Search for intracranial aneurysm susceptibility gene(s) using Finnish families

Background

Cerebrovascular disease is the third leading cause of death in the United States, and about one-fourth of cerebrovascular deaths are attributed to ruptured intracranial aneurysms (IA). Epidemiological evidence suggests that IAs cluster in families, and are therefore probably genetic. Identification of individuals at risk for developing IAs by genetic tests will allow concentration of diagnostic imaging on high-risk individuals. We used model-free linkage analysis based on allele sharing with a two-stage design for a genome-wide scan to identify chromosomal regions that may harbor IA loci.

Methods

We previously estimated sibling relative risk in the Finnish population at between 9 and 16, and proceeded with a genome-wide scan for loci predisposing to IA. In 85 Finnish families with two or more affected members, 48 affected sibling pairs (ASPs) were available for our genetic study. Power calculations indicated that 48 ASPs were adequate to identify chromosomal regions likely to harbor predisposing genes and that a liberal stage I lod score threshold of 0.8 provided a reasonable balance between detection of false positive regions and failure to detect real loci with moderate effect.

Results

Seven chromosomal regions exceeded the stage I lod score threshold of 0.8 and five exceeded 1.0. The most significant region, on chromosome 19q, had a maximum multipoint lod score (MLS) of 2.6.

Conclusions

Our study provides evidence for the locations of genes predisposing to IA. Further studies are necessary to elucidate the genes and their role in the pathophysiology of IA, and to design genetic tests.     

Membrane currents in identified lactotrophs of rat anterior pituitary

Qualitative features of the primary inward and outward current components of identified lactotrophs of the rat anterior pituitary were examined. Identification of lactotrophs in heterogeneous dissociated anterior pituitary cultures was accomplished by application of the reverse hemolytic plaque assay. Currents in lactotrophs were subsequently examined using whole-cell or patch recording techniques. Two components of inward calcium current were observed: a transient component and a sustained component. The transient component activated at voltages as negative as -50 mV and was the major contributor to total lactotroph calcium current. The sustained component activated at voltages above about -10 mV. The 2 currents could be qualitatively separated by differences in inactivation properties and in sensitivity to cadmium. At least 3 components of outward current were distinguished. Either 30 mM TEA or 0 calcium eliminated a major portion of sustained outward current. This is likely to represent primarily calcium- and voltage-activated potassium current. The remaining current could be further differentiated into a transient current component that could be inactivated with conditioning potentials above -60 mV. A slowly activating and deactivating potassium current remained following inactivation of the transient current. Although the time course of the transient current is reminiscent of "A" current, activation of this current required potentials above -30 mV. Candidates for the single-channel currents that underlie the whole-cell outward currents were observed in cell-attached recordings. When combined with patch-clamp electrophysiological methods, the reverse hemolytic plaque assay promises to be a powerful technique for the electrophysiological characterization of specific cell subtypes in heterogeneous dissociated cell populations.     

Comparison of physiological, cytopathogenic and immunological properties between two environmental isolates of Acanthamoeba spp

The aim of this study was to determine whether pathogenic and less-pathogenic isolates of environmental Acanthamoeba exhibit differences in adhesion to human erythrocytes. Based on physiological properties of temperature, tolerance, and rapid growth, Acanthamoeba were divided into pathogenic and less-pathogenic isolates. Acanthamoeba were tested for their ability to produce cytopathic effects (CPE) using two human cell lines, HEp-2 and KB cells. Both ameba isolates caused CPE to both cell lines with the same pattern without significant difference. Human erythrocytes from 20 healthy volunteers were used to study the erythrocyte reactivity of Acanthamoeba by co-incubation with trophozoites. The pathogenic Acanthamoeba exhibited significantly higher erythrocyte adhesion as compared to the less-pathogens (p<0.05). Erythrocyte activity occurred in the presence of plasma in all blood samples, suggesting the role of plasmatic components and contact-dependent mechanisms to produce host cell cytotoxicity. The present results showed correlation between the physiological properties and erythrocyte reactivity of Acanthamoeba.     

Factorial design analysis of parameters for the sorption-enhanced steam reforming of ethanol in a circulating fluidized bed riser using CFD

The sorption-enhanced steam reforming of ethanol (SESRE) has recently been reported as a novel process for hydrogen (H₂) production. SESRE can operate well on a Ni-based catalyst with dolomite as a sorbent in packed-bed reactors. In this study, the circulating fluidized bed (CFB) concept was proposed to obtain higher productivity and continuous operation of SESRE. Particular focus was directed to the design and selection of suitable operating conditions of the CFB riser. Two-dimensional transient models using the Euler–Euler approach and the kinetic theory of granular flows were applied to investigate the H₂ production performance from a pilot-scale riser. The 2^k full factorial design method was utilized to examine the significances of five specific parameters, namely, the riser diameter, inlet temperature, catalyst-to-sorbent ratio, solid flux, and inlet gas velocity on two response variables, namely, H₂ purity and H₂ flux. From the ANOVA results, either the main effect or the interactions of each parameter were shown to be significant on both the H₂ purity and the H₂ flux, particularly the riser diameter and the solid flux. For optimizing the operation and reaction parameters, the best case was the system with riser diameter of 0.2 m, inlet temperature of 600 °C, catalyst-to-sorbent ratio of 2.54 kg kg⁻¹, solid flux of 200 kg m⁻² s⁻¹, and gas velocity of 3 m s⁻¹, obtaining H₂ purity of 91.30% on a dry basis with a significantly high H₂ flux of 0.147 kg m⁻² s⁻¹. The hydrodynamics showed that SESRE reached breakthrough within the bottom dense zone. However, incomplete conversion occurred in the core of the riser because of the very dilute bed.

The sorption-enhanced steam reforming of ethanol (SESRE) has recently been reported as a novel process for hydrogen (H₂) production.     

High prevalence of equine-like G3P[8] rotavirus in children and adults with acute gastroenteritis in Thailand

Group A rotavirus (RVA) is a major cause of acute gastroenteritis in infants and young children worldwide. This study aims to clarify the distribution of G/P types and genetic characteristics of RVAs circulating in Thailand. Between January 2014 and September 2016, 1867 stool specimens were collected from children and adults with acute gastroenteritis in six provinces in Thailand. RVAs were detected in 514/1867 (27.5%) stool specimens. G1P[8] (44.7%) was the most predominant genotype, followed by G3P[8] (33.7%), G2P[4] (11.5%), G8P[8] (7.0%), and G9P[8] (1.3%). Unusual G3P[9] (0.8%), G3P[10] (0.4%), G4P[6] (0.4%), and G10P[14] (0.2%) were also detected at low frequencies. The predominant genotype, G1P[8] (64.4%), in 2014 decreased to 6.1% in 2016. In contrast, the frequency of G3P[8] markedly increased from 5.5% in 2014 to 65.3% in 2015 and 89.8% in 2016. On polyacrylamide gel electrophoresis, most (135/140; 96.4%) of the G3P[8] strains exhibited a short RNA profile. Successful determination of the nucleotide sequences of the VP7 genes of 98 G3P[8] strains with a short RNA profile showed that they are all equine-like G3P[8] strains. On phylogenetic analysis of genome segments of two representative Thai equine-like G3P[8] strains, it was noteworthy that they possessed distinct NSP4 genes, one bovine-like and the other human-like. Thus, we found that characteristic equine-like G3P[8] strains with a short RNA electropherotype are becoming highly prevalent in children and adults in Thailand.     

Calcium-dependent epileptogenesis in an in vitro model of stroke-induced "epilepsy"

PURPOSE: Stroke is the most common cause of acquired epilepsy. The purpose of this investigation was to characterize the role of calcium in the in vitro, glutamate injury-induced epileptogenesis model of stoke-induced epilepsy. METHODS: Fura-2 calcium imaging and whole-cell current clamp electrophysiology techniques were used to measure short-term changes in neuronal free intracellular calcium concentration and long-term alterations in neuronal excitability in response to epileptogenic glutamate injury (20 microM, 10 min) under various extracellular calcium conditions and in the presence of different glutamate-receptor antagonists. RESULTS: Glutamate injury-induced epileptogenesis was associated with prolonged, reversible elevations of free intracellular calcium concentration during and immediately after injury and chronic hyperexcitability manifested as spontaneous recurrent epileptiform discharges for the remaining life of the cultures. Epileptogenic glutamate exposure performed in solutions containing low extracellular calcium, barium substituted for calcium, or N-methyl-d-aspartate (NMDA)-receptor antagonists reduced the duration of intracellular calcium elevation and inhibited epileptogenesis. Antagonism of non-NMDA-receptor subtypes had no effect on glutamate injury-induced calcium changes or the induction epileptogenesis. The duration of the calcium elevation and the total calcium load statistically correlated with the development of epileptogenesis. Comparable elevations in neuronal calcium induced by non-glutamate receptor-mediated pathways did not cause epileptogenesis. CONCLUSIONS: This investigation indicates that the glutamate injury-induced epileptogenesis model of stroke-induced epilepsy is calcium dependent and requires NMDA-receptor activation. Further, these experiments suggest that prolonged, reversible elevations in neuronal free intracellular calcium initiate the long-term plasticity changes that underlie the development of injury-induced epilepsy.     

Anticonvulsant activities of the FS-1 subfraction isolated from roots of Delphinium denudatum

Delphinium denudatum Wall. (Ranunculaceae) is a medicinal herb used for the treatment of epilepsy in the subcontinent. The present study reports the anticonvulsant activities in the maximal electroshock test (MEST) and subcutaneous pentylenetetrazole (PTZ), bicuculline (BIC), picrotoxin (PIC)-induced seizures of the FS-1 subfraction (FS-1) that was obtained by purification of an aqueous fraction isolated from the roots of D. denudatum. In CF 1 mice, FS-1 (600 mg/kg i.p.) exhibited very potent anticonvulsant activity that was comparable to the effects of the well-known antiepileptic drug phenytoin (20 mg/kg) in MEST and protected 100% animals from hind limb tonic extension phase of this model. FS-1 also suppressed PTZ-induced threshold seizure and the loss of the righting reflex with tonic fore and hind limb extension by 100%, similar to the antiepileptic drug valproic acid (350 mg/kg). BIC-induced seizures were suppressed in 80% of the animals. FS-1 exhibited weak anticonvulsant effect on PIC-induced seizures, however, it significantly reduced mortality and delayed the onset of seizures. FS-1 had no effect on strychnine (STN)-induced extensor seizures. The results demonstrate the broad and potent anticonvulsant activity of the compounds in FS-1 of D. denudatum.     

In vitro inhibitory effects of plant-based foods and their combinations on intestinal ¿-glucosidase and pancreatic ¿-amylase

ABSTRACT: BACKGROUND: Plant-based foods have been used in traditional health systems to treat diabetes mellitus. The successful prevention of the onset of diabetes consists in controlling postprandial hyperglycemia by the inhibition of alpha-glucosidase and pancreatic alpha-amylase activities, resulting in aggressive delay of carbohydrate digestion to absorbable monosaccharide. In this study, five plant-based foods were investigated for intestinal alpha-glucosidase and pancreatic alpha-amylase. The combined inhibitory effects of plant-based foods were also evaluated. Preliminary phytochemical analysis of plant-based foods was performed in order to determine the total phenolic and flavonoid content. METHODS: The dried plants of Hibiscus sabdariffa (Roselle), Chrysanthemum indicum (chrysanthemum), Morus alba (mulberry), Aegle marmelos (bael), and Clitoria ternatea (butterfly pea) were extracted with distilled water and dried using spray drying process. The dried extracts were determined for the total phenolic and flavonoid content by using Folin-Ciocateu's reagent and AlCl3 assay, respectively. The dried extract of plant-based food was further quantified with respect to intestinal alpha-glucosidase (maltase and sucrase) inhibition and pancreatic alpha-amylase inhibition by glucose oxidase method and dinitrosalicylic (DNS) reagent, respectively. RESULTS: The phytochemical analysis revealed that the total phenolic content of the dried extracts were in the range of 460.0-230.3 mg gallic acid equivalent/ g dried extract. The dried extracts contained flavonoid in the range of 50.3-114.8 mg quercetin equivalent/g dried extract. It was noted that the IC50 values of chrysanthemum, mulberry and butterfly pea extracts were 4.24+/-0.12 mg/ml, 0.59+/-0.06 mg/ml, and 3.15+/-0.19 mg/ml, respectively. In addition, the IC50 values of chrysanthemum, mulberry and butterfly pea extracts against intestinal sucrase were 3.85+/-0.41 mg/ml, 0.94+/-0.11 mg/ml, and 4.41+/-0.15 mg/ml, respectively. Furthermore, the IC50 values of roselle and butterfly pea extracts against pancreatic alpha-amylase occurred at concentration of 3.52+/-0.15 mg/ml and 4.05+/-0.32 mg/ml, respectively. Combining roselle, chrysanthemum, and butterfly pea extracts with mulberry extract showed additive interaction on intestinal maltase inhibition. The results also demonstrated that the combination of chrysanthemum, mulberry, or bael extracts together with roselle extract produced synergistic inhibition, whereas roselle extract showed additive inhibition when combined with butterfly pea extract against pancreatic alpha-amylase. CONCLUSIONS: The present study presents data from five plant-based foods evaluating the intestinal alpha-glucosidase and pancreatic alpha-amylase inhibitory activities and their additive and synergistic interactions. These results could be useful for developing functional foods by combination of plant-based foods for treatment and prevention of diabetes mellitus.     

Host-specific genetic variation of highly pathogenic avian influenza viruses (H5N1)

The complete genome sequences of two isolates A/chicken/Egypt/CL6/07 (CL6/07) and A/duck/Egypt/D2br10/07 (D2br10/07) of highly pathogenic avian influenza virus (HPAI) H5N1 isolated at the beginning of 2007 outbreak in Egypt were determined and compared with all Egyptian HPAI H5N1 sequences available in the GenBank. Sequence analysis utilizing the RNA from the original tissue homogenate showed amino acid substitutions in seven of the viral segments in both samples. Interestingly, these changes were different between the CL6/07 and D2br10/07 when compared to other Egyptian isolates. Moreover, phylogenetic analysis showed independent sub-clustering of the two viruses within the Egyptian sequences signifying a possible differential adaptation in the two hosts. Further, pre-amplification analysis of H5N1 might be necessary for accurate data interpretation and identification of distinct factor(s) influencing the evolution of the virus in different poultry species.     

Development of pharmacoresistance to benzodiazepines but not cannabinoids in the hippocampal neuronal culture model of status epilepticus

Status epilepticus (SE) is a life-threatening neurological disorder associated with a significant morbidity and mortality. Benzodiazepines are the initial drugs of choice for the treatment of SE. Despite aggressive treatment, over 40% of SE cases are refractory to the initial treatment with two or more medications. It would be a major advance in the clinical management of SE to identify novel anticonvulsant agents that do not lose their ability to treat SE with increasing seizure duration. Cannabinoids have recently been demonstrated to regulate seizure activity in brain. However, it remains to be seen whether they develop pharmacoresistance upon prolonged SE. In this study, we used low Mg(2+) to induce SE in hippocampal neuronal cultures and in agreement with animal models and human SE confirm the development of resistance to benzodiazepine with increasing durations of SE. Thus, lorazepam (1 microM) was effective in blocking low Mg(2+) induced high-frequency spiking for up to 30 min into SE. However, by 1 h and 2 h of SE onset it was only 10-15% effective in suppressing SE. In contrast, the cannabinoid type-1 (CB1) receptor agonist, WIN 55,212-2 (1 microM) in a CB1 receptor-dependent manner completely abolished SE at all the time points tested even out to 2 h after SE onset, a condition where resistance developed to lorazepam. Thus, the use of cannabinoids in the treatment of SE may offer a unique approach to controlling SE without the development of pharmacoresistance observed with conventional treatments.