Domestic dogs as reservoir hosts for Leishmania donovani in the southernmost Western Ghats in India.
The peripheral blood samples from domestic dogs (n=47) and wild rats (n=25) in the Kani Tribe settlements, located southernmost part of the Western Ghats, Thiruvananthapuram district, Kerala, India were examined for Leishmania infection. This area is known for cases of leishmaniasis with cutaneous manifestations and sandfly abundance. The tribes domesticate dogs to protect them from untoward activities of wild animals. Leishmania donovani parasite DNA was detected only from 6.4% (n=3) of the blood samples collected from the domestic dogs by amplification of the diagnostic kinetoplast mini-circle DNA and PCR-RFLP analysis of the UTR region of heat shock protein 70 (hsp70) gene. None of the blood samples collected from rats was positive. Through sequencing, L. donovani infection among dogs was confirmed. The DNA sequences generated for hsp70 were deposited with the GenBank. The GenBank accession numbers of these samples are KR905363, KR905364 and KR905365 for hsp70 genes. The results indicated that the DNA isolates from dog blood samples matched precisely with that of our earlier isolates from skin lesions of Kani tribes and also from P. argentipes vector. Thus, the role of dogs as reservoirs for L. donovani parasite in the Kani tribe settlements is confirmed.


Validation of the Bronchiectasis Impact Measure (BIM): a novel patient-reported outcome measure.
Existing quality-of-life and symptom tools used in bronchiectasis trials are either not disease specific or are complex and have not been consistently responsive. We developed a simple patient-reported visual analogue outcome measure, the Bronchiectasis Impact Measure (BIM), for use in clinical research, including clinical trials. Patients with bronchiectasis attending a tertiary referral clinic in the east of Scotland were invited to complete the BIM questionnaire and the quality-of-life bronchiectasis questionnaire at baseline with repeat questionnaires after 2 weeks and 6 months. We assessed internal consistency, test-retest reliability, construct validity and responsiveness by evaluating change during an acute exacerbation. 173 patients were included. The eight domains (cough, sputum, breathlessness, tiredness, activity, general health, control, exacerbations) showed excellent internal consistency (Cronbach's α 0.93). The intraclass correlation coefficient demonstrated excellent reliability over a 2-week period: cough (0.79, 95% CI 0.70-0.85), sputum (0.86, 95% CI 0.80-0.90), dyspnoea (0.82, 95% CI 0.74-0.87), tiredness (0.88, 95% CI 0.82-0.91), activity (0.84, 95% CI 0.77-0.89), general health (0.81, 95% CI 0.74-0.87), control (0.83, 95% CI 0.75-0.88) and exacerbation (0.71, 95% CI 0.60-0.79). Domains correlated strongly with bronchiectasis severity and exacerbation history. Both distribution and patient-based methods estimated the minimal clinically important difference for each domain as 1.5 points on a 10-point scale. Statistically significant changes in all BIM domains were observed during an acute exacerbation. The BIM is a simple patient-reported outcome. This study validates the internal consistency, reliability, construct validity and response of the tool at acute exacerbation. Further validation of the tool is now required.


Silencing of Curlin Protein via M13 Phagemid-Mediated Synthetic sRNA Expression Reduces Virulence in the Avian Pathogenic E. coli (APEC).
Curli fimbriae, a virulent factor of the Avian Pathogenic Escherichia coli (APEC), is responsible for adhesion, biofilm formation, and colonization of pathogen. Major curli fimbriae protein is encoded by csgA gene. APEC is one of the leading causes of colibacillosis in poultry flocks and due to excessive use of antibiotics and vaccines in poultry, the emergence of various multi-drug resistant (MDR) bacterial strainsare is frequently reported. The growing concern of MDR bacterial strains necessitate novel antibacterial approaches to combat colibacillosis in poultry. RNA-based gene silencing is a very specific and robust strategy to target specific bacterial factors involved in pathogenicity and virulence. In this study, a phagemid-mediated sRNA expression system to target a vital gene, csgA, is employed. This comprises an M13 phagemid harboring a sRNA expression cassette and a pre-designed GUIDE sequences for the csgA target gene. To target the csgA gene at the mRNA level, a GUIDE sequence was computationally designed for pre-designed sRNA expression cassette. Online web tools were used to predict the binding energy, secondary structure, and off-target binding potential of the sRNA to optimize its expression. Results showed that the designed sRNA has a binding energy of - 29.60 kcal/mol with zero off-targets. After expression of the sRNA in the APEC cells, ̴ 45% reduction in the csgA level was observed via RT-PCR in the CS-APEC-O1 strains compared to the wt-APEC-O1. Similarly, the biofilm forming ability decreased by 40% in the CS-APEC-O1 strains. The swarming motility and hemagglutination efficiency were not affected by the sRNA expression. Future studies investigating the in vivo efficiency of M13 phagemid delivery are required to evaluate its candidacy in phage therapy.


Reduced expression of FRG1 facilitates breast cancer progression via GM-CSF/MEK-ERK axis by abating FRG1 mediated transcriptional repression of GM-CSF.
Multiple molecular subtypes and distinct clinical outcomes in breast cancer, necessitate specific therapy. Moreover, despite the improvements in breast cancer therapy, it remains the fifth cause of cancer-related deaths, indicating the involvement of unknown genes. To identify novel contributors and molecular subtype independent therapeutic options, we report reduced expression of FRG1 in breast cancer patients, which regulates GM-CSF expression via direct binding to its promoter. Reduction in FRG1 expression enhanced EMT and increased cell proliferation, migration, and invasion, in breast cancer cell lines. Loss of FRG1 increased GM-CSF levels which activated MEK/ERK axis and prevented apoptosis by inhibiting p53 in an ERK-dependent manner. FRG1 depletion in the mouse model increased tumor volume, phospho-ERK, and EMT marker levels. The therapeutic potential of anti-GM-CSF therapy was evident by reduced tumor size, when tumors with decreased FRG1 were treated with anti-GM-CSF mAb. We found an inverse expression pattern of FRG1 and phospho-ERK levels in breast cancer patient tissues, corroborating the in vitro and mouse model-based findings. Our findings first time elucidate the role of FRG1 as a metastatic suppressor of breast cancer by regulating the GM-CSF/MEK-ERK axis.


Novel family of terpene synthases evolved from trans-isoprenyl diphosphate synthases in a flea beetle.
Sesquiterpenes play important roles in insect communication, for example as pheromones. However, no sesquiterpene synthases, the enzymes involved in construction of the basic carbon skeleton, have been identified in insects to date. We investigated the biosynthesis of the sesquiterpene (6R,7S)-himachala-9,11-diene in the crucifer flea beetle Phyllotreta striolata, a compound previously identified as a male-produced aggregation pheromone in several Phyllotreta species. A (6R,7S)-himachala-9,11-diene-producing sesquiterpene synthase activity was detected in crude beetle protein extracts, but only when (Z,E)-farnesyl diphosphate [(Z,E)-FPP] was offered as a substrate. No sequences resembling sesquiterpene synthases from plants, fungi, or bacteria were found in the P. striolata transcriptome, but we identified nine divergent putative trans-isoprenyl diphosphate synthase (trans-IDS) transcripts. Four of these putative trans-IDSs exhibited terpene synthase (TPS) activity when heterologously expressed. Recombinant PsTPS1 converted (Z,E)-FPP to (6R,7S)-himachala-9,11-diene and other sesquiterpenes observed in beetle extracts. RNAi-mediated knockdown of PsTPS1 mRNA in P. striolata males led to reduced emission of aggregation pheromone, confirming a significant role of PsTPS1 in pheromone biosynthesis. Two expressed enzymes showed genuine IDS activity, with PsIDS1 synthesizing (E,E)-FPP, whereas PsIDS3 produced neryl diphosphate, (Z,Z)-FPP, and (Z,E)-FPP. In a phylogenetic analysis, the PsTPS enzymes and PsIDS3 were clearly separated from a clade of known coleopteran trans-IDS enzymes including PsIDS1 and PsIDS2. However, the exon-intron structures of IDS and TPS genes in P. striolata are conserved, suggesting that this TPS gene family evolved from trans-IDS ancestors.


Experimental modelling studies on the removal of dyes and heavy metal ions using ZnFe2O4 nanoparticles.
The presence of dyes and heavy metals in water sources as pollutants is harmful to human and animal health. Therefore, this study aimed to evaluate the efficacy of zinc ferrite (ZnFe2O4) nanoparticles (ZF-NPs) due to their outstanding properties including cost-effectiveness, availability, and applicability for removal of auramine O (AO), methylene blue (MB), and Cd (II). The effect of the main operating parameters such as AO concentration, MB concentration, Cd (II) concentration, adsorbent amount, solution pH, and sonication time was optimized by the response surface methodology (RSM). Optimal conditions were obtained at adsorbent amount of 0.25 g, pH = 6, sonication time of 15 min, and concentration of 15 mg L-1, and more than 91.56% were removed from all three analytes. The adsorption of AO, MB, and Cd (II) onto ZF-NPs followed pseudo-second-order kinetics and the equilibrium data fitted well with Langmuir isotherm. The maximum adsorption capacities of ZF-NPs for AO, MB and Cd (II) were as high as 201.29 mg g-1, 256.76 mg g-1 and 152.48 mg g-1, respectively. Also, the reuse of the adsorbent was investigated, and it was found that the adsorbent can be used for up to five cycles. Based on the results of interference studies, it was found that different ions do not have a significant effect on the removal of AO, MB, and Cd (II) in optimal conditions. The ZF-NPs was investigated successfully to remove AO, MB, and Cd (II) from environmental water samples. The results of this study showed that ZF-NPs can be used as a suitable adsorbent to remove AO, MB, and Cd (II) from aqueous solution.


Long noncoding RNA 00976 promotes pancreatic cancer progression through OTUD7B by sponging miR-137 involving EGFR/MAPK pathway.
Accumulation evidence indicates the vital role of long non-coding RNAs (lncRNAs) in tumorigenesis and the progression of malignant tumors, including pancreatic cancer (PC). However, the role and the molecular mechanism of long non-coding RNA 00976 is unclear in pancreatic cancer. In situ hybridization (ISH) and qRT-PCR was performed to investigate the association between linc00976 expression and the clinicopathological characteristics and prognosis of patients with PC. Subsequently, linc00976 over-expression vector and shRNAs were transfected into PC cells to up-regulate or down-regulate linc00976 expression. Loss- and gain-of function assays were performed to investigate the role of linc00976 in proliferation and metastasis in vitro and vivo. ITRAQ, bioinformatic analysis and rescue assay were used to illustrate the ceRNA mechanism network of linc00976/miR-137/OTUD7B and its downstream EGFR/MAPK signaling pathway. linc00976 expression was overexpressed in PC tissues and cell lines and was positively associated with poorer survival in patients with PC. Function studies revealed that linc00976 knockdown significantly suppressed cell proliferation, migration and invasion in vivo and in vitro, whereas its overexpression reversed these effects. Based on Itraq results and online database prediction, Ovarian tumor proteases OTUD7B was found as a downstream gene of linc00976, which deubiquitinated EGFR mediates MAPK signaling activation. Furthermore, Bioinformatics analysis and luciferase assays and rescue experiments revealed that linc00976/miR137/OTUD7B established the ceRNA network modulating PC cell proliferation and tumor growth. The present study demonstrates that linc00976 enhances the proliferation and invasion ability of PC cells by upregulating OTUD7B expression, which was a target of miR-137. Ultimately, OTUD7B mediates EGFR and MAPK signaling pathway, suggesting that linc00976/miR-137/OTUD7B/EGFR axis may act as a potential biomarker and therapeutic target for PC.


LncRNA RP4-639F20.1 interacts with THRAP3 to attenuate atherosclerosis by regulating c-FOS in vascular smooth muscle cells proliferation and migration.
The aberrant proliferation and migration of vascular smooth muscle cells (VSMCs) play an essential role in the pathogenesis of atherosclerosis (AS). Long noncoding RNAs (lncRNAs) have been reported as important regulators in a number of diseases. However, very little is known regarding the functional role of lncRNAs in governing proliferation and migration of VSMCs and AS development. Both in vitro and in vivo assays were performed to investigate the role of lncRNA in the pathophysiology of AS. Our previous lncRNA arrays revealed that lncRNA RP4-639F20.1 was significantly decreased in atherosclerotic plaques. Lentivirus overexpressing RP4-639F20.1 and lncRNA RP4-639F20.1 silencing vectors (Si-lnc-RP4-639F20.1) were constructed and transfected in VSMCs. The in vitro functions of lncRNA were analyzed by CCK-8 assays, EdU assays, scratch wound assays, transwell assays, qRT-PCR and Western blot analyses. RNA fluorescence in situ hybridization, immunoprecipitation and mRNA microarrays were used to explore the underlying mechanism. Adeno-associated-virus-9 (AAV9) overexpressing RP4-639F20.1 was constructed and injected intravenously into ApoE-/- mice to explore the role of lncRNA in vivo. In vitro experiments showed that lncRNA RP4-639F20.1 interacted with THRAP3 and downregulated c-FOS expression. Both increase of lncRNA RP4-639F20.1 expression and knockdown of c-FOS inhibited the expression of MMP10 and VEGF-α in VSMCs and suppressed VSMCs proliferation and migration. In vivo experiments using ApoE-/- mice fed a high-fat diet demonstrated that lncRNA RP4-639F20.1 overexpression deterred atherosclerosis and decreased lipid levels in atherosclerotic lesions. Patients with coronary artery disease were found to have higher c-FOS levels than healthy individuals and c-FOS expression was positively correlated with the SYNTAX score of patients. Overall, these data indicated that lncRNA RP4-639F20.1/THRAP3/c-FOS pathway protects against the development of atherosclerosis by suppressing VSMCs proliferation and migration. LncRNA RP4-639F20.1 and c-FOS could represent potential therapeutic targets to ameliorate atherosclerosis-related diseases.


Direct determination of phosphate sugars in biological material by (1)H high-resolution magic-angle-spinning NMR spectroscopy.
The study aim was to unambiguously assign nucleotide sugars, mainly UDP-X that are known to be important in glycosylation processes as sugar donors, and glucose-phosphates that are important intermediate metabolites for storage and transfer of energy directly in spectra of intact cells, as well as in skeletal muscle biopsies by (1)H high-resolution magic-angle-spinning (HR-MAS) NMR. The results demonstrate that sugar phosphates can be determined quickly and non-destructively in cells and biopsies by HR-MAS, which may prove valuable considering the importance of phosphate sugars in cell metabolism for nucleic acid synthesis. As proof of principle, an example of phosphate-sugar reaction and degradation kinetics after unfreezing the sample is shown for a cardiac muscle, suggesting the possibility to follow by HR-MAS NMR some metabolic pathways. Graphical abstract Glucose-phosphate sugars (Glc-1P and Glc-6P) detected in muscle by 1H HR-MAS NMR.


Urinary 1-hydroxypyrene in pregnant women in a Northeastern U.S. city: socioeconomic disparity and contributions from air pollution sources.
Maternal exposure to polycyclic aromatic hydrocarbons (PAHs), ubiquitous constituents of air pollution, has been associated with adverse birth outcomes. Yet it remains unclear whether and how socioeconomic status (SES) affects gestational PAH exposure. To examine whether there are socioeconomic disparities in PAHs exposure among pregnant women from Rochester, NY, and if so, to what extent disproportionate proximity to air pollution sources, measured by residential distance to transportation-related sources, contributed to the exposure disparity. We measured 1-hydroxypyrene concentrations in 726 urine samples collected from 305 pregnant women up to three samples throughout pregnancy. Residential distances to transportation-related sources were calculated based on participants' home addresses. We used linear mixed-effects models with random intercepts of participants to examine associations between 1-hydroxypyrene, SES indicators, and distance to transportation-related sources. We used structural equation modelling to assess to what extent distance to transportation-related sources contributes to the socioeconomic disparity in 1-hydroxypyrene concentrations. Reduced household income and maternal education level were both significant SES predictors of 1-hydroxypyrene concentrations, after the adjustment for other maternal demographic characteristics. Each interquartile range (IQR) increases in residential proximity to the airport (from 14.3 to 6.0 km), the railroad yard (from 22.3 to 6.0 km), and annual average daily traffic within 300 m (from 3796 to 99,933 vehicles/year) were associated with 15.0% (95%CI: 7.0-22.2%), 15.4% (95%CI: 6.5-23.5%), and 13.6% (95%CI: 4.7-23.3%) increases in 1-hydroxypyrene concentrations, respectively. Proximity to these sources jointly explained 10% (95%CI: 1.6-18.4%) of the 1-hydroxypyrene concentration change associated with decreases in SES as a latent variable defined by both household income and education level. Our findings suggest that efforts to address disproportionate residential proximity to transportation-related sources may reduce the socioeconomic disparity in PAH exposure.


Experience of directing a national laboratory animals centre.
The challenges of managing a facility serving a new and developing science are outlined. Basic to success are the appropriate handling of aspects such as staff selection, line management, financial support, changing demands, training and relations with staff unions, welfare organisations and at the international level.


Pertussis toxin-mediated ADP-ribosylation of rabbit luteal Gi uncouples enkephalin inhibition of adenylyl cyclase.
1. Some of the actions of pertussis toxin on the rabbit luteal adenylyl cyclase system were analyzed. 2. Incubation of luteal membranes with pertussis toxin and [32P]NAD resulted in the [32P]ADP-ribosylation of a 40,000 Da protein that is distinct from the proteins ADP-ribosylated by cholera toxin. 3. Pertussis toxin specific [32P]ADP-ribosylation was time-dependent and dependent upon the concentration of pertussis toxin present during the incubation. 4. Pertussis toxin mediated [32P]ADP-ribosylation was enhanced by ATP, ADP, adenylyl imidodiphosphate, GTP, guanosine-5'-O-(2-thiodiphosphate), guanosine-5'-O-(3-thiotriphosphate), and NaF but not AMP or guanylyl imidodiphosphate [GMP-P(NH)P]. 5. Treatment of luteal membranes with NAD and pertussis toxin prevents GTP and enkephalin but not GMP-P(NH)P mediated inhibition of forskolin stimulated adenylyl cyclase, demonstrating the existence of a functional Gi in the rabbit corpus luteum.


The Impact of Herbal Additives for Poultry Feed on the Fatty Acid Profile of Meat.
Researchers often found that herbal additives to chicken feed can favorably alter the fatty acid profile of the meat. The most desirable effects of diet modification comprise an increased content of polyunsaturated fatty acids (PUFA) and monounsaturated fatty acids (MUFA) and a reduced content of saturated fatty acids (SFA) in the breast and thigh muscles. A modified fatty acid profile contributes to improvement in the quality of poultry meat, which is reflected in its increased consumption. However, it may be problematic that PUFAs are oxidized easier than other lipids, which can have a negative impact on the sensory traits of meat. By contrast, herbs and herbal products contain antioxidants that can prevent the oxidation of unsaturated fatty acids and cholesterol present in animal-origin products and increase the antioxidant potential of the consumer's body. This paper aims to review the influence of herbal additives for broiler chicken diets on the fatty acid profile of poultry meat. Special attention was paid to changes in the content of SFAs, MUFAs, and PUFAs, but also alterations in the omega-6:omega-3 ratio. The presented reference literature supports the statement that herbs and bioactive components of herbs added to chicken diets can improve the quality of broiler chicken meat by altering the content of fatty acids.


Genetics and genomics of primary biliary cirrhosis.
The etiologic and pathogenic factors contributing to primary biliary cirrhosis (PBC) development, progression, response to treatment, and outcome remain a mystery. Recognition of the genomic regions harboring risk factors is hindered by the rarity and late onset of PBC. Recent advancements in genomics hold promise for understanding, prevention, and therapy of PBC. Large registries and biospecimen repositories of patients who have PBC, their family members, and controls are needed. Haplotype mapping-based association studies are necessary for defining genetic predisposition. Experimental data will provide the means for fine mapping studies, resequencing efforts, functional experimentation, and elucidation of gene-environment and gene-gene interaction.


Type 2 diabetic rats are sensitive to thioacetamide hepatotoxicity.
Previously, we reported high hepatotoxic sensitivity of type 2 diabetic (DB) rats to three dissimilar hepatotoxicants. Additional work revealed that a normally nonlethal dose of CCl4 was lethal in DB rats due to inhibited compensatory tissue repair. The present study was conducted to investigate the importance of compensatory tissue repair in determining the final outcome of hepatotoxicity in diabetes, using another structurally and mechanistically dissimilar hepatotoxicant, thioacetamide (TA), to initiate liver injury. A normally nonlethal dose of TA (300 mg/kg, ip), caused 100% mortality in DB rats. Time course studies (0 to 96 h) showed that in the non-DB rats, liver injury initiated by TA as assessed by plasma alanine or aspartate aminotransferase and hepatic necrosis progressed up to 48 h and regressed to normal at 96 h resulting in 100% survival. In the DB rats, liver injury rapidly progressed resulting in progressively deteriorating liver due to rapidly expanding injury, hepatic failure, and 100% mortality between 24 and 48 h post-TA treatment. Covalent binding of 14C-TA-derived radiolabel to liver tissue did not differ from that observed in the non-DB rats, indicating similar bioactivation-based initiation of hepatotoxicity. S-phase DNA synthesis measured by [3H]-thymidine incorporation, and advancement of cells through the cell division cycle measured by PCNA immunohistochemistry, were substantially inhibited in the DB rats compared to the non-DB rats challenged with TA. Thus, inhibited cell division and compromised tissue repair in the DB rats resulted in progressive expansion of liver injury culminating in mortality. In conclusion, it appears that similar to type 1 diabetes, type 2 diabetes also increases sensitivity to dissimilar hepatotoxicants due to inhibited compensatory tissue repair, suggesting that sensitivity to hepatotoxicity in diabetes occurs in the absence as well as presence of insulin.


Lung cancer metabolomic data from tumor core biopsies enables risk-score calculation for progression-free and overall survival.
Metabolomics has emerged as a powerful method to provide insight into cancer progression, including separating patients into low- and high-risk groups for overall (OS) and progression-free survival (PFS). However, survival prediction based mainly on metabolites obtained from biofluids remains elusive. This proof-of-concept study evaluates metabolites as biomarkers obtained directly from tumor core biopsies along with covariates age, sex, pathological stage at diagnosis (I/II vs. III/VI), histological subtype, and treatment vs. no treatment to risk stratify lung cancer patients in terms of OS and PFS. Tumor core biopsy samples obtained during routine lung cancer patient care at the University of Louisville Hospital and Norton Hospital were evaluated with high-resolution 2DLC-MS/MS, and the data were analyzed by Kaplan-Meier survival analysis and Cox proportional hazards regression. A linear equation was developed to stratify patients into low and high risk groups based on log-transformed intensities of key metabolites. Sparse partial least squares discriminant analysis (SPLS-DA) was performed to predict OS and PFS events. Univariable Cox proportional hazards regression model coefficients divided by the standard errors were used as weight coefficients multiplied by log-transformed metabolite intensity, then summed to generate a risk score for each patient. Risk scores based on 10 metabolites for OS and 5 metabolites for PFS were significant predictors of survival. Risk scores were validated with SPLS-DA classification model (AUROC 0.868 for OS and AUROC 0.755 for PFS, when combined with covariates). Metabolomic analysis of lung tumor core biopsies has the potential to differentiate patients into low- and high-risk groups based on OS and PFS events and probability.


Selenium Health Benefit Values: Updated Criteria for Mercury Risk Assessments.
Selenium (Se)-dependent enzymes (selenoenzymes) protect brain tissues against oxidative damage and perform other vital functions, but their synthesis requires a steady supply of Se. High methylmercury (CH3Hg) exposures can severely diminish Se transport across the placenta and irreversibly inhibit fetal brain selenoenzymes. However, supplemental dietary Se preserves their activities and thus prevents pathological consequences. The modified Se health benefit value (HBVSe) is a risk assessment criterion based on the molar concentrations of CH3Hg and Se present in a fish or seafood. It was developed to reflect the contrasting effects of maternal CH3Hg and Se intakes on fetal brain selenoenzyme activities. However, the original equation was prone to divide-by-zero-type errors whereby the calculated values increased exponentially in samples with low CH3Hg contents. The equation was refined to provide an improved index to better reflect the risks of CH3Hg exposures and the benefits provided by dietary Se. The HBVSe provides a biochemically based perspective that confirms and supports the FDA/EPA advice for pregnant and breast-feeding women regarding seafoods that should be avoided vs. those that are beneficial to consume. Since Se can be highly variable between watersheds, further evaluation of freshwater fish is needed to identify locations where fish with negative HBVSe may arise and be consumed by vulnerable subpopulation groups.


Prothrombin-antibody coexistent with lupus anticoagulant (LA): clinical study and immunochemical characterization.
The patient is a 23 y.o. man with acute nephritis and bleeding at presentation. Laboratory data consistent with the diagnosis of systemic lupus erythematosus. A lupus anticoagulant was found: tissue thromboplastin inhibition test (TTIT) ratio 3.4; diluted Russell viper venom (DRVV) ratio 2.6. Hypoprothrombinemia (FII:C less than 1%; FIIR:Ag 5%) was present; prothrombin survival time (FII concentrate infusion 60 U/kg): t1/2 approximately to 9 hours. A prothrombin antibody was identified: it is not neutralizing but forms an immunecomplex with prothrombin. The antibody was characterized as IgG2, IgA, k, lambda. The prothrombin survival time indicates that the hypoprothrombinemia is due to the clearance of the prothrombin-antiprothrombin complex in vivo.


Sonographic signs of nephritis in children. A comparison of renal echography with clinical evaluation, laboratory data and biopsy.
In 40 children with histologically confirmed diagnosis of nephritis 82 sonographic examinations were performed for evaluation of echographic signs of nephritis. Increased cortical renal echointensity in comparison with the normal liver and enlarged parenchymal volume proved to be the most specific and reliable signs of nephritis, particularly if both were present. Accentuated corticomedullary differentiation was too insensitive to be of additional value. Twenty-one patients had follow up examinations in the course of their nephritis. In 15 patients sonographic signs, serum creatinine and clinical classification showed parallel changes. In two cases sonographic improvement preceded clinical and laboratory normalization by several weeks, giving valuable prognostic information.


In vitro evidence for CCl4 metabolites covalently bound to lipoprotein micelles.
CCl4-induced impairment of the lipoprotein secretion pathway of intact rat hepatocytes was carried out using 14CCl4 to check the possibility of binding to lipoproteins by CCl4 metabolites. After separation of different cell suspension fractions by means of ultracentrifugation and chemical precipitation procedures, a significant amount of the radioisotope was found covalently bound to the lipid and protein components of low density lipoproteins. Suitable experiments demonstrated that the bound radioisotope was represented by CCl4 metabolites and not by unactivated CCl4.


Injectable gellan gum-based nanoparticles-loaded system for the local delivery of vancomycin in osteomyelitis treatment.
Infection spreading in the skeletal system leading to osteomyelitis can be prevented by the prolonged administration of antibiotics in high doses. However systemic antibiotherapy, besides its inconvenience and often low efficacy, provokes numerous side effects. Thus, we formulated a new injectable nanoparticle-loaded system for the local delivery of vancomycin (Vanc) applied in a minimally-invasive way. Vanc was encapsulated in poly(L-lactide-co-glycolide) nanoparticles (NPs) by double-emulsification. The size (258 ± 11 nm), polydispersity index (0.240 ± 0.003) and surface potential (-25.9 ± 0.2 mV) of NPs were determined by dynamic light scattering and capillary electrophoresis measurements. They have a spherical morphology and a smooth topography as observed using atomic force microscopy. Vanc loading and encapsulation efficiencies were 8.8 ± 0.1 and 55.2 ± 0.5 %, respectively, based on fluorescence spectroscopy assays. In order to ensure injectability, NPs were suspended in gellan gum and cross-linked with Ca(2+); also a portion of dissolved antibiotic was added to the system. The resulting system was found to be injectable (extrusion force 11.3 ± 1.1 N), reassembled its structure after breaking as shown by rheology tests and ensured required burst release followed by sustained Vanc delivery. The system was cytocompatible with osteoblast-like MG-63 cells (no significant impact on cells' viability was detected). Growth of Staphylococcus spp. reference strains and also those isolated from osteomyelitic joints was inhibited in contact with the injectable system. As a result we obtained a biocompatible system displaying ease of application (low extrusion force), self-healing ability after disruption, adjustable drug release and antimicrobial properties.


Endogenous opioid-dopamine neurotransmission underlie negative CBV fMRI signals.
Previous studies showed noxious unilateral forepaw electrical stimulation surprisingly evoked negative blood-oxygenation-level-dependent (BOLD), cerebral blood flow (CBF), and cerebral blood volume (CBV) fMRI responses in the bilateral striatum whereas the local neuronal spike and c-Fos activities increased. These negative responses are associated with vasoconstriction and appeared to override the increased hemodynamic responses that typically accompanied with increased neural activity. The current study aimed to investigate the role of μ-opioid system in modulating vasoconstriction in the striatum associated with noxious stimulation on a 4.7-Tesla MRI scanner. Specifically, we investigated: i) how morphine (a μ-opioid receptor agonist) affects the vasoconstriction in the bilateral striatum associated with noxious electrical forepaw stimulation in rats, and ii) how naloxone (an opioid receptor antagonist) and eticlopride (a dopamine D(2)/D(3) receptor antagonist) modulates the morphine effects onwards. Injection of morphine enhanced the negative striatal CBV responses to noxious stimulation. Sequential injection of naloxone in the same animals abolished the stimulus-evoked vasoconstriction. In a separate group of animals, injection of eticlopride following morphine also reduced the vasoconstriction. Our findings suggested that noxious stimulation endogenously activated opioid and dopamine receptors in the striatum and thus leading to vasoconstriction.


A series of novel indazole derivatives of Sirt 1 activator as osteogenic regulators.
A series of indazole derivatives were identified as Sirt 1 activators though high-throughput screening. Optimization of each substituent on the indazole ring led to the identification of compound 13. Compound 13 appeared to give the best Sirt 1 activity of the compounds tested and also showed osteogenesis activity in a cell assay. Sirt 1 activators are therefore potential candidates for the treatment of osteoporosis.


Cyclin dependent kinase 5 and its interacting proteins in cell death induced in vivo by cyclophosphamide in developing mouse embryos.
Activation or inactivation of members of the cyclin-dependent kinase family is important during cell cycle progression. However, Cdk5, a member of this family that was originally identified because of its high structural homology to Cdc2, is activated during cell differentiation and cell death but not during cell cycle progression. We previously demonstrated a correlation between the up-regulation of Cdk5 protein and kinase activity and cell death during development and pathogenesis. We report here that cyclophosphamide (CP) induces massive apoptotic cell death in mouse embryos and that Cdk5 is expressed in apoptotic cells displaying fragmented DNA. During CP-induced cell death, Cdk5 protein expression is substantially increased as detected by immunohistochemistry but not by Western blot, while its mRNA level remains the same as control, and its kinase activity is markedly elevated. The up-regulation of Cdk5 during CP-induced cell death is not due to de novo protein synthesis. We also examined p35, a regulatory protein of Cdk5 in neuronal differentiation. Using a yeast two-hybrid system, we isolated p35, a neuronal differentiation specific protein, as a protein that interacts with Cdk5 in CP-treated embryos. p35 mRNA level does not change, but the protein expression of p25, a truncated form of p35, is elevated during cell death in vivo, as established here, as well as during cell death in vitro. Our results suggest a role for Cdk5 and its regulatory proteins during CP induced cell death. These results further support the view that Cdk5 and its regulation may be key players in the execution of cell death regardless of how the cell dies, whether through biological mechanisms, disease states such as Alzheimer's disease, or induction by CP.


Organ-specific endothelial cell heterogenicity and its impact on regenerative medicine and biomedical engineering applications.
Endothelial cells (ECs) are a key cellular component of the vascular system as they form the inner lining of the blood vessels. Recent findings highlight that ECs express extensive phenotypic heterogenicity when following the vascular tree from the major vasculature down to the organ capillaries. However, in vitro models, used for drug development and testing, or to study the role of ECs in health and disease, rarely acknowledge this EC heterogenicity. In this review, we highlight the main differences between different EC types, briefly summarize their different characteristics and focus on the use of ECs in in vitro models. We introduce different approaches on how ECs can be utilized in co-culture test systems in the field of brain, pancreas, and liver research to study the role of the endothelium in health and disease. Finally, we discuss potential improvements to current state-of-the-art in vitro models and future directions.


[Experimental study on the change and effect of TGF-beta 1 after skin expansion].
To study the injury process of the skin due to expansion. New Zealand rabbits were divided into 4 groups: rapid expansion and slow expansion as well as two control groups. The changes of skin TGF-beta 1 were observed immediately after expansion and at 1, 12, 24 weeks after expansion. Immunohistochemistry and in situ hybridization technique were applied. TGF-beta 1 increased in the skin immediately after expansion. In groups of rapid expansion, TGF-beta 1 increased faster and then decreased also faster than slow expansion groups. The results from immunohistochemistry and in situ hybridization were almost same. Expansion resulted in skin injury. Rapid expansion injured the skin more seriously than slow expansion. TGF-beta 1 may be the main regulating factor to repairing process.


Artificial skyglow disrupts celestial migration at night.
Torres et al. demonstrate that artificial skyglow disrupts nightly migrations undertaken by the amphipod Talitrus saltator, which is normally guided by the sky position of the moon.


Interplay of hydrophobic and electrostatic interactions in biopolymer chromatography. Effect of salts on the retention of proteins.
The effect of salt on the retention behavior of proteins in electrostatic and hydrophobic interaction chromatography is described by a three-parameter equation, log k' = A - B log ms + Cms, where k' is the retention factor and ms is the molality of the salt in the eluent. Parameter B, termed the electrostatic interaction parameter, depends on the characteristic charge of the protein and the salt counterion and governs the change of retention with the salt concentration in ion-exchange chromatography. According to the model the magnitude of the hydrophobic interaction parameter C is determined by the hydrophobic contact area upon protein binding at the stationary phase surface and the properties of the salt as measured by its molal surface tension increment. Retention data measured at different salt concentrations in the eluent on a variety of ion exchangers can be fitted to the above equation which yields U-shaped plots of log k' against log ms. The limiting slopes of the appropriate plots at sufficiently low and high salt concentrations can be used to evaluate the electrostatic and hydrophobic interaction parameters, respectively. The approach, which is based on a combination of established treatments of electrostatic and hydrophobic interactions offers a convenient framework for analyzing retention data in biopolymer high-performance liquid chromatography and for the characterization of stationary phases. Furthermore, it may facilitate some characterization of protein molecules on the basis of their retention behavior as a function of the concentration and nature of the salt in the eluent. In the treatment of electrostatic interactions use is made of the counterion condensation theory that is believed to make possible a more comprehensive analysis than the traditional stoichiometric ion-exchange model which assumes binding of the proteins by coulombic interactions at discrete sites. The treatment of hydrophobic interactions is based on an adaptation of the solvophobic theory which predicts that the hydrophobic portion of the free energy of binding is proportional to the hydrophobic contact area and the microthermodynamic surface tension of the aqueous salt solution. Despite its simplicity the theory was successful in explaining the observed effect of the nature and concentration of salt in the eluent, the pH and the effect of the density of fixed charges at the surface of the stationary phase in the absence of specific salt effects.


Hepatic lipase: a member of a family of structurally related lipases.
Partial amino acid sequence of rat hepatic lipase was obtained by gas-phase microsequence analysis of proteolytic fragments. Sequence comparison to bovine lipoprotein lipase and porcine pancreatic lipase reveals a highly conserved region existing among these three physiologically distinct lipolytic enzymes. In a stretch of 36 amino acid residues previously reported for pancreatic lipase (De Caro, J., Boudouard, M., Bonicel, J., Guidoni, A., Desnuelle, P. and Rovery, M. (1981) Biochim. Biophys. Acta 671, 129-138), nineteen residues are identical for all three enzymes, whereas 27 of 36 are identical in rat hepatic lipase and bovine lipoprotein lipase. The fact that this primary structural conservation extends to three different animal species emphasizes the conclusion that these lipolytic enzymes comprise a protein family originating from a common ancestral gene.


Effect of epidermal growth factor on intestinal adaptation after allogeneic small bowel transplantation in rats.
We reported that epidermal growth factor (EGF) stimulated graft adaptation in a rat model of syngeneic small bowel transplantation. However, graft rejection is a severe problem with clinical small bowel transplantation, because small intestinal wall contains large amounts of lymphoid tissue. Studies were performed to investigate the effect of EGF on allogeneic graft adaptation after small bowel transplantation in rats treated with an immunosuppressant FK506. The transplanted animals received intraperitoneally EGF or saline (untreated) after surgery and were examined for analysis one week later. EGF-treated group markedly enhanced the water absorption and induction of sodium glucose cotransporter (SGLTI) as compared with EGF-untreated group. EGF-treated group also increased the mucosal crypt depth and its cell proliferating rate, although there was no significant difference in the mucosal villus height between the two groups. These results indicate that EGF accelerates intestinal allograft adaptation in part by the recovery of mucosal structure and function after small bowel transplantation in rats. EGF may have relevance to promote graft function in clinical small intestinal transplantation.


Two successive cesarean deliveries through separate posterior and anterior hysterotomy due to asymptomatic uterine torsion. A case report.
Uterine torsion during pregnancy is a rare obstetrical complication that can be life-threatening for both mother and child. Although torsion usually presents with acute, non-specific symptoms, it can also occur without any symptoms and pose no immediate health threat. Ultimately, the diagnosis of torsion is often made only during cesarean section. We present a case of a patient who underwent two successive cesarean sections through separate posterior and anterior hysterotomy due to asymptomatic uterine torsion in both cases. During the first cesarean section an incision was inadvertently made in the posterior segment of the uterus. At the second cesarean section the degree of rotation was very different and an anterior hysterotomy was performed. The patient made an uneventful recovery after both deliveries. If access to the lower anterior uterine segment is not safely available due to uterine torsion, a hysterotomy in the lower posterior uterine segment can be performed. The risk of rupture of a posterior hysterotomy scar in future pregnancies is unclear.


Tension Pneumocephalus: A Potentially Fatal Complication of Expanded Endoscopic Endonasal Approach.
While some volume of pneumocephalus occurs following any surgery entailing dural breach, tension pneumocephalus (TP) is a rare complication of endoscopic endonasal surgery described in less than 1% cases including expanded endoscopic endonasal approaches (EEEA). It is a neurosurgical emergency warranting urgent decompression. Two cases, who developed TP following EEEA are presented. One had sinonasal malignancy (adenoid cystic carcinoma) eroding the anterior skull-base (T4N0M0) and the other was a large olfactory groove meningioma. TP was heralded in both by sudden deterioration in neurological status. Both cases underwent bifrontal craniotomy for decompression with simultaneous skull-base repair incorporating a vascularised pericranial flap. Brief literature review regarding the pathophysiology, contributing factors, diagnosis, management, and prevention of TP following EEEA is presented. TP, a life-threatening neurosurgical emergency, warrants meticulous precautions for its prevention, and vigilant postoperative monitoring for early detection. Urgent decompression with thorough skull-base repair is imperative to prevent complications.


Nanoliquid chromatography-mass spectrometry of oligosaccharides employing graphitized carbon chromatography on microchip with a high-accuracy mass analyzer.
The nanoLC separations of oligosaccharides using microchip-based columns are described. Mixtures of alditols from mucins and human milk are separated on graphitized carbon. The nanoLC-MS device showed high mass accuracy for the oligosaccharides ranging between 1 and 6 ppm on routine analyses. The high mass accuracy readily allowed identification of oligosaccharide peaks and the determination of their compositions. High retention time reproducibility was exhibited by the microchip LC. Little variation was observed for standard sample either alone or in a complex heterogeneous mixture. The nanoLC-MS exhibits excellent capabilities in profiling mixtures of oligosaccharides.


Cardiovascular-specific mortality among multiple myeloma patients: a population-based study.
Multiple myeloma (MM) survival has greatly improved in recent decades. MM is usually diagnosed at a median age of 66-70 years. MM patients do not necessarily die from primary cancer, so cardiovascular health may be a key factor threatening long-term survival. This study was designed to explore the cardiovascular disease mortality (CVM) trends in MM patients and compare them with those in the general population. In total, 88,328 MM patients from the Surveillance, Epidemiology, and End Results (SEER) database (1975-2016) were included. Standardized mortality ratios (SMRs) were used to assess CVM risk. The CVM risk was significantly higher in MM patients than in the general population (SMR, 1.84 (95% CI, 1.78-1.89)). MM patients had the highest CVM SMR, at 2.62 (95% CI, 2.49-2.75), in the first year after diagnosis, and it decreased over the follow-up period. Over the study period, the incidence of CVM continued to decrease in MM patients diagnosed at age 65-74 (APC, -1.2% (95% CI, -1.9% to -0.4%)) and ⩾75 years (APC, -1.9% (95% CI, -2.6% to -1.2%)) but not younger. CVM was the second-most common cause of death in patients ⩾75 years. In only MM case analyses, male sex, Black race, older age at diagnosis, and earlier year of diagnosis were poor prognostic factors for heart-specific mortality. The CVM risk in MM patients was significantly higher than that in the general population. To improve survival, cardiovascular health should receive attention upon diagnosis.


It's a lipid's world: bioactive lipid metabolism and signaling in neural stem cell differentiation.
Lipids are often considered membrane components whose function is to embed proteins into cell membranes. In the last two decades, studies on brain lipids have unequivocally demonstrated that many lipids have critical cell signaling functions; they are called "bioactive lipids". Pioneering work in Dr. Robert Ledeen's laboratory has shown that two bioactive brain sphingolipids, sphingomyelin and the ganglioside GM1 are major signaling lipids in the nuclear envelope. In addition to derivatives of the sphingolipid ceramide, the bioactive lipids discussed here belong to the classes of terpenoids and steroids, eicosanoids, and lysophospholipids. These lipids act mainly through two mechanisms: (1) direct interaction between the bioactive lipid and a specific protein binding partner such as a lipid receptor, protein kinase or phosphatase, ion exchanger, or other cell signaling protein; and (2) formation of lipid microdomains or rafts that regulate the activity of a group of raft-associated cell signaling proteins. In recent years, a third mechanism has emerged, which invokes lipid second messengers as a regulator for the energy and redox balance of differentiating neural stem cells (NSCs). Interestingly, developmental niches such as the stem cell niche for adult NSC differentiation may also be metabolic compartments that respond to a distinct combination of bioactive lipids. The biological function of these lipids as regulators of NSC differentiation will be reviewed and their application in stem cell therapy discussed.


A comparison of CPAP and CPAPFLEX in the treatment of obstructive sleep apnea in World Trade Center responders: study protocol for a randomized controlled trial.
Following the World Trade Center disaster, a large number of individuals involved in rescue and recovery activity were exposed to significant amounts of dust, and reported symptoms of chronic nasal and sinus inflammation. An unusually high prevalence of obstructive sleep apnea (OSA) has also been observed in this World Trade Center Responder population. This project aims to examine the relationship between nasal pathology and OSA. Our hypothesis is that increased nasal resistance due to nasal inflammation predisposes to OSA in this population. Continuous Positive Airway Pressure (CPAP) is the standard therapy for OSA but despite its efficacy has poor adherence. Subjects with high nasal resistance may have greater difficulty in tolerating this therapy than those who do not have high nasal resistance. Reduction of excess expiratory positive pressure by the modality known as Cflex(™) during Continuous Positive Airway Pressure therapy (CPAP(Flex)) has been suggested to improve comfort without compromising efficacy. We will compare CPAP to CPAP(Flex) in subjects with OSA. Subjects with new onset habitual snoring will be screened for OSA using home sleep studies and rhinomanometry will be used to determine nasal resistance. In 400 subjects with OSA we will perform a randomized double blind cross-over study comparing CPAP to CPAP(flex), and relate nasal resistance to adherence to CPAP therapy. This is the first multicenter trial designed to test the hypothesis that adherence to CPAP therapy relates to nasal resistance and CPAP(Flex) will improve adherence to CPAP in those subjects with high nasal resistance. We anticipate the following results from this trial: 1. Increased nasal resistance is associated with decreased adherence to CPAP therapy. 2. Use of CPAP(Flex) improves adherence with CPAP therapy in subjects with high nasal resistance, but not in those with low nasal resistance. 3. The benefit of CPAP(Flex) on adherence is greatest when offered at CPAP therapy initiation rather than as a "rescue" therapy in subjects with high nasal resistance. ClinicalTrials.gov Identifier: NCT01753999, Date: 12 December 2012.


Topological distribution of different forms of neural cell adhesion molecule in the developing chick visual system.
The cell adhesion molecule isolated from neural tissue (N-CAM) is a membrane glycoprotein which is directly involved in calcium-independent adhesion among nerve cells and their processes (for review see refs 1,2). N-CAM has an unusual carbohydrate moiety containing a large and variable amount of sialic acid, the variation reflecting both the type of tissue and its developmental age. N-CAM is believed to be a ligand in the formation of cell-cell bonds and a decrease in sialic acid content from 30% to 10% is associated with a marked enhancement of the molecule's binding activity. Antibodies to N-CAM block its function and inhibit or alter bundling of nerve fibres, retinal cell development and nerve-muscle interaction. Here we use micro-gel electrophoresis to compare N-CAM from several parts of the developing chick visual system. The results indicate that N-CAM from the retina of 5-10-day-old embryos already exists in a relatively sialic acid-poor form, whereas the tectum and optic nerve beyond the eye contain sialic acid-rich N-CAM until much later in development. These studies suggest that the perikaryon and proximal axon shaft of retinoganglion cells have N-CAM with a lower sialic acid content than the distal portion of the axons, and that resulting differences in neurite adhesivity may be an important factor in the formation of the optic system.


First report on isolation of Mucor bainieri from honeybees, Apis mellifera: Characterization and biological activities.
Fungi are potential biocontrol agents and rich sources of secondary metabolites with demonstrated biological activities. This study aimed to isolate and identify fungi from surface-sterilized honeybees (Apis mellifera), as well as to evaluate their biological activities. One fungal isolate was obtained and identified morphologically and genetically as Mucor bainieri MK-Bee-2. Gas chromatography-mass spectroscopy (GC-MS) analysis of fungus crude extract, showed the existence of six major metabolites representing 92.48% of the total peak area. The crude extract of Mucor bainieri MK-Bee-2 was tested for antimicrobial, antioxidant, and antitumor activities. It demonstrated wide antimicrobial activities against human pathogenic Gram-positive and Gram-negative bacterial strains, as well as Candida albicans, with MIC values ranged from 62.5 to 250 µg/ml. The results revealed that the extract exhibited considerable antioxidant activities indicated by strong inhibition of both DPPH and ABTS free radicals. Additionally, the extract exhibited greater potential anticancer activity against both adenocarcinomic human non-small cell lung cancer cells (A549) [IC50 = 6.45 μg/ml], and immortal cell line hepatoma G2 (HepG2) human liver cancer cells [IC50 = 27.48 μg/ml] and higher selectivity in cancer cells than normal cell lines. Furthermore, the extract showed less cytotoxic activity against normal cells with higher IC50 values of 106.99 and 132.57 μg/ml against human lung fibroblast Wistar-38 (Wi-38) and oral epithelial cells (OEC), respectively. Taken together, the Mucor bainieri MK-Bee-2 extract comprises bioactive compounds as promising potential therapeutic candidates for the treatment of lung cancer. Strikingly, the extract sensitizes the lung cancer cells A549 to the ionizing radiation through the pro-apoptotic pathway as indicated by the annexin V flow cytometry analysis which showed that the extract reduced the apoptosis of lung cancer cells.


Nordic walking for people with relapsing-remittent multiple sclerosis: A case series study.
Patients with multiple sclerosis (MS) show fatigue, impaired gait and decreased functional mobility, which lead to a low quality of life (QoL). Low-cost and easy to perform exercise modalities having a positive impact on the aforementioned symptoms are needed. The performance of Nordic Walking (NW) could be a useful rehabilitation strategy but, to our knowledge, no study has been published in this regard as yet. This case series study aims at providing information regarding the usefulness and impact of a three-month NW program on self-perceived fatigue, functional mobility, physical fitness and QoL on a group of people with mild relapsing-remitting MS. A case series study with multiple assessments was performed. Three data collection points at 1-week intervals for three weeks were done at baseline. The intervention consisted of a 12-weeks NW program with a total of four measurement occasions, one every three weeks. The Fatigue Severity Scale (FSS), the Timed up and Go test (TUG), the 6-Minute Walk test (6MWT) and the Multiple Sclerosis Impact Scale (MSIS-29), were used to assess the outcomes of the program on the participants' self-perceived fatigue, functional mobility, physical fitness and QoL, respectively. Of the 14 participants who volunteered for the study, five completed at least 80% of the planned sessions and were included in the final analysis. The intervention did not have a clinical significant impact on the participants' fatigue while its impact on their QoL showed mixed results. Positive trends were found for all the participants in their functional mobility and physical fitness. Regarding functional mobility, clinically important changes were observed in three participants. None of the observed changes in physical fitness achieved clinical significance. A 12-week Nordic walking program did not appear to be particularly feasible and had little impact on the functional levels on a group of people with mild relapsing-remitting MS. Given that beneficial effects were seen in some participants, and taken into account the low adherence rate observed, future studies should explore the feasibility of NW programs performed under different environmental conditions (i.e. outdoors) and including motivational strategies aimed at increasing participation.


Protective role of nuclear factor of activated T cells 2 in CD8+ long-lived memory T cells in an allergy model.
The transcriptional regulation of cytokines released and controlled by memory T cells is not well understood. Defective IFN-gamma production in allergic asthma correlates in human beings with the risk of wheezing in childhood. To understand the role of the transcription factor nuclear factor of activated T cells 2 (NFATc2) in memory and effector T cells in the airways in experimental allergic asthma. We used murine models of allergic asthma and adoptive cell transfer of fluorescence-activated sorted cells in a disease model. Mice lacking NFATc2 developed an increase in airwayhyperresponsiveness (AHR), remodeling, and serum IgE levelson ovalbumin sensitization. This phenotype was associated withCD81CD1222 T cells deficient in IFN-g production in theairways. The origin of this phenotype in NFATc2(2/2) mice wasrelated to an expanded population of lung CD81CD1221(IL-2Rb chain) CD127hi (IL-7 receptor [R] a chain1) long-livedmemory cells. Adoptive transfer of ovalbumin-specific CD81NFATc2(2/2) T cells enhanced the AHR generated byNFATc2(2/2) CD41 T cells in immunodeficient mice, increasedIL-17, and reduced IFN-g production in the reconstituted mice. Depletion of the memory CD81CD1221IL-7Rhigh T-cellpopulation corrected the defect in IFN-g production by lungNFATc2(2/2) CD81CD1222 cells and abrogated the increasedAHR observed in NFATc2(2/2) CD81 T-cell-reconstituted micewith a severe combined immunodeficiency disorder. Taken together, our results suggest that NFATc2 expression in long-lived memory CD8+ T cells controls IL-2 and IFN-gamma production in lung CD8+ T cells, which then limits TH17 and TH2 development in the airways during allergen challenge.


Can whole brain nerve conduction velocity be derived from surface-recorded visual evoked potentials? A re-examination of Reed, Vernon, and Johnson (2004).
Reed, Vernon, and Johnson [Reed, T. E., Vernon, P. A., & Johnson, A. M. (2004). Sex difference in brain nerve conduction velocity in normal humans. Neuropsychologia, 42, 1709-1714] reported that "nerve conduction velocity" (NCV) of visual transmission from retina to the primary visual area (V1) is significantly faster in males than females. The authors estimated the NCV by dividing head length (nasion-to-inion distance) by the latency of the well-known P100 component of the visual evoked potential (VEP). Here, we critically examine these metrics and we contend that knowledge of the underlying physiology of neural transmission across the initial stages of the visual processing hierarchy dictates that a number of their assumptions cannot be reasonably upheld. Alternative, and we believe, more parsimonious interpretations of the data are also proposed.


Pathomorphology of the vascular bed of postinfarction heart in various types of remodeling.
We evaluated pathomorphological changes in the vascular bed of postinfarction heart in various types of remodeling. Dilatational remodeling was characterized by dilation of coronary arteries, increase in their volume density, and regular arrangement. Signs of coronary blood flow reduction and microcirculatory disturbances in the left ventricle were revealed during hypertrophic remodeling. Aneurysmal remodeling was characterized by the presence of small-vascular collateral-anastomotic plexus and reduction of the microcirculatory bed. Endocardial remodeling was accompanied by hypervascularization of the myocardium, formation of new coronary vessels, and reduced capillarization in the left ventricle of postinfarction heart.


Changes in surface sediment carbon compositions in response to tropical seagrass meadow restoration.
Seagrass meadows are declining at a global scale, threatening their capacity as blue carbon sinks. Restoration of seagrasses (via seagrass seeds or plant transplantation) may recover their carbon sequestration capacity. Previous studies have predominantly focused on sediment organic carbon (SOC), while variations in sediment carbon compositions remain poorly understood, limiting our comprehension of the influence of seagrass restoration on sediment carbon stability. Here, we researched the differences in surface (0-3 cm) sediment carbon compositions in response to tropical seagrass transplantation among species (Thalassia hemprichii and Enhalus acoroides); specifically, differences in labile, recalcitrant and refractory SOC, as well as sediment inorganic carbon (SIC) compositions variations under transplanted T. hemprichii and E. acoroides communities. It was found that seagrass transplantation enhanced suspended particle organic matter, and epiphyte and macroalgae input to surface sediment, which recovered the surface SOC concentration and stock rapidly to natural levels (increased ∼1.6-fold) within two years following transplantation. The elevated contribution of epiphyte and macroalgae significantly increased the surface labile sediment organic matter (SOM), but not the recalcitrant and refractory SOM composition after short-term transplantation. Meanwhile, surface SIC was significantly elevated, which might be mainly ascribed to allochthonous carbonate particle trapped under transplanted area with implications for carbon sequestration. The higher canopy and longer leaf seagrass species, E. acoroides, had elevated SOC, SIC and was more labile composition, compared to T. hemprichii transplant. Overall, this research suggests that tropical seagrass transplantation can increase the surface SOC, SIC concentration by increasing the labile organic matter and allochthonous carbonate particle input, respectively, with varying significantly among seagrass species.


miR-124-3p relieves allergic rhinitis by inhibiting dipeptidyl peptidase-4.
MicroRNA-124-3p (miR-124-3p) and dipeptidyl peptidase-4 (DPP4) are closely related to the development of inflammation. Allergic rhinitis (AR) models in mice and HNEpC cells were established. AR progression was assessed assessing by the frequency of nasal rubbing and sneezing, hematoxylin and eosin (HE), and TUNEL staining. Tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), granulocyte-macrophage colony-stimulating factor (GM-CSF), eotaxin, and MUC5AC were assessed by enzyme-linked immunosorbent assay and quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Apoptosis in HNEpC cells was assessed using flow cytometry. DPP4, activated-caspase-3, and pro-caspase-3 protein expression were evaluated by western blotting. In addition, we clarified the influence of miR-124-3p-targeted DPP4 on AR inflammation and cell injury. MiR-124-3p was downregulated in AR nasal mucosa tissue. Upregulation of miR-124-3p reduced the frequency of nasal rubbing and sneezing, pathological changes, eosinophil number, and apoptosis of nasal mucosa, TNF-α and IL-6 protein and mRNA levels in serum and HNEpC cells, and MUC5AC, eotaxin, and GM-CSF levels in HNEpC cells. Downregulation of miR-124-3p has the opposite effect. Therefore, the miR-124-3p /DPP4 axis may be an attractive target for AR therapy.


Minimum joint space width (mJSW) of patellofemoral joint on standing "skyline" radiographs: test-retest reproducibility and comparison with quantitative magnetic resonance imaging (qMRI).
To assess the intraobserver, interobserver, and test-retest reproducibility of minimum joint space width (mJSW) measurement of medial and lateral patellofemoral joints on standing "skyline" radiographs and to compare the mJSW of the patellofemoral joint to the mean cartilage thickness calculated by quantitative magnetic resonance imaging (qMRI). A couple of standing "skyline" radiographs of the patellofemoral joints and MRI of 55 knees of 28 volunteers (18 females, ten males, mean age, 48.5 ± 16.2 years) were obtained on the same day. The mJSW of the patellofemoral joint was manually measured and Kellgren and Lawrence grade (KLG) was independently assessed by two observers. The mJSW was compared to the mean cartilage thickness of patellofemoral joint calculated by qMRI. mJSW of the medial and lateral patellofemoral joint showed an excellent intraobserver agreement (interclass correlation (ICC) = 0.94 and 0.96), interobserver agreement (ICC = 0.90 and 0.95) and test-retest agreement (ICC = 0.92 and 0.96). The mJSW measured on radiographs was correlated to mean cartilage thickness calculated by qMRI (r = 0.71, p < 0.0001 for the medial PFJ and r = 0.81, p < 0.0001 for the lateral PFJ). However, there was a lack of concordance between radiographs and qMRI for extreme values of joint width and KLG. Radiographs yielded higher joint space measures than qMRI in knees with a normal joint space, while qMRI yielded higher joint space measures than radiographs in knees with joint space narrowing and higher KLG. Standing "skyline" radiographs are a reproducible tool for measuring the mJSW of the patellofemoral joint. The mJSW of the patellofemoral joint on radiographs are correlated with, but not concordant with, qMRI measurements.


A metal-free turn-on fluorescent probe for the fast and sensitive detection of inorganic azides.
Sodium azide is toxic and widely used in agricultural, commercial products, and research laboratories. Thus it is of a significant environmental concern and there is a need for the development of a rapid detection method. A fluorogenic dibenzylcyclooctyne derivative (Fl-DIBO) is herein described as a fluorescent probe for the rapid detection of inorganic azide via Strain-Promoted Azide-Alkyne Cycloaddition (SPAAC). Fl-DIBO was found to be highly selective toward NaN3 in comparison to other common anions with good sensitivity and detection limit of 10μM.


Age-dependent variation in the cytosol/membrane distribution of red cell protein kinase-C.
An age-dependent increase in membrane association of protein kinase-c and a decrease in the cytosolic enzyme, especially in the densest fraction rich in senescent red cells was observed in Stractan-gradient-separated normal erythrocytes.


Erratum to "Applications and immobilization strategies of the copper-centred laccase enzyme; a review"[Heliyon Volume 9, Issue 2, February 2023, Article e13156].
[This corrects the article DOI: 10.1016/j.heliyon.2023.e13156.].


Platelet-specific proteins in patients with transient ischemic attacks. (Effects of anti-platelet drugs).
Levels of beta-thromboglobulin and platelet factor 4 in 46 patients with transient ischemic attacks and the influence of treatment with dipyridamole or aspirin plus dipyridamole were evaluated. Patients not receiving therapy showed levels of beta-thromboglobulin significantly higher than controls. However, patients taking only dipyridamole showed values of beta-thromboglobulin significantly lower than untreated patients or those taking aspirin plus dipyridamole. No significant difference was observed with regard to platelet factor 4 between the patients.


Plasma lipids and lipoproteins in young patients with brain infarction.
Thirty-two patients aged 60 or less with brain infarction were examined with regard to possible changes in lipoprotein pattern as compared with a reference group. Except for a lower concentration of total lipids, cholesterol and high-density lipoproteins in the female patients, no difference was demonstrated in the two groups. In all patients a significant reduction in total lipids, cholesterol, triglyceride and all lipoproteins was demonstrated after the acute cerebrovascular accident presumedly due to the stress situation.


Immunological response of lame sheep to clinical interdigital dermatitis and footrot: Procalcitonin, acute phase proteins, and pro-inflammatory cytokines.
In many countries, sheep lameness is a cause of economic concern and a contributing factor to a declining economy. This study aimed to investigate changes in procalcitonin (PCT), acute phase proteins (APPs), and cytokines (CYTs) in response to interdigital dermatitis and footrot in sheep under field conditions, to emphasize their role in the disease pathogenesis, diagnosis, as well as monitoring treatment response. Fifty-three sheep with foot diseases (26 clinical cases with interdigital dermatitis and 27 clinical cases with footrot) and 20 clinically healthy naemi sheep were used in this study. Real time PCR for detection of Fusobacterium necrophorum (F. necrophorum) and Dichelobacter nodosus (D. nodosus) revealed that, all samples collected from lame sheep (N = 53) were positive for D. nodosus (100 %), whereas F. necrophorum was detected in 19 out of 53 samples (35.84 %). The virulent D. nodosus was detected in 48 lameness cases where non-virulent D. nodosus were identified in 5 cases (in concurrent with F. necrophorum). The mean serum levels of PCT, C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (HP), fibrinogen (Fg) and CYTs (IL1-β, IL-1α, IFN-γ, IL-6 and TNF-α) in sheep with clinical interdigital dermatitis and footrot were remarkably higher than those detected in control healthy sheep. The serum levels of PCT, CRP, SAA, HP, Fg, and CYTs markers in lame sheep pre- and post-treatment were measured. A substantial decline was detected in serum levels of tested biomarkers of lame sheep after 14 days of treatment. The ROC curves were created. The AUC was assessed to evaluate the accuracy of each variable in distinguishing diseased and healthy sheep. Based on the ROC curves and AUCs; PCT, CRP, SAA, HP, and CYTs were highly diagnostic and predictive for the treatment response of sheep with clinical interdigital dermatitis and footrot. Moreover, all tested biomarkers had a noteworthy role in disease immuno-pathogenesis. Nevertheless, PCT and CRP are better than other tested APPs and CYTs as diagnostic markers for interdigital dermatitis and footrot. However, PCT only has the ability to differentiate sheep with different lameness score.


Observation of Chinese Hamster Ovary Cells retained inside the non-woven fiber matrix of the CellTank bioreactor.
This data article shows how the recombinant Chinese Hamster Ovary (CHO) cells are located in the interstices of the matrix fibers of a CellTank bioreactor after completion of a perfusion culture, supporting the article entitled "Very high cell density perfusion of CHO cells anchored in a non-woven matrix-based bioreactor" by Zhang et al. [1]. It provides a visualization of the cell distribution in the non-woven fiber matrix in a deeper view.


Identification of a novel protein synthesis inhibitor active against gram-positive bacteria.
In an effort to identify novel antibacterial chemotypes, we performed a whole-cell screen for inhibitors of Staphylococcus aureus growth and pursued those compounds with previously uncharacterized antibacterial activity. This process resulted in the identification of a benzothiazolium salt, ABTZ-1, that displayed potent antibacterial activity against Gram-positive pathogens. Several clinically desirable qualities were demonstrated for ABTZ-1 including potent activity against multidrug-resistant clinical isolates of methicillin-resistant S. aureus (MRSA) and vancomycin-resistant enterococci (VRE), retention of this activity in human serum, and low hemolytic activity. The antibacterial activity of ABTZ-1 was attributed to its inhibition of bacterial translation, as this compound prevented the incorporation of [³⁵S]methionine into S. aureus proteins, and ABTZ-1-resistant strains were cross-resistant to known inhibitors of bacterial translation. ABTZ-1 represents a promising new class of antibacterial agents.


Marijuana, receptors and immunomodulation.
THC, the major psychoactive component of marijuana, has been shown both in humans and experimental animals to have immunomodulatory properties. For example, marijuana smokers may show impaired immunological functions, including deficiency of blood leukocyte blastogenesis to mitogens. Detailed studies with mice have shown that animals given THC can show marked immunomodulation, including suppression of antibody formation, deficient cytokine production, etc. However, recent studies have also shown that lymphoid cells evince enhanced production or release or IL1, but suppression of IL2 and interferon production. Such lymphoid cells treated in vitro with THC also show suppressed blastogenesis to antigens and mitogens, suppressed NK activity, etc. In contrast, it has recently been shown that THC can enhance production or release of pro-inflammatory cytokines. This includes release of these cytokines from macrophages, including augmented release of IL1, TNF alpha, and IL6 activity. Susceptibility of mice to infection with opportunistic organisms such as L. pneumophila has been found and this increased susceptibility can be modulated by THC. A toxic shock-like death to Legionella has been induced by THC treatment given one day before and one day after infection. Receptors to THC have been detected in the brain as well as in peripheral tissues, including lymphoid cells. Thus, immunomodulation induced by THC may be related to receptor effects as well as unrelated to such receptors. It is clear that THC and other cannabinoids are excellent tools for studying the mechanisms of immune modulation, especially altered susceptibility to microbial infection.


Longterm reduction of back pain risk in women with osteoporosis treated with teriparatide compared with alendronate.
To compare the effects on back pain of teriparatide versus alendronate, we analyzed the reporting of back pain in a head to head comparator trial and a followup study. In the comparator trial, women were randomized to receive either daily self-injected teriparatide 40 microg plus an oral placebo (n = 73), or daily oral alendronate 10 mg plus self-injected placebo (n = 73). Treatment was for a median 14 months. After completion of the comparator trial, 72% of these patients enrolled in a nontreatment followup study. Adverse events were recorded at each comparator trial visit and followup study visit, and the incidence of new or worsening back pain in each group was compared. During the comparator trial, compared with women randomized to alendronate 10 mg, women randomized to teriparatide 40 microg had reduced risk for any back pain (relative risk 0.27, 95% CI 0.09-0.82) and moderate or severe back pain (relative risk 0.19, 95% CI 0.04-0.86). The differences in the reporting of back pain between the teriparatide treated women and the alendronate treated women were sustained during an interval including the comparator trial plus 18 additional months. During an interval including the comparator trial plus 30 additional months, teriparatide treated patients had numerically fewer occurrences of back pain and moderate or severe back pain. Compared with women randomized to alendronate 10 mg, women randomized to teriparatide 40 microg had reduced risk of back pain during the trial and 2.5 years of followup.


Inequities in Family Planning in Low- and Middle-Income Countries.
Inequities in reproductive health are widespread, and periodic surveys can trace trends in inequities to guide policies and program implementation. We examined National Composite Index for Family Planning surveys from 2017 and 2021 that assessed inequities in access to 7 contraceptive methods and traced patterns of discrimination involving 5 subgroups in low- and middle-income countries. These surveys use 10-25 informants in each country who are knowledgeable at the national level. Measures are based on questionnaire ratings on a scale of 1-10. Access to contraceptive methods averages about half of the maximum of 100%, with substantial variation across regions and countries for the score levels. Score profiles are similar among high-scoring and low-scoring countries, suggesting that access to each method reflects common determinants in the nature of each method and the influences acting upon national family planning programs. Access to short-term methods (pill, injectable, and condom) is much better than for long-term methods (sterilization, intrauterine device, and implant). Community-based distribution of contraceptives averages low, as it is not part of some programs. Over time the scores have improved modestly. Correlations imply that better access leads to more contraceptive use. Inequity of use across wealth groups is less where overall equity has improved. Measures of discrimination against youth, unmarried women, postabortion clients, HIV carriers, and different wealth groups indicate a need for additional policies and considerable latitude for stronger actions by providers. The surveys in 2017 and 2021 demonstrate both progress and deficiencies for equitable access to contraceptive methods, with highly variable results among regions and countries. Much remains to be done to alleviate discriminatory practices against particular subgroups. Equity has improved for access to contraception and contraceptive use, and it can continue to do so with greater attention to policies and practices in national programs.


Dysplasia in Gallbladder: What Should We Do?
On occasional cholecystectomies, pathologists encounter incidental dysplasia in the gallbladder mucosa in the sections submitted per protocol for histologic examination. If dysplasia is identified, additional sections are taken and/or the gallbladder is entirely submitted to rule out underlying adenocarcinoma. The aim of our study was to assess the incidence of subsequent identification of invasive adenocarcinoma on additional sections, after an incidentally detected dysplasia was noted on a routine cholecystectomy section. We also aimed to study the significance of the incidental detection of dysplasia and adenocarcinoma, as well as showing the association of gallbladder dysplasia to synchronous or metachronous dysplasia/neoplasia in the biliary tract. Our study was approved by the Institutional Review Board. We retrospectively identified 41 consecutive cases of routine cholecystectomies from 1991 to 2017, which had no clinical suspicion of neoplasia, and did not have any identifiable mass lesion, but on histopathologic analysis, had neoplasia (adenocarcinoma in 4 cases, and dysplasia in 37 cases). The pathologies of all cases were reviewed, and the diagnosis and grade of dysplasia were confirmed. The clinical information was obtained from the electronic medical records. Of the 37 cases with dysplasia, 10 (27%) had high-grade dysplasia (HGD) and the remaining showed low-grade dysplasia (LGD). All 4 cases of adenocarcinoma had some gross abnormalities (such as porcelain gallbladder, or ruptured, thickened, and roughened walls, or a granular mucosa). In contrast, none of the 37 cases with dysplasia had any gross abnormality. In 24 (of 37) cases of dysplasia, additional sections were submitted (median 8; ranging from 2 to 29), and in 11 cases, the gallbladder was entirely submitted. None of these cases showed any additional pathologic finding on the extra sections. Interestingly, 7 cases with dysplasia (18.9%; 6 LGD and 1 HGD) were associated with a concomitant pancreatobiliary malignancy. For the remaining 30 cases, follow-up information was available in 16 cases (53.3%) with a mean follow-up of 76.5 months (ranging from 12 to 204 months). None of these showed any subsequent development of pancreatobiliary neoplasms. Incidentally detected gallbladder dysplasia in a cholecystectomy specimen, without any gross abnormality, has almost no risk of a hidden invasive carcinoma. Although cholecystectomy is sufficient treatment for gallbladder dysplasia, in our study cohort, 18.9% of cases with incidental dysplasia in gallbladder had an associated pancreatobiliary carcinoma, which supports the hypothesis of multifocal neoplastic potential in the pancreatobiliary tree (also known as field effect). Although follow-up on 16 cases shows no subsequent development of any other pancreatobiliary neoplasm, this number is probably not enough to rule out a serial imaging follow-up of patients who have reported dysplasia in their gallbladder, to assess for subsequent development of neoplasia elsewhere in the pancreaticobiliary tree.


Digital Gene Expression Analysis on Cytology Smears Can Rule Out Malignancy in Follicular-Patterned Thyroid Tumors.
Patients with indeterminate thyroid nodules (Bethesda III and IV) are often treated with diagnostic lobectomy, which in most cases represents an overtreatment. A reliable rule-out molecular test could spare patients unnecessary surgery. Stained smears of 88 indeterminate thyroid nodules with histologic diagnosis of follicular-patterned tumors were selected: 34 follicular adenomas (FAs), 34 follicular variant papillary thyroid carcinomas (FVPTCs), and 20 noninvasive follicular neoplasms with papillary-like nuclear features (NIFTPs). The expression level of 126 genes was measured by digital counting. Mutation testing was performed for the main gene mutations and fusions. Performance of gene expression and mutation tests was calculated by receiver operating characteristic analysis. The gene expression model showed an area under the curve (AUC) of 88%, with 91% negative predictive value in FAs and FVPTCs only. Part of NIFTPs was labeled as benign, and part was labeled as malignant; thus, the classifier performance worsened. Two FAs (5.9%), eight NIFTPs (40%), and 22 FVPTCs (64.7%) were mutation positive. Mutation testing AUC was 79% in FAs and FVPTCs, and decreased by including NIFTPs. This gene expression-based test was feasible in thyroid-stained smears, showed higher AUC than mutation test, and had a high negative predictive value-making it a good candidate as a rule-out test for indeterminate thyroid cytology. NIFTPs have a heterogeneous phenotype, and their preoperative diagnosis requires further investigation.


Simple strategy for single-chain fragment antibody-conjugated probe construction.
A combination of biomarker and instrument technology diagnosis methods, especially antigen-targeted imaging methods, is required to increase the accuracy of the diagnosis of cancer. Currently, the targeting efficiency is limited by the conjugation methods used for the conjugation of antibodies and imaging materials. Here, a simple strategy for the conjugation of a probe and a single-chain fragment antibody (scFv) that does not change the characteristics of the antibody was shown. An ScFv was conjugated with superparamagnetic iron oxide (SPIO) or indocyanine green (ICG) via a linker by utilizing the reaction between cysteine and maleimide. The characterization of the probe was performed by flow cytometry, confocal imaging, optical imaging and magnetic resonance imaging (MRI). After conjugation, the scFv retained high affinity, antigen specificity, and strong internalization ability. The application of the conjugated probe was also confirmed by optical imaging and MRI. The proposed strategy provides a simple method for the production of high efficiency antigen-targeted imaging probes for tumor diagnosis.


Foreign Body Aspiration in Children and Emergency Rigid Bronchoscopy: A Retrospective Observational Study.
Foreign body aspiration is potentially life-threatening in paediatric age group. Early recognition and emergency intervention by Rigid bronchoscopy is life-saving. To highlight various difficulties in emergency paediatric bronchoscopy and discuss our experience in 138 patients. < 12 years children with suspected foreign body aspiration were included. Data of 138 patients < 12 years of age were studied. The most common foreign body found was peanut and organic foreign bodies constituted of total foreign bodies removed. Choking, Cough and sudden onset breathlessness were common symptoms. Tachypnoea, asymmetric breath sound, rhonchi, stridor, reduced chest movements were common signs. Obstructive emphysema was commonest radiological findings. Majority of the patients were discharged within 72 h & only two patients expired. History, clinical and radiological findings are highly indicative of foreign body in the airway. Inspite of being a high risk procedure, Rigid bronchoscopy when performed with necessary expertise,trained anaesthesia team and a paediatric ICU, saves majority of lives of children with tracheobronchial foreign bodies.


Predictors of technique failure and mortality on peritoneal dialysis: An analysis of New Zealand peritoneal dialysis registry data.
Technique failure is a major disadvantage associated with peritoneal dialysis (PD). This study aimed to analyse the demographic and risk predictors of technique failure and mortality in patients on PD. All incidental PD patients registered on the New Zealand Peritoneal dialysis registry (NZPDR) from January 1995 to December 2014 were included in the study. The primary outcomes were time to technique failure and its specific causes, while as the secondary outcome was time to death. Risk predictors of technique failure and mortality were analysed using multivariate Cox proportional hazards (PH) regression model. Besides, competitive risk regression analysis was undertaken to analyse the effect of death as a competing event to technique failure. Of 6379 patients, there were 2993 (46.9%) episodes of technique failure and 2684 (42%) deaths. The crude technique failure and mortality rates were 165 ± 5.90 and 147.9 ± 5.50 (mean ± SD)/1000 patient-years, respectively. Hazards of technique failure were lower in older individuals above 60 years, HR 0.72 (95% CI 0.67-0.79), larger centres, HR 0.89 (95% CI 0.79-1.00) and higher with coiled catheters, HR 1.26 (95% CI 1.16-1.37). Early nephrology referral, continuous ambulatory peritoneal dialysis (CAPD) and Asian ethnicities were associated with better technique survival. Infections were the major cause of technique failure (58.4%) with peritonitis being the leading cause (30.2%). There are multiple factors associated with risk of technique failure, therefore it is persuasive to construct a mathematical model for early prediction, for a planned transition to HD.


Characterisation of tone oscillations in placental and myometrial arteries from normal pregnancies and those complicated by pre-eclampsia and growth restriction.
Agonist-induced tone oscillations (rhythmic contractions and relaxations) occur in vascular beds to allow acute regulation of volume flow and thus the delivery of oxygen and nutrients to the tissue. Mechanisms responsible for the control of human placental vasomotor tone and blood flow are poorly characterized. This study aimed to characterise thromboxane-induced tone oscillations in human placental and myometrial arteries. Chorionic plate and myometrial arteries obtained from biopsies at term were mounted for isometric tension measurement. Tone oscillations were observed in chorionic arteries only when exposed to sub-maximal (<1 microM) concentrations of U46619. Slow (mean+/-SEM) frequency (2.6+/-0.5 per hour), large amplitude (39+/-7% of peak contraction) tone oscillations were elicited by 0.03 microM U46619 (n=18). In the presence of the nitric oxide synthase (NOS) inhibitor l-NNA (100 microM) the amplitude was significantly reduced (40+/-13% to 18+/-8%, P<0.05, n=6), frequency was unaltered and the bradykinin-dependent vasodilator response was reduced (68+/-13% to 40+/-19%, P<0.05, n=6). Myometrial arteries exposed to 1 microM U46619 developed tone oscillations within 10 min, which increased in amplitude over 30min occurring at relatively constant frequency. The mean amplitude of oscillations at 30 min (31+/-7%, n=16) was similar to that in chorionic arteries but the occurrence more frequent (42.8+/-9.7 per hour, P<0.001). Inhibition of NOS did not alter tone oscillations in myometrial arteries. Tone oscillations in chorionic arteries from pre-eclamptic and growth restricted (FGR) pregnancies were reduced in amplitude whereas those in myometrial arteries had increased frequency. Inhibition of NOS further reduced oscillation amplitude in chorionic arteries from FGR pregnancies. The alterations may contribute to the vasculopathology of these conditions, or, may represent compensatory mechanisms to maintain a matching of materno-placental blood flow.


Optical detection of atherosclerosis at molecular level by optical coherence tomography: An in vitro study.
There is an urgent need for contrast agents to detect the first inflammation stage of atherosclerosis by cardiovascular optical coherence tomography (CV-OCT), the imaging technique with the highest spatial resolution and sensitivity of those used during coronary interventions. Gold nanoshells (GNSs) provide the strongest signal by CV-OCT. GNSs are functionalized with the cLABL peptide that binds specifically to the ICAM-1 molecule upregulated in the first stage of atherosclerosis. Dark field microscopy and CV-OCT are used to evaluate the specific adhesion of these functionalized GNSs to activated endothelial cells. This adhesion is investigated under static and dynamic conditions, for shear stresses comparable to those of physiological conditions. An increase in the scattering signal given by the functionalized GNSs attached to activated cells is observed compared to non-activated cells. Thus, cLABL-functionalized GNSs behave as excellent contrast agents for CV-OCT and promise a novel strategy for clinical molecular imaging of atherosclerosis.


Examining the Impact of the Golden Compass Clinical Care Program for Older People with HIV: A Qualitative Study.
The combined burden of geriatric conditions, comorbidities, and HIV requires a model of HIV care that offers a comprehensive clinical approach with people 50 years or older with HIV. Golden Compass is an outpatient, multidisciplinary HIV-geriatrics program with an onsite HIV geriatrician, cardiologist, pharmacist, and social worker, offering specialist referrals, care navigation, and classes on improving functional status and cognition. Participants (13 patients and 11 primary care providers) were recruited using a non-probability sampling method to participate in semi-structured interviews on the perceived impact of Golden Compass on care delivered to older people with HIV. Interviews were transcribed verbatim and framework analysis used to analyze the transcripts. The perceived impacts of Golden Compass by patients and providers were organized by the Compass points (Northern: Heart and Mind, Eastern: Bones and Strength, Southern: Navigation and Network, Western: Dental, Hearing, and Vision). Overall, patients valued the focus on functional health and whole-person care, leading to greater trust in the ability of providers. Providers gained new skills through the geriatrics, cardiology and/or pharmacist consultations. The HIV-geriatrics specialty approach of Golden Compass improved functional ability and quality of life for older adults with HIV. Few integrated care programs for older people with HIV have been evaluated. This study adds to the limited literature demonstrating high patient and provider satisfaction with a HIV-care model that incorporated principles of geriatric medicine emphasizing a comprehensive approach to sustaining functional ability and improving quality of life.


Reactive oxygen species inhibit the succinate oxidation-supported generation of membrane potential in wheat mitochondria.
In order to gain a first insight into the effects of reactive oxygen species (ROS) on plant mitochondria, we studied the effect of the ROS producing system consisting of xanthine plus xanthine oxidase on the rate of membrane potential (DeltaPsi) generation due to either succinate or NADH addition to durum wheat mitochondria as monitored by safranin fluorescence. We show that the early ROS production inhibits the succinate-dependent, but not the NADH-dependent, DeltaPsi generation and oxygen uptake. This inhibition appears to depend on the impairment of mitochondrial permeability to succinate. It does not involve mitochondrial thiol groups sensitive to either mersalyl or N-ethylmaleimide and might involve both protein residues and/or membrane lipids, as suggested by the mixed nature. We propose that, during oxidative stress, early generation of ROS can affect plant mitochondria by impairing metabolite transport, thus preventing further substrate oxidation, DeltaPsi generation and consequent large-scale ROS production.


The CRF neuron: immunocytochemical study.
In the central nervous systems of several species belonging to different vertebrate classes, immunocytochemical stainings with an antiserum to ovine CRF 41 show multiple location of CRF perikarya and various areas containing CRF fibres and terminals. These stainings reflect species and interspecies functional adaptations of the CRF neurones which constitute a prominent hypothalamo-infundibular system involved in pituitary gland control, and also interneurone systems as attested to by extrahypothalamic perikarya and by CRF perisomatal endings in several brain areas. Perikarya of the hypothalamo-infundibular system are mainly packed in the paraventricular nucleus (mammals, birds) or in homologous areas, e.g.: paraventricular organ (turtle) where they are CSF-contacting neurones and preoptic nucleus (amphibians, fishes). In all species but fish, CRF fibres end in the median eminence (ME) against portal vessels. In fish, CRF processes terminate in the peripheral areas of proadenohypophyseal neurodigitations, close to corticotrophs. In all species these stainings are abolished by preabsorption of the serum by CRF. In fishes, reptiles and amphibians they are also suppressed by urotensin I, which is thought to be the teleost's CRF. Adrenalectomy experiments in the rat provided evidence for a corticosteroid regulation of ME CRF: short term (12-24 hr) adrenalectomy induces a complete depletion of CRF immunoreactivity followed by a secondary accumulation (5-20 days). This biphasic evolution is prevented by a dexamethasone replacement therapy. Inhibitory role of catecholamines on CRF release was indicated by: disappearance of ME CRF, induced by a single reserpine injection and suppression of this effect by monoamineoxidase inhibitor (pargyline or tranylcypromine) pretreatment. CRF fibres were first observed in the ME at the 16th week of fetal development in the human, and at the 18th day in the rat fetus. Thus, immunoreactive CRF system develops later than pituitary corticotrophs.


Correlation of thoracic aortic atherosclerotic plaque detected by multiplane transesophageal echocardiography and cardiovascular risk factors.
This study of 416 patients identified age, male gender, smoking, diabetes, hypertension, and hypercholesterolemia as independent predictors of thoracic aortic atherosclerotic plaque. Age, smoking, hypercholesterolemia, hypertension, and diabetes were predictors of the severity and extent of thoracic aortic atherosclerosis.


'You are Okay': Effects of a support and educational program for children with mild intellectual disability and their parents with mental health concerns.
Children with mild intellectual disabilities and emotional and behavioural problems whose parents have mental health concerns are at an increased risk of developing problems themselves. This study analysed the effectiveness of the 'You are Okay'-program aimed to support these children and parents and to reduce the risk of problem development in children. A quasi-experimental design was used to compare the experimental group ('care as usual' and 'You are Okay') with the control group ('care as usual') on the main outcome of emotional and behavioural problems and proposed intermediate outcomes. Children in the experimental group reported a significant medium to large decrease in emotional and behavioural problems compared to the control group. The effects on the intermediate outcomes were non-significant. This specific study population comprising a vulnerable group of children seems to benefit from this specially adapted 'You are Okay'-program. More research is necessary to draw further conclusions.


Water addition, evaporation and water holding capacity of poultry litter.
Litter moisture content has been related to ammonia, dust and odour emissions as well as bird health and welfare. Improved understanding of the water holding properties of poultry litter as well as water additions to litter and evaporation from litter will contribute to improved litter moisture management during the meat chicken grow-out. The purpose of this paper is to demonstrate how management and environmental conditions over the course of a grow-out affect the volume of water A) applied to litter, B) able to be stored in litter, and C) evaporated from litter on a daily basis. The same unit of measurement has been used to enable direct comparison-litres of water per square metre of poultry shed floor area, L/m(2), assuming a litter depth of 5cm. An equation was developed to estimate the amount of water added to litter from bird excretion and drinking spillage, which are sources of regular water application to the litter. Using this equation showed that water applied to litter from these sources changes over the course of a grow-out, and can be as much as 3.2L/m(2)/day. Over a 56day grow-out, the total quantity of water added to the litter was estimated to be 104L/m(2). Litter porosity, water holding capacity and water evaporation rates from litter were measured experimentally. Litter porosity decreased and water holding capacity increased over the course of a grow-out due to manure addition. Water evaporation rates at 25°C and 50% relative humidity ranged from 0.5 to 10L/m(2)/day. Evaporation rates increased with litter moisture content and air speed. Maintaining dry litter at the peak of a grow-out is likely to be challenging because evaporation rates from dry litter may be insufficient to remove the quantity of water added to the litter on a daily basis.