Capsular Staining Research Articles

Collagen immunostains can distinguish capsular fibrous tissue from septal fibrosis and may help stage liver fibrosis.

Core-needle biopsy remains essential for diagnosis of cirrhosis; however, evaluation of fibrosis in such biopsies is often challenging due to the fragmented nature of cirrhotic liver specimens. It is also common to see portions of liver capsules present in the biopsy which adds to the diagnostic challenge. The distinction between capsular/subcapsular fibrous tissue and septal fibrosis is critical to avoid potential overstaging of liver fibrosis. We compared the differential immunostaining in liver capsular and septal areas for collagens III, IV, V, VI, vitronectin, laminin, Orcein, and Trichrome in 15 whole sections of explanted cirrhotic livers and 5 simulated liver biopsies. Collagens III, IV, V, VI, Trichrome, and Orcein show distinct staining patterns in capsular fibrous tissue and septal fibrosis. Collagen IV shows strong diffuse septal staining and consistently weak to negative capsular staining. Collagens III and VI stain similar to IV for septal fibrosis, whereas collagen V, Trichrome, and Orcein show strong staining in both areas. Collagen IV, possibly with III or VI in addition to the routine Trichrome and hematoxylin and eosin stain, is useful in differentiating capsular fibrous tissue from septal fibrosis on challenging and fragmented liver biopsies.

Serological versus molecular typing of surface-associated immune evading polysaccharide antigens-based phenotypes of Staphylococcus aureus.

The aim of this study was to compare the performance of serological versus molecular typing methods to detect capsular polysaccharide (CP) and surface-associated polysaccharide antigen 336 phenotypes of Staphylococcus aureus isolates. Molecular typing of CP types 1, 5 and 8 was carried out using PCR, whereas serological typing of CP1, 2, 5, 8 and antigen 336 was carried out by slide agglutination using specific antisera. By genotyping, 14/31 strains were CP8 positive, 12/31 strains were CP5 and the remaining 6/31 isolates were non-typable (NT). One isolate was positive for both CP5 and CP8 by PCR, but was confirmed as CP8 type serologically. Detection of CP2 and type 336 by PCR was not possible because specific primers were either not available or non-specific. Using serotyping, 14/31 strains were CP8 positive, 11/31 CP5 positive and 2/31 positive for antigen 336. The remaining four S. aureus isolates were serologically NT. However, three of four NT and two 336-positive S. aureus isolates were encapsulated as determined by light microscopy after capsular staining. This discovery was surprising and warrants further investigations on the identification and characterization of additional capsular phenotypes prevalent among S. aureus clinical isolates. It was concluded that serological typing was a better method than molecular typing for use in epidemiological investigations based upon the distribution of surface-associated polysaccharide antigens-based phenotypes.

English

  Virulency is the degree of pathogenicity exhibited by most of the pathogens and it is a measure that effectively differentiates pathogenic and non-pathogenic strains. In the present study, culture of bacteria for 10 to 12 h (overnight) at 37°C in 10% India ink included broth that can be used as marker test for initial virulence identification while screening a number of bacterial isolates. These results were further confirmed by the in vivo virulence tests. Although, this is the first report, there is no clear explanation to the degradation of India ink by virulent bacteria which was obtained but this could act as a marker for initial screening of isolates for their virulence. Moreover, India ink is used as a suitable capsular stain and capsule act as a virulence factor for most of the bacteria. So virulence test by 10% India ink including broth acts as a simple, low cost and easy method for initial screening of virulent from non virulent strains.   Key words: Aeromonas hydrophila, bacteria, India ink, pathogens, virulence

Risk factors for intraocular pressure rise following phacoemulsification

Purpose:This study was designed to analyze the risk factors resulting in high intraocular pressure (IOP), which was accepted as IOP higher than 22 mmHg, following uncomplicated phacoemulsification.Materials and Methods:The records of 812 eyes of 584 patients who underwent uncomplicated phacoemulsification were evaluated. There were 330 men and 254 women ranging between the age of 26 and 89 years (65.4 ± 9.8 years). The preoperative, postoperative first day (day 1), first week (day 7), and first month (day 30) IOP values were analyzed. Data on history of diabetes, glaucoma, pseudoexfoliation (PXF), incision site, capsular staining with trypan blue, and surgeon were recorded. A multinomial regression analysis was performed to analyse the relationship of the factors with postoperative high IOP.Results:The mean IOP was 15.6 ± 4.3 mmHg preoperatively. Postoperatively that were changed to 19.7 ± 9.0 mmHg at day 1, 12.7 ± 4.5 mmHg at day 7, and 12.8 ± 3.7 mmHg at day 30. The factors such as surgeon, presence of PXF, diabetes, surgical incision site, and trypan blue were not related to the postoperative high IOP (P > 0.05, in all). The only factor that related to high IOP at all visits was glaucoma (P < 0.005).Conclusion:According to our results, preoperative diagnosis of glaucoma seems to be the only factor to affect the postoperative IOP higher than 22 mmHg.

Unintentional staining of the posterior lens capsule with trypan blue dye during phacoemulsification: case report

We report a complication related to the use of trypan blue during capsular staining of the anterior lens capsule during phacoemulsification surgery. In phacoemulsification surgery of the left eye, trypan blue was injected using an air bubble technique. Unintentionally, trypan blue was administered under high pressure, dispersing the dye through the zonules leading to staining of the posterior capsule. This caused a temporary disturbance of visual acuity during the postoperative period.

Comparative Study of Different Concentration of Trypan Blue for Staining of Capsule in Mature Cataract using RGB Scale in “Adobe Photoshop Image Reader”

Satisfactory capsulorhexis is essential in modern small incision cataract surgery. Visualization of capsule in mature cataract is difficult so staining of capsule with dye is required. Trypan blue is most commonly used dye. Carcinogenic effect of trypan blue is proved in animal study. Aim of this study was to determine minimum concentration of trypan blue that stains the anterior capsule. Methods: The study was conducted in 50 patients with senile mature cataract. Capsular staining was done with different concentration of trypan blue (Group A 0.06%, Group B 0.03%, Group C 0.015%, Group D 0.0075% and Group E 0.00375%) to enhance visualization of capsule for rhexis in cataract surgery. Comparative study of blueness of dye with different concentration was done by RGB scale values in “Adobe Photoshop image reader” Software (Version-7). Results : Trypan blue stains the capsule satisfactorily in group A, B and C. In group D and E capsular staining was not satisfactory. Conclusion: Concentrations of 0.03% and 0.015% were as effective as 0.06% to stain the capsule. Concentration of 0.0075% and 0.00375% could not stain the capsule.

Adjunct devices for managing challenging cases in cataract surgery: capsular staining and ophthalmic viscosurgical devices

Cataract surgeons should employ all reasonable technology to facilitate safe and consistent outcomes. Knowledge of adjunct devices to enhance visualization of the capsular bag and appropriate use of ophthalmic viscosurgical devices allow for enhanced protection of intraocular structures and reduced rates of complication. Trypan blue is US Food and Drug Administration-approved for facilitating visualization of the anterior capsule. Animal data demonstrate Brilliant Blue G to be an effective capsular stain with a superior safety profile. Healon5 is a safe viscosurgical device that is particularly useful in children, poorly dilating pupils, intraoperative floppy-iris syndrome, and intumescent cataracts. The viscoadaptivity of Healon5 enables its utility throughout the procedure of cataract extraction. The recent literature on capsular dyes and advances in ophthalmic viscosurgical technology is reviewed. Inadequate capsular visualization and poor tissue stabilization/protection increase the chance of discontinuous capsulorhexis, retained nuclear material, vitreous loss and corneal decompensation. Utilization of adjunctive tools in the setting of challenging cataract cases can significantly limit adverse intraoperative outcomes and result in reproducible surgical success.

Severe Toxic Effect of Methylene Blue 1% on Iris Epithelium and Corneal Endothelium

To report a case of corneal endothelial decompensation and iris pigment dispersion following the inadvertent use of methylene blue 1% for capsular staining during cataract surgery. Case report. During an otherwise routine phacoemulsification cataract surgery, inadvertent anterior capsule staining with methylene blue 1% instead of trypan blue 0.025% was performed. Copious irrigation of the anterior chamber with balanced salt solution was initiated upon identification of the wrong dye. The operation was completed with minimal ultrasound energy without complications. Iris discoloration and severe corneal edema developed in the early postoperative period, resulting in severe visual loss. The patient developed bullous keratopathy and underwent penetrating keratoplasty 16 months later. In vivo intracameral injection of methylene blue 1% induces extreme cytotoxicity, primarily on the corneal endothelium and iris epithelium.

Staining Ability and Biocompatibility of Brilliant Blue G

To evaluate the effectiveness and biocompatibility of brilliant blue G (BBG) for capsular visualization for continuous curvilinear capsulorrhexis. The capsular staining ability of BBG was evaluated at graded concentrations of 10.0, 1.0, 0.5, 0.25, 0.1, and 0.01 mg/mL in enucleated pig's eyes. The biocompatibility of BBG was assessed in rat's eyes for 2 months. The eyes were analyzed using light, fluorescence, transmission electron, and scanning electron microscopy. TUNEL (terminal deoxynucleotidyl transferase-mediated biotin-deoxyuridine triphosphate nick-end labeling) was used to detect apoptotic cells, and endothelial cell counts were analyzed using scanning electron microscopy. The results were compared using indocyanine green and trypan blue. The BBG improved capsular visualization, and a complete capsulorrhexis could be performed. In the rat model, no apparent toxic effect was observed using biomicroscopy during 2 months. Histologically, BBG showed satisfactory biocompatibility. Apoptotic cell death of the endothelial cells was detected in only the trypan blue group. In contrast to BBG, indocyanine green and trypan blue showed degeneration of corneal endothelial cells using transmission and scanning electron microscopy. The BBG contributed to better capsular visualization and caused no apparent complications to the corneal endothelium.Clinical Relevance The BBG is effective and safe capsular staining for continuous curvilinear capsulorrhexis.

Capsular staining: recent developments

This paper reviews the recent literature regarding anterior capsular staining. Safety and efficacy of dyes used in facilitating visualization of the anterior capsule during phacoemulsification are compared. Advantages of using different techniques for injection of the dyes are discussed. Trypan Blue and Indocyanine Green appear to be most effective in staining the anterior capsule with the highest safety profile. Injection strategies should be aimed at reducing uncontrolled dispersion especially into the vitreous.

Impregnação inadvertida da cápsula posterior do cristalino com azul de tripano durante a facectomia extracapsular: relato de caso

Report of a complication related to the use of 0.1% trypan blue during capsular staining of the anterior crystalline lens capsule in an extracapsular cataract extraction with intraocular lens implantation. The capsular dye was injected using an air-bubble technique, inadvertently, it was done using high pressure dispersing the dye through the zonules leading to a complete lost of red reflex, the following steps of the procedure becoming more difficult after the cataract extraction and causing a temporary disturbance of visual acuity in the postoperative period. In order to avoid this complication, the authors also describe the use of "Modified air-bubble technique" where dispersive viscoelastic is placed into the iridolenticular space 360 degrees to create a protection barrier.

An outbreak of conjunctivitis due to a novel unencapsulated Streptococcus pneumoniae among military trainees.

Bacterial conjunctivitis usually occurs as sporadic cases; outbreaks are uncommon and usually are associated with school campuses. We report an outbreak of Streptococcus pneumoniae conjunctivitis at a military training facility. An outbreak investigation was done. Each case of conjunctivitis was evaluated with an assessment tool including demographic and clinical data. Conjunctival swabs were obtained. Pneumococci underwent standard testing, including serotyping with the Quellung reaction, capsular staining, and multilocus sequence typing. Sequence types were compared with previous reported outbreak strains by construction of dendrograms. Carriage rates of S. pneumoniae were determined among previously undiagnosed case patients with conjunctivitis, and a case-control study was performed. Control measures included education to increase hand washing, distribution of alcohol-based hand gel, and prompt treatment of patients with conjunctivitis. During a 6-week period, 92 cases of conjunctivitis occurred among 3500 persons, with an attack rate of 1.75 cases per 100 person-months. Eighty cases (87%) were due to S. pneumoniae; 45 (49%) were confirmed, and 35 (38%) were probable. Ten percent of recruits surveyed carried the outbreak strain. Twenty-two percent self-reported symptoms consistent with conjunctivitis during the outbreak period; sharing washcloths was associated with conjunctivitis (odds ratio, 11.7; P=.03). The causative organism was resistant to azithromycin but susceptible to telithromycin. The outbreak strain was an unencapsulated S. pneumoniae that has not been previously described; it was most closely related to the sequence type causing the Dartmouth College (Hanover, NH) outbreak of conjunctivitis in 2002. We report a conjunctivitis outbreak among military trainees caused by a novel, unencapsulated strain of S. pneumoniae.

Magnolol stimulates steroidogenesis in rat adrenal cells.

1. This study investigated the effect of magnolol, a compound purified from Magnolia officinalis, on glucocorticoid production by primary adrenal cell culture. 2. Magnolol increased corticosterone secretion in a dose-dependent manner, this effect being maximal at 40 microM. A similar effect was seen in a minced adrenal gland system. 3. In magnolol-treated cells, the number and total area of cytoplasmic lipid droplets were reduced, suggesting a high utilization rate of cholesterol esters stored in lipid droplets. In control cells, the capsule of the lipid droplet was clearly delineated by immunostaining with antibody A2, whereas capsular staining was discontinuous or undetectable following magnolol treatment. The percentage of decapsulated cells increased significantly from 20% in the control group to 80% in the magnolol-treated group. 4. Magnolol-induced steroidogenesis was not mediated either via the traditional ACTH-cyclic AMP-protein kinase A pathway or by protein kinase C, since the intracellular cyclic AMP level did not change and inhibition of protein kinase A or C did not block the action of magnolol. Furthermore, calcium/calmodulin-dependent protein kinase II was not involved in magnolol-induced steroidogenesis. 5. The stimulatory effect of magnolol on steroidogenesis apparently requires new protein synthesis, since cycloheximide inhibited magnolol-induced corticosterone production by 50%. 6. Although other studies have shown that high concentrations of magnolol inhibit acyl-CoA: cholesterol acyltransferase and 11 beta-hydroxysteroid dehydrogenase in a cell-free system, our data show that, in adrenal cell cultures, low concentrations of magnolol have a stimulatory effect on steroidogenesis, and the glucocorticoid produced may explain the effective control of asthma by Magnolia officinalis.

Benign neural tumors of the oral cavity: A comparative immunohistochemical study

To determine if immunohistochemistry can be used as adjunct to the diagnosis and classification of oral benign neural tumors, we stained 77 neurally differentiated tumors with a panel of neural-associated antibodies (S-100 protein, CD57, epithelial membrane antigen, factor XIIIa, CD34, CD68, collagen IV). Using standard histologic criteria, we identified 13 schwannomas, 16 neurofibromas, 23 traumatic neuromas, 16 palisaded and encapsulated neuromas, and 9 granular cell tumors from archived oral pathology specimens. Silver stains showed that neurofibromas, traumatic neuromas, and palisaded and encapsulated neuromas consistently contained axon filaments. Although all neural tumors contained S-100-positive cells, schwannomas and palisaded and encapsulated neuromas contained the most. All tumors expressed CD57; traumatic neuromas were stained intensely and the others stained weakly. The consistent epithelial membrane antigen capsular staining of schwannomas and the absence of factor XIIIa-positive dendritic/spindle cells helped distinguish these tumors from others. Many CD34-positive cells were found in schwannomas, and few were found in palisaded and encapsulated neuromas. Variable numbers CD68-positive cells were seen in all neural tumor types; some of these cells appeared to be macrophages and mast cells, but many were thought to be Schwann cells expressing this antigen. Collagen IV staining, apparently representing basement membrane, was generally a feature of all benign neural tumors. The immunophenotype of the granular cells of the GCTs was S-100+, CD57+, and collagen IV+ supporting the putative neural origin of these tumors. We conclude that neural origin/differentiation of a connective tissue tumor can be confirmed with stains for S-100 protein, epithelial membrane antigen, CD57, and collagen IV. Staining patterns and intensities associated with the panel of antibodies tested can be useful in tumor classification.

Adrenal vasoactive intestinal peptide participates in neonatal corticosteroid production in the rat.

Neonatal rats (3-14 days old) exhibit a period of adrenal hyporesponsiveness characterized by blunted corticosterone (B) responses to stress and reduced adrenal sensitivity to adrenocorticotropic hormone (ACTH). Several adrenomedullary peptidergic systems like vasoactive intestinal peptide (VIP) are postulated to influence cortical function. VIP is known to stimulate corticosterone secretion in vitro and to be released from the adrenal medulla following splanchnic nerve stimulation. Here, we tested whether 1) accelerated sympathetic innervation of the adrenal gland by daily L-thyroxine (T4) treatment modified the ontogeny of adrenal VIP and 2) an increase in VIP synthesis could prematurely increase adrenal sensitivity and corticosteroid output during neonatal life. Immunohistochemical VIP staining revealed a different ontogenetic pattern between adrenal regions from days 2-18 and different sensitivities to T4 treatment. Capsular staining was most abundant at all ages and increased with T4 treatment, whereas medullary staining was seen by day 18 and was not affected by T4. Throughout development, VIP receptors were detected mostly in the capsular region, but not in the adrenal cortex. Although receptor levels were not modified by T4 injections, T4 significantly enhanced VIP mRNA levels in the whole adrenal at all ages. In vivo administration of VIP (0.1-2.0 mg/kg body wt ip) to 9- to 12-day-old neonates increased pituitary ACTH, adrenal B, and aldosterone secretion significantly. Corticotropin-releasing factor immunoneutralization before VIP injection diminished VIP-induced ACTH release but still produced small but significant B and aldosterone secretion. Our results show that 1) VIP innervation of the adrenal capsule is present soon after birth and is increased by sympathetic activity whereas VIP appears only much later in the medulla and does not coincide with the onset of splanchnic innervation and 2) exogenous VIP stimulates ACTH, B, and aldosterone release during development and the effect of VIP on steroidogenic secretion is occurring through ACTH secretion, but also, at least in part, directly at the level of the adrenal gland.

Identification and Serotyping ofSalmonella typhi

A Vi-specific monoclonal antibody was used successfully in a passive latex agglutination test for the serotyping of Salmonella typhi . The Vi-sensitized latex reagent gave strong agglutination with all 42 strains of S typhi tested, but of the 234 other bacteria, which included 66 Salmonella , 93 non- Salmonella enterobacteria,and 75 other gram-negative bacteria,only one strain of Salmonella agona was agglutinated weakly. With immunofluorescence, the Vi-specific monoclonal antibody could also identify broth cultures of S typhi to give characteristic capsular staining but did not stain other bacteria including the strain of S agona , which was weakly agglutinated by the Vimonoclonal antibody. The practicality and further potential applications of using the Vi monoclonal antibody in these two serological tests for the laboratory diagnosis of typhoid fever are discussed.

Effective preoperative irradiation of highly vascular cerebellopontine angle neurinomas

Three cases of large cerebellopontine angle neurinoma with marked vascularity and tumor staining on the angiogram were treated with effective preoperative irradiation. The radiotherapy was given before the second operation in two cases and before the first operation in the other case. Irradiation doses administered with a linear accelerator were 2.34 to 3.0 Gy for 3 to 3.5 weeks, and radical operations were done 1.5 to 2 months after irradiation. After the irradiation, vertebral angiography showed moderate to marked decrease of the hypervascular capsular stain and disappearance of the early draining vein. Computed tomographic scan showed enlargement of the central necrotic area within the heterogeneously enhanced tumor, which was unchanged in size. Radical operations, which had been impossible because of uncontrollable massive bleeding, were successful without any intraoperative bleeding after radiotherapy. Postirradiation radiological findings corresponded well with those of histopathological examination, which showed decrease in cellularity and in vascularity and diffuse coagulation necrosis around the collapsed tumoral vessels as radiation effects. Preoperative irradiation of the hypervascular neurinoma was thought to facilitate radical surgery by abolishing or diminishing the risk of intraoperative bleeding. (Neurosurgery 22:566-573, 1988)

Acoustic Neurinoma with Early Venous Drainage and Caput Medusae-like Vasculature on Angiogram

It has been considered that acoustic neurinomas are hypovascular tumors on angiogram. Recent angiographical analysis, however, revealed that some of them showed tumor stain, capsular stain, and even arteriovenous shunt. The problem with operation on the acoustic neurinoma with a peculiar vasculature on angiogram has not been discussed in previous reports. In this paper, the neuroradiological findings of a unique case are described, in conjunction with the operative problem. A case of huge acoustic neurinoma with early venous drainage and caput medusae-like vasculature on angiogram is presented. The patient was a 46-year-old woman suffering from dizziness and gait disturbance. On admission, she had bilateral choked disc, left fifth and seventh nerve palsy, and left cerebellar signs. The audiogram showed scale out on the left, carolic test and auditory brain stem response were nil on the left side. On computed tomography, a huge, heterogeneously enhanced mass was detected in the left cerebellopontine angle. Vertebral angiography revealed that the tumor was supplied by the anterior inferior cerebellar artery and ascending pharyngeal artery. Caput medusae-like vasculature was visualized in arterial and capillary phases, and abnormal draining veins were also opacified in these phases. They were considered to be an arteriovenous fistula. At operation, the abnormal vasculature looked like an arteriovenous malformation on the tumor capsule, which made the surgical procedure of dissecting the tumor from the pons extremely difficult.

Morphological differences in the capsular polysaccharide of nitrogen-fixingAzotobacter chroococcumB-8 as a function of iron and molybdenum starvation

Azotobacter chroococcum B-8 was nutritionally stressed for iron (Fe) and molybdenum (Mo) by successive transfers on Fe-deficient and Mo-deficient solid-plating media and was subsequently observed with transmission electron microscopy after capsular staining in ruthenium red. Under Fe- and Mo-sufficient conditions, the capsule appeared to be condensed into a thin, electron-dense layer. Under iron stress, and to a lesser extent molybdenum stress, the capsule appeared more diffuse and extensive. This apparent diffusion was irreversible upon the addition of iron before processing. Thus there seems to be a correlation between capsule diffusion and metal starvation, as well as a correlation between capsule extent and metal starvation.

Morphological differences in the capsular polysaccharide of nitrogen-fixing Azotobacter chroococcum B-8 as a function of iron and molybdenum starvation

Azotobacter chroococcum B-8 was nutritionally stressed for iron (Fe) and molybdenum (Mo) by successive transfers on Fe-deficient and Mo-deficient solid-plating media and was subsequently observed with transmission electron microscopy after capsular staining in ruthenium red. Under Fe- and Mo-sufficient conditions, the capsule appeared to be condensed into a thin, electron-dense layer. Under iron stress, and to a lesser extent molybdenum stress, the capsule appeared more diffuse and extensive. This apparent diffusion was irreversible upon the addition of iron before processing. Thus there seems to be a correlation between capsule diffusion and metal starvation, as well as a correlation between capsule extent and metal starvation.