Three-Dimensional “Honeycomb” Culture System that Helps to Maintain the Contractile Phenotype of Vascular Smooth Muscle Cells

Vascular smooth muscle cells (VSMCs) in the normal aorta are described as having a contractile phenotype because they can contract and do not proliferate. VSMCs in pathological conditions such as atherosclerosis and restenosis can proliferate and migrate, but lose their ability to contract, which is referred to as a synthetic phenotype. VSMCs show plasticity by changing their phenotype according to the surrounding environment. When VSMCs are cultured on a plastic plate, which is a normal two-dimensional culture system, they display the synthetic phenotype because they proliferate and migrate without contraction. Recently, we successfully cultured VSMCs that display features similar to the contractile phenotype, using type I collagen three-dimensional matrices, “honeycombs,” in the presence of abundant fetal bovine serum albumin. VSMCs cultured in honeycombs stop proliferating and can contract. The honeycomb culture system can maintain VSMCs in the contractile phenotype for a long period of time. In this chapter, we show the method of this new culture system and the characteristics of VSMCs in honeycombs. It is expected that the use of this culture system will generate new information on the characteristics of VSMCs

ENSINO DE CAMPO: SUA IMPORTÂNCIA NA FORMAÇÃO DO ENFERMEIRO

The authors describe the clinical teaching as an essential element in nurses's training, giving its characteristics, advantages and requirements.As autoras abordam o ensino de campo como elemento essencial na formação de enfermeiros, dando suas características, vantagens e requisitos

Regulation of Pharmaceuticals in Developing Nations: A Case from Nepal

Department of Drug Administration (DDA), medicine regulatory authority of Nepal, is aligned towards achieving Universal Health Coverage through the regulation of medicines, pharmacies, manufacturing facilities and medicine testing laboratories from its offices located in Kathmandu, Birgunj, Biratnagar and Nepalgunj and takes legal or administrative actions as appropriate.  It has its own regulatory laboratory, National Medicines Laboratory for testing and analysis of medicines. Besides, DDA reports suspected ADR to Upshala Monitoring Centre, Sweden, provides drug information and conducts regulatory trainings. Unlike in the developed world, DDA may not be able to fulfill all of its objectives. But then, a mechanism of regulation of pharmaceuticals is functional and is dynamic as per national need.https://doi.org/10.21423/jrs-v04n04p018 (DOI assigned 5/14/2019

Assessment of physical premises of selected pharmacies of Nepal

Background: This work attempts to investigate the standards of physical premises of pharmacies in terms of cleanliness, ventilation, protection from physical environment, spaciousness and storage facilities of two selected districts of Nepal: Kathmandu and Kaski. Methods: A cross-sectional, observational study of 100 randomly selected retail and wholesale pharmacies (55 from Kathmandu and 45 from Kaski) was conducted for their compliance to provisions of physical premises: cleanliness, ventilation, protection from physical environment, spaciousness and storage facilities. Preliminarily, scoring was done by converting 5-point Likert scale to a 100-point scale as follows: 1 = 0–20, 2 = 21–40, 3 = 41–60, 4 = 61–80 and 5 = 81–100. These scores were then reported as poor (score ⩽ 40), below average (score of 41–49), average (score of 50), above average (score of 51–60) and good (score ⩾ 61). Cronbach’s alpha for retails and wholesales obtained was .720 and .757, respectively. Results: In wholesales, protection from physical environment was least scored among other components, with the mean score 34.7 ± 8.3 for Kathmandu and 37.3 ± 13.3 for Kaski. Ventilation was scored 35.3 ± 6.3 for Kathmandu and 39.3 ± 11.6 for Kaski. All other components were obtained below average. However, in retails, the storage facilities were rated ‘poor’ with mean score 36.3 ± 13.9 for Kathmandu and 36.0 ± 11.9 for Kaski. Most other components were scored above average except for ventilation which was scored ‘good’. It was found that there was no significant difference between physical premises of the pharmacies of Kathmandu and Kaski in terms of all the indicators. The findings suggest that physical premise of many pharmacies was not found appropriate as provisioned in the good pharmacy practice requirements of Codes on Sales and Distribution of Drugs. Conclusion: Most pharmacies were found non-compliant to the aforementioned pharmacy premise components. Therefore, key partners should come up with commitment for increasing compliance to the legislative provisions of physical premises for ensuring quality and safety of medicines

OxyStressSens: Quantum dot@Multi-spin-cyclodextrin sensor for mapping of oxidative stress in cells using EPR and optical imaging

The best strategy of fighting “free radical diseases” (as cancer, neurodegeneration, atherosclerosis, etc.) is an effective early stage diagnosis. This study proposes a new concept to realize revolutionary class contrast agents to be applied as sensitive probes for detection of “oxidative stress” during development and progression of different pathologies, by using optical and magnetic resonance imaging techniques.We developed a new “oxidative stress” sensor (OxiStressSens) based on conjugation of quantum dots with multi-nitroxide-functionalized cyclodextrin. The oxidation of nitroxide residues to their radical form was accompanied by strong quenching of quantum dot fluorescence and appearance of high EPR contrast. In opposite, the reduction of nitroxide residues in their hydroxylamine form was accompanied by rapid decay of EPR signal and appearance of strong fluorescence signal. Thus, OxyStressSens allows evaluation of the balance between reducers/oxidizers and level of “oxidative stress”. Besides the large magnetic moment or high quantum yield, the dimension of OxiStressSens and its positive charge ensure enhance permeability and retention effect in cells and tissues. The sensor enters into viable cells, which was visualized by EPR and fluorescent imaging. OxyStressSens was applied for detection of “oxidative stress” in cell suspensions, treated by rotenone and 2-methoxyestradiol – a combination, inducing mitochondrial dysfunction, overproduction of superoxide and mild or severe oxidative damages. The physicochemical characteristics and easy cell permeability give a reason to believe that the sensor will be appropriate for in vivo imaging. Since the T1-weigheted MRI contrast of nitroxides follows the same dynamics as their EPR contrast, the sensor is applicable also for nitroxide-enhanced MRI studies.Acknowledgements: This study was supported by the Japanese Society for Promotion of Science (JSPS), Grant-in-aid “Kakenhi C”.ISER-450th International Conference on Nanoscience, Nanotechnology & Advanced Materials (IC2NAM-2018

Magnetic Resonance Imaging of Very Small Disruptions of Blood Vessel Permeability in Hypercholesterolemia Mice Using Nitroxide Probes

第5回分子イメージング学

Imaging of redox-imbalance and oxidative stress in kidney in vivo, induced by dietary cholesterol

We used a mitochondria-penetrating nitroxide, mito-TEMPO, as a contrast probe for imaging of kidney dysfunction in mice, based on the redox-imbalance and oxidative stress in the renal tissues. Kidney dysfunction was triggered by hypercholesterolemia. The mice were divided in three groups: (i) on normal diet (ND; control); (ii) on cholesterol diet (CD); (iii) on cholesterol plus cholestyramine diet (CC). CD mice showed increased plasma levels of total cholesterol and non-HDL-cholesterol, as well as increased serum levels of blood urea nitrogen, uric acid and creatinine, compared to ND mice. CC mice showed slightly increased plasma levels of total cholesterol and HDL-cholesterol, but not non-HDL-cholesterol, compared to ND mice. The serum levels of blood urea nitrogen, uric acid and creatinine in CC mice were equal to those in ND mice. The MRI signal of mito-TEMPO in the kidneys was characterized by: high intensity and long life-time in CD mice, indicating a high oxidative capacity of renal tissues; poor intensity and short life-time in ND mice, indicating a high reducing capacity of renal tissues; moderate intensity and relatively short life-time in CC mice, which shows the protective effect of lipid-lowering agents against oxidative damage. The data suggest that hypercholesterolemia induces redox-imbalance and oxidative stress in kidneys and this process could be visualized using MRI and mito-TEMPO as a redox-sensitive contrast substance

Imaging of redox-imbalance and oxidative stress in kidney in vivo, induced by dietary cholesterol

We used a mitochondria-penetrating nitroxide, mito-TEMPO, as a contrast probe for imaging of kidney dysfunction in mice, based on the redox-imbalance and oxidative stress in the renal tissues. Kidney dysfunction was triggered by hypercholesterolemia. The mice were divided in three groups: (i) on normal diet (ND; control); (ii) on cholesterol diet (CD); (iii) on cholesterol plus cholestyramine diet (CC). CD mice showed increased plasma levels of total cholesterol and non-HDL-cholesterol, as well as increased serum levels of blood urea nitrogen, uric acid and creatinine, compared to ND mice. CC mice showed slightly increased plasma levels of total cholesterol and HDL-cholesterol, but not non-HDL-cholesterol, compared to ND mice. The serum levels of blood urea nitrogen, uric acid and creatinine in CC mice were equal to those in ND mice. The MRI signal of mito-TEMPO in the kidneys was characterized by: high intensity and long life-time in CD mice, indicating a high oxidative capacity of renal tissues; poor intensity and short life-time in ND mice, indicating a high reducing capacity of renal tissues; moderate intensity and relatively short life-time in CC mice, which shows the protective effect of lipid-lowering agents against oxidative damage. The data suggest that hypercholesterolemia induces redox-imbalance and oxidative stress in kidneys and this process could be visualized using MRI and mito-TEMPO as a redox-sensitive contrast substance