A flow cytometric method for the measurement of phagocytosis by polymorphonuclear leucocytes

A new method for the measurement of phagocytosis of Candida albicans by human polymorphonuclear leucocytes (PMN) is described using a fluorescence activated cell sorter. We have used acridine orange to discriminate between PMN which have internalised yeast particles and those which have not. This method allows accurate measurement of particle phagocytosis as an event distinct from particle adherence. It also permits detailed examination of the kinetics of phagocytosis, the study of which is likely to be of value in the investigation of diseases where abnormalities of PMN function are suspected.     

Secretin dissipates red aridine orange fluoescence from pancreatic duct epithelium

This study was undertaken to elucidate whether duct cells in the pancreas contain acidic cytoplasmic compartments regulated by secretion. Microdissected pancreatic ducts from pigs were examined by acridibe orange (AO) and 2′, 7′-biscarboxyethyl-5(6)-carboxyfluorescein/tetraacetioxymethyl ester (BCECF/AM) epifluorescence microscopy. Estimated cytoplasmic pH using BCECF fluorescence was 7.43pL0.04 and was not changed by altering CO₂ tension in the incubation mdium. The epithelium of acridine orange incbated peripheral interlobular pancreatic ducts exhibited green and fluorescence was sen in resting pancreatic ducts and was greatly accentuated by raising CO₂ in the incubation medium with chloroqyuine or NH₄Cl or the protonophores carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) or carbonyl cyanide M-chlorophenylhydrazone (CCCP), leaving uniform gren fluoresence. These findings suggest that pancreatic duct cells contain CO₂-dependent acidic compartments which vanishduring seceatin stimulation and which may be cytoplasmic tubulovesicles.     

Hypofluorescent spots in indocyanine green angiography of peau d‘orange and fundus

Purpose. Hypofluorescent spots were seen inindocyanine green (ICG) angiography of peau dorangefundus in eyes with angioid streaks. Origin of the hypofluorescentspots were examined with attention to their correlationwith a peau dorange appearance of the central fundususing a computer-assisted image comparison system. Methods. ICG angiography was performed in 5 patientshaving peau dorange appearance of fundus using ascanning laser ophthalmoscope (SLO) and a digitalvideo-fundus camera. The same central fundus areas corresponding to hypofluorescent spots in an ICGangiogram were then digitally identified in afluorescein angiogram and in a red-free picture in all10 eyes of the 5 patients. Monochromatic lightobservation was also performed with a dark fieldobservation using a SLO to see subretinal orintrachoroidal pigment clumping. Results. In no patient, the areas identified withhypofluorescent spots did show relevant changes ina fluorescein angiogram or a red-free picture. SLOexamination revealed not perfusion defect at the sameareas. The dark field observation showed no pigmentclumping at the peripapillary and papillomacularbundle regions where hypofluorescent spots were seen.Conclusions: Hypofluorescent spots seen in ICGangiograms did not show exact consistency with peau dorange changes in their location and shape. Perfusion defects or blocking by pigments were not acause of hypofluorescent spots. The scatteredhypofluorescent spots were considered to be relevantwith irregular affinity of the fundus to ICG dye.     

Characterization of lymphoid cell populations in bovine lymphosarcomas (“leukosis”) by flow-cytofluorometry

Acridine orange stained cell suspensions from ten cases of bovine lymphosarcoma were analyzed by flow-cytofluorometry. The green DNA-AO fluorescence showed in all cases, but one, two distinct separate lymphoid cell subpopulations. One corresponded to (normal) diploid lymphoid cells, the other had variable aneuploid values. The histograms of the latter cell populations also indicated cells in S- and G₂-phase. Low-angle light scatter histograms did not show a distinct separation between the different cell types, although two maxima could be distinguished. The relative proportions between the two cell populations varied between 20 and 80% with domination of one or the other cell type.There was some conformity to the histological and cytological picture in the ordinary light microscope, although neither a clear distinction in separable cell types nor differential counts was possible.The diploid lymphocyte subpopulation in the tumor might represent an immunoreactive process against the BLV-induced lymphosarcoma.     

吖啶橙染色法在膀胱癌无创诊断中的应用

[目的]运用吖啶橙染色荧光显微镜(acridineorenge,AO-F)检查法进行膀胱癌无创性诊断。[方法]采用AO-F检查法及巴氏染色分级(papanicolaousstaingradingsmethod,PG)检查法检测79例膀胱癌患者及80例非肿瘤患者的尿脱落细胞。[结果]两种染色法对诊断膀胱癌均有统计学意义,分层卡方检验,χMH2=173.461,P=0.000。巴氏染色法与吖啶橙荧光染色法比较,后者膀胱癌的检出率优于前者,χ2=19.606,P=0.000。巴氏染色法检出膀胱癌的敏感度为56.6%,特异度为100%。吖啶橙荧光染色法检出膀胱癌的敏感度为85.5%,特异度为100%。80例非膀胱癌患者两种尿脱落细胞检查的阳性检出率为0%。[结论]吖啶橙染色法比单从细胞形态上检测肿瘤细胞更具有重要的临床价值,可作为膀胱癌的辅助诊断方法。     

双波长双指示剂催化动力学光度法测定痕量铬(Ⅵ)

目的:建立测定痕量铬(Ⅵ)的双波长双指示剂催化动力学光度分析的新方法。方法:在稀硫酸介质中,利用痕量铬(Ⅵ)催化溴酸钾氧化甲基橙和结晶紫褪色的指示反应,通过测量510 nm和600 nm下,催化体系和非催化体系吸光度的变化,用双波长双指示剂催化动力学光度测定痕量铬。结果:测定铬的线性范围为0.0010~0.20μg/m l,检出限为1.1×10-10g/m l。结论:该法简单、灵敏度高、选择性好,可用于废水中痕量铬(Ⅵ)的测定。     

Physicochemical and functional properties of micronized jincheng orange by-products (Citrus sinensis Osbeck) dietary fiber and its application as a fat replacer in yogurt

Orange by-products from juice extraction are generally discarded or used in animal feed due to their low market value. However, orange by-products show potential as dietary fiber (DF) and fat replacers in products such as yogurt. This study assessed the benefits of using orange by-products in DF-enriched materials such as DF powders (OP) and micronized DF with ball-milling (MDF). The study also investigated the effects of adding different levels of OP and MDF on the quality of low-fat yogurt. Results show that MDF showed better physicochemical and functional properties than OP, and that 2% MDF as a fat replacer in yogurt retained most of the textural and sensory properties of full-fat yogurt. Therefore, this study showed that MDF is a promising alternative as a fat replacer in low-fat yogurt, without sacrificing good taste and other qualities of full-fat yogurt.     

High‐Fiber Orange Juice as a Nutrition Supplement in Women

Background: The daily consumption of dietary fiber is frequently below suggested recommendations. Using a double‐blind, controlled, randomized study, we assessed the efficiency and tolerance of a fiber‐enriched orange juice to supplement fiber intake in women. Materials and Methods: After 1 week of noninterventional observation, 192 healthy adult women ingested 400 mL of orange juice for 21 days, which either was not (placebo group) or was enriched with fiber (fiber group). Orange juice ingestion was registered daily and controlled for each week during the study period. Macronutrient, fiber, and energy intake were determined using a 3‐day food record, validated food chemical composition databases, and the “Pro Diet” software. Gastrointestinal symptoms were self‐evaluated daily by scoring 4 grades of symptom intensity and using a visual analog scale to grade pain severity. Results: No changes were observed for macronutrient and energy ingestion. For the placebo group (n = 97), the total fiber intake record was under the daily recommended value. In contrast, the fiber group (n = 95) displayed higher comparative values of total and soluble fiber consumption (P ≤ .001), achieving the daily recommended values of fiber intake. Both groups reported an increased frequency of slight bloating and rumbles over time (P ≤ .05). The fiber group also experienced a higher frequency of slight flatulence over time (P = .002). Conclusion: Consumption of fiber‐enriched orange juice was efficient to achieve the daily fiber intake recommendation for women, was not accompanied by intense adverse events, and may represent a suitable method to supplement fiber intake in woman.     

Association of Polyphenols from Oranges and Apples with Specific Intestinal Microorganisms in Systemic Lupus Erythematosus Patients

Our group has recently shown the existence of a gut microbial dysbiosis in systemic lupus erythematosus (SLE), supporting previous evidence involving intestinal bacteria in the initiation and amplification of autoimmune diseases. While several studies have addressed the use of dietary fibres to modify intestinal microbiota, information about other correlated components, such as polyphenols, is scarce. The aim of this work was to identify dietary components able to influence this altered microbiota in 20 SLE women and 20 age-matched controls. Food intake was recorded by means of a food frequency questionnaire. The intake of fibres was calculated from Marlett tables, and Phenol-Explorer was used for polyphenol consumption. Results showed positive associations between flavone intake and Blautia, flavanones and Lactobacillus, and dihydrochalcones and Bifidobacterium in the SLE group. Regarding the controls, dihydroflavonols were directly associated with Faecalibacterium, whereas flavonol intake was inversely associated with Bifidobacterium. From the food sources of these polyphenols related to microbiota, orange intake was directly associated with Lactobacillus and apple with Bifidobacterium in SLE, whilst red wine was the best contributor to Faecalibacterium variation. The association between common foods and particular microbial genera, reported to be decreased in SLE, could be of great importance for these patients.     

Young red blood cells; observations about the cytometric and morphologic alterations in the beginning of their life

Summary A recently described separation technique consisting of a combination of counterflow centrifugation and subsequent density (Percoll) scparation was tested for its ability to enrich red cell populations with young cells in comparison to either separation technique alone. The relative age of every fraction was determined by HbAlc measurements, resulting in the lowest HbAlc for the combination method. Conventional reticulocyte counting and floweytometric counting with thiazole orange indicated that in the youngest fractions the combination method showed the highest reticulocyte counts. There was a good correlation between manual and flowcytometric counting results. Radio-iron studies showed a two-fold enrichment with young cells in the fraction with the lowest HbAIc acquired by the combination technique in comparison to the other two methods. Cytometric measurements showed that the fractions with the lowest HbAlc were the ones with the highest MCV and MCH and the lowest MCHC. Besides loss of their RNA-material, young cells already seem to loose water and haemoglobin like older cells, resulting in a decrease of MCV and MCH and in increase in MCHC. It is concluded that combining counterflow centrifugation with subsequent density fractionation results in superior enrichment with young cells in comparison to the results of each method alone.