C increased the percentage of particle surface area. This concludes that the surface chemistry of these organic coatings on the particles is important for assessing the health impacts of PM.


Moniliophthora perniciosa, the mushroom causing witches' broom disease of cacao: Insights into its taxonomy, ecology and host range in Brazil.
Witches' broom caused by Moniliophthora perniciosa is the main disease of cacao (Theobroma cacao) in Brazil. The fungus is known to occur on other host families and these populations have been addressed in the literature as biotypes: C (Malvaceae); H (Malpighiaceae); L (Bignoniaceae) and S (Solanaceae). No complete elucidation of the phylogenetic relationships of isolates obtained from this disparate host range appears in the literature. One member of H (ex Heteropterys acutifolia) has been described as a distinct species. But should other biotypes be also recognized as distinct taxa? In the present study, a survey yielding 24 isolates of M. perniciosa from ten hosts and covering a wide range of geographic regions in Brazil was undertaken. These isolates were compared with those from T. cacao using three DNA regions for the phylogenetic analyses: ITS, LSU and RPB1. Morphology was also examined. All isolates in this study were found to belong to M. perniciosa, including the population from H. acutifolia, formerly treated as Moniliophthora brasiliensis but reduced here to a synonym of M. perniciosa. This species ranged from pathogenic to a previously unreported occurrence as a non-pathogenic endophyte in the Atlantic rainforest tree Allophylus edulis (Sapindaceae). M. perniciosa was recorded on a range of solanaceous hosts (16 species) over a wide variety of ecosystems. The ecological and evolutionary significance of these novel findings are discussed.


Associations between early in-hospital medications and the development of delirium in patients with stroke.
Patients hospitalized with stroke develop delirium at higher rates than general hospitalized patients. While several medications are associated with existing delirium, it is unknown whether early medication exposures are associated with subsequent delirium in patients with stroke. Additionally, it is unknown whether delirium identification is associated with changes in the prescription of these medications. We conducted a retrospective cohort study of patients admitted to a comprehensive stroke center, who were assessed for delirium by trained nursing staff during clinical care. We analyzed exposures to multiple medication classes in the first 48 h of admission, and compared them between patients who developed delirium >48 hours after admission and those who never developed delirium. Statistical analysis was performed using univariate testing. Multivariable logistic regression was used further to evaluate the significance of univariately significant medications, while controlling for clinical confounders. 1671 unique patients were included in the cohort, of whom 464 (27.8%) developed delirium >48 hours after admission. Delirium was associated with prior exposure to antipsychotics, sedatives, opiates, and antimicrobials. Antipsychotics, sedatives, and antimicrobials remained significantly associated with delirium even after accounting for several clinical covariates. Usage of these medications decreased in the 48 hours following delirium identification, except for atypical antipsychotics, whose use increased. Other medication classes such as steroids, benzodiazepines, and sleep aids were not initially associated with subsequent delirium, but prescription patterns still changed after delirium identification. Early exposure to multiple medication classes is associated with the subsequent development of delirium in patients with stroke. Additionally, prescription patterns changed following delirium identification, suggesting that some of the associated medication classes may represent modifiable targets for future delirium prevention strategies, although future study is needed.


Cost-effectiveness of sibutramine in the treatment of obesity.
This study reports incremental cost-utility of sibutramine compared to diet and lifestyle advice for the treatment of obesity. The model estimates the costs and quality of life benefits associated with weight loss itself and the reduced incidence of coronary heart disease (CHD) and diabetes in the "healthy obese." The key source of effectiveness data is 2 randomized controlled trials over 12 months. Utility gain per kilogram lost is analyzed using Short Form-36 data from sibutramine trials. The impact on CHD is estimated using the Framingham risk equation, which relates age/sex/body mass index to risk of heart disease. The reduced incidence of diabetes due to weight loss is estimated from published literature. A life tables approach was used to calculate the cost per quality-adjusted life year (QALY) of 1 year's treatment with sibutramine compared to diet and lifestyle advice. The incremental cost per QALY of sibutramine is 4,780 UK pounds. Sensitivity analyses show that this result is sensitive to utility associated with weight loss and the frequency of monitoring. Sibutramine is a cost-effective treatment for obesity when combined with diet and lifestyle advice.


The RIO kinases: an atypical protein kinase family required for ribosome biogenesis and cell cycle progression.
Atypical protein kinases (aPKs) include proteins known to be involved in the phosphorylation-mediated regulation of a wide variety of cellular processes, as well as some for which the function is, as yet, unknown. At present, 13 families of aPKs have been identified in the human genome. This review briefly summarizes their known properties, but concentrates in particular on the RIO family of aPKs. Representatives of this family are present in organisms varying from archaea to humans. All these organisms contain at least two RIO proteins, Rio1 and Rio2, but a third Rio3 group is present in multicellular eukaryotes. Crystal structures of A. fulgidus Rio1 and Rio2 have shown that whereas the overall fold of these enzymes resembles typical protein kinases, some of the kinase structural domains, particularly those involved in peptide substrate binding, are not present. The mode of binding of nucleotides also differs from other kinases. While the enzymatic activity of Rio1 and Rio2 has been demonstrated and both have been shown to be essential in S. cerevisiae and required for proper cell cycle progression and chromosome maintenance, the biological substrates of RIO proteins still remain to be identified.


Derivative spectrophotometry of dimer and monomer of enolase.
1. SDS causes significant polar exposure of aromatic amino acids of enolase. The alpha-helix content remains unchanged. The enzyme lost all its activity. 2. The presence of 1 M K Br in enzyme solution results in a smaller increase of polarity of aromatic amino acids residues environment. The amount of alpha-helix does not decrease in comparison to native enzyme. Enzyme lost nearly 80% of its initial activity. 3. The extreme pH values and the presence of 6 M Gnd.HCl influence the whole structure of enolase. It is accompanied by a large polar shift of aromatic amino acids and significant decrease of alpha-helix content of the protein.


Observer-based rather than population-based confidence limits for determining probability of change in visual fields.
Determining confidence intervals on psychophysical thresholds is straight forward if the psychometric function is known. In clinical settings, however, there is only partial information about the psychometric function, hence confidence limits are usually derived from test-retest data collected from many subjects. In this paper, we introduce a computational technique for deriving confidence limits for an individual's endpoint threshold using data typically obtained in a clinical setting, rather than a database of test-retest performance. The technique uses probabilistic analysis of all possible response sequences in a test procedure. We then extend this procedure to allow for levels of typical uncertainty in data measurement.


Seroprevalence and risk factors associated with Leishmania infantum in dogs in Sardinia (Italy), an endemic island for leishmaniasis.
Leishmaniasis is a widespread, vector-borne parasitosis causing clinical manifestations in animals and in humans. In dogs, Canine Leishmaniasis has been reported in as much as 50 countries and the Mediterranean basin is known to be one of the most affected zones. Within these areas, the Island of Sardinia (Italy) has long been considered endemic for leishmaniasis and the presence of two arthropod vectors has recently been reported there. Nevertheless, to date, no epidemiological surveys regarding CanL have been carried out on the island. Hence, for the first time, the seroprevalence and the risk factors were investigated. Blood samples, as well as clinical and general information from 1.147 dogs, were collected and analyzed. Dogs consisted of two distinct populations, namely "owned dogs" and "kennel dogs." Anti-Leishmania IgG antibodies were detected using IFAT and samples were scored as positive at a cut-off dilution of 1:80. Data was analyzed using a Chi-squared test and bivariate and multivariate analyses were performed. Overall, 15.4% of dogs were found to be infected with CanL while only 44.1% of these animals exhibited clinical signs. Owned dogs (27.2%) were found to be infected more often than kennel dogs (10.6%); male dogs were found to be more frequently infected than female dogs and the number of infected animals increases with age. The present survey confirmed the endemic nature of leishmaniasis in Sardinia with a similar seroprevalence as mainland Italy. The results obtained serve as validation for the hypothesis that, in endemic areas, clinical CanL representations constitute only a fraction of the leishmaniasis cases.


Efficacy of topical insulin for recurrent epithelial corneal erosions.
Topical insulin can promote and accelerate corneal regeneration, even in eyes with serious comorbidities, and offers several benefits over other treatment options. The aim of this study is to evaluate the effect of topical insulin in treatment of recurrent epithelial corneal erosion. Patients with recurrent epithelial erosions were included in a prospective non-randomized hospital-based study, divided into two groups, one of them received persistent epithelial defects (PEDs) conventional treatment and the other received the same treatment with insulin eye drops 4 times/day. All patients were examined carefully by slit lamp. Patients during the 1st, 2nd, 3rd, and 4th weeks as well as after 2 months. Demographics, etiology, therapy, comorbidities, and the healing time of PED were performed. Area shows significant improvement after 2 weeks (p = 0.006), 2 months (p = 0.046), and 3 months (p = 0.002) in group II (cornetears gel and topical insulin) as compared to group I (cornetears gel). The recurrence was statistically significant decreased with cornetears gel and topical insulin (group II) by 0.0%, as compared to cornetears gel (group I) by 3 patients (21.4%). Topical insulin can promote corneal reepithelization in recurrent epithelial erosion and decreases recurrence in these cases. Other advantages include excellent tolerance, availability, and cost-effectiveness.


Effect of acute umbilical cord compression on hepatic carbohydrate metabolism in the fetal lamb.
Although the liver plays a central role in glucose homeostasis in the adult, its importance in fetal glucose homeostasis during acute reductions of substrate delivery is unknown. To examine this, we studied eight fetal lambs at 121 +/- 2 d gestation. We placed catheters in the descending aorta, inferior vena cava umbilical vein and the left (n = 6) or right (n = 2) hepatic vein, and a balloon occluder around the umbilical cord. At least 4 d after surgery, before and during umbilical cord compression, we measured blood oxygen saturation, glucose, lactate, and Hb concentrations, and blood flows using the radiolabeled microsphere technique. Gluconeogenesis was assessed by infusion of [U14C]lactate. Reducing umbilical flow by 50-60% from a control value of 181 +/- 20 mL/min/kg (mean +/- SD) caused a dramatic decrease in hepatic blood flow from 332 +/- 99 to 94 +/- 77 mL/min/100 g (p less than 0.05). Oxygen delivery to the fetus fell by 50% and that to the liver by 73%. However, hepatic O2 consumption was maintained by increased extraction. Glucose delivery to the liver fell from 67 +/- 24 to 20 +/- 13 mg/min/100 g (p less than 0.001), but lactate delivery did not change. In spite of the maintenance of lactate delivery, net hepatic lactate uptake fell significantly from 3.3 +/- 1.7 to 1.4 +/- 0.9 mg/min/100 g (p less than 0.05). This could account, in part, for the increase of blood lactate concentration from 16 +/- 4 to 27 +/- 7 mg/dl.(ABSTRACT TRUNCATED AT 250 WORDS)


The European Portuguese adaptation of the Fear of Pain Questionnaire.
In Portugal, it is estimated that chronic pain affects 36.7% of the population, constituting a multifactorial phenomenon with great impact on individual, family, community, and social levels. In the fear-avoidance model of pain, one of the most consistent consensus in the literature, the fear arises as one of the variables that can contribute to the development and maintenance of this condition. Thus, instruments for evaluating the fear of pain, as Fear of Pain Questionnaire (FPQ-III), may be useful in the conceptualization of the subjective experience of pain. Accordingly, this paper aims to describe the adaptation of FPQ-III to the European Portuguese idiom. A total of 1,094 participants (795 female; mean age=25.16, SD=7.72 years old) completed the web based questionnaire. The results point to a different factor model found in the first study of the original scale (five factors: minor pain, severe pain, medical pain, injection pain, and afflicted pain), good internal consistency (0.75 to 0.85) and good correlations (between 0.30 and 0.59) between subscales and (between 0.68 e 0.85) for the total score and subscales. Given the need to meet the various dimensions of the subjective experience of pain, the Fear of Pain Questionnaire is assumed as a useful tool that, in combination with other tools, may contribute to the evaluation and intervention procedures progressively more comprehensive and adjusted to the challenges raised with the issue of chronic pain.


Prognostic factors.
The best predictive models of today can predict no more than 50% of the natural variability of the disease, despite the sophisticated mathematic analyses and the dozens of variables assessed. Clearly, a universe of still unknown prognostic factors remains to be discovered. Analogous to infinity being immeasurable, the fate of the individual patient will never be calculated mathematically. As the discovery of new prognostic factors continues, however, the prediction of the outcome of patients becomes more reliable. Although some prognostic factors (eg, tumor neoangiogenesis and quality of life) are already supported by a large body of evidence, recent research has introduced new groups of prognostic factors (eg, molecular genetic markers and the subclinical activation of coagulation fibrinolysis). Other intriguing factors (eg, a state of mental depression) might also be recognized as important in the near future. A new small portion of the universe has been explored.


Expression of apolipoprotein B epitopes in very low density lipoprotein subfractions. Studies with monoclonal antibodies.
Epitope expression was studied in both denatured apolipoprotein B (apo B) on Western blots and in intact low density lipoprotein (LDL) and very low density lipoprotein subfractions VLDL1 (Sf120-400), VLDL2 (Sf60-120), and VLDL3 (Sf20-60) in competitive binding immunoassays with the aid of six monoclonal anti-LDL antibodies. The apo B in all lipoprotein fractions was shown to bind to all antibodies, but significant differences in apo B epitope expression were observed. On the average, the immunoreactivity of VLDL subfractions (expressed as binding affinity and as relative 125I-LDL displacing potency) decreased with increasing flotation rate. Similarly, VLDL1 was less immunoreactive than lipolyzed "remnants" of VLDL1 after treatment with bovine milk lipoprotein lipase. The results indicate that, even when lipoprotein fractions obtained from the same individual and having the same kind of apo B subspecies were compared, significant differences in immunoreactivity occurred due to the modulating effect of other lipoprotein components on apo B epitope expression.


The prevalence of Gasterophilus intestinalis in horses in northern England and Wales.
The stomachs of 448 horses from northern England and Wales were examined for Gasterophilus larvae, and 237 (52.7%) were found to be infected with G. intestinalis. Larvae were present in stomachs examined during each month of the year except August. Second instar larvae occurred from September through February and third instars were present from November through July. Adult fly activity began in August as indicated by the presence of eggs on horses. The life-cycle of G. intestinalis in northern England and Wales is outlined from the data presented. The mean instar burdens were 15.7 second and 38.0 third instars, and more than 75% of the infections consisted of up to 50 larvae. Prevalence of infection and mean larval burdens declined with increasing age of host. Only one of 258 duodena examined was infected with G. nasalis and this horse originated from the south coast of England, outside of the catchment area of the other horses examined.


Synthesis of scutellarein derivatives to increase biological activity and water solubility.
In order to improve the biological activity and water solubility of scutellarin (1), some derivatives of its main metabolite (scutellarein) were designed and synthesized. All the compounds were tested for their thrombin inhibition activity through the analyzation of thrombin time (TT), activated partial thromboplastin time (APTT), prothrombin time (PT) and fibrinogen (FIB). Their antioxidant activities were assessed by measuring their scavenging capacities toward 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and the ability to protect PC12 cells against H2O2-induced cytotoxicity, their water solubility were also assessed by ultraviolet (UV) spectrophotometer. The results showed that compound 8b demonstrated stronger anticoagulant and antioxidant activity, better water solubility compared with scutellarein (2), which warrants it as a promising agent for the treatment of ischemic cerebrovascular disease.


Ablation of Iqgap2 protects from diet-induced hepatic steatosis due to impaired fatty acid uptake.
Long-chain fatty acids (LCFA) serve as structural components for membrane biogenesis and as primary energy sources during mitochondrial β-oxidation reactions. Hepatic LCFA uptake is complex, with characteristics suggestive of a dual-kinetic model manifested by rapid (carrier-assisted/facilitated) and delayed (passive diffusional) phases. Our previous work using mice deficient of the Iqgap2 gene established a highly novel link between IQGAP2, a putative GTPase-activating protein, and hepatocarcinogenesis. Now we report that Iqgap2 deficiency also results in selective loss of the facilitated phase of hepatocyte LCFA uptake with preservation of the diffusional component. This molecular defect was seen in Iqgap2(-/-) hepatocytes of all ages studied (1-, 4-, 8-months). The loss of facilitated LCFA uptake protected against development of hepatic triglyceride accumulation in Iqgap2-deficient mice fed high-fat diet, consistent with a fundamental role in physiological fat partitioning. These phenotypic changes could not be explained by genetic loss of fatty acid processing proteins known to regulate lipid uptake or metabolic processing pathways. Iqgap2-deficient livers also displayed enhanced insulin sensitivity. These observations identify a novel property of the putative GTPase-activating protein IQGAP2 in LCFA uptake in vitro and in vivo, and implicate IQGAP2 in an intracellular signaling pathway necessary for functional fatty acid uptake, lipid processing, and, possibly, glucose homeostasis.


Inflammation activates self hsp60-specific T cells.
Injection of incomplete Freund's adjuvant (IFA) into the footpads of BALB/c mice induced an acute inflammation. Draining popliteal lymph nodes showed major histocompatibility complex (MHC) class II-restricted proliferation when challenged in vitro with recombinant Mycobacterium bovis 65-kDa heat shock protein (hsp65). alpha beta T cell receptor-positive, CD4+, hsp65-specific T cell lines and clones were generated from these lymph nodes, and 87% of clones responded to a beta galactosidase fusion protein containing residues 238-573 of human hsp60. Seventy percent of these hsp60-responsive clones also responded to a synthetic peptide corresponding to residues 412-423 of the mouse hsp60. This peptide also induced significant responses in IFA-primed lymph node cells but not in lymphoid cells from unimmunized mice. These results demonstrate that T cells specific for epitopes in self hsp60 are activated during inflammatory responses induced in the absence of exogenous bacterial hsp65. The findings of this study may provide a basis for understanding the often reported isolation of mycobacterial hsp65-responsive T cells from inflammatory sites of arthritis patients, and the protective effects of preimmunization with hsp65 in experimental models of arthritis.


RNF213 rs112735431 polymorphism in intracranial artery steno-occlusive disease and moyamoya disease in Koreans.
The rs112735431 polymorphism of the RNF213, a susceptibility variant for moyamoya disease (MMD), may be associated with non-MMD intracranial artery steno-occlusive disease of non-MMD type (non-MMD ICAD) in Asian. We investigated whether the rs112735431 polymorphism of the RNF213 affect the development of non-MMD ICAD in Koreans compared to MMD and control group. We included 31 patients with non-MMD ICAD, 25 patients with MMD, and 100 participants as control group. The rs112735431 polymorphism of the RNF213 was evaluated by polymerase chain reaction amplification of target and detection by restriction fragment length polymorphism analysis. Clinical phenotype was compared between patients with and without the rs112735431 polymorphism in non-MMD ICAD and MMD. The rs112735431 polymorphism of the RNF213 was significantly associated with non-MMD ICAD (p=0.001; odds ratio, 14.3; 95% confidence interval, 2.80-73.2) and MMD (p<0.0001; odds ratio, 126.0; 95% confidence interval, 24.2-656.0). The rate of hypertension was more frequent in MMD with the rs112735431 polymorphism than MMD without polymorphism (p=0.010). The rs112735431 polymorphism of the RNF213 is highly associated not only with MMD but also with non-MMD ICAD in Koreans. Also, our study suggests that the rs112735431 polymorphism of the RNF213 may be linked to the hypertension in MMD. Further studies are needed to clarify the relationship between the rs112735431 polymorphism of the RNF213 and hypertension in patients with MMD.


Author Correction: Calcium mishandling in absence of primary mitochondrial dysfunction drives cellular pathology in Wolfram Syndrome.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


A high fat diet fosters elevated bisretinoids.
High dietary fat intake is associated with metabolic dysregulation, but little is known regarding the effects of a high fat diet (HFD) on photoreceptor cell functioning. We explored the intersection of an HFD and the visual cycle adducts that form in photoreceptor cells by nonenzymatic reactions. In black C57BL/6J mice and albino C57BL/6Jc2j mice raised on an HFD until age 3, 6, or 12 months, chromatographically quantified bisretinoids were increased relative to mice on a standard diet. In vivo measurement of fundus autofluorescence, the source of which is bisretinoid, also revealed a significant increase in the HFD mice. Additionally, mice provided with a diet high in fat presented with elevated retinol-binding protein 4, the protein responsible for transporting retinol in plasma. Vitamin A was elevated in plasma although not in ocular tissue. Bisretinoids form in photoreceptor cell outer segments by random reactions of retinaldehyde with phosphatidylethanolamine. We found that the latter phospholipid was significantly increased in mice fed an HFD versus mice on a control diet. In leptin-deficient ob/ob mice, a genetic model of obesity, plasma levels of retinol-binding protein 4 were higher but bisretinoids in retina were not elevated. Photoreceptor cell viability measured as outer nuclear layer thickness was reduced in the ob/ob mice relative to WT. The accelerated formation of bisretinoid we observed in diet-induced obese mice is related to the high fat intake and to increased delivery of vitamin A to the visual cycle.


Covalent organic framework in situ grown on the metal-organic framework as fiber coating for solid-phase microextraction of polycyclic aromatic hydrocarbons in tea.
A novel MIL-88-NH2@COF composite was produced by in situ growth of covalent organic framework (COF) on the metal-organic framework (MOF) surface. To obtain a coating fiber for solid-phase microextraction (SPME), the MIL-88-NH2@COF composite physically adhered to the stainless steel wire. Combined with gas chromatography-flame ionization detection (GC-FID), various analytes such as chlorophenols (CPs), phthalates (PAEs), and polycyclic aromatic hydrocarbons (PAHs) were extracted and determined to evaluate the extraction performance of MIL-88-NH2@COF coated fibers and explore their extraction mechanism. This composite exhibit excellent extraction performance and adsorption capacity for various analytes, especially for PAHs with enrichment factor up to 9858. The SPME-GC-FID method based on MIL-88-NH2@COF fiber was established for the determination of five PAHs after the main extraction conditions were optimized. Under optimal conditions, the proposed technique showed a wide linear range (1-150 ng mL-1) with a low limit of detection (0.019 ng mL-1) and a high coefficient of determination (R2 > 0.99). The developed SPME-GC-FID method was used to determine PAHs in green tea and black tea samples, with good recoveries of 51.70-103.64% and 68.56-103.64%, respectively. It is worth mentioning that this is the first time MIL-88-NH2@COF composites have been prepared and applied to SPME. The preparation method of the composite provides a new idea in adsorbent preparation, which will contribute to the field of SPME.


Bacillus subtilis SsbA and dATP regulate RecA nucleation onto single-stranded DNA.
Bacillus subtilis RecA preferentially hydrolyzes dATP over ATP and supports an efficient DNA strand exchange reaction in the presence of dATP when compared to ATP. Saturating amounts of SsbA, independently of the order of addition, reduce the single-stranded (ss) DNA-dependent dATPase activity of RecA, and block the ATPase activity. SsbA added prior to RecA slightly stimulates the dATP-dependent DNA strand exchange activity, whereas added after RecA greatly enhances the extent of strand exchange. In the presence of ATP, 10 times more RecA is required to achieve a comparable level of strand exchange than in the presence of dATP. We propose that dATP binding and hydrolysis as well as SsbA provide different levels of regulation of the dynamic RecA nucleoprotein filament.


Encephalocoele as a complication of intranasal sinus surgery: optimal evaluation with magnetic resonance imaging.
We report a case of post-operative frontal basal encephalocoele evaluated using a new magnetic resonance imaging (MRI) sequence, fast inversion recovery for myelin suppression (FIRMS). FIRMS was developed to enhance the differentiation between grey and white matter. In this case, the sequence was beneficial in distinguishing the encephalocoele from adjacent nasal mucosa and secretions.


Involvement of glutamate mutase in the biosynthesis of the unique starter unit of the macrolactam polyketide antibiotic vicenistatin.
The macrolactam antibiotic vicenistatin, produced in Streptomyces halstedii HC34, is biosynthesized by the polyketide pathway, using a unique 3-methylaspartate-derived molecule as starter unit. The vinI gene in the vicenistatin biosynthetic gene cluster encoding glutamate mutase, which rearranges glutamate to 3-methylaspartate, was disrupted. The vinI disruption completely abolished the production of vicenistatin, while the disruptant recovered the production of vicenistatin when 3-methylaspartate was added to the culture. These results indicate that vinI is essential for the 3-methylaspartate formation in the vicenistatin biosynthesis. Furthermore, the mutant accumulated new vicenistatin derivatives (desmethylvicenistatins), which lacked a methyl group in the starter unit. The desmethylvicenistatins were shown by feeding experiments to be derived from aspartate instead of 3-methylaspartate as the starter unit. These results indicate that the vicenistatin polyketide synthase can accept alternative starter units toward the production of novel polyketides.


The cross-talk between STAT1/STAT3 and ROS up-regulates PD-L1 and promotes the release of pro-inflammatory/immune suppressive cytokines in primary monocytes infected by HHV-6B.
Programmed death ligand 1 (PD-L1) up-regulation on antigen presenting cells induces T cell dysfunction, strongly impairing immune response. Human Herpesviruses (HHV) 6B is a β-herpesvirus that, although displays a higher tropism for T cells, can infect other immune cells including monocytes and dendritic cells (DCs) and neuronal cells. We have previously shown that HHV-6B infection of primary monocytes reduced autophagy and induced Endoplasmic Reticulum (ER) stress/ Unfolded Protein Response (UPR), impairing their survival and differentiation into DCs. In this study, we found that PD-L1 expression was up-regulated by HHV-6B on the surface of infected monocytes and that its extracellular release also increased, effects known to lead to an impairment of anti-viral immune response. At molecular level, PD-L1 up-regulation correlated with the activation of a positive regulatory circuit between the increase of intracellular ROS and the activation of STAT1 and STAT3 induced by HHV-6B, accompanied by a high release of pro-inflammatory/immune suppressive cytokines. In conclusion, this study unveils new strategies put in place by HHV-6B to induce immune dysfunction and the underlying molecular pathways that could be targeted to counteract such immune suppressive effects.


Elastic behavior of neodymium based manganites.
With a view to investigate the elastic behavior of Nd(0.67)A(0.33)MnO(3) (where A=Ca, Sr, Ba, Pb) manganite system, the samples were prepared by the sol gel method. After characterizing the samples structurally, a systematic investigation of ultrasonic longitudinal and transverse sound velocities of all the samples was undertaken by pulse transmission technique in the temperature range, 100-300K. It has been found that all the elasticity parameters, including Debye temperature, are found to increase continuously with increasing ionic radii of the dopant ion. All the samples are also found to exhibit anomalies in both the longitudinal and transverse velocities near their ferro to para magnetic transition (T(C)) temperatures. Apart from this, Nd(0.67)Ca(0.33)MnO(3) sample is also found to exhibit, a transition at its charge ordering temperature (T(co)). An explanation for the observed elastic anomalies based on a mean field theory has been given.


miR-888: A Novel Cancer-Testis Antigen that Targets the Progesterone Receptor in Endometrial Cancer.
Cancer-testis (CT) antigens are a large family of genes that are selectively expressed in human testis germ cells, overexpressed in a variety of tumors and predominantly located on the X chromosome. To date, all known CT antigens are protein-coding genes. Here, we identify miR-888 as the first miRNA with features characteristic of a CT antigen. In a panel of 21 normal human tissues, miR-888 expression was high in testes and minimal or absent in all other examined tissues. In situ hybridization localized miR-888 expression specifically to the early stages of sperm development within the testes. Using The Cancer Genome Atlas database, we discovered that miR-888 was predominately expressed in endometrial tumors, with a significant association to high-grade tumors and increased percent invasion. In a separate panel of endometrial tumor specimens, we validated overexpression of miR-888 by real-time polymerase chain reaction. In addition, miR-888 expression was highest in endometrial carcinosarcoma, a rare and aggressive type of endometrial tumor. Moreover, we identified the progesterone receptor (PR), a potent endometrial tumor suppressor, as a direct target of miR-888. These data define miR-888 as the first miRNA CT antigen and a potential mediator of an aggressive endometrial tumor phenotype through down-regulation of PR.


Engaging an HIV vaccine target through the acquisition of low B cell affinity.
Low affinity is common for germline B cell receptors (BCR) seeding development of broadly neutralizing antibodies (bnAbs) that engage hypervariable viruses, including HIV. Antibody affinity selection is also non-homogenizing, insuring the survival of low affinity B cell clones. To explore whether this provides a natural window for expanding human B cell lineages against conserved vaccine targets, we deploy transgenic mice mimicking human antibody diversity and somatic hypermutation (SHM) and immunize with simple monomeric HIV glycoprotein envelope immunogens. We report an immunization regimen that focuses B cell memory upon the conserved CD4 binding site (CD4bs) through both conventional affinity maturation and reproducible expansion of low affinity BCR clones with public patterns in SHM. In the latter instance, SHM facilitates target acquisition by decreasing binding strength. This suggests that permissive B cell selection enables the discovery of antibody epitopes, in this case an HIV bnAb site.


Optimization and SAR research at the benzoxazole and tetrazole rings of JNJ4796 as new anti-influenza A virus agents, part 2.
We have already reported the modification on the piperazine and phenyl rings of JNJ4796, a small-molecule fuse inhibitor targeting hemagglutinin (HA). In this study, we described the structure-activity relationship of the benzoxazole and tetrazole rings of JNJ4796. Many derivatives demonstrated good in vitro activity against IAV H1N1and Oseltamivir-resistant IAV H1N1 stains. Although compounds (R)-1e and (R)-1h exhibited excellent in vitro activity, high drug exposure level and low hERG inhibition, they displayed low oral efficacy. Excitedly, (R)-1a, a representative identified in our previous study, was found to show potent in vivo anti-IAV activity with the survival rates of 100%, 100% and 70% at 15, 5 and 1.67 mg/kg, respectively, comparable to JNJ4796. Currently, we are exploring different ways to ease its gastrointestinal response.


Mediating role of learned helplessness' components in the association between health literacy/social support and self-management among maintenance haemodialysis patients in Changsha, China: a cross-sectional study.
To explore the multiple mediating roles of the learned helplessness's core system in the relationship between health literacy/social support and self-management. Cross-sectional survey design. Changsha, China. 239 Chinese maintenance haemodialysis (MHD) patients. Two multiple mediator models were constructed based on the COM-B (Capacity, Opportunity, Motivation - Behaviour) model. A total of 239 Chinese MHD patients participated in a cross-sectional study, which included surveys on the Learned Helplessness Scale for MHD patients, Dialysis Knowledge Questionnaire, Social Support Scale and Self-Management Scale for Haemodialysis. The PROCESS macro in SPSS was used for mediated effects analysis. Helplessness and internality partially mediated the relationship between health literacy/social support and self-management ((β=-0.212, p<0.01; β=0.240, p<0.01)/(β=-0.331, p<0.001; β=0.376, p<0.001)). The mediation effect size was 0.780 (95% CI (0.373 to 1.218)) in the health literacy model, accounting for 45.29% of the total effect, and 0.286 (95% CI (0.207 to 0.377)) in the social support model, accounting for 57.88% of the total effect. The differences in effect sizes for helplessness and internality in the two models were -0.080 (95% CI (-0.374 to 0.216)) and -0.041 (95% CI (-0.127 to 0.043)), respectively. Health literacy/social support directly affects MHD patients' self-management and indirectly affects it by changing learned helplessness, such as increasing internality while reducing helplessness.


[Syndrome of senile dementia with dilatation of the cerebral ventricles without other lesions (apropos of 18 observations anatomo-clinical)].
Study of 18 anatomo-clinical observations with dilatation of the cerebral ventricles (without vascular, degenerative or other cerebral lesions), and neuropsychiatric syydrome during the senile period of life. In the morphological and pathogenetic point of view, for one half of these cases a traumatic, microhemorrhagic, or inflammatory aetiology could be taken into consideration; for the other half, the hydrocephalus could be considered as idiopathic.


Large negative lymph nodes - a surrogate for immune activation in rectal cancer patients?
The size of regional, tumor draining lymph nodes without metastasis (LNneg) found in rectal cancer resection specimens varies and seems to be related to patient survival. Yet, the histopathological features influencing LNneg size in rectal cancer have not been studied in detail. Our pilot study focused on investigating the relationship between lymph node (LN) size and LNneg microarchitecture in rectal cancer (RC) resection specimens. In this retrospective cohort study, resection specimens from 146 RC patients, treated with either surgery alone (n = 29) or neoadjuvant therapy followed by resection (n = 117), were included in the study. Histology of LNnegs was reviewed to establish number of lymphoid follicles and presence of intranodal fat. Longest long axis and area of each LN were measured digitally. 1830 LNnegs were measured. The microarchitecture was analyzed in a subset of 680 LNnegs. 153 (22.5 %) LNnegs contained intranodal fat. After neoadjuvant treatment, presence of intranodal fat was related to smaller LNneg area (median (range) area of LNneg without intranodal fat: 4.51 mm2 (0.15-46.89 mm2), with intranodal fat: 3.46 mm2 (0.12-27.22 mm2), p = 0.048). A higher number of lymphoid follicles was related to a larger LNneg area in both patient groups (p < 0.001). Our pilot data suggest that in rectal cancer the presence of large regional LNnegs may reflect increased immune activation due to tumor related antigens. Further studies are warranted to investigate whether histologically visible microarchitectural features of LNnegs such as lymphoid follicles translate to particular features in radiological images and hence could potentially help to identify LNneg with more certainty at the time of pre-treatment disease staging.


Swallowing dysfunction as a factor that should be remembered in recurrent pneumonia: videofluoroscopic swallow study.
The swallow function is one of the strong defense mechanism against aspiration. Aspiration and pneumonia are unavoidable in patients with defective mechanism of swallowing. The aim of this study was to evaluate patients with recurrent pneumonia in terms of videofluoroscopic examination results. The study comprised fifty pediatric cases (22 boys, 28 girls) with an average age of 2.9 years (2 months-7.5 years) who were referred to our clinic due to suffering from recurrent pneumonia. The videofluoroscopic swallow study (VFSS) was performed on all patients. The presence of a correlation with pneumonia was investigated. In 45 of the children, VFSS results were not normal. Of the children, 41 had mental-motor retardation. The results of the videofluoroscopic study show that silent aspiration was the most common pathology in participants with the percentage of 40 (27.5% mild, 17.5% severe). Patients in the study had pneumonia with an incidence of 2.6 illnesses per year. Having one than more results on VFSS was found to be associated with more number of annual pneumonia episodes. Children with neurological impairments are at risk of recurrent acute pneumonia due to aspiration. Disturbances of swallowing phases should be remembered as a cause of pneumonia in these patients.


Using a cDNA microarray to study cellular gene expression altered by Mycobacterium tuberculosis.
To examine the global effects of Mycobacterium tuberculosis (M. tuberculosis) infection on macrophages. The gene expression profiling of macrophage U937, in response to infection with M. tuberculosis H(37)R(a), was monitored using a high-density cDNA microarray. M. tuberculosis infection caused 463 differentially expressed genes, of which 366 genes are known genes registered in the Gene Bank. These genes function in various cellular processes including intracellular signalling, cytoskeletal rearrangement, apoptosis, transcriptional regulation, cell surface receptors, cell-mediated immunity as well as a variety of cellular metabolic pathways, and may play key roles in M. tuberculosis infection and intracellular survival. M. tuberculosis infection alters the expression of host-cell genes, and these genes will provide a foundation for understanding the infection process of M. tuberculosis. The cDNA microarray is a powerful tool for studying pathogen-host cell interaction.


Zebrafish yolk-specific not really started (nrs) gene is a vertebrate homolog of the Drosophila spinster gene and is essential for embryogenesis.
By using retroviral insertional mutagenesis in zebrafish, we have identified a recessive lethal mutation in the not really started (nrs) gene. The nrs mutation disrupts a gene located in linkage group 3 that is highly homologous to the spinster gene identified in Drosophila and to spinster orthologs identified in mammals. In flies, spinster encodes a membrane protein involved in lysosomal metabolism and programmed cell death in the central nervous system and in the ovary. In nrs mutant fish embryos, we detect an opaque substance in the posterior yolk cell extension at approximately 1 day after fertilization. This material progressively accumulates and by 48 hr after fertilization fills the entire yolk. By day 3 of embryogenesis, mutant embryos are severely reduced in size compared with their wild-type siblings and they die a few hours later. By in situ hybridization, we show that the nrs mRNA is expressed in the yolk cell at the time the mutant phenotype becomes apparent. In wild-type embryos, nrs message is present maternally and zygotically throughout embryogenesis and is also detected in adult animals. In nrs homozygous mutant embryos, nrs transcripts are undetectable at the time the phenotype becomes apparent, indicating that the retroviral insertion has most likely abolished expression of the nrs gene. Finally, the nrs phenotype can be partially rescued by microinjection of nrs encoding DNA. These results suggest that the nrs mutation affects an essential gene encoding a putative membrane-bound protein expressed specifically in the yolk cell during zebrafish embryogenesis.


Reinnervation of the ventral root L7 from ventral horn neurons following intramedullary axotomy in adult cats.
Through small longitudinal incisions in the ventral funiculus of the adult cat spinal cord the intramedullary portions of motoraxons forming the L7 ventral root were divided. The animals were sacrificed by glutaraldehyde-perfusion 1-15 weeks postoperatively. In some cases this was preceded by HRP injection into large L7 ventral horn neurons. Sections from the lesioned ventral funiculus and the denervated ventral root L7 were examined by light and electron microscopy. During the first postoperative month increasing numbers of unmyelinated and thinly myelinated large-diameter axons coursed through the lesion and entered the denervated ventral root. Occasional neuroma-like formations were also found in the lesion area. After 5 weeks survival clusters of regenerating axons reached the distal end of the L7 ventral root. Light microscopic examination of Vibratome sections from HRP-injected animals showed that at least some regenerated axons come from large neurons, presumably alpha-motoneurons, in the ventral horn motor nuclei. Most of the examined labeled regenerated axons reached the CNS/PNS junction of the ventral root but a few followed a grossly aberrant course and/or terminated within the lesion. These observations show that regeneration of large-diameter axons through a CNS-lesion is possible. Both the intrinsic regenerative capacity of the axotomized neurons and the presence of a denervated ventral root in the immediate vicinity are likely to be contributing factors with respect to this example of successful CNS axon regeneration.


Amiodarone therapy before heart transplantation as a predictor of thyroid dysfunction after transplantation.
In recent years we have observed new cases of thyroid disease occurring after heart transplantation (HTx). In these patients, the presence of this disease complicates their post-transplant course and occasionally results in life-threatening thyrotoxicosis. The present study examines the incidence and risk factors of thyroid disease in these patients with special emphasis on the use of amiodarone before HTx. Recommendations for follow-up of thyroid disease in HTx patients are discussed. A study was performed on a cohort of 380 patients who received HTx in Rotterdam between January 1, 1985 and April 1, 2001. Twenty patients (5%) developed thyroid dysfunction after HTx, 10 of whom had been treated with amiodarone before HTx. Cumulative survival curves showed that most patients (60%) developed thyroid dysfunction within the first year after HTx (median interval between HTx and diagnosis was 9 months). Using univariate Cox regression analysis, amiodarone was a significant predictor for the development of thyroid dysfunction (hazards ratio 5.2, 95% confidence interval 2.5 to 10.8). After adjustment for age, gender, heart disease and Quetelet index, amiodarone remained a significant predictor. Our study clearly shows that, despite discontinuation of amiodarone treatment at the time of the HTx procedure, patients remain at risk for developing thyroid disease. HTx recipients treated with amiodarone before HTx should therefore be monitored carefully for development of thyroid dysfunction, especially within the first post-transplant year.


Perioperative Mortality Rates in Australian Public Hospitals: The Influence of Age, Gender and Urgency.
A decline in surgical deaths has been observed in Australia since the introduction of the Australian and New Zealand Audit of Surgical Mortality (ANZASM). The current study was conducted to determine whether the perioperative mortality rate (POMR) has also declined. This study is a retrospective review of the POMR for surgical procedures in Australian public hospitals between July 2009 and June 2013, using data obtained from the Australian Institute of Health and Welfare. Operative procedures contained in the Australian Refined Diagnosis Related Groups were selected and the POMR was modelled using urgency of admission, age and gender as explanatory covariates. The POMR in Australian public hospitals reduced by 15.4 % over the 4-year period. The emergency admissions POMR dropped from 1.40 to 1.12 %, and the elective admissions POMR from 0.09 to 0.08 %. The binary logistic regression model used to predict patient mortality showed emergency admissions to have a higher POMR than elective, being more evident at older ages. For emergency admissions, the difference in POMR between females and males increased with age, from about 55 years onwards, with females being lower. For elective surgeries, the difference between males and females was of little practical importance across ages. The reduction in the POMR in Australia confirms the reduction in surgical deaths reported to ANZASM. Continuing to monitor POMR will be important to ensure the safest surgery in Australia. Further investigations into case-mix will allow better risk adjustment and comparison between regions and time-periods, to facilitate continuous quality improvement.


Cardiac electrophysiologic effects of R 54718.
We studied the cellular electrophysiologic effects of a lidocaine derivative, R-54718, that has been found effective in treating arrhythmias in experimental animals and in human subjects. R-54718 depresses Vmax, action potential amplitude, and conduction and accelerates repolarization of the canine Purkinje fiber action potential, with an effective concentration range 2 or 3 orders of magnitude lower than that of most class I antiarrhythmic drugs. Its action on Vmax shows no use dependence. In blood superfusion experiments its depressant effects on conduction are more profound than those of other class I compounds and significant toxicity can be seen. R-54718 also depresses delayed afterdepolarizations, as well as the abnormal automaticity occurring at low membrane potentials. However, automaticity occurring in fully polarized Purkinje fibers is not affected. In conclusion, R 54718 is unusual in its lack of use dependent action, in its differential action on normal and abnormal automaticity, and in its potency.


Monolayer culture of cells originating from a preimplantation bovine embryo.
The objective of this study was to establish a method by which trophectodermal cells originating from individual preimplantation bovine embryos could be perpetuated in monolayer culture. A single, Day-11 bovine embryo collected nonsurgically from a mixed-breed beef cow was cultured in Ham's F10 medium supplemented with fetal bovine serum, sodium pyruvate, insulin, and epidermal growth factor. After 13 d in culture the embryo had adhered to the surface of the plastic culture vessel and a monolayer covering 0.3 cm2 had developed in the manner of a tissue explant. The monolayer was successfully dispersed using trypsin-EDTA and the cells were passaged. Expansion to a 25-cm2 flask was achieved by the 4th passage. By passaging cultures at a dilution ratio of 1:2, cells were maintained for 38 passages before growth slowed. Transfers beyond the 44th passage were unsuccessful. The cell line, designated BE-13, was successfully frozen and thawed at the 9th, 12th, 15th, and 20th passages. The cell line contains both mono- and binucleate cells with a prominent rough endoplasmic reticulum characteristic of ruminant trophoblast cells. Susceptibility to eight bovine viruses was demonstrated. Such cell lines may provide inexpensive systems for the study of trophoblast metabolism and for investigation of the role of the trophoblast in the pathogenesis of selected bovine abortifacient diseases. Because of their range of viral susceptibility, these cells might also be useful for diagnostic purposes.


Myocardial injury or infarction associated with fat embolism in sickle cell disease: a report of three cases with survival.
Fat embolism represents a dread complication of sickle cell hemoglobinopathies. We present the details of three cases that, in addition to an acute chest syndrome, had serological and clinical parameters consistent with myocardial damage. A favorable outcome was obtained with judicious use of blood transfusion.


Murine gamma-herpesvirus 68 latency protein M2 binds to Vav signaling proteins and inhibits B-cell receptor-induced cell cycle arrest and apoptosis in WEHI-231 B cells.
The MHV-68 latent protein, M2, does not have homology to any known viral or cellular proteins, and its function is unclear. To define the role played by M2 during MHV-68 latency as well as the molecular mechanism involved, we used M2 as bait to screen a yeast two-hybrid mouse B-cell cDNA library. Vav1 was identified as an M2-interacting protein in two independent screenings. Subsequent yeast two-hybrid interaction studies showed that M2 also binds to Vav2, but not Vav3, and that three "PXXP" motifs located at the C terminus of M2 are important for this interaction. The interactions between M2 and Vav proteins were also confirmed in vivo in 293T and WEHI-231 B-cells by co-immunoprecipitation assays. Rac1/GST-PAK "pull-down" experiments and Western blot analysis using a phospho-Vav antibody demonstrated that expression of M2 in WEHI-231 cells enhances Vav activity. We further showed in WEHI-231 cells that M2 expression promotes proliferation and survival and is associated with enhanced cyclin D2 and repressed p27(Kip1), p130, and Bim expression. Taken together, these experiments suggest that M2 might have an important role in disseminating the latent virus during the establishment and maintenance of latency by modulating B-cell receptor-mediated signaling events through Vav to promote B-cell activation, proliferation, and survival.


Gastrointestinal consequences of lipopolysaccharide-induced lung inflammation.
Respiratory inflammation is the body's response to lung infection, trauma or hypersensitivity and is often accompanied by comorbidities, including gastrointestinal (GI) symptoms. Why respiratory inflammation is accompanied by GI dysfunction remains unclear. Here, we investigate the effect of lipopolysaccharide (LPS)-induced lung inflammation on intestinal barrier integrity, tight-junctions, enteric neurons and inflammatory marker expression. Female C57bl/6 mice (6-8 weeks) were intratracheally administered LPS (5 µg) or sterile saline, and assessed after either 24 or 72 h. Total and differential cell counts in bronchoalveolar lavage fluid (BALF) were used to evaluate lung inflammation. Intestinal barrier integrity was assessed via cross sectional immunohistochemistry of tight junction markers claudin-1, claudin-4 and EpCAM. Changes in the enteric nervous system (ENS) and inflammation in the intestine were quantified immunohistochemically using neuronal markers Hu + and nNOS, glial markers GFAP and S100β and pan leukocyte marker CD45. Intratracheal LPS significantly increased the number of neutrophils in BALF at 24 and 72 h. These changes were associated with an increase in CD45 + cells in the ileal mucosa at 24 and 72 h, increased goblet cell expression at 24 h, and increased expression of EpCAM at 72 h. LPS had no effect on the expression of GFAP, S100β, nor the number of Hu + neurons or proportion of nNOS neurons in the myenteric plexus. Intratracheal LPS administration induces inflammation in the ileum that is associated with enhanced expression of EpCAM, decreased claudin-4 expression and increased goblet cell density, these changes may contribute to systemic inflammation that is known to accompany many inflammatory diseases of the lung.


Sarcoidosis within a renal allograft: a case report and review of the literature.
Sarcoidosis is an unusual disorder of unknown etiology. Clinically apparent renal involvement is rare in sarcoidosis. The incidence of recurrence in transplant recipients is unknown with few cases having been reported previously. Herein we report a case of sarcoidosis involving a renal allograft that occurred 3 years after transplantation and provide a literature review.


The role of antiarrhythmic drug therapy for the prevention of sudden cardiac death.
Sudden cardiac death (SCD) accounts for more than 300,000 deaths annually in the United States alone. The utility of antiarrhythmic drugs in survivors of SCD (secondary prevention) is limited because of their incomplete efficacy and long-term toxicity. Efforts to target primary prevention of SCD have focused on left ventricular dysfunction in conjunction with congestive heart failure. Antiarrhythmic drugs are not able to decrease mortality in this patient population either; in fact, certain drugs may actually increase overall mortality. In both primary and secondary prevention patients, only implantable cardioverter-defibrillator implantation is associated with improved survival. Antiarrhythmic drugs like azimilide, dofetilide, sotalol, and amiodarone can be used as adjunct treatment for management of atrial arrhythmias and to decrease implantable cardioverter-defibrillator shocks. There is an unmet need for more effective and less toxic antiarrhythmic medications.


Seasonal acclimation in the epiphytic lichen Parmelia sulcata is influenced by change in photobiont population density.
CO2 gas exchange, radial growth, chlorophyll (Chl) content and photobiont density of an epiphytic population of Parmelia sulcata were monitored every 2 months during 1 year in a temperate deciduous forest of Central Italy, to verify possible seasonal variations. Light response curves of south-exposed thalli, built up in the laboratory at 6 and 27°C at optimal thallus hydration, showed that CO2 gas exchange changed significantly during the year, with a maximum for gross photosynthesis in December at both temperatures. Photoinhibition phenomena occurred in early spring, immediately before tree leaves sprouted. The principal component analysis of CO2 gas exchange parameters clearly separated the months with from the months without tree canopy cover. Radial growth, measured on marginal lobes of north- and south-exposed thalli, was the highest in December, and the lowest in April. Photobiont density, measured in lobes of south- and north-exposed thalli with a sedimentation chamber, also changed during the year: the number of photobionts was highest in June and December, and lowest in April, although no significant change in cell size and Chl content per cell was evident throughout the year. South-exposed thalli had slightly, but constantly higher photobiont density both on a weight and an area basis. The acclimation of lichen photosynthesis and Chl content to seasonal temperature and light changes should partially be re-visited on the basis of the significant variation in photobiont population density. This phenomenon still awaits, however, a satisfactory explanation, although it is probably related to the seasonal change in nutrient availability.


Drugs--a growing problem in Sri Lanka.
Cannabis, hashish and opium were the drugs generally abused in Sri Lanka. The laws relating to drugs were consolidated in 1935. Heroin is the most widely abused drug today. Tourists are mainly responsible for the introduction of heroin into Sri Lanka. The separatist terrorists of Sri Lanka are engaged in trafficking in drugs to find the money for the purchase of arms and ammunition.


A perspective on LILRBs and LAIR1 as immune checkpoint targets for cancer treatment.
Immunosuppressive myeloid cells in the tumor microenvironment inhibit anti-tumor immunity and support tumor development. The leukocyte Ig-like receptor subfamily B (LILRB) proteins and the related receptor LAIR1 are immune checkpoint receptors that support the immunosuppressive activity of myeloid cells. All LILRBs and LAIR1 have intracellular immunoreceptor tyrosine-based inhibitory motifs in their signaling domains, but the individual proteins have different functions. The determinants of the distinct functions of these inhibitory receptors likely rest in their interactions with different ligands and other surface proteins, characteristic signaling domains, and expression dynamics in different cell types regulated by various extrinsic cues and transcription factors. Significant advancement of immuno-oncology therapeutic products based on targeting or reprogramming of LILRB- and LAIR1-mediated signaling is anticipated.


Intravenous immunoglobulin ameliorates experimental autoimmune encephalomyelitis and reduces neuropathological abnormalities when administered prophylactically.
Immunomodulation with intravenous immunoglobulin (IVIG) represents a way of interfering with the disease process in multiple sclerosis (MS). In this study, the effects of IVIG on neurological symptoms and central nervous system (CNS) pathology were evaluated in experimental autoimmune encephalomyelitis (EAE), an MS animal model. EAE was induced in susceptible Dark Agouti rats by active immunization with a spinal cord homogenate, and infusions of 1 g/kg IVIG were given prophylactically or therapeutically. The administration of IVIG at the time of immunization significantly suppressed the development of neurological symptoms compared with infusions of placebo (mean EAE score 0.6+/-0.3 versus 2.3+/-0.4). Moreover, the prophylactic IVIG administration resulted in a significant inhibition of the inflammatory response in CNS tissue (inflammation score 1.1+/-0.2 versus 1.8+/-0.2 after placebo). No beneficial effects were obtained by therapeutic IVIG infusions as the EAE disease course and the degree of inflammation and demyelination in the CNS were not different from animals receiving treatment with placebo. The present study indicates that IVIG reduces the symptoms of EAE by suppression of the CNS inflammation that characterizes CNS pathology in these animals. Taking into account data from clinical trials of IVIG in MS, the results further suggest that IVIG acts primarily during the induction phase of the immune response thus preventing the development of relapses in MS.


Thrombosis, prevalence and new evidence on current perceptions of risk.
Venous thromboembolism is a significant but unquantified clinical problem in patients with cancer. The FRONTLINE survey was designed to evaluate clinicians' perceptions of thrombosis risk for cancer patients and to examine current clinical practice with regard to thromboprophylaxis and treatment of venous thromboembolism. Over 3800 responses from surgeons, haematologists and other clinicians were received. Their perception of thrombosis risk for patients varied according to the site of primary tumour. Most surgeons thought that patients with brain tumours were at high risk of thrombosis. Most clinicians treating medical (non-surgical) cancer patients perceived pancreatic tumours to carry a high thrombosis risk in the absence of thromboprophylaxis.


The use of graph matching algorithms to identify biochemical substructures in synthetic chemical compounds: Application to metabolomics.
Metabolomics is a rapidly growing field studying the small-molecule metabolite profile of a biological organism. Studying metabolism has a potential to contribute to biomedical research as well as drug discovery. One of the current challenges in metabolomics is the identification of unknown metabolites as existing chemical databases are incomplete. We present a novel way of utilizing known mammalian metabolites in an effort to identify unknown ones. The system relies on a mammalian scaffolds database to aid the classification process. The results show that 96% of the mammalian compounds were identified as truly mammalian in a leave-one-out experiment. The system was also tested with a random set of synthetic compounds, downloaded from ChemBridge and ChemSynthesis databases. The system was able to eliminate 54% of the set, leaving 46% of the compounds as potentially unknown mammalian metabolites.


[Impact of advanced maternal age on fecundity and women's and children's health].
There has been a consistent trend towards delayed childbearing in most Western countries. We present a review of the current epidemiological understanding of the effects of advanced maternal age on fecundity, maternal and child health. A narrative review of the literature based mostly on the key population-based studies on the subject. Delayed childbearing results in a substantial decrease in fecundity and a considerable increase in fetal loss; risk of the latter is doubled between 20-24 and 40-44 years of age. Other risks include those related to maternal health (maternal mortality and morbidity) and adverse pregnancy outcomes (multiple births, preterm delivery, fetal growth retardation and congenital anomalies). Risks associated with delayed childbearing become significant for maternal age greater than 35 years, and are particularly high for women aged 40 years and greater. Knowledge about the risks associated with advancing age can be helpful for couples in their decisions regarding childbearing.


Interaction of Surgeon Volume and Nurse-to-Patient Ratio on Post-operative Outcomes of Medicare Beneficiaries Following Pancreaticoduodenectomy.
We sought to assess the effect of nurse-to-patient ratio on outcomes with a focus on defining whether nurse-to-patient ratio altered outcomes relative to pancreaticoduodenectomy (PD) surgeon specific volume. Medicare SAFs from 2013-2015 were used to identify patients who underwent PD. Nurse-to-patient ratio, PD specific surgeon volume were stratified. Association of factors associated with short term outcomes was evaluated. Overall, 6668 patients (median age 73, IQR 68-77; 52.8% male) were identified. The median annual PD volume of surgeons in the highest volume tier was 24 (IQR 21-29), whereas surgeons in the lowest tier performed 2 PDs annually (IQR 1-3) (p < 0.001). Compared with hospitals that had the highest nurse-to-patient ratio tier, patients at hospitals with the lowest nurse-to-patient ratio tier were 26% more likely to have a complication (OR 1.26, 95% CI 1.02-1.55). Additionally, patients of surgeons in the lowest tier had 43% greater odds of suffering a complication compared to patients of surgeons in the highest tier (OR 1.43, 95% CI 1.11-1.84). However, patients who underwent a PD by a surgeon within the lowest tier had similar odds of a complication irrespective of nurse-to-patient ratio (OR 1.34, 95% CI 0.97-1.86). Compared with patients who underwent an operation by a surgeon in highest PD volume tier, patients treated by surgeons in the lowest tier had higher odds of post-operative complications which was not mitigated by a higher nurse-to-patient ratio.


Use of positron emission tomography (PET) in stereotactic conditions for brain biopsy.
In order to take advantage of the metabolic information provided by positron emission tomography (PET) in cases of brain tumour, we have developed a technique to integrate PET images routinely in the planning of stereotactic brain biopsy. We used stereotactic PET with [18F]-labelled fluorodeoxyglucose (PET-FDG) in 38 patients undergoing brain biopsy. To evaluate the contribution of PET-FDG in guiding brain biopsy, we analyzed the diagnosis provided by the 78 stereotactic trajectories obtained in these patients. We found that stereotactic PET-FDG seemed to provide more information in cases of anaplastic astrocytomas and glioblastomas than in low-grade gliomas. Our results also show that biopsy trajectories performed in areas where increased FDG uptake is found within the lesion boundaries always provide interpretable specimens; this was not the case for trajectories guided by CT only. Therefore, the routine integration PET-FDG in the planning of stereotactic brain biopsy may lead to a reduction in sampling. Recently, we also tested consecutive stereotactic PET with [11C]-labelled methionine (PET-Met) and PET-FDG. This technique allowed us to compare accurately the tumoural glucose metabolism and protein synthesis. Our results suggest that stereotactic PET may increase the diagnostic yield of brain biopsy and may improve the understanding of PET in neuro-oncology.


Bordetella bronchiseptica and toxigenic type D Pasteurella multocida as agents of severe atrophic rhinitis of swine.
Bordetella bronchiseptica and toxigenic type-D Pasteurella multocida were cultured from pigs in each of five herds diagnosed as having severe atrophic rhinitis (AR). B. bronchiseptica alone, P. multocida alone, or both organisms isolated from four herds were inoculated intranasally into 1-week-old gnotobiotic pigs which were necropsied 4 weeks post-inoculation (PI). Nasal turbinate atrophy in B. bronchiseptica-inoculated pigs was moderate to severe, while P. multocida-inoculated pigs had slight to severe atrophy. Pigs inoculated with both organisms had moderate to complete turbinate atrophy. P. multocida was reisolated at necropsy from all pigs receiving the organism except those having no turbinate damage. B. bronchiseptica and P. multocida from a fifth herd were simultaneously inoculated into six naturally farrowed 6-day-old SPF pigs. Necropsy performed 4 weeks PI revealed severe to complete turbinate atrophy. Nasal turbinates were normal for control pigs in both experiments.


Postcards from the EDge project: randomised controlled trial of an intervention using postcards to reduce repetition of hospital treated deliberate self poisoning.
To determine whether an intervention using postcards (postcards from the EDge project) reduces repetitions of hospital treated deliberate self poisoning. Randomised controlled trial. Regional referral service for general hospital treated deliberate self poisoning in Newcastle, Australia. 772 patients aged over 16 years with deliberate self poisoning. Non-obligatory intervention using eight postcards over 12 months along with standard treatment compared with standard treatment alone. Proportion of patients with one or more repeat episodes of deliberate self poisoning and the number of repeat episodes for deliberate self poisoning per person in 12 months. The proportion of repeaters with deliberate self poisoning in the intervention group did not differ significantly from that in the control group (57/378, 15.1%, 95% confidence interval 11.5% to 18.7% v 68/394, 17.3%, 13.5% to 21.0%: difference between groups -2%, -7% to 3%). In unadjusted analysis the number of repetitions were significantly reduced (incidence risk ratio 0.55, 0.35 to 0.87). A postcard intervention reduced repetitions of deliberate self poisoning, although it did not significantly reduce the proportion of individual repeaters.


Differential contribution of Ih to the integration of excitatory synaptic inputs in substantia nigra pars compacta and ventral tegmental area dopaminergic neurons.
The selective vulnerability of substantia nigra pars compacta (SNc) dopaminergic (DA) neurons is an enigmatic trait of Parkinson's disease (PD), especially if compared to the remarkable resistance of closely related DA neurons in the neighboring ventral tegmental area (VTA). Overall evidence indicates that specific electrophysiological, metabolic and molecular factors underlie SNc vulnerability, although many pieces of the puzzle are still missing. In this respect, we recently demonstrated that 1-methyl-4-phenylpyridinium (MPP+), the active metabolite of the parkinsonizing toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), alters the electrophysiological properties of SNc DA neurons in vitro by inhibiting the hyperpolarization-activated current (Ih). Here, we present an electrophysiological investigation of the functional role of Ih in the integration of synaptic inputs in identified SNc and VTA DA neurons, comparatively, in acute midbrain slices from TH-GFP mice. We show that pharmacological suppression of Ih increases the amplitude and decay time of excitatory postsynaptic potentials, leading to temporal summation of multiple excitatory potentials at somatic level. Importantly, these effects are quantitatively more evident in SNc DA neurons. We conclude that Ih regulates the responsiveness to excitatory synaptic transmission in SNc and VTA DA neurons differentially. Finally, we present the hypothesis that Ih loss of function may be linked to PD trigger mechanisms, such as mitochondrial failure and ATP depletion, and act in concert with SNc-specific synaptic connectivity to promote selective vulnerability.


Citrate carrier promoter is target of peroxisome proliferator-activated receptor alpha and gamma in hepatocytes and adipocytes.
Citrate carrier (CiC), a mitochondrial inner membrane protein, is an essential component of the shuttle system which transports acetyl-CoA from mitochondria to the cytosol where lipogenesis occurs. CiC is regulated by SREBP-1, a transcription factor that controls the expression of several lipogenic genes. CiC is also implicated in cholesterol synthesis, glycolysis and gluconeogenesis, suggesting that besides SREBP-1 other transcription factors could modulate the expression of its gene. Here, we provide evidences demonstrating that CiC expression is regulated by peroxisome proliferator-activated receptor (PPAR) alpha and gamma in hepatocytes and adipocytes, respectively. CiC expression increased in rat BRL-3A hepatocytes treated with WY-14,643, agonist of PPARα, and in murine 3T3-L1 adipocytes treated with rosiglitazone, agonist of PPARγ. The overexpression of PPARα/RXRα and PPARγ/RXRα heterodimer enhanced CiC promoter activity in BRL-3A and 3T3-L1, respectively. Luciferase reporter gene and gel mobility shift assays indicated that a functional peroxisome proliferator-activated receptor response element (PPRE), identified in the CiC promoter, conferred responsiveness to activation by PPARs. The binding of PPRE of CiC promoter by PPARα and PPARγin vivo was confirmed by ChIP assay in BRL-3A and 3T3-L1 cells, respectively.


Effect of oral Kohki tea on bladder dysfunction induced by severe partial outlet obstruction.
Extracts of the leaves of Engelhardtia chrysolepis, a subtropical plant that grows wild in southern China, have been used medicinally in east Asia for hundreds of years. A standard extract named Kohki tea (Maruzen Pharmaceuticals, Onomichi City, Japan) is sold over the counter in Japan as a sweet tea shown to confer many beneficial effects on general health and well-being. The tea contains strong antioxidants, including several dihydroflavonol glycosides. The results of previous studies show that natural products with antioxidant activities provide protective effects on the bladder of rabbits with partial outlet obstruction. We determined in vivo and in vitro whether oral pretreatment of rabbits with Kohki tea protects the bladder from dysfunction induced by partial outlet obstruction. A total of 28 New Zealand White rabbits were separated into 4 groups of 7 each. Rabbits in groups 1 and 2 were treated by gavage with 100 mg./kg. Kohki tea daily in distilled water, while those in groups 3 and 4 were given distilled water. After 4 weeks of daily oral administration each rabbit was sedated, the bladder was catheterized and cystometry was performed at a filling rate of 1 ml. per minute. At the completion of cystometry the rabbits were immediately anesthetized. Moderate outlet obstruction was created in groups 1 and 3, and sham surgery was performed in groups 2 and 4. Treatment was continued for an additional 4 weeks, when each rabbit was sedated and cystometry was repeated. After cystometry the bladder was exposed through a midline incision, excised, weighed and 4 strips of bladder body were cut for contractility studies. The balance of the bladder was separated between smooth muscle and mucosa by blunt dissection, frozen in liquid nitrogen and stored at -70C for biochemical analyses. Partial outlet obstruction stimulated similar increases in the bladder weight of all obstructed rabbits. Partial outlet obstruction resulted in a significant decrease in bladder compliance in all obstructed animals. However, the bladder of obstructed rabbits given Kohki tea were significantly more compliant than those given water. Voiding pressures in the control group and the obstructed group given distilled water were approximately equal, while obstructed rabbits given Kohki tea showed significantly higher maximal voiding pressure. The contractile responses to all forms of stimulation were reduced by obstruction to a significantly greater degree in the rabbits not given tea than in those given tea. Sarcoplasmic reticulum Ca2+-adenosine triphosphatase enzyme activity of the bladder was significantly reduced in obstructed rabbits given vehicle but activity was not reduced in obstructed rabbits given Kohki tea. Kohki tea had a significant protective effect on bladder function, contractile responses and bladder biochemistry in rabbits with moderate to severe partial outlet obstruction.


Increasing self-awareness among older drivers: the role of self-screening.
Self-screening by older drivers has shown considerable promise for increasing self-awareness about functional abilities associated with safe driving. The purpose of this study was to improve upon existing self-screening instruments by focusing entirely on "health concerns" that affect driving - that is, the symptoms that people experience due to medical conditions and the medications used to treat them - rather than the medical conditions or medications themselves. A computer-based, easy-to-use self-screening instrument for older drivers was developed to provide individualized feedback intended to increase self-awareness about declines in driving-related abilities, as well as suggestions for behavioral changes or safety tips to maintain safe driving, further evaluation from a physician/health professional, and vehicle modifications to help compensate for driving-related declines. This paper describes the development of the self-screening instrument and summarizes findings relative to increasing self-awareness among older drivers. This research represents an important first step in improving self-awareness among older drivers through self-screening.


Metal-free and atom-efficient protocol for diarylation of selenocyanate by diaryliodonium salts.
We developed an atom- and reaction mass efficient strategy for the preparation of diarylselenides using iodonium salts as reactants. The developed approach allows the obtaining of diarylselenides from the corresponding trimethoxyphenyl-substituted iodonium salts via a two-step one-pot reaction sequence. The proposed metal-free methodology is based on the involvement of both iodonium aryl groups for diarylation.


[Hibernoma of the forehead. A rare tumor of brown fatty tissue in an unusual site].
We report a hibernoma located on the forehead in a 52-year-old female patient. The hibernoma is a rare tumour derived from brown adipose tissue. Location on the forehead is unusual and has not been reported before.


AI-Enhanced PET and MR Imaging for Patients with Breast Cancer.
New challenges are currently faced by clinical and surgical oncologists in the management of patients with breast cancer, mainly related to the need for molecular and prognostic data. Recent technological advances in diagnostic imaging and informatics have led to the introduction of functional imaging modalities, such as hybrid PET/MR imaging, and artificial intelligence (AI) software, aimed at the extraction of quantitative radiomics data, which may reflect tumor biology and behavior. In this article, the most recent applications of radiomics and AI to PET/MR imaging are described to address the new needs of clinical and surgical oncology.


37-kDa laminin receptor precursor promotes lung adenocarcinoma cell invasion and metastasis by epithelial-to-mesenchymal transition.
37-kDa laminin receptor precursor (37LRP) has a crucial role in migration of some human cancers. Epithelial-to-mesenchymal transition (EMT) has received much attention in invasion and metastasis of lung cancer. Nevertheless, the role of 37LRP is not entirely clear in EMT promotion of lung cancer at present. In this study, we firstly examined the possible role of 37LRP in the invasiveness and metastasis process of lung cancer using immunohistochemistry of 80 lung adenocarcinoma cases, western blot and real-time PCR of 12 fresh lung adenocarcinoma tissues. The results showed that 37LRP significantly correlated with clinical stage and were highly expressed in metastatic lung adenocarcinomas compared with nonmetastatic ones. In vitro, we observed that 37LRP significantly increased the adhesive, invasive and metastatic abilities of human lung adenocarcinoma cell lines A549 by 37LRP-lentivirus interference. Furthermore, inoculation of A549 cells transduced with 37LRP-lentivirus in nude mice resulted in multi-metastases including the lung. In addition, western blotting and immunofluorescence were used to detect the significant difference in expression of E-cadherin and fibronectin in A549 by 37LRP-lentivirus interference compared with 37LRP-small interference RNA-lentivirus interference in vitro and vivo. The data indicated that A549 cells of epithelial cell characteristics might be induced to undergo EMT by 37LRP. A549 cells transduced with 37LRP-lentivirus showed marked morphological changes, accompanied by the decrease of epithelial marker E-cadherin and the increase of mesenchymal marker fibronectin. These results indicated that 37LRP may promote lung adenocarcinoma invasion and metastasis via the mechanism of EMT.


Adolescent Δ9-Tetrahydrocannabinol Exposure and Astrocyte-Specific Genetic Vulnerability Converge on Nuclear Factor-κB-Cyclooxygenase-2 Signaling to Impair Memory in Adulthood.
Although several studies have linked adolescent cannabis use to long-term cognitive dysfunction, there are negative reports as well. The fact that not all users develop cognitive impairment suggests a genetic vulnerability to adverse effects of cannabis, which are attributed to action of Δ9-tetrahydrocannabinol (Δ9-THC), a cannabis constituent and partial agonist of brain cannabinoid receptor 1. As both neurons and glial cells express cannabinoid receptor 1, genetic vulnerability could influence Δ9-THC-induced signaling in a cell type-specific manner. Here we use an animal model of inducible expression of dominant-negative disrupted in schizophrenia 1 (DN-DISC1) selectively in astrocytes to evaluate the molecular mechanisms, whereby an astrocyte genetic vulnerability could interact with adolescent Δ9-THC exposure to impair recognition memory in adulthood. Selective expression of DN-DISC1 in astrocytes and adolescent treatment with Δ9-THC synergistically affected recognition memory in adult mice. Similar deficits in recognition memory were observed following knockdown of endogenous Disc1 in hippocampal astrocytes in mice treated with Δ9-THC during adolescence. At the molecular level, DN-DISC1 and Δ9-THC synergistically activated the nuclear factor-κB-cyclooxygenase-2 pathway in astrocytes and decreased immunoreactivity of parvalbumin-positive presynaptic inhibitory boutons around pyramidal neurons of the hippocampal CA3 area. The cognitive abnormalities were prevented in DN-DISC1 mice exposed to Δ9-THC by simultaneous adolescent treatment with the cyclooxygenase-2 inhibitor, NS398. Our data demonstrate that individual vulnerability to cannabis can be exclusively mediated by astrocytes. Results of this work suggest that genetic predisposition within astrocytes can exaggerate Δ9-THC-produced cognitive impairments via convergent inflammatory signaling, suggesting possible targets for preventing adverse effects of cannabis within susceptible individuals.


Taxonomic and functional diversity of land snails reflects habitat complexity in riparian forests.
Habitat complexity affects the structure and dynamics of ecological communities, more often with increased complexity leading to greater species diversity and abundance. Among the terrestrial invertebrate groups, the low vagility of land snails makes them susceptible to react to small-scale habitat alteration. In the current paper we aimed to assess the relationship between taxonomic and functional composition and diversity of land snail communities and habitat structure in the riparian forest habitat. We found that both snail abundance and species richness responded positively to the increase in habitat complexity. The complexity of the riparian forest affected also the snail trait composition. Forest species, species living in woody debris, leaf litter, and root zone and those feeding on detritus were more abundant in complex habitats, while large snails with more offspring, snails having the ability to survive longer periods of dryness, as well as species that prefer arid habitats, were more abundant in less complex habitats. We concluded that habitat complexity promoted functional diversity, with the amount of woody debris as main positive driver, and the adjacent agricultural fields as negative driver of functional diversity.


Progress in non-viral localized delivery of siRNA therapeutics for pulmonary diseases.
Emerging therapies based on localized delivery of siRNA to lungs have opened up exciting possibilities for treatment of different lung diseases. Localized delivery of siRNA to lungs has shown to result in severalfold higher lung accumulation than systemic route, while minimizing non-specific distribution in other organs. However, to date, only 2 clinical trials have explored localized delivery of siRNA for pulmonary diseases. Here we systematically reviewed recent advances in the field of pulmonary delivery of siRNA using non-viral approaches. We firstly introduce the routes of local administration and analyze the anatomical and physiological barriers towards effective local delivery of siRNA in lungs. We then discuss current progress in pulmonary delivery of siRNA for respiratory tract infections, chronic obstructive pulmonary diseases, acute lung injury, and lung cancer, list outstanding questions, and highlight directions for future research. We expect this review to provide a comprehensive understanding of current advances in pulmonary delivery of siRNA.


Immunocytochemical localization of endopeptidase-24.11 in cultured neurons from pig striatum.
Endopeptidase-24.11 ("enkephalinase") appears to play a key role in the metabolism of a number of neuropeptides at cell surfaces. It has been previously mapped in the central nervous system, but some doubt has been expressed concerning the identity of the cell type expressing this peptidase. Primary cell cultures derived from striata of new-born piglets were set up and cells were characterized by immunocytochemistry using antibodies to neurofilament protein, a glial fibrillary acidic protein and a neuronal antigen recognized by a monoclonal antibody BM88 and by histochemistry for acetylcholinesterase. Some cultures were set up in which neurons were selectively enriched. Cells which were thus morphologically defined as neurons were recognized by an affinity-purified polyclonal antibody to endopeptidase-24.11. The staining for the peptidase, which was punctate in appearance, was shown to be at the cell surface and extended to the perikaryon and all neurites. Compared with the number of neurofilament protein-positive cells, relatively few cells were positive for endopeptidase-24.11. No glial cells, immunochemically defined by glial fibrillary acidic protein, were stained by the antibody to endopeptidase-24.11. We conclude that endopeptidase-24.11 is expressed on the surface of a set of neurons derived from the striatum in primary culture and not by any glial cells in these cultures.


Meiotic resumption and intracellular cAMP levels in mouse oocytes treated with compounds which act on cAMP metabolism.
We have studied the effect of agents known to stimulate adenylate cyclase on spontaneous meiotic resumption in vitro by mouse oocytes. We have found that cholera toxin (CT) (up to 1 microM) and prostaglandin E1 (PGE1) (up to 160 microM) are not able to prevent meiotic resumption, but a clear dose-dependent delay in meiosis resumption was observed during the first 3 h of incubation in medium containing CT or PGE1. The effect became clearer when a small concentration of isobutylmethylxanthine (MIX) (1 microM) was added to the medium. We have also measured the cAMP content of the oocyte: the basal content is 2.1 fmol; this value drops to 0.9 fmol during a 2-h culture period. The decrease is partially prevented if CT is present in the incubation medium, while total cAMP content increases to 3.1 fmol in the presence of 1 mM MIX. The results suggest that isolated mouse oocytes contain toxin and prostaglandin sensitive adenylate cyclase and also an active phosphodiesterase system.


Development and validation of a mtDNA multiplex PCR for identification and discrimination of Calicophoron daubneyi and Fasciola hepatica in the Galba truncatula snail.
Paramphistomosis and Fasciolosis caused by Calicophoron daubneyi and Fasciola hepatica, respectively, are frequent and important trematodoses in ruminant livestock worldwide. Both parasites use the same snail, Galba truncatula, as intermediate host. The aim of this study was to develop and validate an analytical method based on a mitochondrial DNA (mtDNA) multiplex PCR technique which would allow the early and specific identification, in one step, of C. daubneyi and F. hepatica infection in G. truncatula. First of all, a 1035 bp fragment of mtDNA from adult C. daubneyi worms was obtained. Then two pairs of specific mtDNA primers, which amplified a DNA fragment of 885 pb in the case of C. daubneyi, and of 425 pb in that of F. hepatica, were designed. By means of the multiplex PCR technique developed, there was always a specific amplification in samples from adult F. hepatica and C. daubneyi, but not from Calicophoron calicophorum, Cotylophoron cotylophorum, Cotylophoron batycotyle or Dicrocoelium dendriticum. Likewise, specific amplifications of the expected DNA fragments happened in all samples from snails harbouring larval stages of C. daubneyi or F. hepatica, previously detected by microscopy. However, amplifications were not seen when DNA from snails harbouring other Digenea (Plagiorchiidae, Notocotylidae and furcocercous cercariae) was analysed. Moreover, DNA from G. truncatula molluscs free from infection was not amplified. The multiplex PCR assay permitted infection in the snails experimentally infected with 4 miracidia to be detected as early as day 1 p.i. in the case of F. hepatica and with only 2 miracidia from day 2 p.i. in both, C. daubneyi and F. hepatica. Nevertheless it was necessary to wait until days 29 and 33 p.i. to see C. daubneyi and F. hepatica immature redia, respectively, using microscope techniques. The detection limit of the PCR technique was very low: 0.1 ng of DNA from C. daubneyi and 0.001 ng of DNA from F. hepatica. This allowed infection by either F. hepatica or C. daubneyi to be detected even when pools made up with only 1 μl (60 ng of DNA) from infected snail plus 99 μl from non-infected ones were analyzed. Moreover, simultaneous detection of both parasites was experimentally possible in pools made up with uninfected (98 μl), C. daubneyi infected (1 μl) and F. hepatica infected (1 μl) snails. The most precise and early diagnosis of the infections using the multiplex PCR technique designed will allow more realistic epidemiological models of both infections to be established and consequently a better strategic control.


Decreased number of asialoglycoprotein receptors in diabetic BB Wistar rat.
The binding of asialoglycoproteins by hepatic binding protein was studied in freshly isolated hepatocytes from genetically diabetic BB Wistar rats. The number of cell surface asialoglycoprotein receptors was dramatically decreased (58,000 +/- 38,000 for diabetic rats compared to 267,000 +/- 70,000 for normal rats), while the association equilibrium constant was not changed. These results parallel those obtained with streptozotocin-diabetic rats and support the hypothesis that insulin deprivation is responsible for the decrease in the receptor number.


Fine structure of muscle and other components of the trunk of Sagitta setosa (Chaetognath).
The histology of the Chaetognath's trunk is largely based upon works of Hertwig (1880) and Burfield (1927) and is revised here essentially on ultrastructural basis, in a study on Sagitta setosa. The trunk is composed of a thick multilayered epidermis, without cuticle, and contains an abundant nervous system. It is separated from the underlying general musculature by a "basement membrane" which is not crossed by nerves fibres in the region of the ventral ganglion. We have been mainly interested in the muscular tissue. It has no close contact with the body cavity but is surrounded by a thin monolayer epithelium. The general musculature is composed of two types of muscles. The first one, forming the primary musculature, is divided by mesenteries and lateral fields into four quadrants. It contains groups of fibres (C then A) alternating with other groups of fibres (B). The second one, constituting the secondary musculature, is divided into four bands. Both types of fibres are readily identifiable by their myofibrils. The fibres of primary musculature, in close contact with their membrane level, have no basement lamina except at the myoepidermic junctions.


Layered material characterization using ultrasonic transmission. An inverse estimation methodology.
This paper presents an inverse methodology with the aim to characterize a layered material through the identification of acoustical and mechanical properties of its layers. The framework to accomplish this objective is provided by the Inverse Problems (IPs) theory. Material characterization refers to the detection and localization of discontinuities, as well as to the identification of physical properties, in order to predict the material behaviour. In this particular case, the IP is solved in the form of a parameter estimation problem, in which the goal is the estimation of the characteristic acoustic impedance, transit time, and attenuation of each layer. These parameters are directly related to relevant material properties, such as the speed of sound, density, elastic modulus and elastic energy dissipation constants. The IP solution is obtained by minimizing a cost functional formulated as the least squares error between the waveform calculated using an equivalent model, and the measured waveform obtained from ultrasonic transmission tests. The applied methodology allowed the accurate estimation of the desired parameters in materials composed of up to three layers. As a second contribution, a power law frequency dependence of the wave attenuation was identified for several homogeneous materials, based on the same ultrasonic transmission experiments.


Global DNA hypomethylation coupled to cellular transformation and metastatic ability.
Global DNA hypomethylation and DNA hypermethylation of promoter regions are frequently detected in human cancers. Although many studies have suggested a contribution to carcinogenesis, it is still unclear whether the aberrant DNA hypomethylation observed in tumors is a consequence or a cause of cancer. Here, we show that the enforced expression of Stella (also known as PGC7 and Dppa3) induced not only global DNA demethylation but also transformation of NIH3T3 cells. Furthermore, overexpression of Stella enhanced the metastatic ability of B16 melanoma cells, presumably through the induction of metastasis-related genes. These results provide new insights into the function of global DNA hypomethylation in carcinogenesis.