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Uniform distribution of epithelial stem cells in the bulbar conjunctiva.

PURPOSE: To determine the distribution of epithelial stem cells in the bulbar conjunctiva by measuring homeostatic movements and mitosis of epithelial cells in this region. METHODS: The ubiquitous GFP mouse was used to monitor movement of conjunctival epithelial cells. Cell movement was determined by histology, analyzing the shape and distribution of GFP cell clusters in flat wholemount specimens, and by in vivo time-lapse microscopy, tracking the movement of GFP-positive cells in the bulbar conjunctiva near the limbus. Mitoses were determined by labeling DNA of adult mice with bromodeoxyuridine (BrdU) for 3 days. Label-retaining cells (LRCs) were determined by a pulse label of newborn mice with BrdU, followed by a chase of 6 to 7 weeks. RESULTS: Similar to the corneal epithelium, only some of the conjunctival epithelial cells expressed a high level of GFP. Histology showed that GFP-positive cells existed as clusters of several to several dozen cells. No stripe pattern of GFP was observed in any part of the conjunctiva, suggesting that directed cell movement was rare or nonexistent. Time-lapse analyses revealed that none of the tracked GFP clusters exhibited a continuous and directed movement and that most GFP clusters were stationary for several weeks and much longer in some occasions. BrdU labeling showed that GFP-positive cells in this region were mitotically active. BrdU pulse-chase experiments demonstrated that LRCs were distributed uniformly in this region. CONCLUSIONS: Epithelial cells of the bulbar conjunctiva near the limbus are mitotically active and yet they are generally immobile in a lateral direction, indicating that these cells are self-sufficient. These results, combined with the uniform distribution of LRCs, suggest that epithelial stem cells are distributed uniformly in this area.

Animals↗

Aquaporin-dependent water permeation at the mouse ocular surface: in vivo microfluorimetric measurements in cornea and conjunctiva.

PURPOSE: Fluorescence methods were developed to quantify membrane and tissue water permeabilities at the ocular surface and to compare water transport in wild-type mice versus transgenic mice lacking each of the water channels, aquaporin (AQP)-1, -3, and -5, normally expressed in cornea or conjunctiva. METHODS: Membrane water permeabilities (P(f)(mem)) of calcein-stained surface epithelial cells were measured from the kinetics of fluorescence quenching in response to rapid (<0.2 seconds) changes in extraocular fluid osmolarity. Tissue water permeabilities (P(f)(tiss)) across intact cornea and conjunctiva--the relevant parameters describing water movement into the hyperosmolar tear film in vivo--were determined by a dye-dilution method from the fluorescence of Texas red-dextran in an anisosmolar solution in a microchamber at the ocular surface. RESULTS: Osmotic equilibration occurred with an exponential time constant (tau) of 1.3 +/- 0.2 seconds (P(f)(mem) = 0.045 cm/s) in calcein-loaded corneal epithelial cells of wild-type mice, slowing 2.1 +/- 0.4-fold in AQP5-deficient mice; tau was 2.4 +/- 0.1 seconds in conjunctiva (P(f)(mem) = 0.025 cm/s), slowing 3.6 +/- 0.7-fold in AQP3-deficient mice. In dye-dilution experiments, P(f)(tiss) of cornea was 0.0017 cm/s and decreased by greater than fivefold in AQP5-deficient mice. P(f)(tiss) in AQP5-null mice was restored to 0.0015 cm/s after removal of the epithelium. P(f)(tiss) of conjunctiva was 0.0011 cm/s and was not sensitive to AQP3 deletion. CONCLUSIONS: These results define for the first time the water-transporting properties of the two principal ocular surface barriers in vivo. The permeability data were incorporated into a mathematical model of tear film osmolarity, providing insights into the pathophysiology of dry eye disorders.

Animals↗

[Study of mycobiota in the healthy conjunctiva of diabetics who reside in the urban area of the city of São Paulo, Brazil].

PURPOSE: To determine the mycobiota of the healthy conjunctiva in diabetic individuals, according to diabetes type, age, sex, disease time, type of treatment, and stage of diabetic retinopathy of the individuals. To identify the anemophilus mycobiota in the sampling rooms. METHODS: A cross-sectional study was carried out on 803 diabetics who reside in the urban area of São Paulo-SP/Brazil. Sabouraud's dextrose agar culture with chloramphenicol was used for primoisolation, and the key of De Hoog was used to identify filamentous fungi. RESULTS: Of the evaluated diabetics, 6.6% (53/803) presented type 1 diabetes and 93.4% (750/803) type 2. The positive cultures for fungi in the conjunctiva of diabetics was 4.2% (34/803), with 1.9% (1/53) in type 1 diabetics and 4.4% (33/740) in type 2 diabetics (p=0.720). With respect to the presence or not of isolated fungi, there was no statistically significant association regarding age (p=0.575), sex (p=0.517), disease time (p=0.633), type of treatment (p=0.422), and diabetic retinopathy stage (p=0.655) of the tested individuals. The identified fungi were all filamentous: Aspergillus spp. represented 59.5% (25/42) of isolations and 47.6% (20/42) of isolated species were Aspergillus niger. Growth of anemophilus fungi occurred in the air of the room and coincidences were observed between the isolated species from the air and those from the conjunctiva. CONCLUSIONS: Presence of mycobiota in healthy conjunctivas of diabetics was identified, with no significant association between the greater number of positive fungi isolations and the type of diabetes, age, sex, disease type, type of treatment, and stage of diabetic retinopathy. In the collection rooms, anemophilus mycobiota was identified.

Adolescent↗

Active chloride transport in the pigmented rabbit conjunctiva.

The present study demonstrates, for the first time, that the excised pigmented rabbit conjunctiva is a tight barrier capable of active Cl- transport. The transepithelial potential difference was 17.7 +/- 0.8 mV (tear-side negative), the short-circuit current was 14.5 +/- 0.7 microA/cm2, and the transconjunctival resistance was 1.3 +/- 0.1 k omega.cm2 for n = 45 tissues. Various inhibitors including ouabain (a Na+/K(+)-ATPase inhibitor), amiloride (a Na+ transport blocker), N-phenylanthranilic acid (a chloride transport inhibitor), bumetanide (an inhibitor of Na(+)-(K+)-Cl- cotransport process), and BaCl2 (a K+ channel blocker) were used on the mucosal and serosal sides of the tissue mounted in Ussing chambers to determine the involvement of the respective ion transport processes in the observed short-circuit current across the conjunctiva. The results suggest that a Cl- conductive pathway is present on the mucosal side of the conjunctiva, whereas Na+/K(+)-ATPase, Na(+)-(K+)-Cl- cotransport process, and K+ conductive pathways are present on its serosal side. Amiloride-sensitive Na(+)-conductive pathways do not appear to be present on either side of the pigmented rabbit conjunctiva.

Animals↗

Contribution of Na(+)-glucose cotransport to the short-circuit current in the pigmented rabbit conjunctiva.

The objective of this study was to determine whether Na(+)-glucose cotransport contributed to the residual short-circuit current (Isc) in the pigmented rabbit conjunctiva not accounted for by Cl- secretion. The Isc of the pigmented rabbit conjunctiva was measured following exposure of its mucosal or serosal side to varying concentrations of D-glucose or phloridzin in a glutathione-bicarbonated Ringer's solution (GBR). Addition of D-glucose to the mucosal side of the conjunctiva bathed in glucose-free GBR elevated Isc up to 26 +/- 6% in a dose-dependent manner (K0.5 = 1.2 mM), while mucosal (but not serosal) addition of phloridzin reduced the Isc (IC50 = 0.05 mM) of the conjunctiva bathed in regular GBR. In a mucosal Na(+)-free medium, neither 20 mM D-glucose nor 0.5 mM phloridzin was effective. In a mucosal glucose-containing medium, deletion of Na+ reduced Isc up to 27 +/- 4%. It was, however, restored to its initial value upon the addition of 17.5-141 mM Na+ to a medium containing 5 mM D-glucose. Hill plot analysis of Isc elevation at 141 mM Na+ in the presence of varying D-glucose concentrations in the mucosal bathing fluid yielded a Hill coefficient of 0.86. There is, therefore, evidence for the apical localization of phloridzin-sensitive Na(+)-coupled D-glucose transport that exhibits apparent 1:1 stoichiometry, being responsible for the maximal 26% increase in the Isc.

Animals↗

Functional characterization of organic cation drug transport in the pigmented rabbit conjunctiva.

PURPOSE: To characterize carrier-mediated organic cation drug transport in the rabbit conjunctiva. METHODS: The transport of [14C]guanidine, the model substrate, in the excised pigmented rabbit conjunctiva was evaluated in the modified Ussing chamber. Tetraethylammonium (TEA) transport also was investigated to determine substrate specificity. RESULTS: The apparent permeability coefficient for guanidine and TEA in the mucosal-to-serosal (ms) direction was 5.4 and 49.6 times greater than that in the serosal-to-mucosal (sm) direction, respectively. Guanidine transport in the ms (but not sm) direction revealed temperature and concentration dependency over 0.02 to 10 mM with an apparent Michaelis-Menten constant of 3.1 mM and a maximal flux of 11.4 nmol/(cm2 x h). Net guanidine transport measured at 0.1 mM across the conjunctiva was decreased by 71% or 82%, respectively, on the addition of 1 microM valinomycin (a K+ ionophore) in both bathing fluids or in a high K+ buffer in the mucosal fluid. Interestingly, net guanidine transport was reduced, rather than enhanced, by 63% upon acidifying the mucosal bathing fluid. By contrast, net guanidine transport was not affected by the serosal presence of 0.5 mM ouabain (a Na+, K+-ATPase inhibitor), by the mucosal and serosal presence of 0.1 microM monensin (a Na+ ionophore) or 0.3 microM carbonyl cyanide p-(trifluoromethoxy)phenyl-hydrazone (FCCP, a H+ ionophore). Guanidine transport in the ms direction was polyspecific, as indicated by the 48% to 82% inhibition by structurally diverse amines. In particular, guanidine ms transport was inhibited by the antiglaucoma drugs dipivefrine (72%), brimonidine (70%), and carbachol (78%). CONCLUSIONS: A carrier-mediated organic cation transport process appears to exist in the conjunctiva, mediating the absorption of organic amines, including certain amine-type ophthalmic drugs. This process may be driven by an inside-negative apical membrane potential difference.

Animals↗

[Changes in the vascular wall in the conjunctiva of patients with diabetes mellitus].

Venous capillary wall in type II diabetic patients' conjunctiva was examined. The study was performed on conjunctiva fragments removed from diabetic patients during the operation of senile cataract. The fragments were fixed in Lillie's solution then studied by using optical microscopy with usual but histochemical stainings, too. Conjunctiva fragments removed from patients of the same age, being operated by senile cataract and one conjunctiva from ten years old patient were examined by using the same techniques as they could be compared. Computer determinations of the external and internal diameter of the same capillary in all three groups were performed. Venous capillary wall thickening by type IV collagen hyperproduction in all the aged patients was noted. This kind of thickening is more revealed in aged diabetic person. Type I-III collagen presence in the capillary wall of the diabetic patients was noted, too. The average value of the differential between the external and internal diameter was increased in the diabetic patients with diabetic retinopathy. Venous capillary wall thickening in the diabetic patients is due to the basal membrana both by means of the type IV collagen hyperproduction and presence of the I-III collagen, too.

Aged↗

Bacterial flora of the conjunctiva and nasal cavity in normal and diseased captive bustards.

A survey was carried out to describe the normal aerobic bacterial flora of the conjunctiva and nasal cavity of captive houbara bustards (Chlamydotis undulata), kori bustards (Ardeotis kori), and white-bellied bustards (Eupodotis senegalensis) maintained at the National Avian Research Center, Abu Dhabi, United Arab Emirates. A total of 58 samples were examined from the nasal cavity and 55 samples from the conjunctiva of healthy bustards. There was no bacterial growth in 45% of conjunctival samples. Bacteria isolated from the conjunctiva of healthy birds included Micrococcus spp., Staphylococcus auricularis, Staphylococcus xylosus, Staphylococcus capitis, Staphylococcus warneri, Bacillus spp., and Enterobacter amigenus. Bacteria isolated from the nasal cavity of healthy birds included Bacillus spp., Micrococcus spp., S. auricularis, S. xylosus, Staphylococcus simulans, Staphylococcus saprophyticus, Staphylococcus hyicus, Staphylococcus cohnii, Staphylococcus sciuri, Aerococcus spp., and Providencia rettgeri. These findings were compared with bacterial isolates from bustards with clinical signs of ocular or upper respiratory tract diseases. Mycoplasma spp., Pseudomonas aeruginosa, Pseudomonas stutzeri, Proteus mirabilis, Escherichia coli, Klebsiella spp., Aeromonas hydrophila, and Staphylococcus aureus were the pathogenic bacteria isolated from the conjunctiva of 34.3% bustards with ocular discharges. Mycoplasma spp., P. aeruginosa, Pseudomonas spp., P. mirabilis, E. coli, Klebsiella pneumoniae, and S. aureus were the pathogenic bacteria isolated from the nasal cavity of 74% bustards with upper respiratory tract diseases.

Animals↗

[Experimental study and application of extracellular matrix of conjunctiva].

OBJECTIVE: To investigate the characteristics and application of extra-cellular matrix (ECM) of conjunctiva. METHODS: Pieces of conjunctival ECM were made, and then the conjunctival defect was repaired with that in an experimental group. Control group 1 received the heterogeneous conjunctiva, and in control group 2, the refrigerated ECM was used for transplantation. All of them were photographed and examined by light microscope, electron microscope (EM), immunohistochemical examination and lymphocyte toxicity test. RESULTS: There were blood vessels growing into the grafts on the 3rd day in the experimental group. The grafts became mildly congestive and similar to normal conjunctival ones in the 4(th) week. But in control group 1, the grafts became white and necrotic in the second week; the conjunctiva was congestive, cicatricial and hyperplastic in the 4(th) week. Under the light microscope, the epithelium covered the most part of the region with the graft in the 2nd week, covered completely in the 4(th) week, and its appearance was as an approximately normal conjunctival epithelium in the experimental group. But the epithelium became ulcerative and a lot of lymphocytes infiltrated in the 2nd week in control group 1. Under the EM, the line of demarcation between the covering region of the regenerative conjunctival epithelium and the naked ECM was quite clear in the 1st week in the experimental group. And the ultrastructure of regenerative conjunctival epithelium was basically the same as an normal one in the 8(th) week. Both before and after transplantation, the immunohistochemical examinations verified that ECM was type I and IV collagen. The lymphocyte toxicity test showed that no obvious humoral immunoreaction existed. CONCLUSION: Conjunctival ECM has the characteristics of tissue activity and low antigenicity. It is an ideal conjunctival succedaneum in the plastic surgery of conjunctiva.

Animals↗

Biochemical and ultrastructural study of human diabetic conjunctiva.

Combined biochemical and ultrastructural study of conjunctival biopsies of 27 normal subjects and 45 diabetics (40 to 60 years old) was made. The "in vitro" incorporation of 14C-glucosamine and 3H-proline in freshly excised conjunctival biopsies was studied. The alterations of the capillary basement membrane of the conjunctiva were studied by electron microscopy. The following results were obtained: 1) A decrease of the specific activity of 14C-glucosmaine incorporation was found in fractions of diabetic conjunctiva. 2) In diabetic conjunctiva the percentages of 3H-proline incorporation in polymeric collagen containing fraction and structural glycoproteins containing fraction were significantly increased with a parallel decrease of 3H-proline incorporation in "crude soluble collagen" fraction expressed as a percentage of total incorporation. 3) Significant thickening of capillary basement membrane was observed with the appearance of collagen-like fibrils within the basement membrane in diabetic conjunctiva. Such fibrils were not seen in normal basement membranes. A relation between the extent of basement membrane thickening and the appearance of collagen-like fibrils is suggested. 4) The higher percentage of incorporation of 3H-proline in polymeric collagen may be related to the appearance of collagen fibrils in thickened basement membranes of the diabetic conjunctival capillaries. 5) These results suggest an abnormal regulation of the relative rate of biosynthesis and/or excretion of intercellular matrix macromolecules (collagen, structural glycoproteins) as part of the metabolic disorders characterising diabetes.

Adult↗

New insights into the surface cells and glands of the conjunctiva and their relevance to the tear film.

BACKGROUND: A number of features of the conjunctiva are highlighted clinically, but there are limited data on the ultrastructure underlying these features. A series of systematic studies have been undertaken to address this, using both scanning electron microscopy (SEM) and transmission electron microscopy (TEM). METHODS: Tissue from pigmented rabbits (2 to 2.5 kg) was prepared for electron microscopy, with special attention being paid to the preparation of the palpebral or bulbar conjunctival tissue by stretching it to its fully extended configuration. The fixative (2% glutaraldehyde in 80 mM cacodylate, pH 7.2 to 7.4, 320 to 340 mOsm/kg) has been previously validated in studies on corneal tissue. RESULTS: Both SEM and TEM reveal the presence of a substantial network of surface pores and underlying ducts associated with the acini of the tarsal accessory gland of Wolfring. These are predominantly found close to the tarsal-orbital boundary of the palpebral conjunctiva. The ductal system is lined with cells containing prominent secretory granules and has features that suggest that it readily collapses if the tissue is not adequately prepared. The same preparative technique also shows the presence of large and open lymphatic vessels in the stroma under the bulbar conjunctiva. Secretory granules and vesicles can also be found associated with the surface ducts and with the surface cells of the palpebral conjunctiva. Studies on the goblet cells located within the conjunctival cell layers also show that the shape of the cells can change substantially with the preparative technique. CONCLUSIONS: The outcome of these studies indicate that there is still much to be learned concerning the contribution of the accessory lacrimal glands, secreted mucins, the goblet cells, and the lymphatic system to the maintenance of a normal healthy ocular surface.

Animals↗

The T cell receptor in normal and inflamed human conjunctiva.

The majority of human peripheral blood and lymphoid tissue T cells express the TCR alpha/beta heterodimer, while the TCR gamma/delta is expressed on only a small subset of T lymphocytes. However, the majority of murine intraepithelial lymphocytes and most Thy-1+ murine dendritic epidermal cells express the TCR gamma/delta. Selective homing of avian TCR gamma/delta bearing lymphocytes to the intestinal epithelium also has been shown. These findings have suggested that these cells play a role against transformation and infection. More recently, a role in autoimmunity also has been proposed. We examined normal human conjunctiva and inflamed conjunctiva from patients with ocular cicatricial pemphigoid (OCP), an autoimmune disorder, and atopic keratoconjunctivitis (AKC). The majority of T cells in the epithelium and substantia propria of normal conjunctiva expressed the TCR alpha/beta. Tropism of TCR gamma/delta-expressing lymphocytes to normal human conjunctiva was not present. However, in OCP, there was a statistically significant increase in the absolute number of TCR gamma/delta cells/mm2 (epithelium, 33.9 +/- 10.5 [mean +/- standard error of the mean] vs. 159.9 +/- 51.5, P = less than 0.0008; substantia propria, 4.1 +/- 0.9 vs. 240.1 +/- 191.3, P less than 0.002) and TCR gamma/delta cells as a percentage of CD3+ cells (epithelium, 0.18 +/- 0.06 vs. 0.39 +/- 0.07, P = less than 0.02; substantia propria, 0.10 +/- 0.05 vs 0.33 +/- 0.08, P = less than 0.03). This was not the case for AKC. These findings suggest that TCR gamma/delta lymphocytes play a specific but undefined role in certain conjunctival inflammatory conditions and may be important in autoimmunity.

Antibodies, Monoclonal↗

In vivo laser and white-light confocal microscopic findings of human conjunctiva.

BACKGROUND AND OBJECTIVE: To report in vivo laser and white-light confocal microscopic findings of human conjunctiva to investigate the potential application of these confocal microscopes as diagnostic devices for normal and pathologic conjunctiva. PATIENTS AND METHODS: Two healthy volunteers, 28- and 35-year-old men, participated in this study. The temporal bulbar conjunctiva, which was approximately 5 mm away from the limbus, was examined by in vivo laser and white-light confocal microscopic analysis. RESULTS: Using laser confocal microscopy, at least two different cell types could be distinguished in both subjects: conjunctival superficial epithelial cells and conjunctival basal epithelial cells. In contrast, conjunctival epithelial cells could not be visualized using white-light confocal microscopy. CONCLUSIONS: These results indicate that laser confocal microscopy, but not white-light confocal microscopy, can be used to visualize in vivo human conjunctiva. Further investigations in a large number of normal subjects and in patients with conjunctival pathologies are required to fully evaluate the usefulness of this device in daily clinical ophthalmology.

Adult↗

[The second, mucus-secreting system of the conjunctiva. Ultrastructural findings].

The second mucus-secreting system (type II cells according to our classification) of the conjunctiva is important for moisturization of the ocular surface epithelia. Ultrastructurally, a merocrine and an apocrine type of secretion can be differentiated. The merocrine secretion (epithelial cells containing homogeneous or inhomogeneous osmiophilic granules) appears more or less in all conjunctival areas. The apocrine mode of secretion is only found in the multilayered cylinder epithelium of the fornix conjunctivae. On the whole, the type II cells with inhomogeneous, osmiophilic vesicles are the most common. In the apical area of these cells secretory vesicles either coalesce with the cell membrane or they are completely released to the cell surface. Within the human conjunctiva about 10% of the superficial epithelial cells have secretory functions. Their number slightly increases in the temporal areas of the conjunctiva and in persons under 20 years and over 60 years of age. These results were obtained by electron microscopical investigations of conjunctival biopsies from 48 persons without conjunctival diseases.

Adolescent↗

Iontophoresis of 5-fluorouracil into the conjunctiva and sclera.

It was investigated in the rabbit eye whether the method of iontophoresis could introduce a sufficient amount of 5-fluorouracil (5-FU) to inhibit fibroblast proliferation into the conjunctiva and sclera, thus considerably reducing the total amount of 5-FU which must be given to the eye. Five-FU was introduced with a simple apparatus which was filled with 5% 5-FU solution and connected to the negative pole of a current source. When the apparatus was placed over the cornea, the 5-FU penetration into the cornea showed a correlation with strength of current (0-0.75 mA). When the apparatus was placed over the bulbar conjunctiva, a sufficient amount of 5-FU to inhibit fibroblast proliferation was introduced into the conjunctiva and sclera with a current of 0.5 mA passed for 30 seconds. Immediately after iontophoresis, mean 5-FU concentrations in the conjunctiva and sclera at the iontophoresis site were 480 and 168 micrograms/g, respectively. They decreased to 0.6 and 1.2 micrograms/g by 10 hr, but were still above the reported ID50 levels for the cultured conjunctival fibroblast. On the other hand, the total amount of 5-FU introduced into the eye averaged only 3.7 micrograms and the current used was much lower than that formerly applied to the patient's cornea.

Animals↗

Synthesis of slow reacting substance-like activity in rabbit conjunctiva and anterior uvea.

The ability of rabbit conjunctiva and anterior uvea to synthesize lipoxygenase products was assessed. Using autoradiographic techniques, we demonstrate that rabbit anterior uvea synthesizes 5 and 12 lipoxygenase products such as 12-HETE, 5-HETE and 5,12-DiHETE and cyclooxygenase product HHT from 14C-arachidonic acid. Indomethacin pretreated conjunctiva and anterior uvea generated slow reacting substance (SRS)-like activity from arachidonic acid in the presence of reduced glutathione and A23187. This SRS-like activity contracted guinea pig ileum. Specific SRS-like activity antagonist FPL-55712 inhibited the contractions of guinea pig ileum induced by SRS-like substance generated by either conjunctiva or anterior uvea. The activity was still present in the sample following extraction with organic solvents. SRS-like activity was destroyed by arylsulfatase and its generation was prevented by either boiling or pretreatment with cyclooxygenase/lipoxygenase inhibitors, BW755 and nordihydroguaiacetic acid. These results indicate that following cyclooxygenase inhibition by indomethacin rabbit conjunctiva and anterior uvea generate SRS-like activity from arachidonic acid via lipoxygenase pathways.

4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyraz↗

Quantitative ocular bacteriology: a method for the enumeration and identification of bacteria from the skin-lash margin and conjunctiva.

Type 1 Calgiswabs were used to collect bacterial specimens from the skin-lash margins and conjunctivae of normal volunteers. The swabs were held in a nonnutritive balanced salt solution during transportation to the laboratory. Alginate swabs were dissolved in a two-step procedure in TC Eagle's medium and 2.5% sodium hexametaphosphate solution. Aliquots of suspended bacteria from the dissolved swab were plated on blood agar. After incubation, bacterial colonies present were enumerated and identified. Cultures from both the skin-lash margin and conjunctiva show significant variation in the number and types of microorganisms isolated. Quantities of bacteria isolated from the lid margin were usually greater than from the conjunctiva. Staphylococcus epidermidis was the most prevalent bacterium isolated. The adsorption of bacteria onto alginate swabs from bacterial suspensions and the recovery of a red pigment-producing organism from the conjunctivae of rabbit eyes previously inoculated show that the method is sensitive and highly reproducible.

Alginates↗

[Comparison between local and systemic administration of cyclosporin A on the effective level in conjunctiva, aqueous humor and serum].

Cyclosporine is an important tool for the therapy of immunological diseases of the cornea and conjunctiva, as well as the treatment of patients with high-risk corneal grafts. However, potentially severe systemic side effects are disadvantageous. The purpose of our study was to determine if topical ocular application leads to about the same concentrations in ocular tissues as systemic application. Therefore, the concentration of cyclosporine in conjunctiva, aqueous humor and blood was measured by radioimmunoassay in six patients with systemic administration of cyclosporine and ten patients who received one drop of topical cyclosporine 2% four times prior to cataract surgery. All patients with systemic application showed measurable concentrations of cyclosporine in blood, as did four patients in the group receiving topical cyclosporine. There was no significant difference between both groups concerning cyclosporine concentration in aqueous humor. The level of cyclosporine in the conjunctiva was significantly higher after topical application (P < 0.02). In conclusion, therapy with cyclosporine eye drops results in levels in the conjunctiva and aqueous humor that are comparable to or even higher than those after systemic application if the last application was no more than 18h prior to the measurement. Therefore, topical ocular application of cyclosporine, which reduces or eliminates the drug's systemic side effects, can be used to induce local immunosuppressive activity, particularly in the treatment of superficial immunological diseases and after limbal allograft transplantation.

Administration, Oral↗