Green Microspheres Research Articles

High-Sensitivity and Environmentally Friendly Humidity Sensors Deposited with Recyclable Green Microspheres for Wireless Monitoring.

The reliable, high-sensitive, wireless, and affordable requirements for humidity sensors are needed in high-precision measurement fields. Quartz crystal microbalance (QCM) based on the piezoelectric effect can accurately detect the mass changes at the nanogram level. However, water-capture materials deposited on the surface of QCM generally show disadvantages in either cost, sensitivity, or recyclability. Herein, novel QCM-based humidity sensors (NQHSs) are developed by uniformly depositing green microspheres (GMs) of natural polymers prepared by the chemical synthesis of the emulsification/inner gel method on QCM as humidity-sensitive materials. The NQHSs demonstrate high accuracy and sensitivity (27.1 Hz/% RH) owing to the various hydrophilic groups and porous nano-3D deposition structure. Compared with the devices deposited with a smooth film, the frequency of the NQHSs shows almost no changes during the cyclic test and exhibits long-term stability. The NQHSs have been successfully applied to non-contact sensing human activities and remote real-time humidity monitoring via Bluetooth transmission. In addition, the deposited humidity-sensitive GMs and QCM substrate are fully recycled and reused (72% of the original value). This work has provided an innovative idea to construct environmental-friendly, high-sensitivity, and wireless humidity sensors.

Improving Techniques to Study Equine Cervical Mucociliary Clearance

Postbreeding bacterial uterine infections inflict major losses on the equine industry. Microcurrents propelled by ciliated cells between the folds of the uterus and cervix have been proposed as a means by which contaminants are expelled. Previous data have shown possible ciliary microcurrents propelling carbon particles, occasionally rotating, through cervical folds. However, adherence to the epithelium may have interfered with movement of carbon in these studies. Therefore, we tested potentially nonadherent substances to reveal ciliary microcurrents on the equine cervix under high magnification videoendoscopy. We hypothesized that polyethylene green microspheres 1–5 and 70 μm in diameter, would be superior to carbon in revealing microcurrents on the cervical epithelium and that 50 μm hemispherically coated bichromal microspheres would display rotation. A suspension containing these microspheres and carbon was deposited onto the cervix of five estrous mares, and movement of each type of particle was recorded under high-magnification videoendoscopy for 10–20 minutes. Particles were subjectively assessed for movement between folds, past stationary points, in opposing directions and at different speeds. Visibility, aggregation, motion, and rotation were scored numerically and analyzed by the Kruskal–Wallis test. Backward rotation of bichromal spheres was interpreted as evidence of ciliary activity. Overall, carbon scored equal to or higher than the microspheres, leading to rejection of the hypothesis. Subjective assessment concluded that cervical movement was closely related to respiratory movements of the mare, and that the constantly moving cervical folds helped clear the deposited particles.

Regional blood flow changes underlying the hypotensive action of 5‐HT:Studies using Doppler and Microsphere technologies

Serotonin (5‐hydroxytryptamine, 5‐HT) causes a sustained decrease in mean arterial pressure (MAP) and elicits a fall in total peripheral resistance (TPR) in normal conscious rats. We hypothesized that the decrease in TPR induced by 5‐HT is caused by relaxation (direct or indirect) of arteries serving one or more major regional vascular beds. Alzet pumps containing vehicle or 5‐HT (25 ug/kg/min) were implanted in male Sprague‐Dawley rats and 24 hours later the rats were anesthetized and underwent: 1) ear and tail blood flow measurements using Doppler to determine cutaneous vascular contribution; or 2) left ventricular injection of fluorescent microspheres (300,000 yellow green microspheres) to determine global organ blood flow. Infusion of 5‐HT increased ear (69.1±20.5% time‐controlled vehicle) and tail (23.5±2.5%) blood flow (Doppler) while elevating small intestinal (50% increase) flow (microsphere) over vehicle‐infused rats. In the microsphere experiments there was no alteration in blood flow by 5‐HT infusion (vs vehicle) in spleen, liver, lung, right or left kidney, leg muscle, abdominal skin, visceral fat or brain, and right and left kidney flows were statistically similar (R; 3.78±0.69 vs L: 4.00±0.74 ml/min/g tissue, p> 0.05). These findings define the cutaneous and splanchnic vasculature as regions of interest to study the mechanism of 5‐HT ability to lower MAP and TPR. NIH HL081115.

Surface Properties of Toxoplasma gondii Oocysts and Surrogate Microspheres

The physical properties that govern the waterborne transmission of Toxoplasma gondii oocysts from land to sea were evaluated and compared to the properties of carboxylated microspheres, which could serve as surrogates for T. gondii oocysts in transport and water treatment studies. The electrophoretic mobilities of T. gondii oocysts, lightly carboxylated Dragon Green microspheres, and heavily carboxylated Glacial Blue microspheres were determined in ultrapure water, artificial freshwater with and without dissolved organic carbon, artificial estuarine water, and artificial seawater. The surface wettabilities of oocysts and microspheres were determined using a water contact angle approach. Toxoplasma gondii oocysts and microspheres were negatively charged in freshwater solutions, but their charges were neutralized in estuarine water and seawater. Oocysts, Glacial Blue microspheres, and unwashed Dragon Green microspheres had low contact angles, indicating that they were hydrophilic; however, once washed, Dragon Green microspheres became markedly hydrophobic. The hydrophilic nature and negative charge of T. gondii oocysts in freshwater could facilitate widespread contamination of waterways. The loss of charge observed in saline waters may lead to flocculation and subsequent accumulation of T. gondii oocysts in locations where freshwater and marine water mix, indicating a high risk of exposure for humans and wildlife in estuarine habitats with this zoonotic pathogen. While microspheres did not have surface properties identical to those of T. gondii, similar properties shared between each microsphere type and oocysts suggest that their joint application in transport and fate studies could provide a range of transport potentials in which oocysts are likely to behave.

ABCA7 expression is regulated by cellular cholesterol through the SREBP2 pathway and associated with phagocytosis

ABCA7 is highly homologous to ABCA1 and mediates cellular cholesterol and phospholipid release by apolipoproteins when transfected in vitro. However, expression of ABCA7 was downregulated by increased cellular cholesterol while ABCA1 was upregulated, and the results were consistent by forced expression or downregulation of sterol-responsive/regulatory element (SRE) binding proteins (SREBPs). We analyzed the promoter of the ABCA7 gene and identified the new exon encoding 96 bp (mouse) and 95 bp (human) of the 5' untranslated region and the transcription start site at 1,122 bp (mouse) and 1,260 bp (human) upstream of the initiation methionine codon. At 5' upstream of this exon is the ABCA7 proximal promoter containing multiple binding sites of transcription factors for hematopoiesis and SRE of 9 bp at 212 bp (mouse) and 179 bp (human) upstream of the new exon. The apolipoprotein A-I-mediated lipid release was not influenced by suppression of the endogenous ABCA7 with small interfering RNA in mouse fibroblasts or by its increase in ABCA1-deficient mouse cells. In contrast, phagocytic activity was altered in parallel to the ABCA7 expression in these cells. When phagocytosis was induced, the messages increased for SREBP2, ABCA7, and other SREBP2-regulated proteins. The ABCA1 message decreased in this condition. We conclude that the ABCA7 gene is regulated by sterol in the opposite direction to ABCA1 through SRE/SREBP2 and that expression of ABCA7 by this regulation is associated with phagocytic activity.

Pancreatic islet blood flow in conscious rats during hyperglycemia and hypoglycemia

Anesthesia affects general hemodynamics and regulation of organ perfusion. We used colored microspheres to measure pancreatic islet blood flow in conscious rats at two time points, during either hyperglycemia or hypoglycemia. This method, using black and green microspheres, was validated by comparison with previous microsphere experiments and by lack of effect of a nonmetabolizable glucose analog, 3-O-methylglucose, on islet perfusion. Basal and glucose-stimulated islet blood flow levels were similar in pentobarbital sodium-anesthetized and conscious rats. However, the basal distribution of pancreatic blood flow was altered by anesthesia (fractional islet blood flow 5.8 +/- 0.4% in conscious rats, 7.9 +/- 0.8% in pentobarbital-anesthetized rats, P < 0.05). Insulin-induced hypoglycemia significantly increased whole pancreatic blood flow in conscious rats, whereas islet blood flow remained unchanged and fractional islet blood flow was decreased (5.8 +/- 0.5% in the basal state, 4.2 +/- 0.4% during hypoglycemia, P < 0.001). Methylatropine pretreatment significantly increased islet blood flow during hypoglycemia by 181%. This result suggests that prevention of hypoglycemia-induced increase in islet perfusion may be mediated, at least in part, by a cholinergic, vagal muscarinic mechanism.

Sources of subcortical projections to the superior colliculus in the ferret

We have identified brainstem and other subcortical afferents to the superior colliculus (SC) in the ferret, by examining the pattern of retrograde labelling that resulted from unilateral injections of red and green fluorescent latex microspheres or of wheat germ agglutinin conjugated to horseradish peroxidase into different regions of this midbrain nucleus. Labelled neurons were found in many structures, including somatosensory, visual and auditory nuclei. These subcortical inputs are almost all bilateral, although most are primarily ipsilateral. Despite some differences in organization, the subcortical projections to the ferret SC broadly resemble those described in other species. Following injections of red and green microspheres in either the rostral and caudal or the medial and lateral regions of the SC, double-labelled cells usually accounted for less than 10% of the total number of retrogradely labelled cells, indicating that most of the subcortical neurons project to discrete regions of the SC. Many of these afferents show some topographic organization, which is much more evident along the rostrocaudal axis of the SC than in the medial-lateral dimension. The intercollicular projection is restricted mainly to the rostral SC and demonstrates a point-to-point organization. Most of the subcortical structures caudal to the central region of the SC were labelled mainly by the tracers injected in caudal SC, while those rostral to this region, including the zona incerta and the pretectal nuclei, were labelled largely by injections in rostral SC. These findings suggest that projections originating from nuclei posterior to the central region of the SC terminate principally in caudal SC whereas afferents from structures anterior to the central region project largely to rostral SC.

A method for labeling embryonic rat medial septal region projection neurons, in vitro, using fluorescent tracers

A retrograde labeling method is described in which rat embryonic (E18, E21) and postnatal (P7) medial septal neurons were labeled with succinyl wheat germ agglutin-fluorescein, fluorescent green microspheres, or 1,1′-dioctadecyl-3,3,3′,3′-tetramethyl-indocarbocyanine perchlorate (Dil) following in vitro hippocampal injections. The brains were removed and immediately immersed in oxygenated Tyrode solution. Dye was pressure injected into the hippocampus bilaterally. After incubating the brain in oxygenated Tyrode, the medial septal region was removed. The neurons were dissociated and cultured at medium density in 35 mm dishes with a hole in the bottom covered by a coverslip with a grid. The neurons were observed with a low light system, and cell counts were made at 5, 24, and 48 h. Labeled and unlabeled neurons showed considerable neurite outgrowth and acetylcholinesterase activity in culture. Highly reproducible labeling was obtained, with Dil giving the best results. Dil labeled the neurons in vitro, was retained during culture for 1 week, and was compatible with cell survival.

Coronary artery occlusion extends perfusion territory boundaries through microvascular collaterals.

Simultaneous in vivo infusions of two different colored 10 microns microsphere suspensions into the left anterior descending (LAD; red spheres) and left circumflex (LCx; blue spheres) coronary arteries of nine anesthetized dogs identified a specific region of canine myocardium perfused by both arterial branches. Subsequently, the LAD was ligated and a third (green) set of micropheres was infused into the patent LCx artery. Analysis of 40 microns serial sections of tissue revealed interface zones with capillaries perfused by both arteries. The first zone, defined as the Interface Transistion Zone (ITZ) was formed by an intermingling of microvessels supplied by the parent arteries of the adjacent perfusion territories; it separated tissue containing only one or the other colored microspheres. Another zone, defined as the Boundary Watershed Zone was located within the ITZ and had capillaries containing both red and blue microspheres. The width of ITZ was 53377 +/- 817 microns (mean +/- SD), and the width of the BWZ was 3358 +/- 618 microns. Green microspheres, infused into the LCx following coronary occlusion were also found in the ITZ and BWZ. Furthermore, capillaries perfused exclusively by the LAD before occlusion (tissue with red but not blue microspheres) adjacent to the perfusion interface contained green microspheres as well as red/green aggregates, indicating lateral extension of the LCx perfusion territory. This extension of the LCx territory was quantitated by comparing the location at which densities of green microspheres or green/red aggregates decreased abruptly compared to the location of the original ITZ and BWZ boundaries, respectively. Results showed that LAD occlusion caused a 24% expansion of the ITZ and a 48% expansion of the BWZ. In addition, all expansions were significantly greater in subepicardial compared to subendocardial regions (p < 0.001). These results clearly demonstrate the capability of microvascular anastomoses in providing blood flow to the periphery of an ischemic region. Furthermore, the perfusion interface is labile and might be amenable to manipulation.

The role of macrophages in particle translocation from lungs to lymph nodes.

Red fluorescent and green fluorescent microspheres were instilled into separate but adjacent areas of dog lung lobes. After 7 days, the tracheobronchial lymph nodes that drained both of the instilled areas contained many macrophages with all red or all green microspheres but rarely both. This indicates that the particles did not translocate passively and that lung macrophages phagocytized the microspheres in the lung and carried them to the tracheobronchial lymph nodes. In addition, two populations of pulmonary alveolar macrophages (PAM's), one that had phagocytized red microspheres in vivo and one that had phagocytized green microspheres, were lavaged from the lungs of dogs, mixed into one population, and instilled back into a previously unexposed lung lobe of the same dogs. As in the first experiment, the tracheobronchial lymph nodes that drained the instilled area contained numerous macrophages with either all red or all green microspheres. This suggested that the instilled PAM's had migrated to the tracheobronchial lymph nodes. Thus, lung macrophages, including PAM's probably play a critical role in the induction of lung immunity and in protection from disease by determining particle translocation.