Activation of Wnt/β-Catenin Pathway in Monocytes Derived from Chronic Kidney Disease Patients

Patients with chronic kidney disease (CKD) have significantly increased morbidity and mortality resulting from infections and cardiovascular diseases. Since monocytes play an essential role in host immunity, this study was directed to explore the gene expression profile in order to identify differences in activated pathways in monocytes relevant to the pathophysiology of atherosclerosis and increased susceptibility to infections. Monocytes from CKD patients (stages 4 and 5, estimated GFR <20 ml/min/1.73 m²) and healthy donors were collected from peripheral blood. Microarray gene expression profile was performed and data were interpreted by GeneSpring software and by PANTHER tool. Western blot was done to validate the pathway members. The results demonstrated that 600 and 272 genes were differentially up- and down regulated respectively in the patient group. Pathways involved in the inflammatory response were highly expressed and the Wnt/β-catenin signaling pathway was the most significant pathway expressed in the patient group. Since this pathway has been attributed to a variety of inflammatory manifestations, the current findings may contribute to dysfunctional monocytes in CKD patients. Strategies to interfere with this pathway may improve host immunity and prevent cardiovascular complications in CKD patients.     

Intra-rater repeatability of gait parameters in healthy adults during self-paced treadmill-based virtual reality walking

Self-paced treadmill walking is becoming increasingly popular for the gait assessment and re-education, in both research and clinical settings. Its day-to-day repeatability is yet to be established. This study scrutinised the test-retest repeatability of key gait parameters, obtained from the Gait Real-time Analysis Interactive Lab (GRAIL) system. Twenty-three male able-bodied adults (age: 34.56 ± 5.12 years) completed two separate gait assessments on the GRAIL system, separated by 5 ± 3 days. Key gait kinematic, kinetic, and spatial-temporal parameters were analysed. The Intraclass-Correlation Coefficients (ICC), Standard Error Measurement (SEM), Minimum Detectable Change (MDC), and the 95% limits of agreements were calculated to evaluate the repeatability of these gait parameters. Day-to-day agreements were excellent (ICCs > 0.87) for spatial-temporal parameters with low MDC and SEM values, <0.153 and <0.055, respectively. The repeatability was higher for joint kinetic than kinematic parameters, as reflected in small values of SEM (<0.13 Nm/kg and <3.4°) and MDC (<0.335 Nm/kg and <9.44°). The obtained values of all parameters fell within the 95% limits of agreement. Our findings demonstrate the repeatability of the GRAIL system available in our laboratory. The SEM and MDC values can be used to assist researchers and clinicians to distinguish ‘real’ changes in gait performance over time.     

Alcohol Enhances Acinetobacter baumannii-Associated Pneumonia and Systemic Dissemination by Impairing Neutrophil Antimicrobial Activity in a Murine Model of Infection

Acinetobacter baumannii (Ab) is a common cause of community-acquired pneumonia (CAP) in chronic alcoholics in tropical and sub-tropical climates and associated with a >50% mortality rate. Using a murine model of alcohol (EtOH) administration, we demonstrated that EtOH enhances Ab-mediated pneumonia leading to systemic infection. Although EtOH did not affect neutrophil recruitment to the lungs of treated mice, it decreased phagocytosis and killing of bacteria by these leukocytes leading to increased microbial burden and severity of disease. Moreover, we determined that mice that received EtOH prior to Ab infection were immunologically impaired, which was reflected in increased pulmonary inflammation, sequential dissemination to the liver and kidneys, and decreased survival. Furthermore, immunosuppression by EtOH was associated with deregulation of cytokine production in the organs of infected mice. This study establishes that EtOH impairs immunity in vivo exacerbating Ab infection and disease progression. The ability of Ab to cause disease in alcoholics warrants the study of its virulence mechanisms and host interactions.     

Determination of enalapril maleate and atenolol in their pharmaceutical products and in biological fluids by flow‐injection chemiluminescence

A chemiluminescent method using flow injection (FI) was investigated for rapid and sensitive determination of enalapril maleate and atenolol, which are used in the treatment of hypertension. The method is based on the sensitizing effect of these drugs on the Ce(IV)–sulfite reaction. The different experimental parameters affecting the chemiluminescence (CL) intensity were carefully studied and incorporated into the procedure. The method permitted the determination of 0.01–3.0 µg mL^?1 of enalapril maleate in bulk form with correlation coefficient r = 0.99993, lower limit of detection (LOD) 0.0025 µg mL^?1 (S/N = 2) and lower limit of quantitation (LOQ) 0.01 µg mL^?1. The linearity range of atenolol in bulk form was 0.01–2.0 µg mL^?1 (r = 0.99989) with LOD of 0.0003 µg mL^?1 (S/N = 2) and LOQ of 0.01 µg mL^?1. In biological fluids the linearity range of enalapril maleate was 0.1–2.0 µg mL^?1 in both urine and serum, and for atenolol the linearity range was 0.1–1.0 µg mL^?1 in both urine and serum. The method was also applied to the determination of the drugs in their pharmaceutical preparations. Copyright © 2009 John Wiley & Sons, Ltd.     

Exome sequencing reveled a compound heterozygous mutations in RTTN gene causing developmental delay and primary microcephaly

RTTN (Rotatin) (OMIM 614833) is a large centrosomal protein coding gene. RTTN mutations are responsible for syndromic forms of malformation of brain development, leading to polymicrogyria, microcephaly, primordial dwarfism, seizure along with many other malformations. In this study we have identified a compound heterozygous mutation in RTTN gene having NM_173630 c.5225A > G p.His1742Arg in exon 39 and NM_173630 c.6038G > T p.Cys2013Phe in exon 45 of a consanguineous Saudi family leading to brain malformation, seizure, developmental delay, dysmorphic feature and microcephaly. Whole exome sequencing (WES) techniques was used to identify the causative mutation in the affected members of the family. WES data analysis was done and obtained data were further confirmed by using Sanger sequencing analysis. Moreover, the mutation was ruled out in 100 healthy control from normal population. To the best of our knowledge the novel compound heterozygous mutation observed in this study is the first report from Saudi Arabia. The identified compound heterozygous mutation will further explain the role of RTTN gene in development of microcephaly and neurodevelopmental disorders.     

Pregnancy-specific protein B (PSPB), progesterone and some biochemical attributes concentrations in the fetal fluids and serum and its relationship with fetal and placental characteristics of Iraqi riverine buffalo (Bubalus bubalis)

This study was carried out to demonstrate the pregnancy-specific protein B (PSPB), progesterone and some biochemical parameters concentrations in amniotic fluid, allantoic fluid and fetal serum collected from slaughtered Iraqi riverine pregnant buffaloes at three different months of gestation (6th, 7th and 8th). Ten out of 22 adult buffaloes of 4.6 ± 0.97 years old were used in this study. The buffaloes were mated naturally by monitoring the estrus cycles via appearance of vaginal fluids and mounting by bulls. Pregnancy was checked for these buffaloes by non-returning to estrus for three estrus cycles and assured by rectal palpation on day 61 post-mating (PM). Buffaloes were slaughtered at three different periods of gestation (three at 6th month, four at 7th month and three at 8th month of gestation) to verify the progesterone and PSPB as well as some blood attributes levels (glucose, cholesterol, total protein, albumin, globulins and albumin: globulins ratio) in amniotic fluid (AF), allantoic fluid (LF) and fetal serum (FS). Progesterone was higher (P<0.01) in LF at the 8th month of gestation and lower in FS during the 7th and 8th months of pregnancy. PSPB concentrations were greater in FS (6th and 8th months in particular) than in both AF and LF. The overall mean of cholesterol concentration was higher in FS (P<0.05) followed by AF and LF that had the lowest concentration. The FS exhibited higher total protein during the three gestation periods. Most of fetal and placental measurements increased as the pregnancy advanced. In conclusion, these results described, for the first time, the PSPB and progesterone concentrations and blood characteristics in fetal fluids and serum in water riverine buffaloes during different stages of pregnancy. Progesterone concentrations were greater in allantoic fluid than in other fluids. In contrast, PSPB and other blood attributes were higher in fetal serum than other fluids of Iraqi riverine buffaloes. These findings reflect the changes in hormones, proteins and other metabolites during different gestation periods.     

Test of colonisation scenarios reveals complex invasion history of the red tomato spider mite Tetranychus evansi

The spider mite Tetranychus evansi is an emerging pest of solanaceous crops worldwide. Like many other emerging pests, its small size, confusing taxonomy, complex history of associations with humans, and propensity to start new populations from small inocula, make the study of its invasion biology difficult. Here, we use recent developments in Approximate Bayesian Computation (ABC) and variation in multi-locus genetic markers to reconstruct the complex historical demography of this cryptic invasive pest. By distinguishing among multiple pathways and timing of introductions, we find evidence for the "bridgehead effect", in which one invasion serves as source for subsequent invasions. Tetranychus evansi populations in Europe and Africa resulted from at least three independent introductions from South America and involved mites from two distinct sources in Brazil, corresponding to highly divergent mitochondrial DNA lineages. Mites from southwest Brazil (BR-SW) colonized the African continent, and from there Europe through two pathways in a "bridgehead" type pattern. One pathway resulted in a widespread invasion, not only to Europe, but also to other regions in Africa, southern Europe and eastern Asia. The second pathway involved the mixture with a second introduction from BR-SW leading to an admixed population in southern Spain. Admixture was also detected between invasive populations in Portugal. A third introduction from the Brazilian Atlantic region resulted in only a limited invasion in Europe. This study illustrates that ABC methods can provide insights into, and distinguish among, complex invasion scenarios. These processes are critical not only in understanding the biology of invasions, but also in refining management strategies for invasive species. For example, while reported observations of the mite and outbreaks in the invaded areas were largely consistent with estimates of geographical expansion from the ABC approach, historical observations failed to recognize the complex pathways involved and the corresponding effects on genetic diversity.     

Immobilization of wood-rotting fungi laccases on modified cellulose and acrylic carriers

Extracellular laccases produced by three different wood-rotting fungi, Cerrena unicolor, Heterobasidion annosum and Trametes versicolor, were immobilized via covalent bonds formation on DEAE-Granocel 500, CM-Granocel 500, and acrylic carriers. Out of the tested carriers, only the DEAE-Granocel 500, which was activated by divinyl sulphone appeared to be a suitable matrix for the expression of enzymic activity. Only one laccase of all the tested enzymes produced by C. unicolor showed the best binding to the carrier and a satisfactory enzymic activity. The immobilized laccase exhibited the highest enzymic activity at pH 5.2 and it was more resistant to thermal denaturation than the native enzyme. At 90 °C, it retained 75% activity compared to the free enzyme. It was also more stable during storage at 4 °C: after 4 months the immobilized laccase retained 98% of initial activity. Immobilized C. unicolor laccase was active in 10–60% concentration of methanol, acetone, isopropanol or acetonitrile. The best enzymic activity was observed in 20% solution of acetonitrile in buffer.