Copyright 2004 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved.

For permission to reproduce any abstract, contact the ARVO Ofce at pubs@arvo.org.
Commercial Relationships are noted at the end of each abstract by None or with codes.
2935 - B570
Upregulated expression of -defensins in the tears of pterygium patients
L.Zhou
1
, R.W. Beuerman
1,2A
, L.Q. Huang
2B
, S.K. Koh
1
, L.Ang
3
, F.T. Chew
2C
, D.Tan
1,3
.
1
Singapore Eye Research Institute, Singapore, Singapore;
A
Dept. Ophthalmology,
B
Dept.
Chemistry,
C
Dept. Biological Sciences,
2
National University of Singapore, Singapore,
Singapore;
3
Singapore National Eye Centre, Singapore, Singapore.
Purpose: To determine if defensins are found in the tear lm of patients with pterygium.
Methods: Tear samples were collected with 10 l pipettes with re polished tips from patients
(n= 15, 12 male, 3 female, average age: 53) prior to pterygium surgery from diseased eyes and
fellow normal control eyes. Tear protein proles were compared using surface enhanced laser
desorption/ionization-time of ight (SELDI-TOF) ProteinChip technology. Conrmation of the
identity of the -defensins in human tears was made by liquid chromatography (LC) purication,
trypsin digestion and electrospray (ESI) tandem mass spectrometry (MS) analysis of tryptic
digests. LC-ESI-MS was used to quantify the levels of human -defensins (HNP-1 and HNP-2)
in human tears by monitoring the ions at m/z = 861.5 for HNP-1 and m/z = 844.0 for HNP-2.
Results: With this proteomic approach, one of the obvious changes we found was that a
group of human -defensins (HNP-1, HNP-2 and HNP-3) was signicantly up-regulated
in the tear uid of pterygium affected eyes when compared with the fellow normal
eyes. The identity of these three human -defensins was conrmed by sequencing the
peptides using tandem mass spectrometry. The mean concentration (n = 5) of -defensins
for pterygium eyes and fellow control eyes were 1.33 0.47 g/ml (HNP-1, pterygium
eye, p < 0.015), 0.61 0.23 g/ml (HNP-2, pterygium eye, p < 0.012) and 0.17 0.12
g/ml (HNP-1, control eye), 0.02 0.02 g/ml (HNP-2, control eye) respectively.
Conclusions: The upregulated expression of the -defensins in tear uids of pterygium patients
indicates that this class of peptides may be part of the response of the ocular surface to the
formation of this brovascular tissue or the accompanying inammation.
CR: L. Zhou, None; R.W. Beuerman, None; L.Q. Huang, None; S.K. Koh, None; L. Ang,
None; F.T. Chew, None; D. Tan, None.
Support: NMRC PG002, NMRC IBG
2936 - B571
Involvement of Endothelial Precursors from Bone Marrow in Pterygium
Y.Song
1A
, M.Kim
1B
, J.Kim
1A
.
A
Ophthalmology,
B
Pathology,
1
Chung-Ang Univ.Yong-San
Hosp., Seoul, Republic of Korea.
Purpose: We evaluated the involvement of endothelial precursors and its
related activating factors in pterygium, then measured chemotactic factors both
locally and systemically before and after operation. And also we confirmed
i ncreased mobi l i zat ion of endot hel ial progenit ors f rom bone mar row.
Methods: Primary (n=15), recurrent pterygium (n=10) and normal control cases (n=5)
were selected for this research. Immunohistochemical staining with endothelial precursor
markers (CD31, CD34, c-kit, VEGF-R1(Flt-1) and VEGF-R2(Flk-1)) and related factors
(hypoxia inducible factor-1 (HIF-1), NF-B and VEGF) were performed. Their
producing site was examined, too. Chemotactic factors (IL-1, GM-CSF, VEGF and
substance-P) were measured in patients tear and serum by using ELISA on pre operation
and post operation 1
st
, 3
rd
and 5
th
day. To conrm the mobilization of endothelial precursors
systemically, owcytometry of peripheral blood with CD34 and c-kit was also done.
Results: The CD31, c-kit and Flt-1 positive endothelial precursor cells were revealed
coincidently in stroma, which were clustered around small vessels and formed rosette pattern.
These foci were increased remarkably in recurrent pterygium compared to in primary
pterygium. HIF-1, as producing factor of VEGF was observed mainly in epithelium. NF-B
was expressed co-localized with HIF-1 in nucleus. Substance-P was increased on 1
st
day and
VEGF was elevated from 3
rd
day of operation in ELISA with serum and tear. These activating
factors were increased more signicantly in recurrent pterygium. The CD34 and c-kit positive
endothelial precursors were increased on 3
rd
~5
th
day of operation especially in recurrent cases.
Conclusions: Co-expressed CD31, c-kit and Flt-1 could be applied as marker of endothelial
precursor. Above results demonstrate that endothelial precursors from bone marrow were
involved in pathogenesis and recurrence of pterygium. This involvement was mediated by
chemotactic factors in both locally and systemically.
CR: Y. Song, None; M. Kim, None; J. Kim, None.
Support: research funds (code : stem cell 13) from the 21C frontier research program
2937 - B572
The Use of Subconjunctival Mitomycin C Preoperatively for Management of
Pterygium
L.Thompson
1
, M.Adenwalla
1
, J.C. Whelchel
1
, A.Elbiaadi
1,2
, P.B. Dray
1,2
, R.M.
Ahuja
1,2
.
1
Ophthalmology, John H. Stroger, Jr. Hospital of Cook County, Chicago, IL;
2
Ophthalmology, FUHS/ The Chicago Medical School, North Chicago, IL.
Purpose: To evaluate the efficacy and safety of using of 0.10 ml of 0.10
mg/ml mitomyci n C as adjunct ive t herapy before pt er ygium surger y.
Methods: A retrospective review of 30 eyes of 22 consecutive patients with no
prior history of pterygium surgery treated with subconjunctival injection of 0.10
ml of 0.10 mg/ml mitomycin C one month prior to bare sclera pterygium excision.
Our main outcome measures were recurrence of pterygium and patient satisfaction.
Results: The pterygia resolved in 96% (29) of patients with an average follow
up time of 2.2 months (range of 1.3 to 3.2 months). All but one patient expressed
satisfaction with the postoperative results. Four (12%) patients suffered from
transient post-injection blepharoconjunctivitis, and one patient complained of
transient diplopia on extreme lateral gaze following excision at 2 months follow-up.
Conclusions: Based on previous studies using higher doses of mitomycin C with successful
outcomes, we conducted this study to determine whether a lower dose could prove equally
effective but with lower adverse effects such as post-injection blepharoconjunctivitis in
addition to other side effects. Based on a limited mean follow up time of 2.2 months, the
lower dose of preoperatively injected subconjunctival mitomycin C is thus far effective in
the treatment of pterygium with regards to recurrence and patient satisfaction. However, the
adverse effect of blepharoconjunctivitis remains similar to prior studies.
CR: L. Thompson, None; M. Adenwalla, None; J.C. Whelchel, None; A. Elbiaadi, None; P.
B. Dray, None; R.M. Ahuja, None.
Support:
2938 - B573
The role of substance P and its receptor in the pathogenesis of pterygium
J.J. Y. Chui
1
, M.T. Coroneo
2
, D.Wakeeld
1
.
1
Department of Pathology, University of New
South Wales, Kensington, Australia;
2
Department of Ophthalmology, Prince of Wales
Hospital, Randwick, Australia.
Purpose: The sensory neuropeptide substance P (SP) participates in inammation, wound
healing and maintenance of corneal health. Pterygium is a corneal disease characterized by
inammation and abnormal wound healing and since its shape parallels the distribution of
sensory corneal nerves, we choose to investigate the role of SP in the pathogenesis of pterygium.
Methods: Primary pterygium epithelial and broblast cultures were established from surgical
tissue and characterised by ow cytometry. Migration of pterygium epithelium, broblasts
and a human microvascular endothelial cell line (HMEC-1) was investigated using a modied
Boyden chamber method. Briey, gelatin-coated polycarbonate lters (10 to 12-m pore size)
separated the top well containing 2.5x10
4
cells from the bottom well containing a range of SP
concentrations (10
-16
to 10
-4
M). After 6 hours, the migrated cells were stained and counted.
Results: Of the cell lines tested, SP induces chemotaxis of pterygium broblasts
(peak concentration of 10
-8
M) and in microvascular endothelial cells (peak range
of 10
-8
to 10
-6
M) but no migration in pterygium epithelial cells (despite incubation
to 24 hours). Cell migration to SP was mediated by the neurokinin 1 receptor
(NK1R) as suggested by blockade with a specific NK1R antagonist (L732138).
Conclusions: The sensory neuropeptide SP may contribute to pterygium formation by acting
as a chemoattractant to pterygium broblasts and vascular endothelium and is a possible
explanation for the triangular shape of pterygium.
CR: J.J.Y. Chui, None; M.T. Coroneo, None; D. Wakeeld, None.
Support:
2939 - B574
The Effect of Doxycycline on the Migration of Pterygial Epithelial Cells grown in Culture
C.A. Cox
1A
, M.John-Aryankalayil
1A
, C.Jaworski
1A
, P.Russell
1B
, T.Gray
2
, K.Freedman
2
,
N.Dushku
3
, T.W. Reid
2
, D.Carper
1A
.
A
Section of Molecular Therapeutics,
B
Aging and
Ocular Diseases,
1
National Eye Institute, Bethesda, MD;
2
Ophthalmology and Visual
Sciences and Cell Biology and Biochemistry, Texas Tech University Health Sciences
Center, Lubbock, TX;
3
Ophthalmology, Kaiser Permanente Medical Center, Sacramento,
CA.
Purpose: To determine if doxycycline, a matrix metalloproteinase inhibitor
(MMPI), i nhibit s t he migrat ion of cult ured pt er ygial epit hel ial cel ls.
Methods: The migration of third-passage pterygial epithelial cells grown in culture was
measured with and without doxycycline of different concentrations using the quantitative
bronectin cell migration Boyden Chamber assay. The control cells were grown under the same
conditions as the treated cells (serum-free Dulbeccos medium containing 5% bovine serum
albumin) except without doxycycline. The extent of migration was measured colorimetrically,
and the data were analyzed using the students t-test and analysis of variance (ANOVA).
Results: Doxycycline (500 g/ml) signicantly decreased migration when compared to
control (P < 0.0002). Doxycycline at the lower concentrations (50 g/ml and 5 g/ml) showed
a trend toward decreasing migration. There was no signicant difference between control sets.
Conclusions: Doxycycline signicantly decreased pterygial cell migration when compared
to the control when used at the 500 g/ml concentration. This may eventually be benecial
to humans in reducing the angiogenesis and growth of pterygia, and may therefore prove
to be vision-saving.
CR: C.A. Cox, None; M. John-Aryankalayil, None; C. Jaworski, None; P. Russell,
None; T. Gray, None; K. Freedman, None; N. Dushku, None; T.W. Reid, None; D. Carper,
None.
Support:
2940 - B575
A Comprehensive System for Pterygium Classication
S.C. Johnston
1
, P.B. Williams
2
, J.D. Sheppard, Jr
2
.
1
Ophthalmology, Eastern Virginia Med
Sch, Norfolk, VA;
2
EyeRx Research Inc., Norfolk, VA.
Introduction: An adequate standardized classication system for pterygium has not been identied. Clinicians currently measure radial and limbal dimensions of corneal involvement and assess serial progression. Other
characteristics, however, may better predict future behavior and response to treatments such as topical steroids, topical anti-metabolites, beta irradiation, anti-VEGF drugs, laser and photodynamic therapy, and surgical excision.
Methods:
Stage 0 Pingeuculum, posterior to the Limbus
Stage I Tissue involvement to the Limbus
Stage II Tissue just on to the Limbus
Stage III Tissue between the Limbus and Pupillary Margin
Stage IV Tissue central to the Pupillary Margin
Conj u nc t i va l a nd
Corneal Tissue Surface
Vascularity (V)
Conj unct i val Ti ssue
Thickness (C)
Corneal Tissue Thickness
(K)
Cor neal Leadi ng Edge
Pigment at ion or Fer r y
Line (P)
V0 - No discernable
vascularity
C0 - Conjunctival ectasia
or thinning
K0 - Corneal ectasia
or thinning with f lat
overlying tissue
P0 - No discernable pigment
line
V1 - Minimal papillary
response without visible
intact vessels
C1 - Flat tissue K1 - Flat tissue
P1 - Fai nt i nter r upted
pigment line
V2 - No r ma l
Vascularity
C2 - Minimally elevated
tissue
K2 - Minimally elevated
tissue
P2 - Minimal pigment
line similar to ipsilateral
Hudson-Staheli line
V3 - Mo de r at e
vascularity and vessel
congestion, more dense
than conjunctiva
C3 - Tissue elevation up to
1 mm with minimal or no
epithelial staining
K3 - Tissue elevation up
to 1 mm with minimal or
no epithelial staining
P3 - Moderate, intensely
colored pigment line
V4 - Severe vascularity
with vessel congestion
and dilation
C4 - Tissue elevation over 1
mm with epithelial staining
or erosion
K4 - Tissue elevation
over 1 mm with epithelial
staining or erosion
P4 - Dense, deeply colored
or elevated pigment line

Results: A typical pterygium was described as Stage II, V2, C2, K2, P2, or an irritated red pingeuculum described as Stage O, V3, C3 from standardized photographs and slit lamp. Reproducibility was assessed by three
knowledgeable graders. Results over a three month period were compared to the standard length, measurement grading system. This system was used to analyze progression or response to treatment and descriptive accuracy.
Conclusions: This unique semiquantitative approach provides uniform standards that provide an overall rating based upon the progression (or in the case of therapy, regression) of the pterygium as well as a more detailed description of
important characteristics of each pterygium. The detailed descriptors may be important predictors of response to a specic therapy.
CR: S.C. Johnston, None; P.B. Williams, None; J.D. Sheppard, Jr, None.
Support:
Tuesday, April 27, 8:30 - 10:30 AM Hall BC Poster Session Program Number/Board # Range: 2935-2956 / B570-B591
367. Clinical Studies: Pterygium Organizing Section: CO
2935-2940
Copyright 2004 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Ofce at pubs@arvo.org.
Commercial Relationships are noted at the end of each abstract by None or with codes.
2941 - B576
Efcacy of the newly developed 3 surgical techniques: 2 conjuncival autograft transplantation
techniques and folded conjunctival edge suturingtechnique for pterygium.
T.Yamaguchi. Dept. of Ophthalmology, St Lukes International Hosp, Chuo-Ku, Tokyo,
Japan.
Purpose: To determine the efcacy of newly developed 3 types of surgical technique: 2 conjunctival
autograft transplantation techniques and folded conjunctival edge suturing technique for pterygium.
Methods: Newly developed surgical techniques were 1) extending of conjunctival ap from
superior conjunctiva 2) transposition of conjunctival ap from temporal conjunctiva (non-
free ap) 3) suturing of folded conjunctival edge on sclera. From Jan. 1988 to Dec. 2001, 218
eyes including 27 recurred eyes (1-10 times) were operated. Combination of technique 1) +
3) was mainly performed and combination of technique 2) + 3) was performed on the cases
with wide conjunctival defect more than 3:30 oclock position at limbus after the excision of
head of pterygium. Combination of procedure 1) + 3) was applied on 190 eyes (15 recurred)
and combination of procedure 2) + 3) was applied on 28 eyes (12 recurred). 8-0 Dexon
S
(R)
(polyglycolic acid) was used for suturing. 0.1%uorometholone eye drops was used
for primary cases for 3 months and 0.1% betamethasone eye drops was used for recurred
cases from 3 to 6 months after surgery. Mean follow-up period was 19.2 months (3-136M).
Results: Combination of technique 1) + 3) developed 1.0% (2/190) recurrence
rate (primary case: 1.1% (2/175), recurred case: 0% (0/15)). Combination of
technique 2) + 3) developed 3.6%(1/28) of recurrence rate (primary case: 0%(0/16),
recurred case: 8.3%(1/12)). In total, 1.4% (3/218) of recurrence rate was obtained.
Conclusions: Combination of newly developed 3 types of surgical techniques, such as 2
types of conjunctival ap autograft transplantation techniques and folded conjunctival
edge suturing technique demonstrated extremely low recurrence rate (1.4%). No severe
complications were observed.
CR: T. Yamaguchi, None.
Support:
2942 - B577
Sutureless Conjunctival Autograft
V.Panday
1A
, S.B. Hannush
1B
.
1
Wills Eye Hospital, Philadelphia, PA;
B
Cornea Service,
1
Wills Eye Hospital, Philadelphia, PA.
Purpose: To evaluate the effectiveness of Tisseel VH f ibrin sealant in
securing a free autologous conjunctival graft without the use of sut ures.
Methods: Fifteen eyes underwent uneventful excision of primary pterygium followed
by autologous conjunctival grafting. The grafts were secured in place with Tisseel
VH brant sealant and 4 sutures (3 eyes), 2 sutures (3 eyes), and no sutures (9 eyes).
Results: All conjunct ival graf t s remai ned secure i n place t hroughout
t he fol l ow- up per i od (mean: 7 mont hs; r ange 1-13 mont hs) .
Conclusion: Tisseel VH brin sealant is an effective method for securing a conjunctival graft
at the time of pterygium surgery. Advantages include rapid healing, increased comfort, and
a lower inammatory prole.
CR: V. Panday, None; S.B. Hannush, None.
Support:
2943 - B578
Under-expression of endothelin 1 RNA in pterygium blood vessels may be related to
pterygium pathogenesis
E.Rechtman
1,2
, N.Levi
3
, A.L. Marcovich
1
, R.Braw-Tal
4
, A.Pollack
1
, A.Harris
2
, R.Meidan
3
.
1
Ophthalmology, Kaplan Hospital / The Hadassa School of Medicine, Hebrew University,
Rehovot, Israel;
2
Ophthalmology, Indiana University School of Medicine, Indianapolis,
IN;
3
Animal Sciences, Faculty of Agricultural, Food and Environmental Quality Sciences
/ The Hebrew University, Rehovot, Israel;
4
Agricultural Research Organization, The
Volcani Center, Institute of Animal Science- Bet Dagan, Rehovot, Israel.
Purpose: To evaluate the role of endothelin 1 (ET1) in the pathogenesis of pterygium by comparing the levels of
ET1 and endothelin converting enzyme 1 (ECE1) between pterygium and autologous conjunctival tissues.
Methods: All procedures were approved by an IRB, and informed consent obtained. In a prospective
study, 7 pairs of pterygia and superior bulbar conjunctival tissues taken from the same eyes during a
pterygium surgery with free conjunctival autograft transplantation were immediately put in liquid
nitrous (-196C) and later stored at -80C. Reverse transcriptase polymerase chain reaction (RT-PCR)
process was then applied to quantify the levels of ET1, ECE1, and platelet endothelial cell adhesion
molecule-1 (PECAM1, a marker of blood vessels density). Total RNA was extracted from the tissues
and was later converted to complementary DNA (cDNA). cDNA was multiplied using PCR with the
specic primers for ET1, ECE1, PECAM1 and GPDH (house keeping gene, which serves as a reference
for the DNA quantity). The DNA product was put on an agarose gel for electrophoresis and each band
was read for intensity. ET1, ECE1 and PECAM1 bands intensity were divided by that of the GPDH.
Pairedt tests were used to compare between the ratios of ET1/GPDH, ECE1/GPDH and PECAM1/
GPDH in the pterygia and conjunctival tissues. P<0.05 was considered statistically signicant.
Results: ET1/GPDH, ECE1/GPDH and PECAM1/GPDH ratios were 1.15 0.1 (mean standard
error), 1.54 0.1, and 2.7 0.22 in the pterygia tissues, as compared to 1.32 0.045, 1.33 0.13 and
1.15 0.06 in the conjunctival tissues, respectively. While no differences were found in the ratios
of ET1/GPDH and ECE1/GPDH between pterygia and conjunctival tissues (P>0.1), PECAM1/
GPDH ratio was 2.35 times greater in the pterygia as compared to the conjunctivas (P<0.01).
Conclusions: Since endothelial cells are the main producers of ET1 and ECE1, these ndings suggest
that the concentrations of ET1 and ECE1 RNA in pterygium blood vessels are signicantly lower
than those in the vessels of the conjunctiva. Further studies are needed.
CR: E. Rechtman, None; N. Levi, None; A.L. Marcovich, None; R. Braw-Tal, None; A.
Pollack, None; A. Harris, None; R. Meidan, None.
Support:
2944 - B579
Signicance of VEGF in the pathogenesis of pterygium
F.P. Paulsen
1
, M.Gebhardt
1
, U.Schaudig
2A
, B.Nlle
3
, T.Pufe
1
, K.Al-Samir
2B
, G.Geerling
4
,
R.Mentlein
1
.
1
Inst of Anatomy, Christian Albrecht Univ of Kiel, Kiel, Germany;
A
Dep of
Ophtalmology,
B
Depart. of Ophtalmology,
2
UKE Univ of Hamburg Eppendorf, Hamburg,
Germany;
3
Depart. of Ophtalmology, Christian-Albrecht-Univ of Kiel, Kiel, Germany;
4
Depart.of Ophtalmology, UKL Univ of Lbeck, Lbeck, Germany.
Purpose: To evaluate the role of vascular endothelial growth factor (VEGF)
and its receptors VEGFR1 and VEGFR2 in the pathogenesis of pterygium.
Methods: Expression of mRNA for VEGF, its different splice forms and its receptors
(VEGFR1 and 2) was analyzed by reverse transcription polymerase chain reaction (RT-PCR)
in pterygium and conjunctiva. Deposition of VEGF and VEGFR1 and 2 was determined by
Western blot and immunohistochemistry. Quantication of VEGFR mRNA was done by real-
time RT-PCR, and of VEGF by ELISA in pterygium, conjunctiva, limbus, cornea and lens.
The proliferative effects of different tumor necrosis factor alpha (TNF) concentrations were
tested on human corneal epithelial cells, conjunctival epithelial cells and endothelial cells.
Results: In analyses of samples from pterygia patients and conjunctiva samples from
cadavers, VEGF
121
and VEGF
165
were identied as the only VEGF splice forms expressed.
In addition to VEGF, VEGFR1 and VEGFR2 were detectable by RT-PCR and Western
blot in pterygium and conjunctiva and immunostained within the epithelium of pterygia
as well as conjunctiva and on intrapterygial and intraconjunctival endothelial cells.
Real-time RT-PCR revealed more VEGFR1 and 2 mRNA in conjunctiva samples than
in pterygium samples, but comparable amounts in limbus and pterygia samples. ELISA
analysis revealed higher VEGF levels in pterygia compared with conjunctiva, cornea and
lens, but comparable levels in limbal samples. Cell cultures showed no effects of TNF on
proliferation of human corneal and conjunctival epithelial cells, but on human endothelial cells.
Conclusions: The results strongly support the assumption that pterygia arise from limbal
epithelial cells and that human conjunctiva is not a suitable control in analysis of pterygia.
Moreover, the results suggest that VEGF does not play such an important role in the
pathogenesis of pterygia.
CR: F.P. Paulsen, None; M. Gebhardt, None; U. Schaudig, None; B. Nlle, None; T. Pufe,
None; K. Al-Samir, None; G. Geerling, None; R. Mentlein, None.
Support: DFG Grant PA 738/6-1
2945 - B580
Safety and Efcacy of Intraoperative Mitomycin C During Conjunctival Autograft
to Prevent Recurrence in Patients with Primary Pterygium and High Risk of
Recurrence
G.L. Andrade, M.A. Di Pascuale, C.Leizaola, L.Gimenez, M.Pezonaga, A.Barrientos,
M.Estribi. Ophthalmology, Hospital Dr. Miguel Prez Carreo, I.V.S.S, Universidad
Central De Venezuela., Caracas, Venezuela.
Purpose: Patients with eshy Pterygium have high risk of recurrence. Mitomycin C (MMC) inhibits
the growth of broblast and therefore reduces the brovascular tissue and might prevent recurrence of
pterygium. We would like to demonstrate whether if the use of intraoperative MMC with conjunctival
autograft is safe and effective to prevent recurrence in patients with primary eshy pterygium.
Methods: Fifty eight patients (58 eyes) with primary nasal pterygium and severe brovascular tissue (eshy
appearance) were prospectively included. Twenty eight eyes received conjunctival autograft alone and 30 eyes
received MMC (0.02 % for 1 minute) with conjunctival autograft. Both groups were matched according to sex,
age and severity of corneal invasion of pterygium. Pterygium invasion to cornea were graded from 1 to 3. Grade
1 were pterygium that reached less one quarter of the cornea, grade 2 the pterygium invasion were between
quarter and center of the corneal and grade 3 the pterygium invasion were more than corneal central axis.
Results: Twenty seven women and 31 men with mean age of 40.3 years old (ranging from 25 to 85 years
old) were included. Nineteen eyes and 9 eyes had pterygium grade 1 and 2, respectively and were treated
with conjunctival autograft alone. Twenty three eyes and 7 eyes had pterygium grade 1 and 2, respectively
and were treated with MMC/conjunctival autograph. There were no patients with pterygium grade 3. The
mean follow- up period was 12 months (ranging from 9 to 20 months). Six eyes (20%) recurred in the
conjunctival autograft group and 2 eyes (6.2%) in the MMC/conjunctival autograft group, but it was not
statistically signicant (Fishers exact Test, P= 0.13). All recurrences occurred before 3 months. There
was no statistically difference in the grade of invasion of pterygium in both groups (Fishers exact Test,
P=0.6). In patients treated with MMC the conjunctival autograft became the invading pterygium with
minimal brovascular tissue, compared with recurrence pterygium in cojunctival autograft alone in which
the eshy and inamed appearance persisted. There were minimal complications in both groups such as
foreign body sensation, photophobia and epithelial cyst that resolved spontaneously in less than 15 days.
Conclusions: The use of MMC reduces the brovascular component in all primary pterygium and it was as
safe and effective as conjunctival autograft alone to prevent the recurrence of pterygium.
CR: G.L. Andrade, None; M.A. Di Pascuale, None; C. Leizaola, None; L. Gimenez,
None; M. Pezonaga, None; A. Barrientos, None; M. Estribi, None.
Support:
2946 - B581
Beta-microseminoprotein is a prostate protein associated with pterygium
R.W. Beuerman
1,2
, V.Viryanti
3
, J.Chew
1
, L.Zhou
3
, L.Ang
3,4
, D.Tan
3,2
.
1
Singapore Eye
Reseach Institute, Singapore, Singapore;
2
Department of Ophthalmology, National
University of Singapore, Singapore, Singapore;
3
Singapore Eye Research Institute,
Singapore, Singapore;
4
Singapore National Eye Center, Singapore, Singapore.
Purpose: BMSP or PSP94 is a unique protein associated with prostate cancer, but more generally
mucosal tissues. This study has examined the association of this protein with primary pterygium.
Methods: All patient procedures received IRB approval and patients signed consent forms.
Patient selection required that the pterygium be unilateral, and primary. Tears were collected
from both eyes prior surgery, pterygia and control conjunctiva (CC) were collected at the
time of surgery and immediately frozen. The analysis consisted of the following: DNA
array (Affymetrix U133A, 8 pterygia, controls (3 groups of ve)), RT-PCR and real-time
PCR using TaqMan Assay on Demand probes employed the same 8 pterygia and pooled
(ve conjunctiva) controls. Tear samples (2-4 l from both eyes of 8 patients) were analyzed
with SELDI-TOF Protein Chip technology. A USBio polycolonal-antibody was used
for immunohistochemistry, and western blots (same 8 pterygia tissues used for both).
Results: Mass spectrometry of tears prior to surgery revealed a protein (10.7-10.8 Kda)
the levels of which were higher in eyes with pterygium (6/8), than in the contralateral
eyes. Analysis of expression levels of genes and ESTs that were represented in pterygia
and control conjunctiva showed that 28 genes were put into the category of up-regulated.
BMSP was found in all samples and up-regulated in 75% of pterygia over CC with an
average of 3.45 fold. Density values from gels of RT-PCR products found that BMSP
was up-regulated over control CC by 0.6 to 20.74 times and all samples were positive.
Real-time PCR showed that in 5/8 pterygium tissues BMSP was up-regulated 3.04 to 5.33
times when compared to control. Western blots showed that the protein was found in all
pterygia tissues (n=8). Immunohistochemistry of pterygia tissue showed the presence of
the protein in the epithelium, whereas in the control conjunctival tissue little or no positive
staining was found. When the antibody was pre-absorbed there was no positive staining.
Conclusions: BMSP may be a bio-marker for pterygium or for persistent inammatory
conditions of the ocular surface.
CR: R.W. Beuerman, None; V. Viryanti, None; J. Chew, None; L. Zhou, None; L. Ang,
None; D. Tan, None.
Support: NMRC IBG, BMRC 03/1/35/19/231, NMRC PG002
Tuesday, April 27, 8:30 - 10:30 AM Hall BC Poster Session Program Number/Board # Range: 2935-2956 / B570-B591
367. Clinical Studies: Pterygium Organizing Section: CO
2941-2946
Copyright 2004 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Ofce at pubs@arvo.org.
Commercial Relationships are noted at the end of each abstract by None or with codes.
2947 - B582
DETERMINATION OF HUMAN PAPILLOMA VIRUS IN PTERYGIUM BY PCR
L.Rodriguez-Cabrera, J.Tovilla-Canales, A.Nava Castaeda, E.Ramirez-C, V.Bosch,
J.Cancino, M.Reyes Lopez. Oculoplastica, Conde de Valenciana, Mexico city, Mexico.
Purpose: To det er mi ne t he prevalence of di fferent t ypes of human
papilloma virus (HPV) in pterygium by polymerase chain reaction (PCR).
Methods: We used PCR to identify the presence of the VPH in 32 pterygiums
assessing the following features of the patients: age, gender, occupation, place
of birth and living family history of pterigyum and the characteristics of the
pterygium (location nasal or temporal- evolution time, size primary or recurrent)
Results: We identied the presence of HPV in 4 of the 32 pterygiums (12.5%). No predilection
was found for HPV regarding sex and age. Of the 4 cases, three patients worked indoors. Three
patients were born and had residence in Mexico city. The four cases were nasal in topography, and
one was a recurrent pterygium. No statistical signicance was observed with size and recurrences .
Conclusions: Many theories have been proposed regarding the pathogenesis of the pterygium,
being exposure to UVA and UVB rays the most commonly accepted. Recently, a possible
viral implication has been proposed, in particular HPV. In our study we found 4 cases of
HPV in pterygiums. We could not determine a possible etiologic association. This study is
different to that reported by Gallagher, because he used parafn samples. We believe PCR
should be considered more routinely in patients with pterygium; however it is necessary to
have a larger number of patients to determine the possible inuence of HPV in etiology,
evolution and behavior of pterygiums.
CR: L. Rodriguez-Cabrera, None; J. Tovilla-Canales, None; A. Nava Castaeda, None; E.
Ramirez-C, None; V. Bosch, None; J. Cancino, None; M. Reyes Lopez, None.
Support:
2948 - B583
Limbal stem cell autograft transplantation in treatment of pterygium
M.-H.Nghiem-Buffet, G.Chaine. Ophthalmology, Hpital Avicenne, Bobigny, France.
Purpose: High rate of recurrence is a major concern in the surgical treatment of pterygium. To observe
the therapeutic effects of limbal stem cell autograft transplantation in treatment of pterygium.
Methods: A noncomperative retrospective case series. Corneal limbal stem cell
autograft and conjunctiva was performed in 14 patients (16 eyes) with primary
pterygium (14 eyes) or recurrence (2 eyes). In all cases a free transplant of the
superotemporal limbus with an adjacent piece of thin conjunctiva was placed in
the excision area. Recurrence of the pterygium and complications were analysed.
Results: Mean follow-up was 17 months (range, 7 - 28). Recurrence occured in two eyes with
primary pterygium ( 12 %) and develloped in the rst year following surgery (3 and 9 months).
Conclusions: Limbal stem cell autograft is an effective technique for pterygium treatment
with a low recurrence rate. This surgical procedure should be proposed in primary care of
evolutive pterygium.
CR: M. Nghiem-Buffet, None; G. Chaine, None.
Support:
2949 - B584
Supression of brotic responses by Mycophenolic Acid in Human Tenon Fibroblasts
in Vitro
R.Amer
1
, I.Puxeddu
2
, I.Anteby
1
, F.Levi-Schaffer
2
, A.Solomon
1
.
1
Ophthalmology, Hadassah
University Hospital, Jerusalem, Israel;
2
Pharmacology, School of Pharmacy, Faculty of
Medicine, The Hebrew Univeristy, Jerusalem, Israel.
Purpose: To evaluat e t he ef fect s of mycophenol ic acid ( MPA) on
human t enon f ibroblast proliferat ion, apoptosis and necrosis i n vit ro.
Methods: Human tenon broblasts were obtained from tissue explants from young healthy
patients during strabismus surgery. Proliferation of subconuent broblasts was assessed
by using the (H) thymidine-incorporation assay. Fibroblast proliferation was assessed
following TGF-1 stimulation at the optimal concentration of 10 ng/ml. MPA was added
(10
-3
M to 10
-8
M in DMEM with 2% fetal calf serum) to the cultures for 24 hours. Fibroblast
apoptosis and necrosis induced by MPA was assessed by the addition of annexin-V and
propidium iodide stains, and by evaluating positively stained cell populations with FACS.
Results: MPA signicantly inhibited TGF-1-induced broblast proliferation with
optimal concentrations ranging from 10
-3
to 10
-6
M. The addition of MPA at 10
-3
M
suppressed TGF-1-induced broblast proliferation by 10% (p<0.001). MPA did not cause
apoptosis nor did it induce signicant necrosis of the broblast populations under study.
Conclusions: Mycophenolic acid demonstrated a signicant antiproliferative effect on human
tenon broblasts in vitro. This effect is not the result of cytotoxic or apoptotic effect of the
drug. This effect can be utilized to modify brotic responses of the ocular surface in various
cicatrizing and allergic disorders, as well as in postoperative brosis in pterygium excision
and in glaucoma ltering surgey.
CR: R. Amer, None; I. Puxeddu, None; I. Anteby, None; F. Levi-Schaffer, None; A.
Solomon, None.
Support: Israel Ministry of Health - Chief Scientist Grant
2950 - B585
Combined excimer laser PTK and amniotic membrane transplantation for palliative
treatment of painful bullous keratopathy
R.Meiller, R.Sauer, C.Hofmann-Rummelt, A.Langenbucher, B.Seitz. Ophthalmology,
Univ. Erlangen-Nuernberg, Erlangen, Germany.
Purpose: To determine the therapeutic potential of combined excimer laser PTK and
amniotic membrane transplantation in patients with painful bullous keratopathy, who are not
suited for penetrating keratoplasty due to poor visual potential or severe general diseases.
Methods: This retrospective study included 10 eyes (7 pseudophakic, 3 aphakic) of six females and
four males treated for painful bullous keratopathy between November 2001 and August 2003. Three
eyes had multiple vitreoretinal surgical procedures before, three eyes severe perforating trauma of
the globe and two eyes previous penetrating keratoplasty. Best corrected preoperative visual acuity
ranged from amaurosis to 20/400. Treatment was done under retrobulbar anesthesia in eight patients
and general anesthesia in two patients. After complete corneal epithelial debridement and pannectomy
an aggressive PTK using the excimer laser MEL 60 (CARL-ZEISS-MEDITEC, Jena, Germany)
was performed. A manually guided spot prole was applied in four cases (Repetitionrate 3 20/s;
571 1790 pulses), a scanning slit procedure in six cases (Repetitionrate 25/s; 1157 2730 pulses).
Then a single layer amniotic membrane was xated as a graft (= inlay) inside the limbus by multiple
interrupted 10-0 nylon sutures in three eyes. In seven eyes the amniotic graft (diameter 9 11 mm)
was positioned into a circular lamellar corneal pocket after performing an incision using a 8 or 9 mm
keratoplasty trephine. In four eyes the latter procedure was completed by an additional supercial
amniotic patch (= sandwich). A soft therapeutic contact lens was applied at the end of surgery.
Results: No intra- or postoperative complications occurred during mean postoperative follow-up
period of 31 20 (range 5 54) weeks. All patients noticed considerable or complete pain relief. The
therapeutic contact lens was removed after 6 2 (range 3 9) weeks postoperatively. The epithelium
remained stable in all eyes during follow-up. The amniotic membrane graft was present at the end
of follow-up completely in 57% and partly in 43%. No further surgical procedures were required.
Conclusions: Combined aggressive excimer laser PTK and amniotic membrane transplantation
seems to be a safe and successful new therapeutic option for long-term epithelial stability and palliative
pain relief in patients with bullous keratopathy not suited for penetrating keratoplasty.
CR: R. Meiller, None; R. Sauer, None; C. Hofmann-Rummelt, None; A. Langenbucher,
None; B. Seitz, None.
Support:
2951 - B586
HIV Related Central Spheroidal Corneal Degeneration
K.P. Han, J.C. Affeldt. Ophthalmology, Loma Linda Univ Med Ctr, Loma Linda, CA.
Purpose: To describe for the rst time development of central spheriodal corneal degeneration
in association with HIV immune suppression. Methods: Observational case report.
Results: A 49-year old African-American male with a 12-year history of HIV immune
suppression (CD4 count 677) presented with complaints of progressive visual loss OU
associated with increasing photophobia and ocular irritation. Best-corrected visual acuity
was 20/200 OU. Slit-lamp exam revealed quiet conjunctiva OU. The corneas were signicant
for multiple bilateral focal brownish translucent supercial elevated deposits limited to the
inferocentral cornea OU. The overall impression was that of multiple surface oil droplets. There
was no staining with uorescen, and no corneal neovascularization. Schirmer test with topical
anesthesia resulted in 40 mm wetting OU. Corneal scraping performed OD histopathologically
revealed fragments of benign stratied squamous epithelium with amorphous basement
(Bowmans) membrane. Focal areas of granular degeneration within Bowmans membrane
were noted which strained as a basophilic plaque with H+E, stained positive for Masons
trichrome, and negative for PAS, Alcian blue, Congo red, and Dahls calcium stain.
Conclusions: A limited number of noninfectious corneal changes have been identied in
immuno-compromised HIV patients including keratoconjunctivitis sicca, peripheral corneal
ulceration, and endothelial deposits related to CMV retinitis and rifabutin therapy. To this
group may now be added an unusually symptomatic and centralized variant of spheroidal
corneal degeneration.
CR: K.P. Han, None; J.C. Affeldt, None.
Support:
2952 - B587
Hereditary mucoepithelial dysplasia.
S.Hakiki
1A
, F.Audren
1A
, P.Rapp
1A
, C.Orssaud
1A
, O.Roche
1A
, J.-L.Duer
1A
, S.Hadj-Rabia
1B
.
A
Ophthalmology,
B
Dermatology,
1
Hopital Necker Enfants-Malades, Paris, France.
Purpose: Here we report a new case of hereditary mucoepithelial dysplasia (HMD)
which is a multiepithelial disorder involving mucosae, skin, hair, eyes, and lungs. This
disease is caused by an abnormality in desmosomes and gap junctions. Methods:
Results: A 3 years old girl was hospitalised for a recurrent severe bilateral keratitis with decreased
visual acuity initially diagnosed as herpetic keratitis. No improvement was observed under
systemic acyclovir. Under general anaesthesia there was a bilateral, severe central keratitis with
a hazy and vascularized cornea. The physical examination made by the dermatologist showed a
non scarring alopecia, an angular cheilitis and a well-demarcated pink-red plaque covered with
ne relatively inapparent small scales extending through the perineum to the superior portion
of the inter gluteal cleft. Zinc blood level was normal. Her mother reports surgical extraction
of a bilateral cataract in chilhood and an unilateral corneal graft after a severe keratitis.
Conclusions: HMD is a rare autosomal, dominantly inherited disease with no identied gene.
HMD should be considered in the differential diagnosis of childhood keratoconjunctivitis, and/
or juvenile cataracts associated to multi-epithelial lesions. A complete physical examination
by the dermatologist was necessary in this case for the diagnosis. The familial history of
keratoconjonctivitis and cataract was also helpful. HMD is a severe disease that may led
to blindness.
CR: S. Hakiki, None; F. Audren, None; P. Rapp, None; C. Orssaud, None; O. Roche,
None; J. Duer, None; S. Hadj-Rabia, None.
Support:
Tuesday, April 27, 8:30 - 10:30 AM Hall BC Poster Session Program Number/Board # Range: 2935-2956 / B570-B591
367. Clinical Studies: Pterygium Organizing Section: CO
2947-2952
Copyright 2004 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Ofce at pubs@arvo.org.
Commercial Relationships are noted at the end of each abstract by None or with codes.
2953 - B588
A new surgical procedure to prevent limbus insufciency after proton beam therapy
for iris melanoma.
F.Majo, L.Zografos, P.Othenin-Girard. Ophthalmology, Hpital Ophtalmique Jules
Gonin, Lausanne, Switzerland.
Purpose: Ocular surface complication is the most important challenge after proton beam
therapy for iris melanoma. It usually appears two or three weeks after the irradiation, it
looks like limbus insufciency. Clinical signs are observed from a few months to a couple of
years. We describe a new surgical procedure to prevent those ocular surface complications.
Methods: Five patients were enrolled in this study. Preoperatively, ocular surface was examined:
redness, quality of the tear lm (BUT, Shirmer) and transparency of the cornea. We removed
half of the limbus before the irradiation and conserved it in an organoculture medium at +4C
for ve days. Then, proton beam therapy was performed during four days four time a day. At the
end of the treatment, the patient was anaesthetized and the limbus put back where it belonged.
Results: Ocular surface of our ve patients was dramatically healthy with this technique
compared with the patients treated for the same disease without radioprotection of the
limbus area. Current complications like corneal melting, chronic epithelial defect,
chronic inammation and in few cases, limbus insufciency - have not been noticed.
Conclusions: This new procedure seems to be useful to prevent ocular surface complication
or limbus insufciency after eye irradiation. Those data precise the keyplace of the limbus in
the ocular surface, not only as a stem cell storage tank for the cornea but also as a regulator
of the inammation in ocular surface.
CR: F. Majo, None; L. Zografos, None; P. Othenin-Girard, None.
Support:
2954 - B589
Noninfectious Pseudodendritic Keratitis
K.K. Chang
1
, J.C. Affeldt
2
, L.Labree
3
, M.Agarwal
2
.
1
Ophthalmology, Loma Linda
University, Loma Linda, CA;
2
Ocular Surface Center, Doheny Eye Institute, Los Angeles,
CA;
3
Dept of Preventive Medicine, Keck School of Medicine of USC, Los Angeles, CA.
Purpose: To document for the rst time the clinical characteristics of a distinctive
variant of neurotrophic keratitis (termed here noninfectious pseudodendritic keratitis)
which masquerades as dendritic herpes keratitis. Methods: Observational case series.
Results: Ten eyes of the 9 patients presented with a branching epitheliopathy closely
resembling dendritic herpes keratitis. Five patients were undergoing topical and/or systemic
antiviral therapy at the time of initial exam. In all cases, the lesions were manifested by
opaque whitish elevated epithelial cell congregations which formed rough branching dendritic
gures without terminal bulbs. All lesions were limited to the inferior or inferocentral
cornea, with the long axis of branches oriented at 180 degrees. In addition, all lesions
stained with uorescein, and were found to be broadly surrounded by a dense eld of
punctate keratopathy. Average central cornal sensation (Cochet-Bonnet esthesiometer)
was signicantly reduced as compared to controls (6.3 mm versus 56.3 mm; P=0.0001),
reecting a neurotrophic environment. In all cases the lesions rapidly resolved with
discontinuation of antivrial therapy coupled with aggressive lubrication and punctal occlusion.
Conclusions: Noninfectious pseudodendritic keratitis appears to represent a neurotrophically
generated variant of vortex keratopathy (cornea verticillata) which can be easily confused
with dendritic herpes simplex or zoster keratitis. It can be distinguished from its infectious
counterparts however by its distincitive clinical presentation including lesion color, location,
orientation, and presence of surrounding punctate keratopathy eld, as well as its rapid
response to aggressive punctal occlusion.
CR: K.K. Chang, None; J.C. Affeldt, None; L. Labree, None; M. Agarwal, None.
Support:
2955 - B590
Clinical Risk Zones for Development and Staging of Neurotrophic Keratitis
N.J. Rudometkin
1
, J.C. Affeldt
2
, L.Labree
3
, M.Agarwal
4
.
1
Loma Linda University
Department of Ophthalmology, Loma Linda, CA;
2
Ophthalmology, Ocular Surface
Center, Doheny Eye Institue, Dept of Ophthalmology, Keck School of Medicine USC, Los
Angeles, CA;
3
Preventive Medicine, Keck School of Medicine of USC, Los Angeles, CA;
4
Ophthalmology, Ocular Surface Center, Doheny Eye Intsitute, Dept of Ophthalmology,
Keck School of Medicine, Los Angeles, CA.
Purpose: To document the existence of meassurable clinical risk zones for
development of neurotrophic keratitis (NTK) and progression of its various
stages utilizing the parameters of quantitated central corneal sensation (CCS)
and aqueous tear production (ATP). Methods: Observational case series
Results: Two hundred twenty-seven eyes manifesting NTK were identied with disease
severity staged according to the Mackie classication system (Stage I = punctate keratitis,
Stage II = epithelial defect, Stage III = stromal melt). CCS was quantitated using the Cochet-
Bonnet esthesiometer, while ATP was measured using the Schirmer method with topical
anesthesia. Mean and standard deviation (SD) of CCS for all affected eyes was signicantly
reduced (5.17.3 mm) as compared to both fellow (34.919.7 mm; p<0.0001) and control eyes
(56.36.7 mm; p<0.0001). Likewise, mean and SD for CCS was signicantly reduced in NTK
stages II and III (3.87.1 mm) as compared to NTK stage I (5.87.5 mm; p=0.004). Graphic
plot of CCS vs. ATP revealed relatively distinct clinical risk zones for not only development
of neurotrophic disease, but also for disease progression from Stage I to Stages II and III.
Conclusions: Utilizing the parameters of quantitated CCS and ATP, risk zones for development
and potential progression of NTK were identied in this series, the largest NTK series ever
reported to the best of our knowledge. These zones can be clinically useful for denitional,
diagnostic, and prognostic purposes, as well as guidance in the treatment of this important
and problematic member of the ocular surface disease family.
CR: N.J. Rudometkin, None; J.C. Affeldt, None; L. Labree, None; M. Agarwal, None.
Support:
2956 - B591
Epithelial debridement and diamond burr polishing of Bowmans layer in the
management of recalcitrant recurrent corneal erosions
M.A. Roberts, A.J. Aldave. Cornea Division, Jules Stein Eye Institute/UCLA, Los
Angeles, CA.
Purpose: Report the safety and efficacy of epithelial debridement with diamond
burr polishing of Bowmans layer for management of recalcitrant corneal erosions.
Methods: Interventional case series of 9 patients (14 eyes) with recurrent corneal epithelial erosions
that had failed to respond to conservative therapy. For each patient, the following information
was recorded: history of ocular trauma, frequency of corneal erosions prior to treatment,
previous medical and surgical treatment, pre-operative and post-operative visual acuities, and
the occurrence of corneal erosions and other complications following epithelial debridement.
Results: The 9 patients had been treated for an average of 4 episodes (range, 1-7 episodes)
of spontaneous corneal erosions prior to presentation. One patient (2 eyes) had a history
of prior corneal trauma and 5 patients (9 eyes) demonstrated clinical ndings of anterior
basement membrane dystrophy. In 1 patient (1 eye), epithelial debridement had been performed
twice, and all patients had been treated conservatively with ocular lubrication, topical
hypertonic agents, and/or bandage soft contact lenses. All patients demonstrated global
epithelial dysadherence at the time of epithelial debridement. Post-operatively, four patients
(6 eyes) developed faint subepithelial haze, rst noted 6-12 days after the procedure, which
was not symptomatic in any patient. All patients were within 1 line of pre-operative best
spectacle corrected visual acuity at an average follow-up period of 140 days (range, 37-353
days). No patient experienced a recurrent corneal erosion during the follow-up interval.
Conclusions: Epithelial debridement with diamond burr polishing of Bowmans layer is a
safe and effective treatment for recurrent corneal erosions in patients with corneal epithelial
dysadherence refractory to more conservative therapy.
CR: M.A. Roberts, None; A.J. Aldave, None.
Support:
Tuesday, April 27, 8:30 - 10:30 AM Hall BC Poster Session Program Number/Board # Range: 2935-2956 / B570-B591
367. Clinical Studies: Pterygium Organizing Section: CO
2953-2956