Two α-L-arabinofuranosidases from Bifidobacterium longum subsp. longum are involved in arabinoxylan utilization.

Arabinoxylan (AX) and arabinoxylooligosaccharides (AXOs) are carbohydrate sources utilized by Bifidobacterium longum subsp. longum. However, their degradation pathways are poorly understood. In this study, we characterized two genes, BLLJ_1850 and BLLJ_1851, in the hemicellulose-degrading gene cluster (BLLJ_1836-BLLJ_1859) of B. longum subsp. longum JCM 1217. Both recombinant enzymes expressed in Escherichia coli exhibited exo-α-L-arabinofuranosidase activity toward p-nitrophenyl-α-L-arabinofuranoside. BlArafE (encoded by BLLJ_1850) contains the glycoside hydrolase family 43 (GH43), subfamily 22 (GH43_22), and GH43_34 domains. The BlArafE GH43_22 domain was demonstrated to release α1,3-linked Araf from AX, but the function of BlArafE GH43_34 could not be clearly identified in this study. BlArafD (encoded by BLLJ_1851) contains GH43 unclassified subfamily (GH43_UC) and GH43_26 domains. The BlArafD GH43_UC domain showed specificity for α1,2-linked Araf in α1,2- and α1,3-Araf double-substituted structures in AXOs, while BlArafD GH43_26 was shown to hydrolyze α1,5-linked Araf in the arabinan backbone. Co-incubation of BlArafD and BlArafE revealed that these two enzymes sequentially removed α1,2-Araf and α1,3-Araf from double-substituted AXOs in this order. B. longum strain lacking BLLJ_1850-BLLJ_1853 did not grow in the medium containing α1,2/3-Araf double-substituted AXOs, suggesting that BlArafE and BlArafD are important for the assimilation of AX. KEY POINTS: • BlArafD GH43 unclassified subfamily domain is a novel α1,2-L-arabinofuranosidase. • BlArafE GH43 subfamily 22 domain is an α1,3-L-arabinofuranosidase. • BlArafD and BlArafE cooperatively degrade α1,2/3-Araf double-substituted arabinoxylan.

Caffeine increases tear volume depending on polymorphisms within the adenosine A2a receptor gene and cytochrome P450 1A2.

PURPOSE: The primary aim of the present study was to examine the effect of caffeine on tear volume. The secondary aim was to investigate the relation between caffeine-induced changes in tear volume and polymorphisms in ADORA2A and CYP1A2. DESIGN: Double-masked, placebo-controlled, crossover study. PARTICIPANTS: Seventy-eight healthy volunteers were recruited for the study. METHODS: Subjects participated in 2 sessions in which they received capsules containing either placebo or caffeine. The caffeine capsules were given to the subjects to keep the caffeine volume per body weight within 5 to 7 mg/kg. After caffeine intake, tear meniscus height (TMH) was measured. Subjects provided a blood sample for genotyping. MAIN OUTCOME MEASURES: Tear meniscus height, single nucleotide polymorphism. RESULTS: The tear volume increased after caffeine consumption. The net increase in TMH was 0.08 mm (95% confidence interval, 0.05-0.10) greater when participants were given caffeine than when given placebo (P<0.0001). In ADORA2A, the difference in the net increase in TMH for participants who were heterozygous at rs5751876 and rs2298383 was 0.07 mm (P = 0.001) and who were minor homozygous was 0.08 mm (P = 0.007). In CYP1A2, the net increase in TMH for participants who were minor homozygous at rs2472304 was lower than for those who were major homozygous; the difference was 0.06 mm (P = 0.039). CONCLUSIONS: Caffeine intake increases tear volume and polymorphisms within ADORA2A, and CYP1A2 is associated with the tear increase after caffeine intake. Genetic polymorphisms had a significant effect on tear meniscus that was of limited clinical significance.

Effects of long-term topical anti-glaucoma medications on meibomian glands.

OBJECTIVE: To examine effects of long-term topical anti-glaucoma medications on meibomian gland morphology and function and assess their relationship with slit-lamp findings. METHODS: This was a cross-sectional observational case series of 31 patients with glaucoma (mean age ± standard deviation, 65.0 ± 13.0 years; mean duration of eye drop use, 7.9 ± 6.0 years) treated with topical anti-glaucoma drugs in only one eye for more than 1 year: 13 receiving prostaglandin analogues (PGs) alone, eight receiving ß-blockers alone, and ten receiving multiple treatments. Untreated contralateral eyes served as controls. Lid margin (lid margin abnormality score: 0-4) and superficial punctate keratopathy (SPK score: 0-1) were observed with a slit lamp. Upper and lower eyelids were turned over to observe meibomian glands using non-contact meibography. Meibomian gland loss was scored for each eyelid from grade 0 (no loss of meibomian glands) through grade 3 (loss >2/3 of total meibomian gland area). Meibomian lipid content (meibum) was scored (meibum score: 0-3). RESULTS: Treated eyes had significantly higher scores for lid margin abnormality (P= 0.001), SPK (P< 0.001), meibo-score (P< 0.001), and meibum (P< 0.001) than control eyes. Tear film break-up time (BUT) was significantly shorter in treated eyes than in control eyes (P= 0.001). Schirmer values were significantly lower in treated eyes than in control eyes (P= 0.0039). Subgroup analysis indicated a significantly higher meibo-score in eyes treated with PGs (P= 0.0046) and in eyes treated with ß-blockers (P= 0.0231) than in the corresponding controls. CONCLUSIONS: Long-term anti-glaucoma eye drop use affects meibomian gland morphology and function.

Fibroblast growth factor receptor 3 mutation in voided urine is a useful diagnostic marker and significant indicator of tumor recurrence in non-muscle invasive bladder cancer.

The fibroblast growth factor receptor (FGFR)-3 gene encodes a receptor tyrosine kinase that is frequently mutated in non-muscle invasive bladder cancer (NMIBC). A sensitive and quantitative assay using peptide nucleic acid-mediated real-time PCR was developed for detecting FGFR3 mutations in the urine samples and evaluated as a molecular marker for detecting intravesical recurrence of NMIBC in patients undergoing transurethral resection of bladder tumor. FGFR3 mutation was examined in tumor tissues and serially taken pre- and postoperative urine sediments in 45 NMIBC patients with a median follow up of 32 months. FGFR3 mutations were detected in 53.3% (24/45) of primary tumor tissues, among which intravesical recurrence developed in 37.5% (9/24) of cases. FGFR3 mutation in the primary tumor was not a significant prognostic indicator for recurrence, while the proportion of FGFR3 mutation (i.e. tumor cellularity was >or=11%) in the preoperative urine sediments was a significant indicator for recurrence in patients with FGFR3 mutations in the primary tumors. FGFR3 mutations were detected in 78% (7/9) of postoperative urine samples from recurrent cases with FGFR3 mutations in the tumor, while no mutations were detected in the urine of 15 non-recurrent cases. Urine cytology was negative in all cases with FGFR3 mutations in the primary tumors, while the sensitivity of cytological examination was as high as 56% (5/9) in cases showing wild-type FGFR3 in the primary tumors. Urine FGFR3 mutation assay and cytological examination may be available in the future as complementary diagnostic modalities in postoperative management of NMIBC.

Proposed diagnostic criteria for obstructive meibomian gland dysfunction.

PURPOSE: To compare clinical findings between patients with obstructive meibomian gland dysfunction (MGD) and normal controls and to propose diagnostic criteria for obstructive MGD. DESIGN: Cross-sectional, observational case series. PARTICIPANTS: Fifty-three eyes of 53 patients (18 men, 35 women; age [mean +/- standard deviation] 71.4 +/- 10.0 years) who were diagnosed with obstructive MGD and 60 eyes of 60 healthy volunteers (22 men, 38 women; 71.0 +/- 9.3 years) as a control group. METHODS: Ocular symptoms were scored from 0 to 14 according to the number of existing symptoms. Lid margin abnormality was scored from 0 to 4 depending on the number of existing abnormalities. Meibomian gland changes were scored from 0 to 6 based on noncontact meibography (meibo-score). Superficial punctuate keratopathy (SPK) was scored from 0 to 3. Meibum was graded from 0 to 3 depending on the volume and quality. Tear film production was evaluated by Schirmer's test. Receiver operating characteristic curves with calculations of area under the curve (AUC) were used to describe the accuracy of each parameter to differentiate obstructive MGD from normal eyes. MAIN OUTCOME MEASURES: Ocular symptom score, lid margin abnormality score, meibo-score, meibum score, SPK score, tear film breakup time (BUT), and the Schirmer value. RESULTS: Ocular symptom score, lid margin abnormality score, meibo-score, meibum score, and SPK score were significantly higher in the obstructive MGD group than in the control group (P<0.0001 for all scores). The BUT was significantly shorter in the obstructive MGD group than in the control group (P<0.0001). The AUC values indicated that the ocular symptom score had the highest diagnostic power as a single parameter, followed by the lid margin abnormality score, meibo-score, and BUT. CONCLUSIONS: Based on these findings, we recommend that physicians use the ocular symptom score, lid margin abnormality score, and meibo-score to diagnose MGD. Obstructive MGD should be suspected when any 2 of the 3 scores are abnormal. Obstructive MGD is very likely when all 3 scores are abnormal.

A newly developed noninvasive and mobile pen-shaped meibography system.

PURPOSE: To examine the usefulness of a newly developed noninvasive mobile pen-shaped meibography system. METHODS: This study evaluated a newly developed noninvasive mobile pen-shaped meibography system comprising an infrared light-emitting diode as the light source and a highly sensitive complementary metal oxide semiconductor image camera. The images were recorded digitally. The utility of this system was compared with that of the previously developed noncontact infrared meibography system for examination of the upper and lower eyelids in 20 healthy volunteers (range, 2-91 years) and 23 patients with meibomian gland dysfunction, 17 patients with dry eyes who wore contact lenses, and 14 patients with allergic conjunctivitis accompanied by itching. RESULTS: Using the newly developed noninvasive mobile pen-shaped meibography system, clear images of the meibomian glands were obtained in all age groups. The quality of the images obtained was similar between the two meibography systems. The quantitative analysis of the images obtained showed no statistically significant difference between the two meibography systems. CONCLUSIONS: The newly developed noninvasive mobile pen-shaped meibography system is a useful tool that provides meibomian gland images of the same quality and quantity as the noncontact meibography system equipped with a slit lamp. This new system is convenient and applicable for examination of meibomian glands in patients of all ages.

Topical diquafosol for patients with obstructive meibomian gland dysfunction.

AIMS: To evaluate the effect of topical diquafosol in patients with meibomian gland dysfunction (MGD) using tear film parameters and quantitatively analyse the meibomian gland morphology. SUBJECTS AND METHODS: The subjects were 19 eyes of 10 patients diagnosed with obstructive MGD. All subjects were given 3% diquafosol ophthalmic solution with instructions to use one drop four times a day. Ocular symptoms were scored from 0 to 14. Lid margin abnormalities were scored from 0 to 4. Changes in the meibomian glands were scored using non-contact meibography (meiboscore). Superficial punctate keratopathy (SPK) was scored from 0 to 3. Meibum was graded from 0 to 3. Tear film production was evaluated by Schirmer's test. Quantitative image analysis of the meibomian glands was performed using the original software. RESULTS: 10 patients completed more than 4 months of therapy. Ocular symptoms, lid margin abnormalities, SPK score and meibum grade were decreased. Break-up time and tear film meniscus were increased. Mean ratio of the meibomian gland area was significantly increased after treatment (p<0.0001). CONCLUSIONS: Quantitative image analysis was useful for evaluating the morphological changes of the meibomian glands. Topical diquafosol therapy was effective for patients with obstructive MGD.

Association of contact lens-related allergic conjunctivitis with changes in the morphology of meibomian glands.

PURPOSE: To observe morphological changes in the meibomian glands of patients with contact lens-related allergic conjunctivitis (CLAC) and to assess the relations between the morphological changes and eyelid and tear film parameters. METHODS: We observed subjects in four groups: 64 eyes of 64 contact lens (CL) wearers with CLAC, 77 eyes of 77 CL wearers without CLAC, 55 eyes of 55 patients with perennial allergic conjunctivitis (perennial AC), and 47 eyes of 47 healthy volunteers. The following tests were performed: slit-lamp examination, measurement of tear film breakup time, grading of morphological changes in meibomian glands (meiboscore) as assessed by noncontact meibography, grading of meibomian gland distortion in meibography, tear production as assessed by Schirmer's I test, and grading of meibum expression. RESULTS: The mean score for meibomian gland distortion was significantly higher in the CL wearers with CLAC than in the CL wearers without CLAC (p < 0.0001); it was also significantly higher in the non-CL wearers with perennial AC than in the non-CL wearers without perennial AC (p < 0.0001). There was no significant difference between the mean scores for meibomian gland distortion of the non-CL wearers with perennial AC and the CL wearers with CLAC (p = 0.27). The score for meibomian gland distortion was significantly positively correlated with the meibum score in the CL wearers with CLAC and with the meiboscore in the CL wearers without CLAC. CONCLUSION: CLAC is associated with an increase in meibomian gland distortion. Allergic reaction, rather than CL wear, appears to be responsible for the increase in meibomian gland distortion in patients with CLAC.

Comparison of the long-term effects of various topical antiglaucoma medications on meibomian glands.

PURPOSE: To examine the effects of long-term antiglaucoma eye drop treatment on meibomian glands. METHODS: The subjects were 71 eyes of 71 glaucoma patients (group 1) receiving one type of antiglaucoma eye drops, 61 eyes of 61 glaucoma patients (group 2) receiving two types of antiglaucoma eye drops, and 30 eyes of 30 glaucoma patients (group 3) receiving three types of antiglaucoma eye drops. Controls comprised 75 eyes of 75 healthy volunteers. Subjective symptoms were evaluated by questionnaire, and lid margin and superficial punctate keratopathy were evaluated by slit lamp examination. Meibomian glands of upper and lower eyelids were observed and scored using noncontact meibography (meiboscore). Tear film break-up time (BUT) was measured and meibum was graded. RESULTS: Lid margin abnormality, superficial punctate keratopathy, meiboscore, and meibum scores were significantly higher in glaucoma patients than in controls (P < 0.001). BUT and Schirmer scores were significantly lower in glaucoma patients than in controls (P < 0.001). Subgroup analysis of the parameters in group 1 revealed no significant difference between patients receiving prostaglandin and those receiving ß-blockers, or among groups 1, 2, and 3. Multivariate regression analysis demonstrated that meiboscore significantly correlated with lid margin abnormality score (P = 0.007) and BUT (P = 0.045) in group 1; with BUT (P = 0.004), symptom score (P = 0.003), and age (P = 0.026) in group 2; and with lid margin abnormality score (P = 0.001) in group 3. CONCLUSIONS: Long-term use of antiglaucoma eye drops was associated with alterations in meibomian gland morphology and function.

Efficacy of diagnostic criteria for the differential diagnosis between obstructive meibomian gland dysfunction and aqueous deficiency dry eye.

PURPOSE: To evaluate diagnostic criteria for obstructive meibomian gland dysfunction (MGD) using three parameters (symptom score, lid margin abnormality score, and meibomian gland morphologic change scores) for differentiating obstructive MGD from aqueous deficiency dry eye (ADDE). METHODS: Twenty-five eyes of 25 patients (mean age, 66.6 years) diagnosed with obstructive MGD and 15 eyes of 15 patients (mean age, 61.3 years) diagnosed with ADDE were analyzed. Ocular symptoms were scored from 0 to 14 according to the number of symptoms. Lid margin abnormality was scored from 0 to 4 according to the number of abnormalities. Meibomian gland changes were scored from 0 to 6 using noncontact meibography (meibo-score). Superficial punctate keratopathy was scored from 0 to 3. Meibum was graded from 0 to 3 according to volume and quality. Tear film break-up time was measured consecutively three times after instillation of fluorescein, and the median value was adopted. Tear film production was evaluated using the Schirmer test. RESULTS: Ocular symptom and lid margin abnormality scores and tear film break-up time did not differ significantly between the obstructive MGD and ADDE groups. The meibum score and meibo-score were significantly higher in the obstructive MGD group than in the ADDE group. The Schirmer value was significantly lower in the ADDE group than in the obstructive MGD group. When obstructive MGD was diagnosed on the basis of three scores (ocular symptom score, lid margin abnormality score, and meibo-score) all being abnormal, the sensitivity and specificity for differentiating between obstructive MGD and ADDE were 68.0% and 80%, respectively. CONCLUSIONS: Although the criteria were moderately reliable for differentiating patients with obstructive MGD from those with ADDE when the diagnosis of obstructive MGD was made on the basis of three abnormal scores, they do not provide comprehensive diagnostic tools for differentiating MGD, ADDE, and healthy individuals. We need to add other parameters such as the Schirmer test value and the meibum score to the diagnostic criteria to enhance their reliability for differentiating MGD and ADDE.

Proposed diagnostic criteria for seborrheic meibomian gland dysfunction.

PURPOSE: To compare clinical findings between patients with seborrheic meibomian gland dysfunction (MGD) and normal controls and to propose diagnostic criteria for seborrheic MGD. METHODS: Thirty eyes of 30 patients [13 men and 17 women; age (mean +/- SD) 73.9 +/- 9.9 years] diagnosed with seborrheic MGD and 60 eyes of 60 healthy volunteers (22 men and 38 women; age: 71.0 +/- 9.3 years) as a control group were included in this study. Ocular symptoms were scored from 0 to 14 according to the number of symptoms present. Lid margin abnormality was scored from 0 to 4 depending on the number of abnormalities present. Meibomian gland changes were scored from 0 to 6 on the basis of noncontact meibography (meiboscore). Superficial punctate keratopathy was scored from 0 to 3. Tear film production was evaluated by Schirmer test. Receiver operating characteristic curves with calculations of the area under the curve were used to describe the accuracy of each parameter to differentiate patients with seborrheic MGD from normal eyes. RESULTS: Ocular symptom score and lid margin abnormality score were significantly higher in the seborrheic MGD group than in the control group (P < 0.0001 for both scores). Area under the curve values indicated that the lid margin abnormality score had the highest diagnostic power as a single parameter followed by the ocular symptom score. When the diagnosis for seborrheic MGD was made on the basis of the 2 scores (ocular symptom score and lid margin abnormality score) being abnormal, the sensitivity was 100% and the specificity was 98.3%. CONCLUSIONS: On the basis of these findings, we recommend that physicians use ocular symptom score and lid margin abnormality score in the diagnosis of seborrheic MGD. Seborrheic MGD should be considered very likely when both of the 2 scores are abnormal.

Meibomian gland duct distortion in patients with perennial allergic conjunctivitis.

PURPOSE: To observe morphologic changes of meibomian glands in patients with and without perennial allergic conjunctivitis (AC) and to assess the relation between morphologic changes of the meibomian glands of both eyelids and tear film parameters. METHODS: In this study, 55 eyes of 55 patients with perennial AC and 47 eyes of 47 healthy volunteers as controls were included. The following tests were performed: a slit-lamp examination, measurement of tear film breakup time, grading of meibomian gland morphologic changes (meibography score) assessed with a noncontact meibography, meibomian gland duct distortion in meibography, tear production by the Schirmer I test, and grading of meibum expression. RESULTS: The frequency of meibomian gland duct distortion was significantly greater in patients with AC (45%) than that in controls (8.5%; P < 0.0001). The meibum (P = 0.049) and superficial punctate keratopathy scores (P = 0.0076) were significantly higher in patients with AC than those in controls. There was no significant difference in meibography score, breakup time, or Schirmer value between the 2 groups. The meibomian expression score was significantly higher in patients with AC with meibomian gland duct distortion than in patients with AC without meibomian gland duct distortion (P = 0.0012). CONCLUSION: Perennial AC is associated with increased meibomian gland duct distortion.