1: Ophthalmic Res. 2010 Jun 3;44(2):125-130 [Epub ahead of print] 

Isolation and Characterization of Fetus Human Retinal Microvascular Endothelial
Cells.

Xiaozhuang Z, Xianqiong L, Jingbo J, Shuiqing H, Jie Y, Yunbin C.

Guangdong Province Maternal and Children's Hospital, Guangzhou, PR China.

Purpose: To isolate and characterize primary fetus retinal microvascular
endothelial cells (RMECs) in order to facilitate the study of their properties
in vitro. Methods: Human RMECs were isolated from abortive fetuses whose
gestational ages were between 20 and 28 weeks by mechanical morcel and
trypsin-collagenase digestion, plated on fibronectin-coated wells and purified
gradually in high-selective condition. The RMECs were characterized for
expression and localization of endothelial cell markers by indirect
immunocytochemical staining of von Willebrand factor (vWF) and CD34, the
distribution of cell markers was observed by confocal immunofluorescence
microscope. The ability of these cells to form capillary-like networks was
assessed subsequently. Cobalt dichloride (CoCl(2)) was used to simulate hypoxia
in the culture condition and Western blot was used to detect the alternation of
VEGF protein level under normoxia and hypoxia. Results: Isolation of fetus RMECs
has been very difficult and primary culture of these special premature RMECs has
not been previously reported due to many reasons. Here we established a simple
and convenient method for the primary culture of fetus human RMECs. Our results
show that delicate RMECs could be obtained readily and grow very fast, and the
purity of RMECs was up to 95% demonstrated by specific staining for vWF in the
cytoplasm. Our research also indicates that nearly 100% of cells could express
CD34. The cells were successfully passaged and maintained in culture for 5-7
passages without a significant loss in expression of endothelia cell markers.
The fetus RMECs formed capillary-like networks on the Matrigel well. Expression
of VEGF protein increased significantly under hypoxia. Conclusions: Pure fetus
RMECs could be obtained rapidly and easily by our method.Fetus RMECs have some
specific properties. Hypoxia could induce expression of VEGF. Cell properties
related with age of the donors should be considered while studying in vitro,
especially in some age-specific diseases such as retinopathy of prematurity and
diabetic retinopathy. Copyright (c) 2010 S. Karger AG, Basel.

PMID: 20523104  [PubMed - as supplied by publisher]

2: Ophthalmic Res. 2010 Jun 3;44(2):119-124 [Epub ahead of print] 

Inducing Ocular Allergy with beta-Lactoglobulin: A New Experimental Model.

Sari A, Dinc E, Yilmaz BC, Adiguzel U, Oztuna S, Tasdelen B.

Department of Ophthalmology, Mersin University School of Medicine, Mersin,
Turkey.

Purpose: To develop a new experimental ocular allergy animal model induced by
beta-lactoglobulin (BLG), a major cow's milk allergen, and to discuss the
clinical, histopathological and immunohistochemical findings. Methods: Forty
Balb/c mice were randomized and separated into groups of 10. Groups were
determined according to the different concentrations of BLG drops used. Study
groups were immunized with 2.5, 5 or 10 mg/ml topical BLG (groups 2, 3 and 4,
respectively) following intraperitoneal injection for the systemic immunization.
The control group (group 1) was immunized with aluminium hydroxide (alum) alone
within the same intervals. After ocular challenge, all the animals were
evaluated clinically, histopathologically (mast cell and eosinophil
infiltration) and immunhistochemically in terms of both T helper type 1-
(IFNgamma, TNFalpha) and T helper type 2- (IL-3, IL-4, IL-5, IL-10) specific
cytokines. Results: Both clinical and immunohistochemical findings showed that
an allergic conjunctivitis was induced in all study groups, with an optimized
dosage of topical 5 mg/ml BLG. The conjunctivitis was associated with both Th1
and Th2 response, with a slight predominance of Th1 reaction. Conclusion: We
describe a new murine model of acute allergic conjunctivitis induced by BLG. We
believe that this new preliminary model has the immune parameters of the late
phase of acute allergic conjunctivitis and it provides an alternative means for
studying the pathogenesis and future treatments of ocular allergy. Our results
should be enhanced with more detailed cellular and humoral parameters. Copyright
(c) 2010 S. Karger AG, Basel.

PMID: 20523103  [PubMed - as supplied by publisher]

3: Ophthalmic Res. 2010 Jun 2;44(2):140 [Epub ahead of print] 

Impact of Once Daily versus Twice Daily Application of Adjunctive Timolol on the
Intraocular Pressure-Lowering Effect of Latanoprost.

Nowroozzadeh MH.

Department of Ophthalmology, Khalili Hospital, Shiraz University of Medical
Sciences, Shiraz, Iran.

Publication Types:
    Letter

PMID: 20516726  [PubMed - as supplied by publisher]

4: Ophthalmic Res. 2010 Jun 2;44(2):131-139 [Epub ahead of print] 

Macular Pigment Optical Density in an Ageing Irish Population: The Irish
Longitudinal Study on Ageing.

Nolan JM, Kenny R, O'Regan C, Cronin H, Loughman J, Connolly EE, Kearney P,
Loane E, Beatty S.

Macular Pigment Research Group, Department of Chemical and Life Sciences,
Waterford Institute of Technology, and Institute of Vision Research, Whitfield
Clinic, Waterford, Ireland.

Purpose: The 3 carotenoids lutein, zeaxanthin, and meso-zeaxanthin, which
account for the 'yellow spot' at the macula and which are referred to as macular
pigment (MP), are believed to play a role in visual function and protect against
age-related macular degeneration (AMD) via their optical and antioxidant
properties. This study was undertaken to compare MP optical density (MPOD) in a
population aged >/=50 years with MPOD values from a normative database of
subjects aged 18-60 years. Methods: Seventy-nine subjects were recruited into
this pilot study (The Irish Longitudinal Study on Ageing-TILDA). MPOD was
measured using heterochromatic flicker photometry. Retinal fundus photographs,
lifestyle data and general health data, were also obtained. Results: The mean
+/- SD age of the 79 subjects recruited into this study was 65 +/- 11 years.
There was a moderate, but statistically significant, age-related decline in MPOD
at 0.5 degrees in the TILDA data (r = -0.251, p = 0.045), which remained upon
merging with a normative database of an additional 462 subjects aged between 18
and 67 years (r = -0.179, p = 0.000). Conclusions: We report an inverse
association between MPOD and increasing age. Longitudinal data in a larger
cohort of participants are required to satisfactorily investigate the
relationship between the optical density of this pigment and age, and with risk
for development and/or progression of AMD. This pilot study represents a first
step in this endeavour. Copyright (c) 2010 S. Karger AG, Basel.

PMID: 20516725  [PubMed - as supplied by publisher]

5: Ophthalmic Res. 2010 Jun 2;44(2):113-118 [Epub ahead of print] 

Correlation of Proteinase Production with Amphotericin B Resistance in Fungi
from Mycotic Keratitis.

Nayak N, Satpathy G, Prasad S, Vajpayee RB, Pandey RM.

Department of Ocular Microbiology and Cornea Services, Dr. Rajendra Prasad
Centre for Ophthalmic Sciences, New Delhi, India.

Two hundred fungal isolates (Aspergillus and Fusarium species) from mycotic
keratitis were tested for in vitro susceptibilities to amphotericin B and
proteinase production. Geometric mean MICs for all fungal species increased
fourfold with thousandfold increase in the inoculum. The MIC(50) and MIC(90)
values ranged between 3.12-6.25 and 3.12-12.5 mug/ml, respectively. Proteinase
production was noted in 113 (56.5%) isolates. Ninety-eight (49%) showed MICs of
>/=1.56 mug/ml that was above the criteria of >/=1 mug/ml for amphotericin B
resistance (CLSI). Seventy-three (74.5%) of these 98 isolates were proteinase
producers, whereas only 40 (39.2%) of the remaining 102 with low MICs (<1.56
mug/ml) were proteinase producers (p < 0.001). Proteinase seems to be an
important virulence marker of filamentous fungi in mycotic keratitis,
correlating significantly with amphotericin B resistance. Copyright (c) 2010 S.
Karger AG, Basel.

PMID: 20516724  [PubMed - as supplied by publisher]

6: Ophthalmic Res. 2010 May 13;44(2):105-112 [Epub ahead of print] 

Taurine Buffers Glutamate Homeostasis in Retinal Cells in vitro under Hypoxic
Conditions.

Chen F, Mi M, Zhang Q, Wei N, Chen K, Xu H, Yuan J, Zhou Y, Lang H, Yu X, Wang
B, Wang J, Tang Y, Chang H.

Department of Nutrition and Food Hygiene, Chongqing Key Laboratory of Nutrition
and Food Safety, Third Military Medical University, Chongqing, China.

Aims: We investigated whether taurine indirectly protects neurons under hypoxia
by affecting retinal Muller cells, which are known to play important roles in
the regulation of retinal glutamate content. Methods: Retinal cells isolated
from rats were exposed to hypoxia for 24 h. We evaluated the retinal neuron
survival, glutamate content in cultures with and without taurine under hypoxic
conditions. The glutamate clearance function correlated with the expression of
glutamine synthetase (GS) mRNA and L-glutamate/L-aspartate transporter (GLAST)
mRNA. Immunohistochemical staining of glial fibrillary acidic protein (GFAP),
vimentin and S-100 protein was performed to examine cytoskeletal changes in
retinal Muller cells. Results: Retinal neurons treated with taurine exhibited
significantly higher survival rates than those without taurine under hypoxia.
Taurine inhibited the upregulation of GFAP and vimentin, and inhibited the
downregulation of GLAST, GS and the nuclear-cytoplasmic ratio of S-100 under
hypoxia. In addition, taurine inhibited the upregulation of the glutamate
content in neurons and retinal Muller cells upon hypoxic exposure. Conclusion:
These data suggest that hypoxic damage to cultured retinal cells is decreased by
taurine. The neuroprotection by taurine may relate to buffering glutamate
homeostasis via modulation of the glutamate clearance by retinal Muller cells.
Copyright (c) 2010 S. Karger AG, Basel.

PMID: 20484951  [PubMed - as supplied by publisher]

7: Ophthalmic Res. 2010 May 13;44(2):82-104 [Epub ahead of print] 

Retinal and Ocular Toxicity in Ocular Application of Drugs and Chemicals - Part
I: Animal Models and Toxicity Assays.

Penha FM, Rodrigues EB, Maia M, Dib E, Fiod Costa E, Furlani BA, Nunes Moraes
Filho M, Dreyfuss JL, Bottos J, Farah ME.

Vision Institute, Department of Ophthalmology, Federal University of Sao Paulo,
Sao Paulo, Brazil.

Aims: Experimental retinal research has gained great importance due to the
ophthalmic pharmacotherapy era. An increasing number of drugs are constantly
released into the market for the treatment of retinal diseases. In this review,
animal species, animal models and toxicity assays in retinal research are
discussed. Methods: An extensive search of the literature was performed to
review various aspects of the methods of investigation of drug toxicity. The
different types of animal species, as well as single animal models available for
the evaluation of safety and efficacy of retinal pharmacotherapy, were
identified. In addition, a large variety of reported laboratory techniques were
critically examined. Results: In vitro studies are the first-line experiments
for the development of a new drug for retinal diseases, using retinal pigment
epithelial cells and other cell lines. The next step involves in vivo animal
studies where nonhuman primates are considered the gold standard. However, cost
and legal issues make their use difficult. Mice and rats provide genetically
controlled models for investigations. Pigs, dogs and cats represent good
large-size animal models, while rabbits are one of the most used species for
retinal toxicity evaluations. Various laboratory methods were identified,
including light microscopy, electron microscopy, electroretinography and new
emerging methods, such as optical coherence tomography and scanning laser
ophthalmoscopy for experimental purposes. Conclusions: A great number of animal
species and models are available that simulate retinal diseases and provide
experimental data for further human use. Work with animal models should include
properly designed toxicity assays to obtain reliable results for safety and
efficacy. Copyright (c) 2010 S. Karger AG, Basel.

PMID: 20484950  [PubMed - as supplied by publisher]