Nurses Practices Regarding Medication Administration through a Central Venus Catheter

Objective:  The purpose of this study was to examine nurses’ practices related to medication administration through the CVC. Material and Methods:  A cross-sectional observational study was conducted at Tertiary Hospital. The study subjects were the female Nurses aged between 21 – 60 years (n = 133). A convenient sampling technique was used.Data collected through an observational checklist which contain two sections; demographic and 19 items of their practices. A pilot study was done on 20 participants to assess the reliability of the observational checklist. Data analyzed by using SPSS version 25. Results:   82.7% participants’ ages were between 21 – 30 years, 13.5% were between 31 – 40 years and 3.8% were between 41 – 50 years. Participants’ education was as follows: general nursing diploma (84.2%), BS Nursing Post RN (registered nurse) (11.3%), BSN -Bachelor Science in Nursing (Generic) (4.5%). Only 41.4% participants performed hand hygiene before procedure and 58.6% did not perform. 34.6% participants remove gloves and perform hand hygiene; remaining 65.4% did not perform hand hygiene correctly. All participants 100% performed documentation on drug chart. Conclusion:  The significance of quality care, combined with good practices, particularly when medicines are administered through the Central Venous Catheter. Incorrect practices of medication administration through CVC increase the risk of blood stream infection. This research study shows 50% unsatisfactory practices

Synthesis and structural features of copper(II) complexes of N,N,N′,N′-tetramethylethylenediamine with 2-chlorobenzoate1− and 2-hydroxybenzoate1−

Published online 22 Jan 2019.Peer reviewedPostprin

A CEP104-CSPP1 Complex Is Required for Formation of Primary Cilia Competent in Hedgehog Signaling

CEP104 is an evolutionarily conserved centrosomal and ciliary tip protein. CEP104 loss-of-function mutations are reported in patients with Joubert syndrome, but their function in the etiology of ciliopathies is poorly understood. Here, we show that cep104 silencing in zebrafish causes cilia-related manifestations: shortened cilia in Kupffer's vesicle, heart laterality, and cranial nerve development defects. We show that another Joubert syndrome-associated cilia tip protein, CSPP1, interacts with CEP104 at microtubules for the regulation of axoneme length. We demonstrate in human telomerase reverse transcriptase-immortalized retinal pigmented epithelium (hTERT-RPE1) cells that ciliary translocation of Smoothened in response to Hedgehog pathway stimulation is both CEP104 and CSPP1 dependent. However, CEP104 is not required for the ciliary recruitment of CSPP1, indicating that an intra-ciliary CEP104-CSPP1 complex controls axoneme length and Hedgehog signaling competence. Our in vivo and in vitro analyses of CEP104 define its interaction with CSPP1 as a requirement for the formation of Hedgehog signaling-competent cilia, defects that underlie Joubert syndrome

Cholesterol transfer via endoplasmic reticulum contacts mediates lysosome damage repair

Lysosome integrity is essential for cell viability, and lesions in lysosome membranes are repaired by the ESCRT machinery. Here, we describe an additional mechanism for lysosome repair that is activated independently of ESCRT recruitment. Lipidomic analyses showed increases in lysosomal phosphatidylserine and cholesterol after damage. Electron microscopy demonstrated that lysosomal membrane damage is rapidly followed by the formation of contacts with the endoplasmic reticulum (ER), which depends on the ER proteins VAPA/B. The cholesterol-binding protein ORP1L was recruited to damaged lysosomes, accompanied by cholesterol accumulation by a mechanism that required VAP-ORP1L interactions. The PtdIns 4-kinase PI4K2A rapidly produced PtdIns4P on lysosomes upon damage, and knockout of PI4K2A inhibited damage-induced accumulation of ORP1L and cholesterol and led to the failure of lysosomal membrane repair. The cholesterol-PtdIns4P transporter OSBP was also recruited upon damage, and its depletion caused lysosomal accumulation of PtdIns4P and resulted in cell death. We conclude that ER contacts are activated on damaged lysosomes in parallel to ESCRTs to provide lipids for membrane repair, and that PtdIns4P generation and removal are central in this response.Peer reviewe

Generation of stable transgenic hTERT-RPE1 cell lines for imaging analysis of cilia tip proteins at high temporal and spatial resolution

The primary cilium is a signaling organelle present on the cell surface of most eukaryotic cells. The organelle integrates several signaling pathways important for biological processes such as tissue homeostasis and embryonic development. Disruption of the primary cilium structure and/or function leads to a wide range of developmental diseases termed ciliopathies. Joubert syndrome is an autosomal recessive ciliopathy with mutations found in several ciliary protein encoding genes, including CSPP1 and CEP104. CSPP-L (predominant isoform of CSPP1) and CEP104 interact biochemically and are involved in cilia formation, but their ciliary function is not clarified at molecular level. In the present study, hTERT-RPE1 cell lines were generated that stably express mNeonGreen-CSPP-L or mNeonGreen-CEP104, respectively, at levels not interfering with cell cycle progression or cilia formation. The mNeonGreen-CEP104 cell line is of particular importance since immunofluorescence compatible CEP104 specific antibodies are no longer commercially available. Study of the ciliary localization of CSPP-L and CEP104 fusion proteins by 3D-Structured Illumination Microscopy resolved previously undetermined structural details of CEP104 and CSPP-L at the centrosome and the ciliary tip, and identified co-localization of endogenous CSPP-L and mNeonGreen-CEP104 at the cilia tip. Ciliary entry of mNeonGreen-CEP104 was found to be CSPP-L independent, supporting the hypothesis that intra-ciliary interaction of CEP104 and CSPP-L is required for formation of cilia of normal length. Finally, this thesis lays foundation for advanced live cell imaging studies of the potential dynamic behavior of these important proteins at the centrosome and the cilia tip

3D-Structured Illumination Microscopy of Centrosomes in Human Cell Lines

The centrosome is the main microtubule-organizing center of animal cells, and is composed of two barrel-shaped microtubule-based centrioles embedded in protein dense pericentriolar material. Compositional and architectural re-organization of the centrosome drives its duplication, and enables its microtubule-organizing activity and capability to form the primary cilium, which extends from the mature (mother) centriole, as the cell exits the cell cycle. Centrosomes and primary cilia are essential to human health, signified by the causal role of centrosome- and cilia-aberrations in numerous congenic disorders, as well as in the etiology and progression of cancer. The list of disease-associated centrosomal proteins and their proximitomes is steadily expanding, emphasizing the need for high resolution mapping of such proteins to specific substructures of the organelle. Here, we provide a detailed 3D-structured illumination microscopy (3D-SIM) protocol for comparative localization analysis of fluorescently labeled proteins at the centrosome in fixed human cell lines, at approximately 120 nm lateral and 300 nm axial resolution. The procedure was optimized to work with primary antibodies previously known to depend on more disruptive fixation reagents, yet largely preserves centriole and centrosome architecture, as shown by transposing acquired images of landmark proteins on previously published transmission electron microscopy (TEM) images of centrosomes. Even more advantageously, it is compatible with fluorescent protein tags. Finally, we introduce an internal reference to ensure correct 3D channel alignment. This protocol hence enables flexible, swift, and information-rich localization and interdependence analyses of centrosomal proteins, as well as their disorder-associated mutations

CSPP1 stabilizes growing microtubule ends and damaged lattices from the luminal side

Microtubules are dynamic cytoskeletal polymers, and their organization and stability are tightly regulated by numerous cellular factors. While regulatory proteins controlling the formation of interphase microtubule arrays and mitotic spindles have been extensively studied, the biochemical mechanisms responsible for generating stable microtubule cores of centrioles and cilia are poorly understood. Here, we used in vitro reconstitution assays to investigate microtubule-stabilizing properties of CSPP1, a centrosome and cilia-associated protein mutated in the neurodevelopmental ciliopathy Joubert syndrome. We found that CSPP1 preferentially binds to polymerizing microtubule ends that grow slowly or undergo growth perturbations and, in this way, resembles microtubule-stabilizing compounds such as taxanes. Fluorescence microscopy and cryo-electron tomography showed that CSPP1 is deposited in the microtubule lumen and inhibits microtubule growth and shortening through two separate domains. CSPP1 also specifically recognizes and stabilizes damaged microtubule lattices. These data help to explain how CSPP1 regulates the elongation and stability of ciliary axonemes and other microtubule-based structures.</p

Synthesis and structural features of copper(II) complexes of N,N,N′,N′-tetramethylethylenediamine with 2-chlorobenzoate¹⁻ and 2-hydroxybenzoate¹⁻

<p>Two copper(II) coordination complexes, formulated as [Cu(tmen)(Clba)₂] (<b>1</b>) and [Cu(tmen)(Hsal)₂·H₂O] (<b>2</b>) (where tmen = N,N,N′,N′-tetramethyl ethylenediamine (C₆H₁₆N₂), Clba¹⁻ = 2-chlorobenzoate (C₇H₄ClO₂¹⁻), and Hsal¹⁻ (C₇H₅O₃¹⁻ = monoanion of <i>o</i>-hydroxybenzoic acid (salicylic acid)), have been synthesized and characterized by elemental combustion analysis, spectroscopic techniques, thermal studies, and single crystal X-ray analyses. Complex <b>1</b> consists of two distinct monomeric units in which the coordination environment around the central copper(II) ion is a distorted octahedron with a CuN₂O₄ chromophore, constituted by a chelating tmen molecule, and two 2-chlorobenzoate¹⁻ anions coordinated through their carboxylate-O atoms in an asymmetrical bidentate fashion. Complex <b>2</b> is also a monomer and consists of an CuN₂O₃ chromophore, in which tmen is coordinated to Cu(II) through its two N atoms in a chelating bidentate fashion, and an aqua-O and the two <i>o</i>-hydroxybenzoate¹⁻ (HSal¹⁻) anions are coordinated through one of their carboxylate-O atoms in a monodentate mode, forming a square pyramidal structure. Hydrogen bonding interactions especially of O–H…O, N–H…O, and C–H…Cl types interweave monomeric units and stabilize the overall crystal structures in both complexes. Thermal analysis and antibacterial activities of <b>1</b> and <b>2</b> against various bacterial strains were also investigated.</p