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PubMed2006

Alterations of the ocular surface epithelial mucins 1, 2, 4 and the tear functions in patients with atopic keratoconjunctivitis.

BACKGROUND: An increased understanding of the ocular surface alterations at the cellular level in the conjunctiva and the cornea, may help explain the pathogenesis and the subsequent clinical appearance of atopic ocular allergies, which may be potentially blinding. PURPOSE: To investigate MUC 1, 2 and 4 alterations, tear function and the ocular surface disorder in patients with atopic keratoconjunctivitis. METHODS: Twenty-eight eyes of 14 atopic keratoconjunctivitis patients as well as 22 eyes of 11 age-and sex-matched normal subjects were studied. The subjects underwent corneal sensitivity measurements, Schirmer's test, tear film break-up time (BUT), fluorescein and Rose Bengal staining of the ocular surface, conjunctival impression cytology and brush cytology. Impression cytology samples underwent periodic acid-Schiff and immunohistochemical staining with MUC 1, 2 and 4 antibodies. Brush cytology specimens underwent evaluation for inflammatory cell numbers and quantitative real-time-PCR for MUC 1, 2 and 4 mRNA expression. Patient eyes with fluorescein and Rose Bengal scores greater than four points were regarded to have significant epithelial disease in this study. RESULTS: The mean corneal sensitivity and BUT values were significantly lower in atopic patients with significant epithelial disease, compared with patients with insignificant epithelial disease and controls (P < 0.01). Brush cytology specimens from patients with significant epithelial disease revealed significantly higher numbers of inflammatory cells (P < 0.01). Specimens from patient eyes showed positive staining for MUC 1, 2 and 4. MUC 1, 2 and 4 mRNA expressions were significantly higher in eyes with significant epithelial disease compared with eyes with insignificant epithelial disease and eyes of control subjects. CONCLUSION: Ocular surface inflammation, decline in corneal sensitivity, tear film instability, changes in conjunctival epithelial MUC 1, 2 and 4 mRNA expressions were thought to be important in the pathogenesis of atopic ocular surface disease.

Adolescent↗
PubMed1981

A triple-stain technique for evaluating normal acrosome reactions of human sperm.

A triple-stain technique has been developed to score normal acrosome-reacted human sperm in fixed smears. Live and dead sperm are first differentiated using the vital stain trypan blue. Sperm are then fixed in glutaraldehyde, dried onto slides, and the postacrosomal region and acrosome are differentiated using Bismark brown and Rose Bengal. Slides are examined at 1,000 X with a bright-field microscope and assessed for 1) the percentage of sperm that were alive at the time of fixation and 2) the percentage of sperm that had undergone normal acrosome reactions. Experiments are included that show that trypan blue is a reliable stain for dead sperm and that Rose Bengal stains only sperm having intact acrosomes. This technique may have applications in experimental and clinical studies on sperm capacitation, acrosome reactions, and fertilization in laboratory and domestic animals as well as in man.

Acrosome↗
PubMed2001

The treatment of severe trachomatous dry eye with canalicular silicone plugs.

PURPOSE: To evaluate the effects of temporary canalicular occlusion with silicone plugs on trachomatous dry eye patients who were on maximal tolerable medical therapy. METHODS: Forty-four trachomatous dry eye patients who had Schirmer testing with topical anaesthetic measuring 5 mm or less and a tear film break-up time of 5 s or less were included. After the lacrimal efficiency test with dissolvable collagen punctal plugs, silicone canalicular plugs were placed in 22 trachomatous dry eye patients. The other 22 patients in the untreated control group were allowed to continue their medical therapy. Pretreatment and post-treatment evaluations included subjective patient assessment, rose Bengal and fluorescein staining, tear film break-up time, Schirmer testing, conjunctival impression cytology and goblet cell counting. RESULTS: Six months after plug placement, 18 eyes (82%) of 22 patients had subjective improvement and all these patients successfully wore plugs for at least 6 months. There were statistically significant differences between the pretreatment and post-treatment test results including rose Bengal and fluorescein staining scores, tear film break-up times and Schirmer testing measurements. Impression cytology showed improvement of squamous metaplasia in 17 eyes (77%). Eight of the patients (36%) were able to decrease dependency on topical therapy. Ten of the patients (45%) completely stopped using artificial tears. There were statistically significant differences between the two groups in the total symptom scores, staining scores, tear film break-up time, Schirmer testing, impression cytology scores and goblet cell counts. CONCLUSION: In cases where topical tear supplementation is insufficient to relieve the signs and symptoms of severe dry eye and the lacrimal puncta have not already been closed by the trachomatous cicatrising process, occlusion of the canaliculi may be useful to prevent drainage of both natural and artificial tears. Canalicular occlusion improves the objective signs and subjective symptoms and may significantly decrease dependency on tear supplements in selected patients.

Aged↗
PubMed1990

Photoactivation of porphyrins: studies of reactive oxygen intermediates and arrhythmogenesis in the aerobic rat heart.

STUDY OBJECTIVE: In situ production of reactive oxygen intermediates (singlet oxygen and superoxide) during the photoactivation of rose bengal can induce arrhythmias in the aerobically perfused rat heart. The present study was undertaken (1) to assess whether these effects occur with other photosensitizers; (2) to identify the injurious intermediates; (3) to probe the site of action of these phenomena. DESIGN - The study involved the use of meso-tetra-(4-sulphonatophenyl)-porphine (TPPS), a porphyrin which, in contrast to rose bengal, promotes the production of singlet oxygen alone when illuminated. After 10 min of TPPS free perfusion, rat hearts (n = 6 per group) were perfused aerobically with TPPS (1, 5, 10 or 50 mumol.litre-1) for 25 min; during the last 20 min, the hearts were illuminated (3600 lux). In additional studies, TPPS (50 mumol.litre-1) was washed out before illumination. EXPERIMENTAL MATERIAL: Hearts from 30 male Wistar rats, weighing 220-280 g, were excised and perfused retrogradely. MEASUREMENTS AND MAIN RESULTS: Cardiac function was unaffected with TPPS alone. Upon illumination, electrocardiographic changes (increase in QT interval and/or T wave changes) and arrhythmias developed in a dose dependent manner. At the highest dose, electrocardiographic changes occurred within 7.0(SEM 0.4) s; all hearts exhibited ventricular premature beats and complete atrioventricular block; 67% developed ventricular tachycardia and 17% ventricular fibrillation. During illumination, hearts also exhibited a dose and time dependent decrease in coronary flow. In additional studies, despite the absence of TPPS in the perfusate, all hearts exhibited complete atrioventricular block, 67% developed ventricular premature beats and 33% ventricular tachycardia; none exhibited ventricular fibrillation. CONCLUSIONS: The results suggest that singlet oxygen, as opposed to superoxide, is responsible for the injury which occurs at tissue surfaces to which photosensitizer is bound.

Animals↗
PubMed2001

Outbreak of polyarthritis with pyrexia in Western Rajasthan.

AIM: To establish the etiology of recent out break of polyarthritis which occurred in Kanvari village of Churu district of Rajasthan in August, 1999. METHODOLOGY: Forty eight patients of polyarthritis were studied by Hb, TDLC, ESR, CRP, throat swab Gram's stain and culture, blood culture, ASO titer, rheumatoid factor, Rose Bengal plate agglutination test, standard tube agglutination test for brucellosis, widal test, urine examination, X-ray chest, ECG and X-ray of the affected joint. RESULTS: Forty eight patients presented with acute polyarthritis with low grade fever of 1-2 week duration. Most common joint involved was sacroiliac joint (52.08%). Most of patients had multiple joint involvement (93.75%). The Rose Bengal plate agglutination test and standard tube agglutination test for brucella were positive in high titres in 44 (91.60%) patients. All the patients were treated with therapy for brucellosis and followed up for 12 weeks and responded well without complications. CONCLUSION: In case of polyarthritis possibility of brucellosis should always be kept in mind.

Acute Disease↗
PubMed2006

Brucellar spondylo-discitis with rapidly progressive spinal epidural abscess presenting with sciatica.

STUDY DESIGN: Case report. OBJECTIVE: To present a patient with spinal brucellosis, which was initially presented with sciatica and misdiagnosed as a lumbar disc herniation owing to nonspecific neurological and radiological findings. The delay in diagnosis led to rapid progression of the disease and complications. SETTING: Department of Neurosurgery at a tertiary university teaching hospital (Sutcu Imam University Medical Center in Turkey). CASE REPORT: A 57-year-old woman with a history of low-back pain for 6 months, fatigue, and severe left-sided sciatica for the last 3 months presented to our hospital. Three months earlier, at another hospital, she had had a negative Rose-Bengal test for brucellosis and a lumbar computed tomography performed at that time showed only minimal L4-5 annular bulging. For 2 months, she was treated with analgesics for 'lumbar disc herniation' without relief of pain. On presentation to our department, her magnetic resonance imaging (MRI) examination showed edema and minimal annular bulging at L3-4 and L4-5. When her Rose-Bengal test returned positive, she was started on triple antibiotics for presumed Brucella infection. When symptoms and neurologic signs worsened while taking antibiotics, repeat MRI scan showed a spinal epidural abscess at the L4-5 level. Emergent surgery and 8 weeks of antibiotics resulted in cure. CONCLUSION: In areas endemic for brucellosis, subtle historical and exam features should be sought to exclude an infection such as brucellar sponylo-discitis. Appropriate serological tests should be readily available to confirm or exclude this diagnosis in selected patients, to avoid delays in antibiotic treatment.

Brucellosis↗
PubMed1996

Characterization of a monoclonal antibody specific for Brucella smooth lipopolysaccharide and development of a competitive enzyme-linked immunosorbent assay to improve the serological diagnosis of brucellosis.

The reactivity of monoclonal antibody (MAb) 12G12 was analyzed in regard to the main biovars of Brucella species and some members of the families Enterobacteriaceae and Vibrionaceae which present serological cross-reactions with the smooth lipopolysaccharide (S-LPS) of Brucella species. This MAb was strictly directed against the common specific epitope of the Brucella S-LPS. It recognized all of the smooth Brucella strains and biovars except B. suis biovar 2. In order to improve the specificity of the serological diagnosis of brucellosis, a competitive enzyme-linked immunosorbent assay (cELISA) was developed with the horseradish peroxidase-conjugated MAbs 12G12 and S-LPS of B. melitensis Rev1. The specificity of the cELISA was analyzed with 936 serum samples from healthy cattle. The assay was evaluated with sera from heifers (n = 18) experimentally infected with B. abortus 544. After infection, the performance of the cELISA was in agreement with those of the complement fixation test and the rose Bengal plate test. Finally, the specificity of the assay was also evaluated in regard to false-positive serological reactions by using sera from heifers experimentally infected with Yersinia enterocolitica 0:9 (n = 4) and with field sera presenting false-positive reactions (n = 74). The specificity of the cELISA was greater than the specificities of the complement fixation test and the rose Bengal plate test. Indeed, the new assay detected only 31 of the 101 false-positive serum samples detected by at least one serological test.

Animals↗
PubMed1991

[Ocular manifestations of Yusho 22 years after the onset].

Ocular manifestations were studied in 91 Yusho patients who took the contaminated rice oil 22 years ago. Tear film dysfunction was examined using the rose bengal test. Eighty-seven percent of the patients complained eye discharge, although edema of the eyelid and pigmentation of the conjunctiva were rarely seen. Cystic swelling of the tarsal glands was seen in 16 patients (18%) and white cheese-like contents in the tarsal glands were detected in 7 patients (8%). Twenty-five patients showed positive staining with the rose bengal test, although the intensity of the staining was slight. From this study it appears that there may not be tear film dysfunction in Yusho patients.

Adult↗
PubMed2005

Memory-switching phenomenon in acceptor-rich organic molecules: impedance spectroscopic studies.

We have studied memory-switching phenomena in terms of impedance spectroscopy in a series of xanthene-class fluorescein molecules. Frequency response studies have revealed that the electronic conduction mechanism was largely tuned by increasing the number of acceptor groups attached to the molecular backbone. In molecules with weak intermolecular interactions, the transition between a low- and a high-conducting state has been associated with a change in bulk resistance and dielectric properties. The devices in the two states remained a parallel combination of a resistor and a capacitor (C(P)-R(P)) network. In devices based on Rose Bengal, which exhibits a strong dipole-dipole interaction due to its acceptor groups, the switching between the states has been modeled as a transition between a C(P)-R(P) network in the off-state and a distributed resistor-capacitor (R-C) line network in the on-state. Instead of isolated clusters, a highly coupled network of reduced Rose Bengal molecules with strong dipole-dipole interaction yielded the high-conducting state. Under a "write-read-erase-read" sequence, the device switched between the two networks reversibly for many cycles.

Journal Article↗
PubMed1991

Association of dry eye signs and symptoms with tear lactoferrin concentration.

Diagnoses of dry eye made with the Lactoplate Immunoassay Test (which assesses tear lactoferrin concentration) were compared to diagnoses based on dry eye symptoms, tear break-up times, and rose bengal staining. For a population of 49 subjects with normal and mild to moderate dry eyes, contingency tables showed that the lactoferrin-based diagnoses were not significantly related to the diagnoses made using any of the other factors. Lactoferrin concentrations were statistically correlated with symptom scores, but there was no significant correlation between lactoferrin concentrations and either tear break-up time or rose bengal staining. These results suggest that: 1) the dry eye problems in the subject population may not have been associated with lacrimal gland dysfunction (which the Lactoplate assesses); 2) Lactoplate tests are not sensitive enough to allow accurate diagnoses for the population of mild to moderate dry eye subjects used; 3) signs and symptoms associated with dry eyes occur before lactoferrin changes are manifest so lactoferrin changes had not yet appeared in the subjects; or 4) in mild to moderate dry eye subjects, lactoferrin changes occur only in reflexive tears and not in the basal tears that were tested in this study. In summary, measurement of tear lactoferrin concentration alone has not been shown to be a sufficiently sensitive and specific test for the diagnosis of mild to moderately dry eye as defined by more common clinical techniques.

Adult↗
PubMed2003

Risk factors for ocular surface disorders in patients with diabetes mellitus.

OBJECTIVE: To evaluate risk factors for ocular surface disorders and tear dysfunction in patients with type 2 diabetes. STUDY DESIGN AND METHODS: This prospective case controlled study included 41 patients with type 2 diabetes and 20 healthy subjects as the control group. All subjects underwent routine ophthalmic examination, tear film break-up time (BUT) test, Schirmer test, fluorescein dye test, rose bengal staining test, and a questionnaire for subjective complaints. The relationship of metabolic control of diabetes mellitus, duration of diabetes, severity of diabetic retinopathy, and argon laser photocoagulation (ALP) to tear dysfunction was evaluated. RESULTS: Tear film BUT and Schirmer test values were significantly lower in diabetic patients compared with control subjects (P < 0.001). In the diabetic group, significantly more subjects had abnormal fluorescein and rose bengal staining than in the control group (P < 0.001). Abnormal tear function tests were associated with poorer metabolic glucose control, panretinal ALP, and proliferative diabetic retinopathy (P < 0.05), but not with duration of diabetes (P > 0.05). CONCLUSIONS: The results of the study indicate that poor metabolic control, panretinal ALP, and proliferative diabetic retinopathy are high risk factors for ocular surface disorders in type 2 diabetes. These patients should be followed more carefully, and should be referred to an ophthalmologist when required.

Adult↗
PubMed2000

[Comparison of primary and secondary Sjögren's syndrome].

PURPOSE: We studied the difference in severity between primary and secondary Sjögren's syndrome (SS). SUBJECTS AND METHODS: Two groups of patients (all females, mean age: 58 years), 31 with primary SS and 18 with secondary SS were studied. We performed the following dry eye tests: fluorescein score and Rose Bengal staining, grading of tear lipid layer interference patterns, measurement of fluorescein break up time, cotton thread test, and Schirmer-I test. Auto antibodies were also investigated. RESULTS: There was no significant difference between primary and secondary SS with respect to any dry eye tests or auto antibodies. In primary SS, however, the presence of anti SS-A antibody was significantly correlated with Rose Bengal scores (p = 0.044). CONCLUSION: The severity of SS is independent of the primary or secondary type. In primary SS, the presence of anti SS-A antibody may be correlated with the severity.

Adult↗
PubMed1989

A new cholescintigraphic agent: ruthenium-97-DISIDA.

These are the first human experiments with 97Ru-DISIDA, a potentially new alternative to 131I-rose bengal where delayed imaging is indicated. 97Ru has a convenient half life. A DISIDA labeling kit was utilized to prepare the radiotracer for 17 patients (age 6 weeks-84 years). The cholescintigraphic data correlated well with other imaging procedures and with clinical findings. Dosimetric calculations were carried out and were compared with the radiation burden associated with the use of 99mTc-DISIDA and 131I-rose bengal.

Biliary Tract Diseases↗
PubMed1996

Reversibility of compound action potential during the acute phase after transitory local ischemia.

We investigated the reversibility of compound action potential (CAP) changes induced after transitory local ischemia induced by rose bengal photochemically induced thrombosis of the rat anterior inferior cerebellar artery (AICA). Cochlear blood flow (CBF) was measured with a laser-Doppler flowmeter positioned on the lateral bony wall of the basal turn of the cochlea, and the CAP to an 8-kHz half-wave of sinusoid sound at 100 dB sound pressure level was monitored. The irradiation was started 5 minutes before the rose bengal administration and continued through the thrombosis formation. Tissue-type plasminogen activator (t-PA, 1 mg/kg) dissolved in saline was injected intravenously 2 minutes after complete photothrombotic blockade of the AICA in the rats presenting complete abolition of CAPs just after the vascular occlusions. Nineteen of the 51 rats presented complete abolition of the CAP just after the AICA occlusion, and the thrombosed AICA was successfully reperfused by t-PA administration in 11 of the 19 rats. The result showed that duration of ischemia was the more important determinant for the reversibility of CAPs during the acute phase (p = .00001), and the residual level of cochlear blood flow during ischemia was also an important factor (p = .066). It appeared that the critical time of ischemia for the complete recovery of CAPs was around 5 minutes, and that the critical limit for the irreversibility of CAPs within the acute phase was between 20 and 25 minutes.

Animals↗
PubMed1997

Photochemically-induced vascular damage in brain cortex. Transmission and scanning electron microscopy study.

Morphological changes of microvessels of cerebral cortex were evaluated in a model of cerebral infarction initiated by a photochemical reaction. Rats were treated with intravenous injection of rose Bengal and irradiated from a halogen lamp source through an intact cranium to precipitate microvascular damage. Investigations in transmission and scanning electron microscopy revealed platelet aggregation on endothelial cells preceded by its early ultrastructural damage. Other typical microscopic features of brain ischaemic injury were present suggesting that the present method may be used as a model for investigating ischaemic brain damage. Since the photochemical activation of the rose Bengal dye results in formation of reactive oxygen species this model may be particularly useful to elucidate the role of free radical-mediated endothelial damage in the formation of microthrombi and blood-brain-barrier integrity.

Animals↗
PubMed2001

[Adverse effects of beta-blocker eye drops on the ocular surface].

PURPOSE: We investigated the adverse effects of beta-blocker eye drops on tears and ocular surface epithelium. METHODS: We studied twenty-three eyes of twenty three glaucoma patients [10 males, 13 females: 53.8 +/- 12.2 (yrs; mean +/- standard deviation)] treated with beta-blocker eye drops for more than three months and thirty two control subjects (16 males, 16 females: 50.4 +/- 10.9). The parameters described below were compared between the groups: 1. the radius of tear meniscus curvature, 2. grades for tear lipid layer interference patterns, 3. non-invasive breakup time (N-BUT), 4. cotton thread value, 5. scores of fluorescein staining, 6. fluorescein breakup time (F-BUT), 7. scores of rose bengal staining, 8. and Schirmer I value. RESULTS: The glaucoma group showed a significant decrease in the radius of tear meniscus curvature (p = 0.0007), a significantly lower distribution in the grades for tear lipid layer interference patterns (p = 0.0270), a significant difference in the scores of fluorescein staining (p < 0.0001), a significant shortening in F-BUT (p = 0.0050), a significantly higher distribution in the scores of rose Bengal staining (p = 0.0010), and a significantly smaller value in Schirmer I value (p = 0.0042). However, there was no significant difference in N-BUT and cotton thread value. CONCLUSIONS: These results clearly demonstrate that the ocular surface in glaucoma patients treated with beta-blocker eye drops show dry-eye-like changes in terms of tears and ocular surface epithelium.

Adrenergic beta-Antagonists↗
PubMed1993

Effect of ibudilast on microcirculation thrombosis in rat inner ear.

The effect of ibudilast (0.1, 0.3 mg/kg), which has cerebral vasodilating and antiplatelet effects, was evaluated in two models of rat inner ear microcirculation thrombosis by using the photochemical reaction between green light (wave length: 540 nm) and intravenous injection of rose bengal. Furthermore, the inner ear blood flow was measured by a laser-Doppler flowmeter. In the hearing disturbance model, under anesthesia, the compound action potential of the cochlear nerve (AP) was measured by an electrocochleogram. The sound stimulus was an 8-kHz sine wave at 80 dB SPL. The AP was calculated 128 times. In the controls, the AP disappeared about 4 min after the intravenous injection of rose bengal (20 mg/kg). The time required to completely suppress the AP in the animals treated with ibudilast (0.1, 0.3 mg/kg) was significantly prolonged as compared with that in the controls. In the equilibrium dysfunction model, ibudilast (0.1, 0.3 mg/kg) reduced the time of abnormal swimming in the swimming test 24 hr after the completion of photo-illumination. Ibudilast (0.3 mg/kg) increased the inner ear blood flow during the 10-min observation period as compared with the controls, while it did not affect the mean blood pressure. In conclusion, ibudilast increased the inner ear blood flow and was effective in two models of rat inner ear microcirculation thrombosis.

Animals↗
PubMed2004

Prophylactic effect of IL-10 gene transfer on induced autoimmune dacryoadenitis.

PURPOSE: To evaluate the effect of viral IL-10 on the lacrimal gland immunopathologic response in the ocular surface disease, induced autoimmune dacryoadenitis. METHODS: Disease was induced in rabbits by injecting inferior lacrimal glands with peripheral blood lymphocytes activated by 5 days of coculture with autologous acinar cells in a mixed-cell reaction. In the treated group, an adenoviral vector carrying the vIL-10 gene was concurrently injected with activated lymphocytes. Tears were collected periodically for quantitation of IL-10 by ELISA. Two weeks after disease induction, tear production, tear film breakup time, and rose bengal staining score were determined. Sectioned glands were immunostained for expression of CD4, CD8, rabbit thymic lymphocyte antigen (RTLA), CD18 and major histocompatibility complex class II. RESULTS: The titer of vIL-10 in tears was at its maximum on day 3, started to decline by day 7, and was undetectable by day 14. In the diseased group, the tear production rate and tear film breakup time were significantly decreased, and rose bengal staining was significantly increased. Diseased glands had immune cell infiltrates containing CD4+, RTLA+, and CD18+ cells, and major histocompatibility complex class II expression was increased. These changes were significantly ameliorated by expression of vIL-10. CONCLUSIONS: In vivo transduction of the lacrimal gland with AdvIL-10 resulted in the transient appearance of vIL-10 in tears. The presence of vIL-10 partially suppressed the appearance of Sjögren-syndrome-like features of reduced tear production, accelerated tear breakup, ocular surface disease, and immunopathologic response. Anti-inflammatory cytokine gene expression may offer a therapeutic modality for the treatment of autoimmune dacryoadenitis, once suitable vectors become available.

Adenoviridae↗