In vivo Cell CFSE Fluorescence Negative Staining for Detection of Super Paramagnetic Iron Oxide Nanoparticles Phagocytosed by Mouse Mononuclear Macrophage Leukemia Cells-RAW264.7 / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the feasibility and fluorescence characteristics of CFSE negative staining for in vivo cell imaging of super paramagnetic iron oxide nanoparticles (SPIO) phagocytosed by mouse mononuclear macrophage leukemia cells-RAW264.7.</p><p><b>METHODS</b>After labeled with SPIO, the RAW264.7 macrophages were stained with Prussian blue stain and CFSE fluorescence negative stain step by step. Furthermore, trypan blue staining was used to evaluate cell viability of cells which stained with CFSE. At last, laser scanning confocal microscope was used to measure SPIO in cells through CFSE fluorescence negative stain method.</p><p><b>RESULTS</b>SPIO within RAW264.7 macrophages showed blue in Prussian's blue staining, while showed negative area in CFSE negative staining. Good consistencies between Prussian's blue staining and CFSE negative staining were observed. In addition, RAW264.7 macrophages showed high viability after SPIO/CFSE dual-labeled method, proved by typan stain.</p><p><b>CONCLUSION</b>The CFSE fluorescence negative staining may be used for detecting SPIO that phagocytosed by RAW264.7 macrophages and it is showed good consistency that confirmed one another when compared to classic Prussian' blue staining.</p>

Amperometric determination of ascorbic acid in pharmaceutical formulations by a reduced graphene oxide-cobalt hexacyanoferrate nanocomposite

Investigation of the redox properties of drugs and their determination are performed by electrochemical techniques. Data obtained from electrochemical techniques are often correlated with molecular structure and pharmacological activity of drugs. In this regard, different modified electrodes were applied as sensors for quantification of different drugs. A nanocomposite of reduced graphene oxide-cobalt hexacyanoferrate was synthesized by a simple precipitation route. Scanning electron microscopy revealed that the nanocomposite comprised nanoparticles of cobalt hexacyanoferrate attached to the reduced graphene oxide nanosheets. A nanocomposite-modified carbon paste electrode was then fabricated. It represented prominent activity toward the electrocatalytic oxidation of ascorbic acid, and the kinetics of the electrooxidation process was evaluated. Finally, an amperometric method was developed for the quantification of ascorbic acid in different pharmaceutical formulations

Development of bioconjugate from Streptomyces tyrosinase and gold nanoparticles for rapid detection of phenol constituents.

Most of the phenol compounds are toxic and have been considered as hazardous pollutants. Several physicochemical and biological methods are available to detect and monitor the phenol pollutants in water and soil. In the present study, phenol constituents of winery, paper and plastic industrial effluents were successfully detected employing tyrosinase-gold nanoparticles bioconjugate. The synthesis of extracellular tyrosinase and gold nanoparticles was achieved by a single isolate of Streptomyces sp. DBZ-39. Enhanced production (369.41 IU) of tyrosinase was produced in submerged bioprocess employing response surface method with central composite design. Extracellular gold nanoparticles synthesized (12-18 nm) by Streptomyces sp. DBZ-39 were characterized with TEM, EDAX and FTIR analysis. A rapid detection (within 10 min) of phenol constituents from winery effluents was achieved by bioconjugate, when compared to tyrosinases and gold nanoparticles independently. Streptomyces tyrosinase could exhibit relatively a better performance than commercially available mushroom tyrosinase in the detection of phenol constituents. Winery effluent has shown much higher content (0.98 O.D) of phenol constituents than paper and plastic effluents based on the intensity of color and U.V absorption spectra.

Serial MR Imaging of Magnetically Labeled Humen Umbilical Vein Endothelial Cells in Acute Renal Failure Rat Model

PURPOSE: To evaluate the usefulness of in vivo magnetic resonance (MR) imaging for tracking intravenously injected superparamagnetic iron oxide (SPIO)-labeled human umbilical vein endothelial cells (HUVECs) in an acute renal failure (ARF) rat model. MATERIALS AND METHODS: HUVECs were labeled with SPIO and poly-L-lysine (PLL) complex. Relaxation rates at 1.5-T MR, cell viability, and labeling stability were assessed. HUVECs were injected into the tail vein of ARF rats (labeled cells in 10 rats, unlabeled cells in 2 rats). Follow-up serial T2*-weighted gradient-echo MR imaging was performed at 1, 3, 5 and 7 days after injection, and the MR findings were compared with histologic findings. RESULTS: There was an average of 98.4+/-2.4% Prussian blue stain-positive cells after labeling with SPIO-PLL complex. Relaxation rates (R2*) of all cultured HUVECs at day 3 and 5 were not markedly decreased compared with that at day 1. The stability of SPIO in HUVECs was maintained during the proliferation of HUVECs in culture media. In the presence of left unilateral renal artery ischemia, T2*-weighted MR imaging performed 1 day after the intravenous injection of labeled HUVECs revealed a significant signal intensity (SI) loss exclusively in the left renal outer medulla regions, but not in the right kidney. The MR imaging findings at days 3, 5 and 7 after intravenous injection of HUVECs showed a SI loss in the outer medulla regions of the ischemically injured kidney, but the SI progressively recovered with time and the right kidney did not have a significant change in SI in the same period. Upon histologic analysis, the SI loss on MR images was correspondent to the presence of Prussian blue stained cells, primarily in the renal outer medulla. CONCLUSION: MR imaging appears to be useful for in vivo monitoring of intravenously injected SPIO-labeled HUVECs in an ischemically injured rat kidney.

Aflatoxin contamination in stored rice variety PAU 201 collected from Punjab, India.

Background & objectives: The present study was carried out on stored rice variety PAU 201 in Punjab that was not permitted for milling and public distribution due to the presence of damaged grains at levels exceeding the regulatory limits of 4.75 per cent. The aim of the study was to determine fungal and aflatoxin contamination in the rice samples to assess hazard from the presence of damaged grains. Presence of iron in discoloured rice grains was also assessed. Methods: Stored samples of paddy of PAU 201 rice variety were collected from six districts of Punjab, milled and analysed for presence of fungal and aflatoxin contamination. Scanning electron microscopy (SEM), energy dispersive X-ray (EDX) analysis and Prussian blue staining was used to determine fungal spores and presence of iron, respectively. Results: Aflatoxin analysis of rice samples indicated that none exceeded the Food Safety and Standards (Contaminants, Toxins and Residues) Regulations, 2011 tolerance limit of 30 μg/kg and majority of the samples had levels <15 μg/kg. The proportion of damaged grains exceeding the limit of 5 per cent was observed in 85.7 per cent of the samples. SEM and Prussian blue staining and EDX analysis of black tipped and pin point damaged rice grains did not show presence of fungal structures and presence of iron. Interpretation & conclusions: The results of the study indicated that the stored rice samples did not pose any health concern with respect to aflatoxin contamination as per the criteria laid down by the Food Safety and Standards Authority of India.

In vitro MRI and Characterization of Rat Mesenchymal Stem Cells Transduced with Ferritin as MR Reporter Gene

PURPOSE: This study was performed to evaluate the characteristics of rat mesenchymal stem cells (RMSCs) transduced with human ferritin gene and investigate in vitro MRI detectability of ferritin-transduced RMSCs. MATERIALS AND METHODS: The RMSCs expressing both myc-tagged human ferritin heavy chain subunit (myc-FTH) and green fluorescence protein (GFP) were transduced with lentiviurs. Transduced cells were sorted by GFP expression using a fluorescence-activated cell sorter. Myc-FTH and GFP expression in transduced cells were detected by immunofluorescence staining. The cell proliferative ability and viability were assessed by MTT assay. The RMSC surface markers (CD29+/CD45-) were analyzed by flow cytometry. The intracellular iron amount was measured spectrophotometically and the presence of ferritin-iron accumulation was detected by Prussian blue staining. In vitro magnetic resonance imaging (MRI) study of cell phantoms was done on 9.4 T MR scanner to evaluate the feasibility of imaging the ferritin-transduced RMSCs. RESULTS: The myc-FTH and GFP genes were stably transduced into RMSCs. No significant differences were observed in terms of biologic properties in transduced RMSCs compared with non-transduced RMSCs. Ferritin-transduced RMSCs exhibited increased iron accumulation ability and showed significantly lower T2 relaxation time than non-transduced RMSCs. CONCLUSION: Ferritin gene as MR reporter gene could be used for non-invasive tracking and visualization of therapeutic mesenchymal stem cells by MRI.

Suppressive Effect of Zinc on the Formation of Colonic Preneoplastic Lesions in the Mouse Fed High Levels of Dietary Iron

We investigated the effect of zinc on the formation of colonic aberrant crypt foci induced by azoxymethane (AOM) followed by dextran sodium sulfate (DSS) in mice with high iron diet (HFe; 450 ppm iron). Six-week old ICR mice were fed on high iron diets with combination of three different levels of zinc in diets, low-zinc (LZn; 0.01 ppm), medium-zinc (MZn; 0.1 ppm), and high-zinc (HZn; 1 ppm) for 12 weeks. Animals were received weekly intraperitoneal injections of AOM (10 mg/kg B.W. in saline) for 3 weeks followed by 2% DSS (molecular weight 36,000~50,000) in the drinking water for a week. To confirm the iron storage in the body, the hepatic iron concentration has been determine chemically and compared with histological assessment visualized by Prussian blue reaction. Aberrant crypt (AC) and aberrant crypt foci (ACF) were analyzed in the colonic mucosa of mouse fed high dietary iron. Superoxide dismutase (SOD) activity and thiobarbituric acid-reactive substances (TBARS) level were also investigated. Apoptosis in the preneoplastic lesion was determined by terminal deoxynucleotidyl transferase-mediated dUTP nickend labeling (TUNEL). In addition, immunohistochemistry of beta-catenin was also performed on the mucous membrane of colon. The number of large ACF (> or = 4 AC/ACF), which possess greater tumorigenic potential, was significantly lower in MZn and HZn groups compared with LZn group. Cytosolic SOD activity in the liver was significantly higher in HZn group compared with LZn group. Hepatic MDA level was decreased significantly in HZn group compared with MZn and LZn groups. Apoptotic index was significantly higher in HZn group. Taken together, these findings indicate that dietary zinc might exert a protective effect against colonic preneoplastic lesion induced by AOM/DSS in ICR mice with high iron status, and suggest that dietary supplement of zinc might play a role in suppressing colon carcinogenesis in mice.

Monitoring Transplanted Human Mesenchymal Stem Cells in the Penile Cavernosal Tissues of Streptozotocin-induced Diabetic Rats Using Molecular Magnetic Resonance Imaging / 대한남성과학회지

PURPOSE: A molecular magnetic resonance (MR) imaging technique using superparamagnetic iron oxide (SPIO) nanocrystals was developed for monitoring stem cells noninvasively. This study was performed to investigate whether the presence of transplanted human mesenchymal stem cells in the penile cavernosum of a diabetic rat model could be evaluated noninvasively using molecular MR imaging. MATERIALS AND METHODS: SPIO nanocrystals (Feridex; AMI, Cambridge, USA) were transferred to human mesenchymal stem cells (hMSCs) using GenePORTER. The SPIO-transferred hMSCs were examined with Prussian blue staining. SPIO-labeled hMSCs were transplanted to the penile cavernosum of a diabetic rat model and MR images were examined in vivo using 1.5 T MR. RESULTS: SPIO was transferred to hMSCs successfully. MR signal intensity at the areas of the SPIO-transferred hMSCs decreased in the penile cavernosum of the diabetic rat model. SPIO particles were confirmed in the transplanted penile cavernosum with Prussian blue staining. CONCLUSIONS: The SPIO-labelled hMSCs in the penile cavernosum of a diabetic rat model can be monitored noninvasively using molecular MR imaging.

Osmium [VIH]-catalyzed oxidation of indoles by potassium hexacyanoferrate [III] in alkaline media: a kinetic and mechanistic study

The reaction of indoles with Os [VIII] catalyzed hexacyanoferrate [III] in alkaline media to produce the corresponding oxindole has been studied at constant temperature and ionic strength. The reaction followed first order kinetics with respect to [indole], [OH[-] and [Os [VIII]], fractional order in [Fe[CN][6][-3]. The effects of added electrolytes, potassium hexacyanoferrate [II], relative permitivity and temperature have also been studied. A mechanism consistent with the kinetic data is proposed. Furthermore, the structural influence of the indoles on the reactivity has been discussed

Kinetics and mechanism of the oxidation of indoles by alkaline potassium hexacyanoferrate [III]

The reaction of indoles with hexacyanoferrate [III] in alkaline media to produce the corresponding oxindole has been studied at constant temperature and ionic strength. The reaction followed first order kinetics with respect to [indole] and [OH[-], whereas fractional order with respect to [Fe[CN][6][-3]] was found. The effects of added electrolytes, potassium hexacyanoferrate [II], relative permitivity and temperature have also been studied. On the basis of experimental observations, a probable reaction mechanism has been proposed

Thallium poisoning, Iraq

Two Iraqi families developed symptoms of thallium poisoning after consuming a meal on January 21st. Patients have been transferred to Amman for care at the Jordanian Hospitals. The Poison Laboratory in Baghdad had detected thallium in specimens taken from patients and foods. Ten family members were identified as cases; all presented with abdominal pain, vomiting and dysphagia. Within 4 days, 9 [90%] case-patients developed lower extremity pain and paresthesis and hair loss. Three case-patients died and one is clinically dead [case fatality rate=40%]. Median onset of illness was 24 hours [range: 12-72 hours]. WHO has sent blood, urine and cake specimens to Birmingham; Toxicology Laboratory to confirm and measure the concentration of Thallium in different specimens [according to IATA regulations to ensure the safe delivery of the specimens]; and facilitated procurement of Prussian blue antidote through contacting the UK Department of Health as it was not available in Jordan and neighboring countries. Intentional contamination is suspected and a legal investigation by authorities in Baghdad is underway. Information and results from this investigation are still pending. [Source: A report from WHO Office, Iraq]

Presence of Iron in Colorectal Adenomas and Adenocarcinomas

BACKGROUND/AIMS: Iron overload reportedly increases the risk of colorectal neoplasms, but the distribution of tissue iron in a colorectal neoplasm remains controversial. In this study, we attempted to determine the significance of tissue iron in colorectal adenomas and adenocarcinomas. METHODS: This study investigated 138 colorectal neoplasms (54 adenocarcinomas, 25 adenomas with high-grade dysplasia, and 59 adenomas with low-grade dysplasia) that were removed by surgical or endoscopic resection in Konkuk University Hospital between August 2005 and August 2006. Adjacent normal colon tissues and colorectal neoplasms were stained with Perls' Prussian blue to reveal ferric compounds. RESULTS: Positive Perls' staining was evident in 35.2% (19/54) of the adenocarcinomas and 22.6% (19/84) of the adenomas, and in only 2.2% (3/138) of the samples of adjacent normal colon tissue (p<0.001). Iron appears to reside exclusively in the stroma and outside the gland, rather than in the epithelial cells. Iron expression was strong in larger (p=0.004) and pedunculated (p<0.001) adenomas, and in all types of adenocarcinomas regardless of their size, shape, and location. CONCLUSIONS: The frequent presence of iron in the stroma of large adenomas, pedunculated adenomas, and adenocarcinomas indicates that iron deposition is a secondary phenomenon to intralesional hemorrhage rather than a consequence of epithelial-cell carcinogenesis.

Instrumentaçäo sônica X Canal Finder System: eficiência da remoçäo dentinária / Sonic instrumentation X Canal Finder System: removal efficiency

Os autores avaliaram, qualitativamente, a uniformidade da remoçäo dentinária em canais radiculares impregnados com ferrocianeto de prata, quando da utilizaçäo de duas técnicas mecânicas de instrumentaçäo: a sônica e com o Canal Finder System. Analisando o canal radicular por terços e cotejando a impregnaçäo remanescente em cada parede (vestibular, lingual, mesial, distal), verificaram que diferença estatisticamente significante ocorreu na parede mesial do terço médio e parede lingual do terço apical para o Canal Finder System e apenas na parede lingual do terço cervical tal diferença foi significativa para o aparelho sônico MM 1500

Estudo qualitativo da uniformidade da remoçäo dentinária em canais radiculares impregnados com ferrocianeto de prata: instrumentaçäo manual x canal "Finder System" / The studies the quality of root canal preparation by menas of impregnation with silver ferrocyanide using the conventional technique for root canal preparation and the Canal Finder System

Os autores avaliaram, qualitativamente, a uniformidade da remoçäo dentinária em canais radiculares impregnados com ferrocianeto de prata, quando da utilizaçäo de técnica manual e do emprego de aparato mecânico, o Canal Finder System. Analisando o canal radicular por terços e cotejando a impregnaçäo remanescente em cada parede (vestibular, lingual, mesial, distal), verificaram que o CFS mostrou-se mais eficiente do que a instrumentaçäo manual, especialmente nas paredes distal e lingual do terço cervical e vestibular, e lingual do terço médio

Role of caudate nucleus on modulation of blood pressure responses in cat.

Changes in blood pressure, heart rate and respiration were studied in 15 cats on bilateral stimulation of rostral, middle and caudal regions of caudate nucleus. Hypotensive response was obtained on stimulation of anterior caudate nucleus, which changed to pressor response when posterior caudate nucleus was stimulated. Biphasic blood pressure responses were recorded on stimulation of middle caudatal area. No detectable changes were observed on heart rate and respiration. This suggests a role for caudate nucleus as a modulator in blood pressure regulation.

Aplicaçäo trimestral do ferrocianeto de prata como agente cariostático em cárie incipiente de primeiros molares permanentes inferiores / Trimestrial application of silver ferrocyanide as a cariostatic agent in initial caries of the first inferior permanent molars inferiores

Estudo referente a eficácia da aplicaçäo trimestral do ferrocianeto de prata como agente cariostático em cárie incipiente de primeiros molares permanentes inferiores, foi realizado em alunos, na faixa etária de 05 a 07 anos, de ambos os sexos, de três escolas públicas, na cidade do Recife-Pernambuco, durante o período de abril a dezembro de 1988. O exame clínico odontológico efetuado em 390 alunos, evidenciou a presença, em 60 alunos, dos dois primeiros molares permanentes inferiores irrompidos e com cárie incipiente. Foram constituídos dois grupos de 30 alunos; efetuadas as aplicaçÖes do ferrocianeto de prata, em dois períodos trimestrais sucessivos e, procedidas as duas avaliaçÖes de progressäo da cárie nos dentes experimental e controle. Os resultados demonstram que os índices percentuais de cárie sem progressäo e com progressäo foram diferentes em se tratando do dnete experimental ou do dente controle. Os valores percentuais, no dente experimental, permaneceram estáveis comprovando a açäo cariostática do ferrocianeto de prata e, a manutençäo dos seus efeitos nas duas avaliaçÖes atestou a validade da metodologia aplicada. A observaçäo clínica, näo evidenciou reaçÖes pulpares nos dentes tratados com o ferrocianeto de prata

Penetração da prata no esmalte de sulcos e fissuras após tratamento preventivo com ferrocianeto de prata / Penetration of silver into the enamel through pits and fissures after preventive treatment with silver ferrocyanide

Desde o século passado, substâncias químicas são utilizadas para prevenir e paralisar cáries de sulcos e fissuras. Visando contribuir para o assunto, os autores realizaram, "in vitro", um estudo com micro-sonda acoplada a microscópio eletrônico de varredura, no qual observaram a penetração da prata no esmalte, em regiões de sulcos e fissuras de molares humanos após tratamento dos mesmos com ferrocianeto de prata, condicionando ou não o esmalte previamente com ácido

Nickel nitrate and K-ferrocyanide in fixative for electron microscopy as tracer of extracellular space

El agregado de nitrato de niquel al glutaraldehido o al glutaraldehido-paraformaldehido, y su posterior refijación en osmio-ferrocianuro de potasio, tiene la facultad de funcionar como trazador de espacios intercelulares y favorecer la coloración posterior. Se realizaron observaciones con el microscopio electrónico de transmisión: en testículo, músculo cardíaco estriado esquelético, riñon e hígado. La fijación se realiza por perfusión vía vacular o por inmersión en el fijador. Asimismo se logra preservación de los detalles ultraestructurales

Preparo do canal radicular: aspectos clínicos relacionados com o acesso / Root canal preparation: clinical aspects related to the acces

Foram estudadas as conseqüências de acessos errôneos no preparo do canal radicular. Com os canais previamente impregnados com ferrocianeto de prata e através de tomadas radiográficas feitas antes e após o preparo, as áreas nao tocadas pelos instrumentos foram identificadas. A análise dos resultados permitiu concluir que acessos errôneos, sem obedecer formas de contorno, conveniência e sem o preparo adequado da entrada do canal, conduziram a uma instrumentaçao inadequada, com maior porcentagem de defeito nos terços cervical e médio, justamente os que apresentam o maior índice de contaminaçao

Acute thallium poisoning: prussian blue therapy in 9 cases

Thallium poisoning produces a complex and serious toxicity with multi-organ involvement. In Guyana, this condition has not been previously diagnosed. the present communication describes nine cases of acute thallium poisoning which were treated with Prussian Blue which is currently recommended by most authorities