Publications by authors named "Mongkol Uiprasertkul"

43 Publications

Intravitreal autologous mesenchymal stem cell transplantation: a non-randomized phase I clinical trial in patients with retinitis pigmentosa.

Stem Cell Res Ther 2021 01 9;12(1):52. Epub 2021 Jan 9.

Department of Ophthalmology, Faculty of Medicine Siriraj Hospital, Mahidol University, 2 Wanglang Road, Bangkok Noi, Bangkok, 10700, Thailand.

Background: Retinitis pigmentosa (RP) is a progressive inherited retinal disease with great interest for finding effective treatment modalities. Stem cell-based therapy is one of the promising candidates. We aimed to investigate the safety, feasibility, and short-term efficacy of intravitreal injection of bone marrow-derived mesenchymal stem cells (BM-MSCs) in participants with advanced stage RP.

Methods: This non-randomized phase I clinical trial enrolled 14 participants, categorized into three groups based on a single dose intravitreal BM-MSC injection of 1 × 10, 5 × 10, or 1 × 10 cells. We evaluated signs of inflammation and other adverse events (AEs). We also assessed the best corrected visual acuity (BCVA), visual field (VF), central subfield thickness (CST), and subjective experiences.

Results: During the 12-month period, we noticed several mild and transient AEs. Interestingly, we found statistically significant improvements in the BCVA compared to baseline, although they returned to the baseline at 12 months. The VF and CST were stable, indicating no remarkable disease progression. We followed 12 participants beyond the study period, ranging from 1.5 to 7 years, and observed one severe but manageable AE at year 3.

Conclusion: Intravitreal injection of BM-MSCs appears to be safe and potentially effective. All adverse events during the 12-month period required observation without any intervention. For the long-term follow-up, only one participant needed surgical treatment for a serious adverse event and the vision was restored. An enrollment of larger number of participants with less advanced RP and long-term follow-up is required to evaluate the safety and efficacy of this intervention.

Trial Registration: ClinicalTrials.gov, NCT01531348 . Registered on February 10, 2012.
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http://dx.doi.org/10.1186/s13287-020-02122-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7796606PMC
January 2021

GD2-specific chimeric antigen receptor-modified T cells targeting retinoblastoma - assessing tumor and T cell interaction.

Transl Oncol 2021 Feb 13;14(2):100971. Epub 2020 Dec 13.

Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida, United States; Shenzhen Geno-Immune Medical Institute, 2nd FL. 6 Yuexing 2nd Rd., Nanshan Dist., Shenzhen, China; School of Medicine, University of Electronic Science and Technology of China, Sichuan, China. Electronic address:

A novel disialoganglioside 2 (GD2)-specific chimeric antigen receptor (CAR)-modified T cell therapy against retinoblastoma (RB) were generated. GD2-CAR consists of a single-chain variable fragment (scFv) derived from a monoclonal antibody, hu3F8, that is linked with the cytoplasmic signaling domains of CD28, 41BB, a CD3ζ, and an inducible caspase 9 death fusion partner. GD2 antigen is highly expressed in Y79RB cell line and in several surgical RB tumor specimens. In vitro co-culture experiments revealed the effective killing of Y79RB cells by GD2-CAR T cells, but not by control CD19-CAR T cells. The killing activities of GD2-CAR T cells were diminished when repeatedly exposed to the tumor, due to an attenuated expression of GD2 antigen on tumor cells and upregulation of inhibitory molecules of the PD1 and PD-L1 axis in the CAR T cells and RB tumor cells respectively. This is the first report to describe the potential of GD2-CAR T cells as a promising therapeutic strategy for RB with the indication of potential benefit of combination therapy with immune checkpoint inhibitors.
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http://dx.doi.org/10.1016/j.tranon.2020.100971DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7745061PMC
February 2021

Phenotypic Characterization of Corneal Epithelium in Long-Term Follow-Up of Patients Post-Autologous Cultivated Oral Mucosal Epithelial Transplantation.

Cornea 2021 Jul;40(7):842-850

Research Division, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.

Purpose: To analyze the phenotype of the corneal epithelium in patients with long-term follow-up who underwent autologous cultivated oral mucosal epithelial transplantation (COMET) using in vivo confocal microscopy (IVCM) and impression cytology with immunofluorescence staining (ICIF).

Methods: Thirteen eyes from patients with severe limbal stem cell deficiency, who underwent COMET at least 48 months before, were recruited in this noncomparative cohort study. After eye examination, IVCM and ICIF were performed. Clinical manifestations of the cornea were evaluated and compared with epithelial findings detected by IVCM and ICIF [cytokeratin (CK) 3, CK7, and CK12]. Two corneal buttons derived from patients receiving the corneal transplantation post-COMET were sent for immunohistochemistry (CK3, CK6, CK7, CK12, paired box gene 6, p63, zonula occludens-1, and integrin β -1).

Results: The mean age of patients was 51.2 ± 20.6 years, and the mean follow-up time since COMET was 78.7 ± 16.3 months. Six of 13 eyes showed clinically successful COMET. In these eyes, IVCM demonstrated predominant cornea-like epithelium and ICIF reported positivity for CK3 and CK12, confirming the presence of oral mucosal and corneal epithelium. Meanwhile, 7 eyes showed total conjunctivalization, corresponding with substantial conjunctival epithelium detected by IVCM and positivity for conjunctival (CK7) and oral mucosal epithelial (CK3) markers detected by ICIF. The immunohistochemistry of corneal buttons stained positive for oral mucosal, corneal epithelial, and stem cell markers (CK3, CK12, and p63).

Conclusions: In long-term follow-up of COMET, epithelium of successful patients demonstrated cornea-like phenotype, whereas failed cases revealed mainly conjunctival phenotype. However, there were evidences that oral mucosal epithelial cells remained across the cornea in both successful and failed COMET as detected by IVCM and ICIF.
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http://dx.doi.org/10.1097/ICO.0000000000002498DOI Listing
July 2021

Orbital Infection by Saksenaea vasiformis in an Immunocompetent Host.

Case Rep Ophthalmol Med 2020 30;2020:8827074. Epub 2020 Sep 30.

Department of Ophthalmology and Visual Sciences, South Australian Institute of Ophthalmology, University of Adelaide, Adelaide, South Australia, Australia.

Orbital mucormycosis caused by is extremely rare. Herein, we report an immunocompetent 22-year-old Thai female who presented with two months of progressive right upper eyelid mass, associated with swelling, redness, and ptosis. She failed to improve despite multiple courses of antibiotic and steroid treatment. Computed tomography (CT) scan showed infiltration involving the upper eyelid and lacrimal gland. Fungal hyphae were revealed by histopathological study. Polymerase chain reaction (PCR) was positive for (GenBank: accession number FR687327.1). The patient was successfully treated with surgical debridement, amphotericin B, and oral posaconazole.
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http://dx.doi.org/10.1155/2020/8827074DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7547347PMC
September 2020

Large Dacryops Mimicking Lacrimal Gland Tumor With Secondary Corneal Ulcer.

Ophthalmic Plast Reconstr Surg 2021 May-Jun 01;37(3S):S167

Mettapracharak Eye Center, Mettapracharak (Wat Rai Khing) Hospital, Nakhon Pathom, and.

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http://dx.doi.org/10.1097/IOP.0000000000001735DOI Listing
May 2021

Unusual non-nanophthalmic uveal effusion syndrome with histologically normal scleral architecture: a case report.

BMC Ophthalmol 2020 Jul 29;20(1):311. Epub 2020 Jul 29.

Department of Ophthalmology, Faculty of Medicine Siriraj Hospital, Mahidol University, 2 Wanglang Road, Bangkoknoi, Bangkok, 10700, Thailand.

Background: To report an unusual case of non-nanophthalmic uveal effusion syndrome (UES) with histologically normal sclera but responsive to scleral resection.

Case Presentation: A73-year-old man presented with a bullous retinal detachment without ciliochoroidal detachment on funduscopic examination of the right eye. The axial length of both eyes was normal. Extensive investigations for possible causes of exudative retinal detachment were performed with unremarkable results except for choroidal hyperpermeability on indocyanine green angiography (ICGA). Ultrasound biomicroscopy (UBM) revealed scleral thickening with peripheral choroidal elevation leading to the diagnosis of UES. Partial thickness sclerectomy and sclerotomy was performed resulting in complete retinal reattachment, reduction of choroidal hyperpermeability on ICGA and improvement of visual acuity. However, histological studies of the excised sclera revealed no scleral architectural changes or abnormal deposits.

Conclusions: The diagnosis of UES in non-nanophthalmic eyes is challenging. Thorough systemic and ocular investigations are critical to rule out other etiologies. UBM can be helpful to evaluate scleral thickness and anterior choroid in equivocal cases. Our case was unique in that, although the sclera was thick, no abnormal microscopic scleral architecture could be identified. Misdiagnosis may lead to different surgical procedures such as vitrectomy resulting in unfavorable outcomes.
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http://dx.doi.org/10.1186/s12886-020-01581-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7391591PMC
July 2020

Comparative analysis of a Thai congenital-Zika-syndrome-associated virus with a Thai Zika-fever-associated virus.

Arch Virol 2020 Aug 30;165(8):1791-1801. Epub 2020 May 30.

Institute of Molecular Biosciences, Mahidol University, Salaya, Nakhon Pathom, Thailand.

In this study, we compared the characteristics of two strains of Zika virus (ZIKV) isolated in Thailand, one isolated from a febrile patient and one isolated from tissues of a fetus medically terminated due to congenital Zika syndrome (CZS). Replication profiles showed that the isolate from the fetal tissues replicated significantly more slowly than the fever-associated isolate in human lung A549 cells during the first 24 hours postinfection but showed a similar growth profile over longer-term infection. A much smaller difference was observed in Aedes albopictus C6/36 cells. In a quasispecies analysis, a high proportion (approximately 20%) of nonfunctional genomes was identified, caused by an adenine insertion in the prM gene. This insertion was found to be present in two Thai fever strains and as such may represent a common feature of Thai endemic ZIKV. Comparison between viral RNA copy number and viral titer showed that the isolate from fetal tissues was produced more efficiently than the fever-associated isolate. Together, these results suggest that different ZIKV isolates differ in their replication capacity, and this might contribute to the fetotropic potential of a particular strain.
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http://dx.doi.org/10.1007/s00705-020-04667-7DOI Listing
August 2020

Characterization of limbal explant sites: Optimization of stem cell outgrowth in in vitro culture.

PLoS One 2020 14;15(5):e0233075. Epub 2020 May 14.

Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.

Simple limbal epithelial transplantation (SLET) and cultivated limbal epithelial transplantation (CLET) are proven techniques for treating limbal stem cell deficiency (LSCD). However, the precise regions that are most suitable for preparing explants for transplantation have not been identified conclusively. Accordingly, this in vitro study aimed at determining ideal sites to be selected for tissue harvest for limbal stem cell culture and transplantation. We evaluated cell outgrowth potential and the expression of stem cell markers in cultures from 48 limbal explants from five cadaveric donors. The limbal explants were generated from the three specific sites: Lcor (located innermost and adjacent to the cornea), Lm (middle limbus), and Lconj (located outermost adjacent to the conjunctiva). We found that explants from the Lconj and Lm sites exhibited higher growth potential than those from the Lcor site. Transcript encoding the stem cell marker and p63 isoform, ΔNp63, was detected in cells from Lm and Lconj explants; expression levels were slightly, though significantly (p-value < 0.05), higher in Lm than in Lconj, although expression of ΔNp63α protein was similar in cells from all explants. Differential expression of ATP-Binding Cassette Subfamily G Member 2 (ABCG2) did not reach statistical significance. Immunohistochemistry by indirect immunofluorescence analysis of limbus tissue revealed that the basal layer in explant tissue from Lconj and Lm contained markedly more stem cells than found in Lcor explant tissue; these findings correlate with a higher capacity for growth. Collectively, our findings suggest that explants from the Lconj and Lm sites should be selected for limbal cell expansion for both CLET and SLET procedures. These new insights may guide surgeons toward specific limbal sites that are most suitable for stem cell culture and transplantation and may ultimately improve treatment outcomes in the patients with LSCD.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0233075PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7224544PMC
August 2020

Complete Genomic Sequences of Highly Pathogenic H5N1 Avian Influenza Viruses Obtained Directly from Human Autopsy Specimens.

Microbiol Resour Announc 2018 Dec 6;7(22). Epub 2018 Dec 6.

Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.

The complete genomic sequences of H5N1 highly pathogenic avian influenza (HPAI) viruses were directly obtained from lung, trachea, and colon tissues and an intestinal fecal sample of a patient by using the next-generation sequencing technique. This is the first report on complete H5N1 viral genomes from human autopsy specimens.
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http://dx.doi.org/10.1128/MRA.01498-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6284082PMC
December 2018

Rare presentation of intractable tuberculous panophthalmitis with intraocular and intraorbital abscesses: a case report.

J Med Case Rep 2017 Jul 4;11(1):180. Epub 2017 Jul 4.

Department of Ophthalmology, Faculty of Medicine, Siriraj Hospital, Mahidol University, 2 Wanglang Road, Bangkoknoi, Bangkok, 10700, Thailand.

Background: We report a rare presentation of extrapulmonary tuberculosis.

Case Presentation: A 29-year-old Burmese woman with human immunodeficiency virus infection and known pulmonary tuberculosis who had been treated for 5 months presented to our hospital with unilateral progressive painful visual loss of 1 month's duration. She was diagnosed with tuberculous panophthalmitis with subretinal and intraorbital abscesses, conjunctival abscess, and extraocular muscle tuberculoma. The diagnosis was confirmed by a conjunctival pus swab with a positive result for acid-fast bacilli and a positive result for a mycobacterial culture. There was high suspicion of multidrug-resistant tuberculosis. Despite receiving ongoing aggressive treatment with conventional antituberculous medications, this patient required subtotal orbital exenteration to control her infection and prevent further progression. Second-line antituberculous medications were added to the first-line therapy, with satisfactory results achieved.

Conclusions: Tuberculous panophthalmitis with intraocular and intraorbital abscesses is a rare presentation of extrapulmonary tuberculosis. Patients who do not respond to first-line antituberculous therapy might be infected with either single-drug or multidrug-resistant Mycobacterium tuberculosis. Patient compliance is one of the key factors that can alter the course of treatment. Careful patient monitoring can improve disease progression, outcome, and prognosis.
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http://dx.doi.org/10.1186/s13256-017-1353-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5496312PMC
July 2017

Long-term result of autologous cultivated oral mucosal epithelial transplantation for severe ocular surface disease.

Cell Tissue Bank 2016 Sep 9;17(3):491-503. Epub 2016 Aug 9.

Department of Ophthalmology, Faculty of Medicine Siriraj Hospital, Mahidol University, 2 Prannok Road, Bangkoknoi District, Bangkok, 10700, Thailand.

The present study aimed to investigate the clinical outcomes of autologous cultivated oral mucosal epithelial transplantation (COMET) on human amniotic membrane (AM) for corneal limbal stem cell deficiency (LSCD). In this prospective, noncomparative case series, 20 eyes (18 patients) with bilateral severe ocular surface disease were chosen to undergo COMET on human AM. The primary outcome was clinical success, and the secondary outcomes were the best-corrected visual acuity difference, corneal opacification, symblepharon formation, and complications. The mean patient age was 48.2 ± 15.5 years. The mean follow-up time was 31.9 ± 12.1 months (range 8-50 months). All except one eye exhibited complete epithelialization within the first postoperative week. A successful clinical outcome, defined as a stable ocular surface without epithelial defects, a clear cornea without fibrovascular tissue invasion at the pupillary area, and no or mild ocular surface inflammation, was obtained in 15 of 20 eyes (75 %). The clinical success rate at 1 year was 79.3 %, and that at 4 years (end of follow-up) was 70.5 %. Fourteen of 20 (70 %) eyes exhibited improvement in visual acuity after COMET, and some required subsequent cataract surgery (2 eyes), penetrating keratoplasty (3 eyes), or keratoprosthesis implantation (1 eye). Preoperative symblepharon was eliminated in most eyes (8 of 13, 61.5 %) after COMET combined with eyelid reconstruction when needed. The only complication was corneal perforation (1 eye) induced by a severe eyelid abnormality; treatment with a tectonic corneal graft was successful. COMET can successfully restore ocular surface damage in most eyes with corneal LSCD.
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http://dx.doi.org/10.1007/s10561-016-9575-4DOI Listing
September 2016

Expression of importin-α isoforms in human nasal mucosa: implication for adaptation of avian influenza A viruses to human host.

Virol J 2016 Jun 4;13:90. Epub 2016 Jun 4.

Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand.

Background: Transportation into the host cell nucleus is crucial for replication and transcription of influenza virus. The classical nuclear import is regulated by specific cellular factor, importin-α. Seven isoforms of importin-α have been identified in human. The preference of importin-α3 of avian influenza virus and -α7 isoform of human strains during replication in human cells was previously identified. In addition, both avian and human influenza viruses were shown to use importin-α1 isoform for their replication.

Finding: The mRNA levels of importin-α1, -α3, and -α7 isoforms in human respiratory tract was determined by real-time RT-PCR. The results indicate that mRNA level of importin-α7 was significantly higher than that of importin-α1 (p-value < 0.0001) and importin-α3 (p-value < 0.0001) isoforms in human nasal mucosa while importin-α1 was detected as the highest expression importin-α isoform in lung tissues.

Conclusions: These results may explain the preference of importin-α7 isoforms in seasonal influenza viruses in human upper respiratory tract and may suggest a selective pressure toward importin-α7 in human respiratory tract infection of an avian virus.
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http://dx.doi.org/10.1186/s12985-016-0546-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4893243PMC
June 2016

Neuraminidase Activity and Resistance of 2009 Pandemic H1N1 Influenza Virus to Antiviral Activity in Bronchoalveolar Fluid.

J Virol 2016 May 14;90(9):4637-4646. Epub 2016 Apr 14.

Department of Microbiology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand

Unlabelled: Human bronchoalveolar fluid is known to have anti-influenza activity. It is believed to be a frontline innate defense against the virus. Several antiviral factors, including surfactant protein D, are believed to contribute to the activity. The 2009 pandemic H1N1 influenza virus was previously shown to be less sensitive to surfactant protein D. Nevertheless, whether different influenza virus strains have different sensitivities to the overall anti-influenza activity of human bronchoalveolar fluid was not known. We compared the sensitivities of 2009 pandemic H1N1, seasonal H1N1, and seasonal H3N2 influenza virus strains to inhibition by human bronchoalveolar lavage (BAL) fluid. The pandemic and seasonal H1N1 strains showed lower sensitivity to human BAL fluid than the H3N2 strains. The BAL fluid anti-influenza activity could be enhanced by oseltamivir, indicating that the viral neuraminidase (NA) activity could provide resistance to the antiviral defense. In accordance with this finding, the BAL fluid anti-influenza activity was found to be sensitive to sialidase. The oseltamivir resistance mutation H275Y rendered the pandemic H1N1 virus but not the seasonal H1N1 virus more sensitive to BAL fluid. Since only the seasonal H1N1 but not the pandemic H1N1 had compensatory mutations that allowed oseltamivir-resistant strains to maintain NA enzymatic activity and transmission fitness, the resistance to BAL fluid of the drug-resistant seasonal H1N1 virus might play a role in viral fitness.

Importance: Human airway secretion contains anti-influenza activity. Different influenza strains may vary in their susceptibilities to this antiviral activity. Here we show that the 2009 pandemic and seasonal H1N1 influenza viruses were less sensitive to human bronchoalveolar lavage (BAL) fluid than H3N2 seasonal influenza virus. The resistance to the pulmonary innate antiviral activity of the pandemic virus was determined by its neuraminidase (NA) gene, and it was shown that the NA inhibitor resistance mutation H275Y abolished this resistance of the pandemic H1N1 but not the seasonal H1N1 virus, which had compensatory mutations that maintained the fitness of drug-resistant strains. Therefore, the innate respiratory tract defense may be a barrier against NA inhibitor-resistant mutants, and evasion of this defense may play a role in the emergence and spread of drug-resistant strains.
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http://dx.doi.org/10.1128/JVI.00013-16DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4836313PMC
May 2016

Distinct pathways in the pathogenesis of sebaceous carcinomas implicated by differentially expressed microRNAs.

JAMA Ophthalmol 2015 Oct;133(10):1109-16

Orbital Oncology and Ophthalmic Plastic Surgery, Department of Plastic Surgery, University of Texas MD Anderson Cancer Center, Houston.

Importance: The molecular-genetic alterations contributing to the pathogenesis of sebaceous carcinoma and sebaceous adenoma remain poorly understood. Given that sebaceous carcinoma is associated with substantial morbidity and mortality, there is a critical need to delineate the pathways driving sebaceous carcinoma and candidate molecules for targeted therapy.

Objective: To describe differentially expressed microRNAs (miRNAs) in a series of periocular sebaceous carcinomas compared with sebaceous adenomas in order to identify pathways driving the pathogenesis of sebaceous carcinoma.

Design, Setting, And Participants: Thirty sebaceous carcinomas and 23 sebaceous adenomas (including 11 that were confirmed to be related to Muir-Torre syndrome and 6 that were confirmed to be sporadic) were obtained from archives (from 48 patients) of 2 institutions (University of Texas MD Anderson Cancer Center, Houston, and Siriraj Hospital, Mahidol University, Bangkok, Thailand) and profiled.

Main Outcomes And Measures: Expression of miRNAs was determined using total RNA from formalin-fixed, paraffin-embedded tissue and real-time reverse transcription-polymerase chain reaction performed in a microfluidics card containing 378 unique miRNAs. Fold change was determined using the ΔΔCt method (reference probe, RNU48). Median centering was used to normalize the data. Two-sample t tests were used to identify differentially expressed miRNAs. The false discovery rate was assessed by β-uniform mixture analysis of P values from the t statistics. Significance was defined by this estimated false discovery rate.

Results: Serial testing and validation confirmed overexpression of 2 miRNAs previously reported to be oncogenic, miR-486-5p (4.4-fold; P = 2.4 × 10-8) and miR-184 (3.5-fold; P = 1.7 × 10-6), in sebaceous carcinoma compared with sebaceous adenoma and downregulation of 2 miRNAs previously reported to have tumor-suppressive properties, miR-211 (-5.8-fold; P = 2.3 × 10-9) and miR-518d (-4.5-fold; 6.7 × 10-5), in sebaceous carcinoma compared with sebaceous adenoma.

Conclusions And Relevance: Sebaceous carcinoma exhibits an miRNA expression profile distinct from that of sebaceous adenoma, implicating dysregulation of NF-κB and PTEN (targets of miR-486-5p) and TGF-β signaling (target of miR-211) in the pathogenesis of sebaceous carcinoma. The identification of miRNAs whose expression is altered in sebaceous carcinoma compared with sebaceous adenoma provides a novel entry point for a more comprehensive understanding of the molecular-genetic alterations pivotal to the development of sebaceous carcinoma.
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http://dx.doi.org/10.1001/jamaophthalmol.2015.2310DOI Listing
October 2015

The N-linked glycosylation site at position 158 on the head of hemagglutinin and the virulence of H5N1 avian influenza virus in mice.

Arch Virol 2015 Feb 11;160(2):409-15. Epub 2014 Dec 11.

Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand.

N-linked glycosylation of the influenza virus hemagglutinin (HA) protein plays crucial roles in HA structure and function, evasion of neutralizing antibodies, and susceptibility to innate soluble antiviral factors. The N-linked glycosylation site at position 158 of highly pathogenic H5N1 virus was previously shown to affect viral receptor-binding preference. H5N1 viruses show heterogeneity with respect to the presence of this glycosylation site. Clade 1 viruses that caused outbreaks in Southeast Asia in 2004 contained this glycosylation site, while the site is absent in the more recent clade 2 viruses. Here, we show that elimination of this glycosylation site increases viral virulence in mice. The mutant lacking the glycosylation site at position 158 showed unaltered growth kinetics in vitro and a comparable level of sensitivity to a major antiviral protein found in respiratory secretions, surfactant protein D (SP-D).
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http://dx.doi.org/10.1007/s00705-014-2306-xDOI Listing
February 2015

Evaluation of nested pcr technique for detection of Pythium insidiosum in pathological specimens from patients with suspected fungal keratitis.

Southeast Asian J Trop Med Public Health 2014 Jan;45(1):167-73

Diagnosis of Pythium keratitis is problematic due to the difficulty in obtaining a culture report resulting in unnecessarily prolonged usage of antimicrobial medication due to misdiagnosis. This study evaluated and compared nested PCR technique with culture and immunoperoxidase staining assays of Pythium insidiosum in paraffin-embedded corneal tissues from patients with suspected fungal keratitis. Six of 51 pathological reports compatible with fungal infection and 6 of 48 culture-proven fungal keratitis were identified as Pythium. Twenty-seven specimens were PCR-positive for Pythium insidiosum. In comparison with fungal culture for P. insidiosum, PCR had 83% sensitivity and 77% specificity with fair agreement (Kappa score of 0.227, p = 0.001). The mean age of PCR-positive is younger than PCR-negative group and there is a female preponderance in Pythium-infected group (p = 0.002 and p = 0.004, respectively). Nineteen specimens had positive results using immunoperoxidase staining assay with fair agreement to culture method (Kappa 0.340, p < 0.001), and 83% sensitivity, 85% specificity and 85% accuracy (95% CI: 76.7-90.7). PCR-based technique compared with culture and/or immunoperoxidase staining assay had 91.7% sensitivity, 81.8% specificity and 83% accuracy (95% CI: 74.5-89.1) with moderate agreement (Kappa 0.477, p < 0.001). Thus nested PCR detection of P. insidiosum should be employed in preliminary diagnosis of Pythium keratitis in order to initiate proper management.
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January 2014

NK/T-cell lymphoma of the nasal cavity causing contralateral dacryoadenitis.

Orbit 2013 Aug 10;32(4):250-2. Epub 2013 May 10.

Department of Ophthalmology, Siriraj Hospital, Mahidol University, Bangkok, Thailand.

Purpose: To report a case of NK/T-cell lymphoma of the nasal cavity with contralateral lacrimal gland involvement.

Methods: Observational case report.

Results: A 39-year-old woman with a 5-month history of right fungal rhinosinusitis was referred to our hospital. A nasal mucosal biopsy performed before referral was consistent with Aspergillus sp. Despite surgical and parenteral antifungal treatment, the symptoms continued to deteriorate. On admission, the ophthalmic evaluation showed inflammation over the left lacrimal gland area. The fundus examination revealed bilateral subretinal infiltration. Computed tomography scans of the orbits and sinuses showed mucosal thickening of the right nasal mucosa and sinuses. There was heterogeneous enhancement and infiltration of the left lacrimal gland. Lacrimal gland biopsy and repeated biopsies of the nasal cavity and sinus tissue were performed. Immunohistopathology of the lacrimal glands and nasal mucosa showed extranodal nasal-type NK/T-cell lymphoma. The patient was treated with cyclophosphamide, vincristine, adriamycin, prednisolone (3 cycles), and intrathecal methotrexate. The patient developed sepsis and died 2 months after initiation of chemotherapy.

Conclusion: Dacryoadenitis can be a clinical manifestation of NK/T-cell lymphoma. To our knowledge, this is the first reported case of nasal NK/T-cell lymphoma with contralateral dacryoadenitis.
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http://dx.doi.org/10.3109/01676830.2013.788665DOI Listing
August 2013

Chronic recalcitrant bacterial infection in steroid modified interstitial (stromal) keratitis: presentation and management.

J Med Assoc Thai 2012 Nov;95(11):1425-32

Department of Ophthalmology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.

Objective: To report histopathologically proven bacterial infection manifested multifocal interstitial (stromal) keratitis (IK) with definite previous history of prolong topical steroid use. Standard managements of bacterial keratitis did not provoke enough benefit.

Material And Method: A retrospective analysis of 19 eyes in 15 patients referred to Siriraj Hospital between 2004 and 2010.

Results: Multifocal intrastromal infiltration, with relatively quiet ocular reaction and mild inflammation were initially presented in all eyes. They all previously had been diagnosed of presumed viral keratitis, and had been given topical corticosteroid treatment for a prolonged period of time without healing. Autoimmune disease workups were all negative. Corneal scrapings showed negative culture results in all eyes. However, bacteria within stromal lamellae with absent or minimal inflammatory cells were demonstrated in all eyes by corneal biopsies. In addition, cytology results obtained from 16S rDNA sequencing revealed Stenotrophomonas maltophilia in one eye and coagulase-negative staphylococci in two eyes. No case responded well to intensive topical and systemic antibiotics. However they were successfully treated with penetrating keratoplasty (11 eyes, 57.9%) or intrastromal antibiotic injections (8 eyes, 42.1%).

Conclusion: Bacterial infection should be a concern in prolonged chronic IK. This was considered as primary bacterial IK or bacterial superinfection in immunocompromised cornea. Early recognition and appropriately aggressive managements contribute to successful outcome. Corneal biopsy is always essential and 16S rDNA sequencing is useful in this distinct clinical entity.
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November 2012

Efficacy of cultivated corneal epithelial stem cells for ocular surface reconstruction.

Clin Ophthalmol 2012 11;6:1483-92. Epub 2012 Sep 11.

Department of Ophthalmology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.

Purpose: To investigate the clinical outcomes of cultivated corneal limbal epithelial transplantation (CLET) using human amniotic membrane for corneal limbal stem-cell deficiency.

Methods: Prospective, noncomparative case series. Eighteen patients (19 eyes) with severe ocular surface diseases were chosen to undergo CLET using human amniotic membrane. Twelve eyes received auto-CLET, and seven eyes received allo-CLET. Clinical outcomes of corneal surface epithelialization, conjunctivalization, inflammation, visual acuity, graft status, and complications were observed.

Results: Corneal epithelium cultivated on amniotic membrane (two to four layers) was positive for molecular markers p63, ABCG2, CK3, and CK12. The mean patient age was 44.7 ± 15.2 years. A successful clinical outcome, defined as corneal epithelialization without central conjunctivalization or severe inflammation, was obtained in 14 (73.7%) of 19 eyes (mean follow-up 26.1 ± 13.5 months; range 6-47). A histopathologic success, defined as absence of goblet cells at the central cornea, was achieved in 12 (63.2%) eyes. Clinical failures occurred in five (26.3%) of 19 eyes, and histopathologic failures occurred in seven (36.8%) of 19 eyes. Survival analysis at 1 year showed that the clinical success rate was 77.9% and the pathological success rate was 72.3%. Fourteen of 19 (73.7%) eyes had visual acuity improvements after CLET. Six cases underwent penetrating keratoplasty; five of these grafts remained clear after 20.4 ± 6.9 months (range, 12-31) of follow-up. Complications included infectious keratitis (three cases) and recurrent symblepharon (one case). All complicated cases had lid abnormalities. Factors affecting the final clinical outcomes were lid abnormalities, abnormal corneal stromal beds, and complications.

Conclusion: CLET can successfully restore ocular surface damage in most cases with corneal limbal stem cell deficiency.
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http://dx.doi.org/10.2147/OPTH.S33951DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3460717PMC
October 2012

The effect of coenzyme Q10 and curcumin on chronic methanol intoxication induced retinopathy in rats.

J Med Assoc Thai 2012 Apr;95 Suppl 4:S76-81

Department of Ophthalmology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.

Background: The retinal pathophysiology of methanol intoxication is that formate inhibits retinal mitochondrial function and increases oxidative stress.

Objective: To investigate the effect of coenzyme Q10 and curcumin on chronic methanol intoxication causing retinopathy in rats.

Material And Method: The authors designed an experimental study of chronic methanol intoxication in rats depleted of folate with methotrexate. The studied group received methanol (2 mg/kg body weight in saline by intraperitoneal injection) and methotrexate (0.1 mg/kg body weight in saline by subcutaneous injection) every other day for ten weeks to induce chronic methanol intoxication, while another group received saline as vehicle and served as control group. The studied rats were confirmed to develop significant retinopathy after 10 weeks and then assigned to three treatment arms: either corn oil (as control) or coenzyme Q10 (20 mg/kg/day) or Curcuma longa extract (2.5 mg/kg/day) for four weeks. Eyes were enucleated and the retinal tissue was prepared for histological examination. The sections were evaluated by an experienced pathologist and blinded to the experimental conditions.

Results: Histological analysis revealed that animals treated with both methanol and methotrexate showed vacuolation of photoreceptor inner segment and disaggregation of cells in the inner and outer nuclear layers of the retina compared to a normal histological appearance in control animals. The retinal histology in the experimental animals with administration of Coenzyme Q10 or Curcuma longa extract appeared essentially normal and this was not found in the experimental animals which received corn oil.

Conclusion: Coenzyme Q10 and curcumin administration improves retinal histology by reversing the pathological changes due to chronic methanol and establish a morphologically normal retina.
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April 2012

Case report: Increased viral receptor expression associated with high viral load and severe pneumonia in a young patient infected with 2009 H1N1 influenza a with no pre-existing conditions.

J Med Virol 2012 Mar;84(3):380-5

Faculty of Medicine, Department of Microbiology, Siriraj Hospital, Mahidol University, Bangkok, Thailand.

A case of unusually high severity of influenza pneumonia leading to acute respiratory distress syndrome and death was investigated. This was a previously a healthy 28-year-old man with no underlying conditions, admitted to a hospital during the first wave of influenza pandemic in Thailand in July 2009. He had experienced high fever and influenza-like illness for 5 days before coming to the hospital. He developed acute respiratory distress syndrome and expired on day 7 after admission. In comparison to three other cases of influenza pneumonia in the same outbreak with known risk factors for severe influenza, such as pregnancy and diabetes mellitus, a much higher viral load was detected in the lungs of this patient despite antiviral treatment. In agreement with the high viral load, the lung specimens from this patient, but not the other three patients, showed a high expression of α-2,6-linked sialic acid by lectin staining. The gene responsible for the synthesis of this sialic acid was also found to be upregulated. The data indicated overexpression of the viral receptor as a potential mechanism for severe disease in some patients.
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http://dx.doi.org/10.1002/jmv.23201DOI Listing
March 2012

Viral load of the highly pathogenic avian influenza H5N1 virus in infected human tissues.

J Med Virol 2011 Aug;83(8):1418-23

Department of Microbiology, Mahidol University, Bangkok, Thailand.

The highly pathogenic avian influenza A (H5N1) virus is a virulent virus that causes an acute febrile respiratory disease with high mortality in humans. To gain a better insight of H5N1 viral distributions in infected human tissues, the levels of viral RNA were determined in the autopsy tissues from two patients who were infected with H5N1 virus by using real-time reverse transcription-polymerase chain reaction. In one patient who died on day 6 of the illness, the viral load in the lung was extremely high, whereas the levels of viral RNA in the other organs were more than 6 log lower. In the other patient who died on day 17 of the illness, the viral load was similar in the lung and other organs, and was comparable to the viral load in the extra-pulmonary tissues of the first patient. These results suggested that while the H5N1 virus can cause disseminated infection in humans, the lung is still the major site of viral replication, and viral replication in the lung in the later stages may decrease as a result of the depletion of the available target cells. In addition, the mRNA levels of the tumor necrosis factor-α (TNF-α) were found to be associated with the viral titers.
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http://dx.doi.org/10.1002/jmv.22146DOI Listing
August 2011

Decreased expression of surfactant protein D mRNA in human lungs in fatal cases of H5N1 avian influenza.

J Med Virol 2011 Aug;83(8):1410-7

Department of Microbiology, Mahidol University, Bangkok, Thailand.

Microarray analysis of gene expression profile of lungs from two fatal H5N1 influenza cases identified 3,435 genes with higher than twofold changes in mRNA levels as compared to those of normal lung. One thousand nineteen genes and 2,416 genes were up-regulated and down-regulated commonly, respectively. Gene ontology analysis identified several ontology terms with significant association with these genes, most of which are related to cellular metabolism and regulation of cellular process including apoptosis and chemotaxis. Pulmonary surfactant protein D (SP-D) was found to be down-regulated. Quantitative RT-PCR confirmed the levels of SP-D mRNA in the lungs infected with H5N1 to be lower than those of normal lungs and lungs from patients with acute respiratory distress syndrome. SP-D plays multiple roles in respiratory innate defense against various pathogens, regulation of inflammatory responses, and maintenance of alveolar integrity. Reduction of SP-D in H5N1 influenza may play important roles in the pathogenesis of the disease.
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http://dx.doi.org/10.1002/jmv.22105DOI Listing
August 2011

Entomophthoramycosis: a rare fungal orbital infection presenting with dacryocystitis.

Orbit 2011 Jan;30(1):21-3

Department of Ophthalmology, Siriraj Hospital, Mahidol University, Bangkok, Thailand.

We report a case of a rare fungal orbital infection in an infant presenting with dacryocystitis. The causative organism was Conidiobolus sp. of the order Entomophthorales. There is no standard treatment for entomophthoramycosis. Our patient responded well to combined antifungal therapy without aggressive surgical débridement.
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http://dx.doi.org/10.3109/01676830.2010.535640DOI Listing
January 2011

Mouse DNA contamination in human tissue tested for XMRV.

Retrovirology 2010 Dec 20;7:108. Epub 2010 Dec 20.

Section of Infectious Diseases, Jefferiss Research Trust Laboratories, Imperial College London, St Mary's Campus, London, W2 1PG, UK.

Background: We used a PCR-based approach to study the prevalence of genetic sequences related to a gammaretrovirus, xenotropic murine leukemia virus-related virus, XMRV, in human prostate cancer. This virus has been identified in the US in prostate cancer patients and in those with chronic fatigue syndrome. However, with the exception of two patients in Germany, XMRV has not been identified in prostate cancer tissue in Europe. Most putative associations of new or old human retroviruses with diseases have turned out to be due to contamination. We have looked for XMRV sequences in DNA extracted from formalin-fixed paraffin- embedded prostate tissues. To control for contamination, PCR assays to detect either mouse mitochondrial DNA (mtDNA) or intracisternal A particle (IAP) long terminal repeat DNA were run on all samples, owing to their very high copy number in mouse cells.

Results: In general agreement with the US prevalence, XMRV-like sequences were found in 4.8% of prostate cancers. However, these were also positive, as were 21.5% of XMRV-negative cases, for IAP sequences, and many, but not all were positive for mtDNA sequences.

Conclusions: These results show that contamination with mouse DNA is widespread and detectable by the highly sensitive IAP assay, but not always with less sensitive assays, such as murine mtDNA PCR. This study highlights the ubiquitous presence of mouse DNA in laboratory specimens and offers a means of rigorous validation for future studies of murine retroviruses in human disease.
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http://dx.doi.org/10.1186/1742-4690-7-108DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3019155PMC
December 2010

Endoscopic treatment of a large colonic polyp as a cause of colocolonic intussusception in a child.

World J Gastrointest Endosc 2010 Jul;2(7):268-70

Nutnicha Suksamanapun, Ravit Ruangtrakool, Division of Pediatric Surgery, Department of Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.

Colocolonic intussusception is an uncommon cause of intestinal obstruction in children. The most common type is idiopathic ileocolic intussusception. However, pathologic lead points occur approximately in 5% of cases. In pediatric patients, Meckel's diverticulum is the most common lead point, followed by polyps and duplication. We present a case of recurrent colocolonic intussusception which caused colonic obstruction in a 10-year-old boy. A barium enema revealed a large polypoid mass at the transverse colon. Colonoscopy showed a colonic polyp, 3.5 centimeters in diameter, which was successfully removed by endoscopic polypectomy.
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http://dx.doi.org/10.4253/wjge.v2.i7.268DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2999144PMC
July 2010

Villous adenoma of the urinary bladder: a case report.

J Med Assoc Thai 2010 Nov;93(11):1336-9

Department of Pathology, Faculty of Madicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.

Villous adenomas of the urinary tract are rare, in contrast to urothelial neoplasms. Most reports were scattered individual cases. Only two case series of this entity have been published. The histopathology is identical to that of the much more common villous adenoma of the gastrointestinal tract. The authors reported a case of urinary bladder villous adenoma in a 41-year-old Thai patient who complained of hematuria for one day without any other symptom. Cystoscopic examination revealed a papillary growth at the bladder neck associated with marked degree of bullous edema and bilateral mild hydroureters. The clinical diagnosis was urothelial carcinoma. Transurethral resection was performed Histologic examination revealed typical features of villous adenoma. The tumor showed identical immunohistochemical profile to colonic villous adenoma. The patient has been well for more than a year after tumor removal.
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November 2010

Enhanced susceptibility of nasal polyp tissues to avian and human influenza viruses.

PLoS One 2010 Sep 24;5(9):e12973. Epub 2010 Sep 24.

Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.

Background: Influenza viruses bind and infect respiratory epithelial cells through sialic acid on cell surface. Differential preference to sialic acid types contributes to host- and tissue-tropism of avian and seasonal influenza viruses. Although the highly pathogenic avian influenza virus H5N1 can infect and cause severe diseases in humans, it is not efficient in infecting human upper respiratory tract. This is because of the scarcity of its receptor, α2,3-linked sialic acid, in human upper airway. Expression of sialic acid can be influenced by various factors including inflammatory process. Allergic rhinitis and nasal polyp are common inflammatory conditions of nasal mucosa and may affect expression of the sialic acid and susceptibility to influenza infection.

Methodology/principal Finding: To test this hypothesis, we detected α2,3- and α2,6-linked sialic acid in human nasal polyp and normal nasal mucosal tissues by lectin staining and infected explants of those tissues with avian influenza viruses H5N1 and seasonal influenza viruses. We show here that mucosal surface of nasal polyp expressed higher level of α2,3- and α2,6-linked sialic acid than normal nasal mucosa. Accordingly, both H5N1 avian influenza viruses and seasonal influenza viruses replicated more efficiently in nasal polyp tissues explants.

Conclusions/significance: Our data suggest a role of nasal inflammatory conditions in susceptibility to influenza infection, especially by avian influenza viruses, which is generally inefficient in infecting human upper airway. The increased receptor expression may contribute to increased susceptibility in some individuals. This may contribute to the gradual adaptation of the virus to human population.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0012973PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2945772PMC
September 2010

Inhibition of H5N1 highly pathogenic influenza virus by suppressing a specific sialyltransferase.

Arch Virol 2010 Jun 10;155(6):889-93. Epub 2010 Apr 10.

Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.

Avian influenza viruses preferentially use alpha2,3-linked sialic acid as a receptor for binding and entry into target cells. The sialic acid is the terminal residue of various types of glycan. There are two major types of alpha2,3-linked sialic acid differing in the penultimate bond: Neu5Acalpha2-3Galbeta1-3GalNAc and Neu5Acalpha2-3Galbeta1-4GlcNAc. In the human airway, while Neu5Acalpha2-3Galbeta1-3GalNAc is present only in alveolar epithelial cells, the Neu5Acalpha2-3Galbeta1-4GlcNAc is expressed in both the upper and lower airway. Previous data showed preferential binding of hemagglutinin from H5N1 highly pathogenic influenza virus to Neu5Acalpha2-3Galbeta1-4GlcNAc. We further show here that suppression of this sialic acid by siRNA against a sialyltransferase, ST3GAL4, can inhibit H5N1 avian influenza virus infection and that this gene is abundantly expressed in human pharynx, trachea and bronchus. These data suggest that the ST3GAL4 gene is responsible for biosynthesis of the viral receptor and may play a crucial role in infection of H5N1 avian influenza virus in humans.
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http://dx.doi.org/10.1007/s00705-010-0658-4DOI Listing
June 2010

Cyclooxygenase-2 expression in retinoblastoma: an immunohistochemical analysis.

Curr Eye Res 2010 Mar;35(3):242-7

Department of Ophthalmology, Siriraj Hospital Mahidol University, Bangkok, Thailand.

Purpose: Increased level of cyclooxygenase-2 (COX-2) plays a significant role in the pathogenesis of cancers. High expression of COX-2 has been demonstrated in several cancer types including retinoblastoma. However, the in vivo study did not confirm the anti-proliferative effect of COX-2 inhibitor, celecoxib, on a murine transgenic retinoblastoma model. We, therefore, aim to investigate COX-2 expression in paraffin-embedded retinoblastoma specimens in a larger study group.

Methods: We reviewed 55 retinoblastoma specimens obtained during 1995 to 2005. Clinical and histopathological data were recorded. Immunohistochemical evaluation of COX-2 expression was performed using a rabbit monoclonal antibody to human cyclooxygenase-2.

Results: Forty-four of 55 specimens (80%) showed negative immunoreactivity for COX-2 expression. For the 11 specimens (20%, 95% CI = 11.6-32.4%) with positive COX-2, all immunostainings were less than 50% of tumor area. Demographic data and treatment details were available in 53 specimens. Enucleation was performed as a primary treatment in 43 specimens (81%). Other treatments, mainly systemic chemotherapy, were given prior to enucleation in 10 specimens (19%). There was no statistical difference in COX-2 expression between the specimens identified as primary and secondary enucleation (p = 0.66). Regarding the histopathological findings, there were no significant differences between COX-2 negative and COX-2 positive groups.

Conclusions: It appears that COX-2 is not overexpressed in our retinoblastoma specimens, which is different from previous studies. This conflicting data reduces the possibility of introducing Cox-2 inhibitors in the treatment of retinoblastoma.
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http://dx.doi.org/10.3109/02713680903477832DOI Listing
March 2010