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Evaluation of A newly developed DOT-ELISA kit for the diagnosis of human brucellosis.

A new dot-ELISA kit for detection of Brucella antibodies in human sera was developed and compared with that of serum agglutination test, Rose Bengal plate test, rapid slide agglutination and Coomb's antiglobulin test. Following testing of 120 human sera from suspected patients of occupational risk, 25 gave positive reaction in Rose Bengal plate test, 25 in rapid slide agglutination test, 26 in serum agglutination test, 27 in Coomb's antiglobulin test and 28 in dot-ELISA kit. Dot-ELISA kit picked up more positive than any other Serological test, indicating its superiority over the other laboratory tests for the diagnosis of brucellosis.

Agglutination Tests↗

[Human brucellosis in Benin: results of a serological survey among exposed workers].

A serological survey was carried out in Bénin in order to assess the rate of brucellosis infection among exposed workers (workers in slaughtering-houses and breeders). 221 sera were tested with rose Bengale test, Wright sero-agglutination test, indirect immunofluorescence test and counter-immuno electrophoresis (brucelline). The percentage of positive sera among exposed workers is 17,7%. The rose Bengale and immunofluorescence tests combination permits complete detection of positive sera. These results suggest the existence of human brucellosis in Bénin and shows the necessity of a national control programme adapted to the socio-economic problems of this country.

Abattoirs↗

Systemic suppression of contact hypersensitivity by UVB radiation is unrelated to the UVB-induced alterations in the morphology and number of Langerhans cells.

Exposure of mice to UVB (280-320 nm) ultraviolet radiation reduces contact hypersensitivity (CHS) reactions to chemicals that are applied subsequently to unirradiated skin. It also decreases the number and alters the morphology of Langerhans cells at the site of irradiation. We addressed the question of whether the systemic suppression of CHS was related to these modifications of Langerhans cells by UVB radiation. In mice treated on the dorsum with UVB radiation, the number and morphology of Langerhans cells in the unexposed areas of skin used for inducing and eliciting CHS appeared normal. Therefore, the depression of CHS could not be attributed to a depletion of Langerhans cells at the sites of application of the sensitizing agent. We also examined the correlation between alterations in Langerhans cells and systemic suppression of CHS after treatment with various types of nonionizing radiation. Treatment of mice with UVA (320-400 nm) radiation eliminated detectable Langerhans cells from the exposed skin, based on ATPase staining and electron microscopy, but did not reduce CHS; in fact, CHS was enhanced in these animals. Neither rose bengal nor eosin, in combination with visible (greater than 400 nm) radiation, affected the number or appearance of Langerhans cells, even though microscopic evidence of phototoxicity was present. However, rose bengal plus visible radiation depressed CHS reactions that were induced and elicited through unexposed skin. Depletion of Langerhans cells from dorsal skin by exposure to UVA radiation did not prevent suppression of CHS by subsequent exposure of the Langerhans cell-depleted skin to UVB radiation. We conclude that systemic suppression of CHS by UVB irradiation is not related to the numerical and morphological alterations in Langerhans cells that occur locally at the site of irradiation.

Animals↗

Low-concentration homogenized castor oil eye drops for noninflamed obstructive meibomian gland dysfunction.

OBJECTIVE: We developed low-concentration homogenized castor oil eye drops for the treatment of patients with noninflamed obstructive meibomian gland dysfunction (MGD), a major cause of lipid-deficiency dry eye, and assessed the safety, stability, and efficacy of the eye drops. DESIGN: Randomized, double-masked, placebo-controlled crossover clinical trial. PARTICIPANTS: Forty eyes of 20 patients with noninflamed MGD. METHODS: After a preliminary study of eye drops containing castor oil, 2% castor oil and 5% polyoxyethylene castor oil (emulsifier) were mixed to formulate homogenized oil eye drops. The patients were assigned randomly to receive oil eye drops or placebo six times daily for 2 periods of 2 weeks each. MAIN OUTCOME MEASURES: At the end of each treatment period, we assessed symptoms, tear interference grade, tear evaporation, fluorescein and rose bengal scores, tear break-up time (BUT), and meibomian gland orifice obstruction. Safety and stability tests were also performed. RESULTS: Symptom scores, tear interference grade, tear evaporation test results, rose bengal scores, tear BUT, and orifice obstruction scores after the oil eye drop period showed significant improvement compared with the results after the placebo period. No complications attributable to the eye drops were observed. The oil eye drops were stable when stored at 4 degrees C. CONCLUSIONS: The results indicate that castor oil eye drops are effective and safe in the treatment of MGD. The possible mechanisms of this treatment are improvement of tear stability as a result of lipid spreading, ease of meibum expression, prevention of tear evaporation, and the lubricating effect of the oil eye drops.

Castor Oil↗

Decreased tear lactoferrin concentration in patients with chronic hepatitis C.

BACKGROUND/AIMS: Decreased tear volume in patients with chronic hepatitis C has been reported in the literature. Lactoferrin is abundantly present in human tears, the main source of which is the acini of the lacrimal glands. In this study tear lactoferrin levels were measured to investigate the dry eye condition of patients with chronic hepatitis C. METHODS: Lactoferrin in tears/fluid was measured by a radial immunodiffusion assay in 42 patients with chronic hepatitis C. The rate of lacrimal secretion was determined by the cotton thread test. Rose bengal staining of the ocular surface was also performed. RESULTS: Only three patients out of 42 complained of dry eye sensation and, in 31 patients, six showed positive results on the rose bengal staining test of the ocular surface. The lactoferrin concentration of tear fluid in the chronic hepatitis C group (1.42 (SD 0.56) mg/ml) was significantly lower than in the control group (1.90 (0.62) mg/ml; p <0.00048). The cotton thread test results in the chronic hepatitis C group (12.9 (5. 5) mm) were significantly lower than in the control group (17.9 (5. 3) mm; p<0.00048). Also, in the chronic hepatitis C group, tear lactoferrin concentration correlated with the results of the cotton thread test (r = 0.35, p<0.05). CONCLUSION: Chronic hepatitis C patients showed both decreased tear volume, and decreased tear lactoferrin concentration. These findings suggest that there may be dysfunction of the lacrimal glands in patients with chronic hepatitis C, which may account for the mild dry eye.

Adult↗

The efficacy of eye care for ventilated patients: outline of an experimental comparative research pilot study.

The aim in this study was to investigate the efficacy of eye care given to mechanically ventilated and unconscious patients in an intensive care unit; and to answer the research question: "Is eye surface integrity maintained after the application of a "Geliperm" dressing to the closed eye of a ventilated patient?' Six patients formed the sample group. In each patient one eye received standard eye care, forming the control group, and the other eye received a 'geliperm' dressing, forming the treatment group. Tear production, using the Schirmer test, and the degree of eye surface staining by rose bengal were used as the measurement indicators. As a pilot study this project was primarily concerned with establishing a sound methodology for further enquiry. There were some difficulties in validating the degree of rose bengal staining; no statistically significant difference could be demonstrated between outcomes of the two forms of eye care after a 24-hour test period, with a P = 0.05 significance level, using the Mann-Whitney-U test. It should be noted, however, that the sample size was too small to measure a statistically significant change or its absence, in either the treatment or control eye between pre- and post-test results, with the same significance level, and using the Wilcoxon signed rank test. This study does provide evidence to support the need for eye care for critically ill patients. The methodology may offer a way forward for future investigation in the search for optimal eye surface preservation.

Aged↗

[A comparative study of 0.15% sodium hyaluronate versus polyvinyl alcohol in the treatment of dry eyes].

PURPOSE: To evaluate the efficacy of two artificial tears for the treatment of dry eyes: 0.15% sodium hyaluronate and polyvinyl alcohol. METHODS: The study included 60 dry eyes in 30 patients. Fifteen of the patients were treated with 0.15% sodium hyaluronate and 15 with polyvinyl alcohol. Five drops a day for one month was given in both groups. Visual acuity, Schirmer's test, tear clearance, dry eye symptoms questionnaire, fluorescein and rose bengal staining, corneal sensibility, bulbar hyperemia, tear detritus, meibomian gland plug, palpebral erythema and edema before and after treatment were evaluated. RESULTS: Symptoms as judged by the dry eye questionnaire, as well as both fluorescein and rose bengal staining, were significantly improved by 0.15% sodium hyaluronate. The polyvinyl alcohol treated patients did not show a significant improvement in any parameter. CONCLUSION: Sodium hyaluronate was more effective than polyvinyl alcohol in the treatment of patients with dry eyes (Arch Soc Esp Oftalmol 2005; 80: 387-394).

Adjuvants, Immunologic↗

[Activity of lacrimal plasmin-like enzymes in the evaluation of the corneal state after photorefraction surgery].

Forty-three patients (58 eyes) were observed, who used contact lenses for a long time before photorefraction operations (photorefraction keratectomy and laser specialized keratomileusis--PRK and LASIK). Measurements of lacrimal plasmin-like enzymes (PLE) were carried out before and after operations on a Uniscan II microreader. Corneal and conjunctival intactness was evaluated by Bengal rose staining. Before surgery PLE activity was 3.5 times higher than normally in the patients using contact lenses. After PRK and LASIK, PLE activity increased and was 25% lower after LASIK than after PRK. Based on the data of preoperative diagnosis (Bengal rose test, measurement of PLE activities, Norn and Schirmer tests), the patients with contact lens intolerance without hypoproduction of tears are recommended rehabilitation treatment with drugs activating the repair processes (Vitacic, etc.), while patients with contact lens intolerance and tear hypoproduction are recommended low concentrations of viscoelastic drugs (Vismed).

Contact Lenses↗

Photodegradation and photosensitization of mycosporine-like amino acids.

The photodegradation and photosensitization of several mycosporine-like amino acids (MAAs) were investigated. The photodegradation of the MAA, palythine, was tested with three photosensitizers: riboflavin, rose bengal and natural seawater. For comparison of degradation rates, the riboflavin-mediated photosensitization of six other MAAs was also examined. When riboflavin was used as a photosensitizer in distilled water, MAAs were undetectable after 1.5h. Palythine showed little photodegradation when rose bengal was added as the photosensitizer (k=0.12x10(-3)m(2)kJ(-1)). Palythine dissolved in natural seawater containing high nitrate concentrations also showed slow photodegradation rate constants (k=0.26x10(-3)m(2)kJ(-1)) over a 24-h period of constant irradiation. Similar experiments in deep seawater with porphyra-334 and shinorine resulted in 75% of the initial MAA remaining after 4h of irradiation and rates of 0.018 and 0.026x10(-3) m(2) kJ(-1), respectively. Experiments conducted in deep seawater with riboflavin additions resulted in photodegradation rate constants between 0.77x10(-3) and 1.19x10(-3)m(2)kJ(-1) for shinorine and porphyra-334, respectively. Photoproduct formation appeared to be minimal with the presence of a dehydration product of the cycloheximine ring structure indicated as well as the presence of amino acids. Evidence continues to build for the role of MAAs as potent and stable UV absorbers. This study further highlights the photostability of several MAAs in both distilled and seawater in the presence of photosensitizers.

Amino Acids↗

[Removal of the vertical portion of the lacrimal canaliculus in dry eye syndrome].

PURPOSE: To demonstrate the efficacy and possible complications of a surgical technique that includes the removal of the vertical portion of the lacrimal canaliculus in patients with dry eye syndrome. METHODS: A study was performed on six canaliculi of six eyes (four patients). Three patients had dry eye, associated with primary Sjögren syndrome. One of the four patients developed keratoconjuntivitis sicca due to lacrimal gland removal. The criteria included: patients with symptoms of dry eye that did not improve even with the continuous use of artificial tears, low results (less than 5 mm) with the Schirmer test, rose bengal staining and cases of recanalization after thermal occlusion with electrocauterium. The lacrimal puncta were examined after 7, 15, 30, 60, 90 and 180 days after surgery. RESULTS: None of the canaliculus recanalized at this time. Punctate keratitis, corneal filaments, Schirmer and rose bengal tests improved in 5 eyes. Diffuse punctate keratitis was present in the patient with lacrimal gland removal even 2 months after of surgery. Eyelid margin complications were not observed. CONCLUSION: This technique is simple and effective in permanent lacrimal canaliculus occlusion. We did not note any complications compared to other procedures.

Adult↗

Lacrimal and salivary immunoglobulins in Sjögren's syndrome.

The diagnosis of Sjögren's syndrome is sometimes difficult. We have previously demonstrated the diagnostic value of salivary immunoglobulins IgG and IgM in Sjögren's syndrome. In the present study, we assessed both lacrimal immunoglobulins (1 Ig) and salivary Ig (s Ig) in Sjögren's syndrome. We studied 112 patients: 71 had rheumatoid arthritis (57 sero positive), 19 had connective tissue diseases or vasculitis (six Sjögren's syndrome alone, six systemic sclerosis, three mixed connective tissue diseases, three polyarteritis, one relapsing polychondritis), and 22 patients had other inflammatory, metabolic or degenerative joint diseases. Lacrimal Ig and salivary Ig were assessed by double immunodiffusion with antisera specific for IgG, IgM and IgA. Each Ig class was scored on a scale ranging from 0 to 3 plus without knowledge of the patient's diagnosis. The results of factorial analysis demonstrated a strong relationship between xerophtalmia, positive Schirmer's test, s IgG, s IgM, 1 IgG, and 1 IgM in patients with seropositive rheumatoid arthritis or other connective tissue diseases. Analysis of individual parameters showed a significant relationship between 1 IgG and ocular complaints (P less than 0.01), positive Schirmer's test (P less than 0.05), positive rose bengal dye test (P less than 0.05), 1 IgM (P less than 0.01), s IgG (P less than 0.01) and s IgM (P less than 0.05). A significant relationship was also found between s IgG and ocular complaints (P less than 0.02), positive rose bengal dye test (P less than 0.01), positive minor salivary gland biopsy (P less than 0.05), s IgM (P less than 0.01), and 1 IgM (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Seroprevalence of brucellosis in mithuns (Bos frontalis) in India.

Studies conducted on the seroprevalence of brucellosis in 98 mithuns maintained at the National Research Centre on Mithun, Nagaland, India revealed that the number of animals found positive for brucellosis in avidin-biotin enzyme-linked-immunosorbent assay, standard tube-agglutination test and Rose-Bengal plate test were 34, 20 and 11%, respectively. Highest prevalence of brucellosis was observed in the Mizoram mithun strain. The relative sensitivity and specificity of the standard tube-agglutination test were 61 and 100%, respectively; the corresponding values of the Rose-Bengal plate test were 33 and 100%, respectively.

Animals↗

Inhibitory effects of total flavones of Hippophae Rhamnoides L on thrombosis in mouse femoral artery and in vitro platelet aggregation.

Total flavones of Hippophae Rhamnoides L (TFH) are extracted from Sea buckthorn, a Chinese herbal medicine. Sea buckthorn has antioxidant, anti-ulcerogenic and hepato-protective actions, and its berry oil is reported to suppress platelet aggregation. Though it is frequently used for patients with thrombosis, the likely mechanism(s) and effects of TFH on thrombogenesis remain unclear. Thus, we have investigated the effect in-vivo of TFH on thrombogenesis and in vitro on platelet aggregation, comparing them to those of aspirin. We measured thrombotic occlusion time in a mouse femoral artery thrombosis model by the photochemical reaction between intravenously injected rose bengal and green light irradiation. In vitro platelet aggregation in whole blood was measured by single platelet counting. Thrombotic occlusion time was 8.5 +/- 0.6 min in the control group. TFH at a dose of 300 micro g/kg, intravenously administered 15 min before the rose bengal injection, significantly prolonged it to 11.6 +/- 1.0 min (P < 0.05), a similar effect on in-vivo thrombogenesis to that of aspirin. TFH at a concentration of 3.0 micro g/ml significantly (P < 0.01) inhibited in vitro platelet aggregation induced by collagen (2 micro g/ml) in a concentration dependent manner, in contrast TFH did not affect aggregation induced by arachidonic acid (80 micro M) and ADP (0.3 micro M). The results of the present study, in which TFH prevented in-vivo thrombogenesis, probably due to inhibition of platelet aggregation, suggest a possible clinical approach for the prevention of thrombosis.

Animals↗

Photochemical brain injury in rats triggers DNA fragmentation, p53 and HSP72.

The aim of the study was to examine whether apoptosis, apoptosis-related protein p53 and heat-shock protein (HSP) 72 participate in the response of the brain to focal injury. Male Sprague-Dawley rats received intravenously a photosensitive dye rose bengal. Unilateral cortical thrombosis was induced by illuminating the skull of rose bengal-treated rats for 10 min with a focused beam of light. Animals were killed and brains were processed for immunohistochemical detection of DNA fragmentation, p53, and HSP72 kD. DNA fragmentation and p53 were increased only in the perifocal area in the cortex ipsilateral to the thrombotic focus, while HSP72 increased throughout the ipsilateral cortex, except in the immediate perifocal area. The results suggest that in response to focal brain injury, some cells die through an apoptotic process that might involve an accumulation of p53.

Animals↗

Effect of cholephilic dyes on hepatic tight junctional permeability in the rat.

Changes in biliary permeability during cholephilic dye-induced choleresis, as assessed by measuring the movement into bile of two permeability probes, [14C]sucrose and horseradish peroxidase, were analyzed following an i.v. infusion (60 nmol/min per 100 g body wt) of the model cholephilic organic anion sulfobromophthalein in rats. Dye infusion led to a progressive increase of the [14C]sucrose bile-to-plasma ratio, which reached a maximum value after 100 min of dye infusion (+97%). Paracellular entry of horseradish peroxidase, as evaluated by the early peak of its biliary appearance curve, was also selectively increased (+69%), without changes in the later (transcytotic) access of the protein. Additional dose-response studies of biliary permeability to [14C]sucrose, using sulfobromophthalein and rose bengal, showed that this effect was dose-dependent and rapidly reversed by interruption of dye administration. The influence of hydrophobic/hydrophilic balance on this effect was also studied by infusing four dyes covering a broad range of hydrophobicity (phenol red, bromocresol green, sulfobromophthalein, and rose bengal), so as to attain a similar value of dye hepatic content at the end of the experiment (approximately 150 nmol/g liver wt). Under these conditions, a strong positive correlation was found between the increase in biliary permeability to [14C]sucrose and dye hydrophobicity. These results suggest that cholephilic dyes increase tight junctional permeability in a reversible and dose-dependent manner, and that this effect depends on the hydrophobic/hydrophilic balance of the dye.

Animals↗

[Seroprevalence of brucellosis in agricultural workers in the coastal zones of Castellón, Spain].

OBJECTIVE: To estimate the seroprevalence of brucellosis in agricultural workers. MATERIAL AND METHODS: This cross-sectional study was conducted between 1996 and 1997 at Unidad de Salud Laboral del Centro de Salud Pública (CSP) in Castellón, España (Public Health Center Occupational Health Unit), among 482 subjects applying for certification to handle pesticides for agricultural use. Serologic testing consisted of rose Bengal staining, Wright's agglutination, and antibrucella Coombs tests: Statistical analysis consisted in estimation of prevalence figures and logistic regression modelling. RESULTS: Fifteen subjects (3.1%, 95% CI 1.8%-5.1%) had titers of 1/40 or higher to Wright and/or Coombs tests; all were negative to rose Bengal staining. No high occupational risk was observed. CONCLUSIONS: A low prevalence of brucellosis was found. The English version of this paper is available at: http://www.insp.mx/salud/index.html.

Adolescent↗

Open-label crossover study of vitamin A ointment as a treatment for keratoconjunctivitis sicca.

The authors evaluated the efficacy of all-trans retinoic acid (vitamin A) ointment as a treatment for keratoconjunctivitis sicca (KCS) in a group of 11 patients selected on the basis of clinical history, slit-lamp examination results, rose Bengal staining, and tear film osmolarity. In this open-label crossover study, vitamin A ointment was no more effective than placebo in increasing tear secretion, as indicated by Schirmer test with proparacaine or tear film osmolarity, or in decreasing ocular surface disease, as indicated by rose Bengal staining. Seven patients stated some preference for the placebo ointment, two patients for the vitamin A ointment, and two patients had no preference.

Adult↗

Vascular effects of singlet oxygen (1O2) generated by photo-excitation on adrenergic neurotransmission in isolated rabbit mesenteric vein.

It has been suggested that reactive oxygen species (ROS) may be involved in the regulation of vascular tone. However, the nature of ROS effects on vascular sensitivity remains to be elucidated. The present study was designed to investigate the effects of ROS, especially 1O2, on neurotransmission at the sympathetic neurovascular junction. Basal noradrenaline (NA) release, release of NA induced by electrical stimulation (ES), and resting NA release at the sympathetic nerve terminals were determined using a superfusion technique. The amount of NA was determined by HPLC; isometric tension changes evoked by ES were also recorded simultaneously. 1O2 was generated from Rose Bengal by photo-activation. The generation of 1O2 in the superfusate was monitored by electron spin resonance (ESR) using the spin trap 2,2,6,6-tetramethyl-4-piperidinol throughout the experimental time course. The ESR results confirmed that 1O2 was generated by photo-activation of Rose Bengal via the formation of 2,2,6,6-tetramethyl-4-hydroxyl-piperidinyloxy. Exposure of helical strips of rabbit mesenteric vein to 1O2 induced a significant increase in tension and NA release during the basal period, but had no effect on ES-induced release. L-histidine, an 1O2 scavenger, significantly inhibited the observed effects on vascular tension and NA release in response to 1O2. These results suggest that 1O2 may induce NA-mediated vasoconstriction at the postjunctional site, and may be associated with Ca(2+)-independent NA release from the prejunctional site of adrenergic neurotransmission.

Animals↗