A Reminder of Methylene Blue's Effectiveness in Treating Vasoplegic Syndrome after On-Pump Cardiac Surgery

The inflammatory response induced by cardiopulmonary bypass decreases vascular tone, which in turn can lead to vasoplegic syndrome. Indeed the hypotension consequent to on-pump cardiac surgery often necessitates vasopressor and intravenous fluid support. Methylene blue counteracts vasoplegic syndrome by inhibiting the formation of nitric oxide.We report the use of methylene blue in a 75-year-old man who developed vasoplegic syndrome after cardiac surgery. After the administration of methylene blue, his hypotension improved to the extent that he could be weaned from vasopressors. The use of methylene blue should be considered in patients who develop hypotension refractory to standard treatment after cardiac surgery.     

Pericardial effusion simulating a tumor. The value of Doppler color echocardiography for its differentiation

We describe the case of a patient aged 64 with aortic valve disease and pericardial effusion. Echocardiographic evaluation showed an intrapericardial mass of about 7 cm of diameter with clotted appearance, adhered to the visceral leaf, at the level of the atrio-ventricular function. Because of this finding we performed color codified Doppler echocardiography, observing that this mass acquired a blue hue during diastole and mosaic hue at the end of the systole, realizing that this phenomenon was due to fluid retention. There are many publications about the value of echocardiography for the identification of pericardial masses and some of them show how an effusion can hide a tumor. The interest of this particular case lies in that the effusion simulated a tumor and by means of the color codified Doppler we could demonstrate that it corresponded to the stream of the pericardial fluid. During the surgical procedure the absence of the mass was corroborated.     

Spectrophotometric measurement of proteoglycans in osteoarthritic synovial fluid.

Cartilage glycosaminoglycans (GAGs) were measured by a spectrophotometric assay in synovial fluid obtained from 30 normal bovine hock joints and 15 osteoarthritic human knee joints. Results were compared with those obtained by radioimmunoassay (RIA). The spectrophotometric method (dimethylmethylene blue (DMB) assay) was found to be simple, safe, and sufficiently reproducible to be of potential value for serial measurement of sulphated GAGs in arthritic joint fluids. The nature of the proteoglycans present in normal bovine and osteoarthritic human synovial fluid was examined by gel chromatography. Whereas normal bovine synovial fluid contained only small molecular weight proteoglycans, osteoarthritic human synovial fluid contained aggregated proteoglycans and predominantly high molecular weight proteoglycan subunits.     

Sonography detection threshold for knee effusion

The aim of this study was to determine the threshold for detecting knee effusion by ultrasound (US). Five knee specimens from embalmed cadavers were studied. Intra-articular injection of saline, blood and synovial fluid was performed under ultrasound control and methylene blue dye instillation. The smallest amount of fluid detectable by US in the knee was 7.4 ml for synovial fluid, 10.1 and 10.4 ml for saline solution and 9.7 for blood. The threshold for detecting knee joint effusion by US in cadaver specimens was 10 ml for saline and blood and 7 ml for synovial fluid.Abbreviations US Ultrasound     

绵羊牦牛棘球蚴囊液及囊壁抗原的SDS—PAGE分析

本文应用十二烷基硫酸钠—聚丙烯酰胺凝胶电泳(SDS-PAGE)对绵羊、耗牛棘球蚴囊液及绵羊棘球蚴囊壁可溶性粗抗原的蛋白组分进行了分析。使用10％凝胶,采用垂直板型电泳、考马斯亮兰R—250染色进行SDS-PAGE的结果表明,绵羊及耗牛棘球蚴囊液分别含有16及11种蛋白组分,分子量范围分别为270～16.5KD及280～17KD;主带均为8条。绵羊棘球蚴囊壁可溶性粗抗原共显示17条蛋白带,分子量范围245～17KD,主带8条。本项研究为进一步分析分离特异性抗原奠定了基础。     

猪囊尾蚴表面蛋白及其抗原性的初步测定分析

本文初步分析了猪囊尾蚴囊壁表面蛋白的特性。根据囊尾蚴漂洗液PAGE和SDS-PAGE考马斯亮蓝染色带分别显示了9条及14条蛋白区带,其中大部分与宿主骨骼肌组织的可溶性蛋白有一致的迁移率,但其中第5、7、10条蛋白带为其独有。漂洗液与囊液兔高免血清IE可出现1条沉淀弧;用漂洗液蛋白抗原对35例囊虫病人血清做ELISA,阳性率为60％。此外,对囊虫表面宿主蛋白的可能作用,做了初步探讨。     

Role of peroxynitrite as a mediator of pathophysiological alterations in experimental pneumococcal meningitis.

This study investigated the role of peroxynitrite in an adult rat model of pneumococcal meningitis. Immunohistochemically, nitrotyrosine residues, as a marker for peroxynitrite formation, were detected perivascularly and in proximity to inflammatory cells in the subarachnoid space. Nitrotyrosine immunoreactivity was colocalized with blood-brain barrier breaching, which was visualized by fluorescence microscopy after intravenous application of Evans blue. Treatment of infected rats with uric acid (300 mg/kg intraperitoneally), a scavenger of peroxynitrite, significantly attenuated intracranial pressure, cerebrospinal fluid white blood cell count, and blood-brain barrier leakage, as indicated by Evans blue concentration in the cerebrospinal fluid (21.6+/-9.3 mm Hg, 5776+/-1790 cells/microL, 9.7+/-6.4 microgram/mL in infected, untreated rats vs. 7.2+/-1.6 mm Hg, 2004+/-904 cells/microL, 1.1+/-1.0 microgram/mL infected, uric acid-treated rats, mean+/-SD, P<.05). These data suggest that peroxynitrite plays a central role in mediating pathophysiological alterations during bacterial meningitis.     

Alternative pathways for drainage of cerebrospinal fluid in the canine brain

Although the brain has no formal lymphatic system, a substantial quantity of cerebrospinal fluid (CSF) has nonetheless been shown to drain via cervical lymphatics. To pursue further the issue of alternative drainage pathways for CSF, we infused a solution of Ringer's lactate (RL) into the cisterna magna of the dog brain and monitored both the flow and concentration of total protein of cervical lymph. This maneuver promoted a nearly three-fold rise in intracranial pressure and was accompanied by a rise in cervical lymph flow and fall in its protein content. In addition, a profuse nasal discharge (11.4 ml/hr) developed with a moderately high protein content of the rhinorrhea fluid (1.8 g/dl), along with similar appearance times of Evans blue dye (instilled in the cisterna magna) in both cervical lymph and the rhinorrhea fluid (48-70 minutes after infusion). These findings suggest alternative drainage pathways for CSF besides the arachnoid villi (Pacchionian bodies) including connections with lymphatics in the neck and along the olfactory nerve, and around the cribiform plate to the nasal submucosa, and with proptosis, perhaps also through the aqueous humor-canal of Schlemm and nasolacrimal duct.     

Hormonal regulation of extracellular plasminogen activators and Mr approximately 54,000 plasminogen activator inhibitor in human neoplastic cell lines, studied with monoclonal antibodies

We have studied the regulation by glucocorticoids and dibutyryl cAMP of the amounts of urokinase-type plasminogen activator (u-PA), tissue-type plasminogen activator (t-PA) and a Mr approximately 54000 plasminogen activator inhibitor accumulated in serum-free conditioned culture fluid by a human fibrosarcoma, a human glioblastoma and a human melanoma cell line (HT-1080, UCT/gl-1 and Bowes). For the quantitation of u-PA and t-PA, we used sandwich-type ELISA with a combination of polyclonal and monoclonal antibodies. For an estimation of variations in the amount of the inhibitor, we used sodium dodecyl sulphate-polyacrylamide gel electrophoresis followed by Coomassie blue staining of conditioned culture fluid proteins, the inhibitor protein band being identified by its selective removal by passage of the conditioned culture fluids through a column with monoclonal antibodies against the inhibitor. The modulation of the 3 proteins by the hormonal agents varied greatly between the cell lines. The proteins were independently regulated, in the sense that the hormonal agents did not concomitantly change their levels in the direction expected either to increase or decrease total extracellular plasminogen activator activity. In conditioned culture fluids containing both t-PA and inhibitor, the two were present in the medium as a Mr approximately 120 000 complex. In contrast, no u-PA inhibitor complexes were found in conditioned culture fluid from any of the cell lines; this is likely to be due to the occurrence of u-PA in the culture fluid in the one-chain proenzyme form, which, unlike active u-PA, does not react with the inhibitor. These findings illustrate the complexity of the regulation of extracellular plasminogen activator activity, and imply that the presumed functional diversity of u-PA and t-PA may be related to their independent regulation.     

An evaluation of OSOM BV blue test in the diagnosis of bacterial vaginosis

ObjectiveTo determine the prevalence of bacterial vaginosis (BV) in patients with vaginal discharge and evaluate the efficacy of OSOM BV blue test in diagnosis.MethodsOSOM BV blue test, a rapid diagnostic test based on the principle of detection of bacterial sialidase activity in vaginal fluid specimens was conducted. A total of 405 patients in the reproductive age group (15–45 years) having vaginal discharge were included in the study along with 10 healthy age-matched controls. Two high vaginal swabs were collected aseptically from each patient. One swab was used to make smear for gram staining, and the other was for OSOM BV blue test. Amine test and vaginal pH test were taken as well.ResultsThe prevalence of bacterial vaginosis was 60.7%. OSOM BV blue test showed good efficacy, as compared with gram staining in diagnosing BV. The sensitivity and specificity of OSOM BV blue test were 97.6% and 97.5% respectively. Amsel's criteria diagnosed 180 (44.4%) cases of BV and had sensitivity and specificity of 67.1% and 90.6% respectively. Thus the performance of OSOM BV blue was better than the methods based on Amsel's criteria.ConclusionsOSOM BV blue test is an efficacious bed side test, helpful in rapidly making an accurate diagnosis of BV in setups lacking laboratory facilities or expert microbiologists.     

A case of arc welder's lung with ground-glass opacities and progressive massive fibrosis]

A 62-year-old man who had worked as a welder for 35 years was admitted with abnormal chest radiograph shadows. Chest CT scan showed ground-glass opacities (GGO) and nodular shadows (progressive massive fibrosis: PMF) with spiculation in both lung fields. Transbronchial lung biopsy (TBLB) findings of a nodule (left segment 8) revealed many iron particles in the alveoli and positive staining for Fe (Berlin blue stain). Moreover, bronchoalveolar lavage (BAL) fluid of a GGO (left segment 4) revealed many iron particles and positive staining for Fe (Berlin blue stain) in macrophages. Serum ferritin was extremely high (6.352 ng/ml) and ferritin in the BAL fluid was 210 ng/ml. Taking the clinical course and pathological findings together, pneumoconiosis (arc welder's lung) was diagnosed. The most common chest CT pattern in arc welder's lung is ill-defined micronodules diffusely distributed in the lung like hypersensitivity pneumonitis. Arc welder's lung rarely presents as PMF. We report a case of arc welder's lung accompanied with PMF.     

Histochemical comparison of mast cells obtained from the airways of mongrel dogs and Basenji-Greyhound dogs by bronchoalveolar lavage

An abnormally large number of mast cells in the airway lumen may be an important factor in the pathogenesis of bronchial hyperreactivity. However, it is unclear just how many mast cells are present in the lumen of normal or hyperreactive airways, in part because of differences in the histochemical techniques that have been used to identify mast cells and questions about the heterogeneity of mast cells. The present study was done (1) to compare the effectiveness of six techniques in the identification of mast cells obtained from dogs by bronchoalveolar lavage (BAL), (2) to compare the mast cells in the airways of normal mongrel dogs with those from a breed of dog (Basenji-Greyhound) known to have bronchial hyperreactivity, and (3) to determine whether the two-type histochemical classification used for rodent mast cells (formaldehyde-resistant or typical and formaldehyde-sensitive or atypical) applies meaningfully to the mast cells in BAL fluid from dogs. Cells obtained by BAL were fixed with Mota's basic lead acetate or formaldehyde. Mast cells were identified by metachromatic staining with toluidine blue or methylene blue, staining of highly sulfated proteoglycans with Alcian blue or berberine sulfate, and a histochemical reaction for chloroacetate esterase (mast cell chymase). After Mota's fixation, the methods were relatively similar in their effectiveness in determining the number of mast cells in lavage fluid from the mongrel dogs, in that all of the values fell within a narrow range: 0.53 to 0.96% of the total number of cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

Mast cells with gonadotropin-releasing hormone-like immunoreactivity in the brain of doves.

Using an antiserum (LR-1) raised against mammalian gonadotropin-releasing hormone (GnRH), we previously identified a nonneuronal cell that was more numerous in the medial habenula (MH) of courting ring doves than in individuals housed in visual isolation. The current studies suggest that they are mast cells. Both acidic toluidine blue and toluidine blue dissolved in water/butanediol revealed metachromatic cells with a distribution and morphology similar to that obtained by immunostaining with the GnRH antiserum in the MH. Some cells had granules reactive to safranin in the presence of alcian blue, indicative of a highly sulfated proteoglycan of the heparin family. Immunocytochemical studies demonstrated that all MH cells containing GnRH-like immunoreactivity contained histamine, another mast cell marker. The GnRH-immunoreactive cells had a unilobular, ovoid nucleus. Secretory granules within the cells were electron dense and displayed a variety of internal structures. Fine filamentous processes appeared evenly distributed on the cell surface whether cells were located on the pial surface or within the brain parenchyma. All of these features are characteristic of mast cells. To test whether the epitope recognized by the GnRH antiserum was produced by the mast cells or endocytosed from the cerebrospinal fluid, an iodinated GnRH analog was injected intracerebroventricularly at the initiation of courtship. Radioautography revealed no radioactive cells in the brain, indicating that the GnRH antibody recognized a molecule synthesized by the nonneuronal cells rather than internalized by a receptor-mediated mechanism. These observations suggest an interaction between a component of the immune network and specific regions of the central nervous system.     

Inflammation as a Major Cause of Fluid Losses in Small-Bowel Obstruction

The importance of inflammation for fluid losses in obstructive ileus was investigated in vivo in the rat. Inflammation was quantified by spectrophotometry of extravasated Evans blue (Eb)-albumin. Net fluid secretion in the obstructed jejunum was measured by a continuous gravimetric technique. The inflammation in the obstructed gut wall was significantly more pronounced than that in the gut distal to the obstruction and the sham-obstructed gut. The inflammation was significantly more pronounced in the serosa and external muscle layer than in the mucosa-submucosa. Acid-base balance in obstructed animals showed a significant metabolic alkalosis, whereas serum albumin and electrolytes were normal. Lumen fluid in obstructed animals showed low levels of albumin and total calcium as compared with serum, whereas fluid from the peritoneal cavity of obstructed rats showed high contents of albumin. Indomethacin and hydrocortisone given intravenously to obstructed animals significantly reduced the degree of extravasated Eb-albumin in the obstructed gut wall. Sham-operated animals showed net fluid absorption, whereas obstructed rats showed net fluid secretion. Secretion in obstructed animals was in all cases reversed into net fluid absorption after intravenous administration of indomethacin and hydrocortisone. These findings suggest that a pronounced inflammation occurs in the wall of the obstructed small intestine and that this inflammation plays an important role in the pathogenesis of the profuse fluid losses of obstructive ileus.     

Trace polypeptides in cellular extracts and human body fluids detected by two-dimensional electrophoresis and a highly sensitive silver stain.

Development of a highly sensitive silver stain permits the characterization of trace cellular and body fluid proteins separated by the two-dimensional electrophoresis technique of O'Farrell. Many of the proteins detected by the silver stain in urine, spinal fluid, amniotic fluid, and cells were undetected with the widely used Coomassie blue stain. Trace polypeptides observed in Escherichia coli cell lysates with this silver stain could be detected previously only by growth in radioactive precursors followed by lengthy autoradiography. In situations that do not permit the use of radioactive labeling, as in human clinical studies, the enhanced ability to detect proteins achieved by the silver stain will facilitate metabolic studies and the screening for protein abnormalities in mutational studies and in genetic diseases.     

Concentrations of glycosaminoglycans in synovial fluids and their relation with immunological and inflammatory mediators in rheumatoid arthritis.

The dimethylmethylene blue assay showed higher concentrations of glycosaminoglycans in many synovial fluids from patients with rheumatoid arthritis (RA) than in autologous sera or sera or synovial fluids from normal subjects. These results were taken to suggest that the glycosaminoglycans in RA synovial fluid were abnormally raised and derived from cartilage. To discover what stimulated such glycosaminoglycan release in RA joints relations were sought between synovial fluid concentrations of glycosaminoglycans and immunological and inflammatory mediators. It was shown that RA synovial fluid glycosaminoglycan concentrations correlated with synovial fluid C3d concentrations but not with synovial fluid rheumatoid factor concentrations, polymorphonuclear leucocyte numbers, myeloperoxidase concentrations, or the ability of the synovial fluids to release free radicals from normal polymorphonuclear leucocytes. A correlation was found between synovial fluid C3d and interleukin 1 concentrations as judged by both lymphocyte activating factor activity and immunoassay, but no significant correlation was detected between interleukin 1 and glycosaminoglycan concentrations. It is suggested that in the rheumatoid joint locally produced cytokines, in addition to interleukin 1, together stimulate glycosaminoglycan release from cartilage and render it vulnerable to attack by other processes.     

A staining sequence for the differentiation of A-, B-, and D-cells of the islets of guinea-pig pancreas

Summary Four μ sections of guinea-pig pancreas that had been fixed in Bouin’s fluid were deparaffinized, hydrated, oxidized in equal volumes of 0.3% KMnO₄ and 0.3% H₂SO₄, decolorized in 2% oxalic acid and stained in 1% alcian blue in 70% alcohol for 10 min. Next they were stained for 5 min in chrome hematoxylin then into a dilute aqueous acid fuchsin solution followed by 2% phosphotungstic acid for 1 min. They were washed in running water until only the A-cells retained the red stain and were then counter stained with alcoholic aurantia for 5 min. Finally, the sections were cleared in xylene and mounted in D.P.X. Within the islets, B-cells stained blue green, A-cells bright red, D-cells pale yellow, while the acinar tissue stained blue grey. The wide separation between these colors represents a considerable advance on previous techniques for individual cell recognition within the islet.     

Fast Blue RR—Siloxane Derivatized Materials Indicate Wound Infection Due to a Deep Blue Color Development

There is a strong need for simple and fast methods for wound infection determination. Myeloperoxidase, an immune system-derived enzyme was found to be a suitable biomarker for wound infection. Hence, alkoxysilane-derivatized Fast Blue RR was immobilized via simple hydrolytic polymerization. The resulting enzyme-responsive siloxane layers were incubated with myeloperoxidase, wound fluid or hemoglobin. The reaction was monitored via HPLC measurements and the color development quantified spectrophotometrically. Myeloperoxidase was indeed able to oxidize immobilized Fast Blue RR leading to a blue colored product. No conversion was detected in non-infected wound fluids. The visible color changes of these novel materials towards blue enable an easy distinction between infected and non-infected wound fluids.     

Unexpected endoscopic full-thickness resection of a duodenal neuroendocrine tumor

A 57-year-old man underwent endoscopy for investigation of a duodenal polyp. Endoscopy revealed a hemispheric submucosal tumor, about 5 mm in diameter, in the anterior wall of the duodenal bulb. Endoscopic biopsy disclosed a neuroendocrine tumor histologically, therefore endoscopic mucosal resection was conducted. The tumor was effectively and evenly elevated after injection of a mixture of 0.2% hyaluronic acid and glycerol at a ratio of 1:1 into the submucosal layer. A small amount of indigo-carmine dye was also added for coloration of injection fluid. The lesion was completely resected en bloc with a snare after submucosal fluid injection. Immediately, muscle-fiber-like tissues were identified in the marginal area of the resected defect above the blue-colored layer, which suggested perforation. The defect was completely closed with a total of 9 endoclips, and no symptoms associated with peritonitis appeared thereafter. Histologically, the horizontal and vertical margins of the resected specimen were free of tumor and muscularis propria was also seen in the resected specimen. Generally, endoscopic mucosal resection is considered to be theoretically successful if the mucosal defect is colored blue. The blue layer in this case, however, had been created by unplanned injection into the subserosal rather than the submucosal layer.     

Massiver, unstillbarer Perikarderguss

This is the report on a case of a 60-year-old male patient who was admitted to our hospital with clinical manifestation of massive pericardial effusion of mild hemodynamic significance. The medical history revealed ascites in conjunction with liver cirrhosis and status after thoracoabdominal trauma followed by surgical treatment. Clinical findings, diagnostic imaging, laboratory values, pericardiocentesis, and consecutive pericardial fluid analysis could not clarify the etiology of the pericardial effusion. Pericardial fluid volume, early recurrence, and a declining ascites fluid volume after pericardiocentesis suggested the likelihood of peritoneopericardial fistula. Intraperitoneal injection of methylene blue solution confirmed the fistula. The treatment of refractory ascites included the implantation of a transjugular intrahepatic portosystemic stent shunt (TIPSS) and medical treatment of the liver cirrhosis. Alternatively surgical treatment for persistent intrapericardial ascites accumulation could be considered.