Golgi-type I and Golgi-type II Neurons in the Ventral Anterior Thalamic Nucleus of the Adult Human: Morphological Features and Quantitative Analysis

The morphological and quantitative features of neurons in the adult human ventral anterior thalamic nucleus were studied in Golgi preparations. Two neuronal types were found and their quantitative features were studied. Golgi-type I neurons were medium to large cells with dense dendritic trees and dendritic protrusions and short hair-like appendages. They have somatic mean diameter of 30.8 μm (±9.4, n = 85). They have an average 100.3 dendritic branches, 48.97 dendritic branching points, and 58.85 dendritic tips. The mean diameters of their primary, secondary, and tertiary dendrites were 3.1 μm (±1, n = 80), 1.85 μm (±0.8, n = 145), and 1.5 μm (±0.4, n = 160), respectively. Golgi-type II neurons were small to medium cells with few sparsely branching dendrites and dendritic stalked appendages with or without terminal swellings. They have somatic mean diameters of 22.2 μm (±5.8, n = 120). They have an average 33.76 dendritic branches, 16.49 dendritic branching points, and 21.97 dendritic tips. The mean diameters of their primary, secondary, and tertiary dendrites were 1.6 μm (±0.86, n = 70), 1.15 μm (±0.55, n = 118), and 1 μm (±0.70, n = 95), respectively. These quantitative data may form the basis for further quantitative studies involving aging or some degenerative diseases that may affect cell bodies and/or dendritic trees of the Golgi-type I and/or Golgi-type II thalamic neurons.     

The role of fluoride and chlorhexidine in preserving hardness and mineralization of enamel and cementum after gamma irradiation

The purpose of this study was to evaluate the effect of 0.05% sodium fluoride and 0.12% chlorhexidine mouthwashes on the micro-hardness of tooth enamel and cementum that was exposed to therapeutic doses of gamma radiation. Sixty extracted human teeth were divided into two groups, one was irradiated, the other was not irradiated. The two groups were further subdivided into three subgroups, which were each treated either with 0.05% sodium fluoride or with 0.12% chlorhexidine; the third subgroup served as a control. After demineralization–remineralization cycling, teeth from the irradiated groups showed a significantly lower micro-hardness when compared to those from the non-irradiated groups. Both in the irradiated and non-irradiated groups, teeth from the control subgroups showed a significantly lower micro-hardness, as compared to teeth treated with sodium fluoride and chlorhexidine. For non-irradiated enamel samples, those treated with chlorhexidine showed a significantly less micro-hardness compared to those treated with sodium fluoride. In contrast, irradiated enamel showed no significant difference in micro-hardness, whatever treatment (chlorhexidine or sodium fluoride) was applied. For cementum, treatment with chlorhexidine resulted in a significantly lower micro-hardness compared to sodium fluoride, both for the irradiated and non-irradiated groups. It is concluded that gamma irradiation with therapeutic doses typically used for head and neck carcinoma treatment has a direct effect in reducing micro-hardness of tooth enamel and cementum. Mouthwash protocols including, for example, application of 0.05% sodium fluoride or 0.12% chlorhexidine three times per day for 6 weeks, can protect enamel and cementum against the reduction in hardness and demineralization caused by gamma irradiation. Sodium fluoride offers more protection compared to chlorhexidine.     

The Mauritian Piracy Act: A Comment on the Director of Public Prosecutions v Ali Abeoulkader Mohamed Decision

In 2011 Mauritius adopted the Piracy and Maritime Violence Act (the Act). The Act does not expressly state that Mauritian Courts have jurisdiction over offenses committed outside of Mauritius. In Director of Public Prosecutions v Ali Abeoulkader Mohamed & Ors, the Mauritian Supreme Court dealt with the issue of whether the Act applied to non-Mauritius citizens where the alleged piracy acts had been committed outside of Mauritius on the high seas. This article assesses the Supreme Court's decision and suggest ways that the Act can be strengthened.     

Detection,isolation, and characterization of chikungunya viruses associated with the Pakistan outbreak of 2016–2017

The chikungunya virus(CHIKV) is a mosquito-transmitted alphavirus, which has infected millions of people in Africa, Asia, Americas, and Europe since it reemerged in India and Indian Ocean regions in 2005–2006. Starting in the middle of November 2016, CHIKV has been widely spread, and more than 4,000 cases of infections in humans were confirmed in Pakistan. Here, we report the first isolation and characterization of CHIKV from the Pakistan outbreak. Eight CHIKV strains were newly isolated from human serum samples using a cell culture procedure. A full-length genome sequence and eight complete envelope(E1) sequences of CHIKV from Pakistan were obtained in this study. Alignment of the CHIKV E1 sequences revealed that the eight new CHIKV isolates were highly homogeneous, with only two nonsynonymous substitutions found at generally conserved sites(E99 and Q235). Based on the comparison of 342 E1 sequences, the two nonsynonymous mutations were located in well-recognized domains associated with viral functions such as the cell fusion and vector specificity, suggesting their potential functional importance. Phylogenetic analysis indicated that the CHIKV strains from Pakistan originated from CHIKV circulating in the Indian region. This study helps elucidate the epidemics of CHIKV in Pakistan and also provides a foundation for studies of evolution and expansion of CHIKV in South Asia.     

Use and limitations of magnetic resonance phase-contrast assessment of coronary flow reserve in a model of collateral dependence

Phase-contrast magnetic resonance imaging (PC-MRI) is useful for assessing coronary artery flow reserves (CFR) in man and acute animal models with intermediate coronary lesions. The present study examines the use of PC-MRI for assessing CFR in a model with critical stenosis and collateral dependence. PC-MRI quantitative flow measurements from the proximal left anterior descending (LAD) and left circumflex (LCX) coronary arteries were compared with myocardial tissue perfusion reserve measurements (microsphere techniques) after placement of a 2.25-mm ameroid constrictor on the proximal LCX in a porcine model; measurements were obtained at implantation (n = 4) and at 3 to 4 weeks (n = 4) and 6 weeks (n = 5) postimplantation. CFR is defined as the ratio of maximal hyperemic flow to baseline flow. Hyperemia was induced using intravenous adenosine (140 mg/kg/min). Collateral dependence in the LCX distri bution was evidenced by angiographic findings of critical stenosis with minimal myocardial histological changes and normal baseline myocardial perfusion (microsphere techniques). In this setting, PC-MRI CFR was correlated with microsphere measures of perfusion reserve. Collateral dependence was confirmed by Evan's blue dye injection. This study provides angiographic, myocardial perfusion, and histological correlates associated with PC-MRI epicardial CFR changes during chronic, progressive coronary artery constriction. It also demonstrates the disparity between epicardial and myocardial measures of coronary flow reserve with collateral dependence and the caveats for PC-MRI use in models of progressive coronary constriction.     

Purification of a synthetic oligonucleotide by anion exchange chromatography: method optimisation and scale-up

A single-step chromatographic method for purification of a synthetic 20-mer oligonucleotide is described. Method optimisation was conducted at laboratory scale where 30 mg crude sample was purified per run with a yield of 17 mg pure oligonucleotide. The protocol was scaled-up in steps to achieve 5-, 58- and a final 230-fold scale-up. At the final scale, 7.0 g of crude material was purified with a yield of 4.1 g product. The purity of the oligonucleotide was in all scales higher than 97%. The cycle time was 110 min, which corresponds to a purification capacity of about 90 g crude oligonucleotide material per 24 h.     

A combination of three distinct trafficking signals mediates axonal targeting and presynaptic clustering of GAD65

The signals involved in axonal trafficking and presynaptic clustering are poorly defined. Here we show that targeting of the gamma-aminobutyric acid-synthesizing enzyme glutamate decarboxylase 65 (GAD65) to presynaptic clusters is mediated by its palmitoylated 60-aa NH(2)-terminal domain and that this region can target other soluble proteins and their associated partners to presynaptic termini. A Golgi localization signal in aa 1-23 followed by a membrane anchoring signal upstream of the palmitoylation motif are required for this process and mediate targeting of GAD65 to the cytosolic leaflet of Golgi membranes, an obligatory first step in axonal sorting. Palmitoylation of a third trafficking signal downstream of the membrane anchoring signal is not required for Golgi targeting. However, palmitoylation of cysteines 30 and 45 is critical for post-Golgi trafficking of GAD65 to presynaptic sites and for its relative dendritic exclusion. Reduction of cellular cholesterol levels resulted in the inhibition of presynaptic clustering of palmitoylated GAD65, suggesting that the selective targeting of the protein to presynaptic termini is dependent on sorting to cholesterol-rich membrane microdomains. The palmitoylated NH(2)-terminal region of GAD65 is the first identified protein region that can target other proteins to presynaptic clusters.     

Involvement of nitric oxide and potassium channels in the bradykinin-induced vasodilatation in the rat kidney perfused ex situ

The role of nitric oxide (NO), K(+) channels, and arachidonic acid metabolism, via cytochrome P450 and cyclooxygenase pathways, in the renal vasodilatory effect of bradykinin was examined in the isolated rat kidney perfused ex situ with a blood-free solution. Bradykinin (BK, 0.25-1.0 microM) induced a dose-dependent reduction of 10-35% in the relative renal vascular resistance (rRVR) of isolated kidneys preconstricted with phenylephrine (PHE, 0.17-0.35 microM). The vasodilating effect of 0.5 microM bradykinin was significantly inhibited by the nitric oxide synthase inhibitors, N(G)-nitro-L-arginine (95% inhibition) and N(G)-nitro-L-arginine methyl ester (45-75% inhibition). Clotrimazole, an inhibitor of cytochrome P450 pathway but not indomethacin, a cyclooxygenase inhibitor, reduced the renal vasodilator response to bradykinin by 84%. The nonspecific K(+) channel inhibitor, tetraethylammonium ion (TEA) and the selective inhibitor of Ca(2+)-activated K(+) channels, charybdotoxin (ChTX) greatly attenuated the vasodilator response to bradykinin by approximately 84% and 79%, respectively. These two K(+) channel inhibitors showed similar effects on vasodilatation induced by S-nitroso-acetyl-D,L-penicillamine (1 microM), a nitric oxide donor. The results suggest that bradykinin releases nitric oxide which, by opening potassium channels specifically the Ca(+)-dependent type, mediates the renal vasodilator response to bradykinin in the isolated kidney perfused ex situ.     

Electrospinning of collagen nanofibers

Electrospinning is a fabrication process that uses an electric field to control the deposition of polymer fibers onto a target substrate. This electrostatic processing strategy can be used to fabricate fibrous polymer mats composed of fiber diameters ranging from several microns down to 100 nm or less. In this study, we describe how electrospinning can be adapted to produce tissue-engineering scaffolds composed of collagen nanofibers. Optimizing conditions for calfskin type I collagen produced a matrix composed of 100 nm fibers that exhibited the 67 nm banding pattern that is characteristic of native collagen. The structural properties of electrospun collagen varied with the tissue of origin (type I from skin vs type I from placenta), the isotype (type I vs type III), and the concentration of the collagen solution used to spin the fibers. Electrospinning is a rapid and efficient process that can be used to selectively deposit polymers in a random fashion or along a predetermined and defined axis. Toward that end, our experiments demonstrate that it is possible to tailor subtle mechanical properties into a matrix by controlling fiber orientation. The inherent properties of the electrospinning process make it possible to fabricate complex, and seamless, three-dimensional shapes. Electrospun collagen promotes cell growth and the penetration of cells into the engineered matrix. The structural, material, and biological properties of electrospun collagen suggest that this material may represent a nearly ideal tissue engineering scaffold.     

Dengue viruses activity in Piauí, Brazil

The present paper reports a laboratory investigation performed between the years of 2000 and 2002 to study a virological surveillance program introduced in the state of Piauí to support an epidemiological survey of the disease. Dengue virus type 3 (DENV-3) existence in the state was detected in May 2002 when a high number of dengue cases due to DENV-1 and DENV-2 were reported. An assessment on the population knowledge about the disease and its transmission showed that almost 50% of the population were still unaware of the epidemiological features of dengue.