Graft Versus Host Disease-Associated Dry Eye: Role of Ocular Surface Mucins and the Effect of Rebamipide, a Mucin Secretagogue

PURPOSE. The present study was designed to investigate the role of ocular surface glycocalyx and mucins in graft versus host disease (GVHD)-associated dry eye. The ameliorative effect of topical rebamipide, a mucin secretagogue, on GVHD-associated dry eye was also tested. METHODS. A mouse model of allogeneic transplantation was used to induce ocular GVHD with C57BL/6 as donors and B6D2F1 as recipient mice. Phenol red thread method and fluorescein staining was used to quantify tear secretion and corneal keratopathy. At 8 weeks after the allogeneic transplantation, corneas were harvested to perform glycocalyx staining and confocal microscopy. Goblet cell staining was performed using periodic acid Schiff’s staining. Corneal and tear film levels of Mucin 1, 4, 16, 19, and 5AC were quantified using ELISA and real-time PCR. Rebamipide was applied topically twice daily to mice eyes. RESULTS. Allogeneic transplantation resulted in ocular GVHD-associated dry eye characterized by a significant decrease in tear film volume and the onset of corneal keratopathy. Ocular GVHD caused a significant decrease in the area and thickness of corneal glycocalyx. A significant decrease in the goblet cells was also noted. A significant decrease in mucin 4 and 5AC levels was also observed. Topical treatment with rebamipide partially attenuated ocular GVHD-mediated decrease in tear film volume and significantly reduced the severity of corneal keratopathy. CONCLUSIONS. Ocular GVHD has detrimental impact on ocular surface glycocalyx and mucins. Rebamipide, a mucin secretagogue, partially prevents ocular GVHD-associated decrease in tear film and reduces the severity of corneal keratopathy

E-commerce protocol supporting automated online dispute resolution

E-commerce now constitutes a significant part of all commercial activity; however the increase in transactions is also leading to more disputes. These disputes are becoming more frequent, more technologically complicated and more difficult in terms of traceability . This thesis focuses specifically on dispute problems related to soft products, i.e. those that are intangible and therefore requiring no physical delivery. With the growing demand for these types of products, e.g. downloadable films, music, software, and prepaid calling time, the prevention of fraudulent transactions is becoming increasingly important. Reasons for the rise in the number of fraudulent transactions include merchants being unable to see the customer to verify an ID or signature and E-commerce enabling soft-products and services to be acquired via soft delivery methods: email, download or logging in. The introductory section provides a critique of current e-commerce fraud detection and prevention techniques and shows that not all are suitable for e-commerce, especially soft-products, and therefore unable to provide complete protection against fraud. The future relating to the detection and prevention of e-commerce fraud is then discussed, leading to suggestions regarding the improvement of the current state-of-the-art technique, the Address Verification Service (AVS), which is used to accommodate the introduction of soft-products. Apart from the exchange process problems, i.e. those involving money and goods, attention is also paid to other important factors such as timing and quality that are usually neglected in these detection and prevention techniques. Dispute scenarios from many different perspectives have been analysed, viz. computer science, business, legal and that of the participants themselves. From the analyses, all possible dispute cases have been formally listed using the 'Truth Table' approach. This analysis has then led to the design of a comprehensive taxonomy framework for dispute in e-commerce. The term Online Dispute Resolution (ODR), is the online technology applied to Alternative Dispute Resolution (ADR) which is resolving disputes other than via litigation in the courts. Current ODR systems and their suitability for the e-commercial world have been examined, concluding that not all are appropriate for e-commerce situations (since most still involve a human element and often make the resolution process more costly than the actual item under dispute). The proposed solution to the problem is by automating the online dispute resolution process. The total solution is described in two parts (i) an E-commerce Transaction Protocol (ETP) forming the infrastructure where the transaction will take place and be able to accommodate any new improvements in the future, and (ii) an Automated Online Dispute Resolution (AODR) system which should automatically resolve any dispute occurring within the proposed e-commerce model. In order for the AODR to resolve any dispute, a product/payment specific plug-in (add-on) has been incorporated into the system. For illustration purposes, credit cards as a payment method has been selected and the appropriate plug-in specification for soft products and credit cards created. The concept of providing every soft product with a quality certificate has also been discussed. A concluding case study of e-commerce in Saudi Arabia has been used to test the viability of both the e-commerce dispute taxonomy and the proposed model. The case study shows the suitability of using ETP with AODR in order to resolve soft-product disputes automatically. Limitations of the work and further research possibilities have then been identified.EThOS - Electronic Theses Online ServiceDepartment of Computing Science, Newcastle UniversityGBUnited Kingdo

Diabetes-Associated Hyperglycemia Causes Rapid-Onset Ocular Surface Damage

Purpose: The metabolic alterations due to chronic hyperglycemia are well-known to cause diabetes-associated complications. Short-term hyperglycemia has also been shown to cause many acute changes, including hemodynamic alterations and osmotic, oxidative, and inflammatory stress. The present study was designed to investigate whether diabetes-associated hyperglycemia can cause rapid-onset detrimental effects on the tear film, goblet cells, and glycocalyx and can lead to activation of an inflammatory cascade or cellular stress response in the cornea. Methods: Mouse models of type 1 and type 2 diabetes were used. Tear film volume, goblet cell number, and corneal glycocalyx area were measured on days 7, 14, and 28 after the onset of hyperglycemia. Transcriptome analysis was performed to quantify changes in 248 transcripts of genes involved in inflammatory, apoptotic, and stress response pathways. Results: Our data demonstrate that type 1 and type 2 diabetes-associated hyperglycemia caused a significant decrease in the tear film volume, goblet cell number, and corneal glycocalyx area. The decrease in tear film and goblet cell number was noted as early as 7 days after onset of hyperglycemia. The severity of ocular surface injury was significantly more in type 1 compared to type 2 diabetes. Diabetes mellitus also caused an increase in transcripts of genes involved in the inflammatory, apoptotic, and cellular stress response pathways. Conclusions: The results of the present study demonstrate that diabetes-associated hyperglycemia causes rapid-onset damage to the ocular surface. Thus, short-term hyperglycemia in patients with diabetes mellitus may also play an important role in causing ocular surface injury and dry eye

Reduction in stiffness of proximal leg muscles during the first 6 months of glucocorticoid therapy for giant cell arteritis: A pilot study using shear wave elastography.

Aim: To investigate muscle stiffness changes in patients treated for giant cell arteritis (GCA) with high‐dose oral glucocorticoids. Methods: Using ultrasound elastography, shear wave velocity (SWV) was measured in the quadriceps, hamstrings and biceps brachii muscles of 14 patients with GCA (4 male, mean age ± SD, 68.2 ± 4.3 years) within the first 2 weeks of initiating glucocorticoid treatment (baseline) and repeated after 3 and 6 months treatment. Muscle strength and performance tests were performed at each visit. Baseline measures were compared with those from 14 healthy controls. Linear mixed models were used to test for change in patient measures over time. Results: At baseline, muscle SWV in patients was not significantly different from controls. With glucocorticoid treatment, there was a reduction in SWV in the leg but not the arm muscles. SWV decreased by a mean of 14% (range 8.3%‐17.3%; P = .001) after 3 months and 18% (range 10.2%‐25.3%; P < .001) after 6‐months in the quadriceps and hamstrings during the resting position. The baseline, 3 and 6 months mean SWV (±SD) for the vastus lateralis were 1.62 ± 0.16 m/s, 1.40 ± 0.10 m/s and 1.31 ± 0.06 m/s, respectively (P < .001). In the patient group as a whole, there was no significant change in muscle strength. However, there were moderate correlations (r = .54‐.69) between exhibiting weaker muscle strength at follow‐up visits and a greater reduction in SWV. Conclusion: Glucocorticoid therapy in patients with GCA was associated with a significant reduction in proximal leg muscle stiffness during the first 6 months. Future research should study a larger sample of patients for a longer duration to investigate if diminished muscle stiffness precedes signs of glucocorticoid‐induced myopathy

Local Renin-Angiotensin System Activation and Myofibroblast Formation in Graft Versus Host Disease–Associated Conjunctival Fibrosis

Purpose: The present study was designed to investigate the role of myofibroblast transdifferentiation and the conjunctival renin–angiotensin system (RAS) in the pathogenesis of graft-versus-host disease (GVHD)–associated conjunctival fibrosis. Methods: A mouse model of major histocompatibility-matched allogeneic transplantation was used to induce GVHD, with male B10.D2 mice as donors and female BALB/c mice as recipients. Male BALB/c to female BALB/c syngeneic transplantation was used as control. Y chromosome staining in the spleen cells obtained from female recipient mice was used to confirm engraftment. The phenol red thread test and fluorescein staining were used to quantify tears and corneal keratopathy. Eyes were harvested at 4 and 8 weeks after the transplant for alpha-smooth muscle actin (α-SMA), angiotensinogen, and angiotensin-converting enzyme (ACE) immunostaining. Conjunctiva was harvested for gene expression quantification of α-SMA, angiotensinogen, and ACE. Results: More than 80% of the spleen cells in the recipient mice were chromosome Y positive, thus conforming successful engraftment. A significant decrease in tear secretion and a marked increase in corneal keratopathy score after allogeneic transplantation indicated the onset of ocular GVHD in these mice. A significant increase in α-SMA gene expression and the presence of a large number of α-SMA–positive cells was noted in the bulbar orbital conjunctiva of mice after allogeneic transplantation. Allogenic transplantation also caused a significant increase in the gene expression and protein expression of angiotensinogen and ACE in the subconjunctival eyelid area. Conclusions: Results of the present study demonstrate that GVHD-associated conjunctival fibrosis is accompanied by myofibroblast formation and activation of the local conjunctival RAS

Effect of High Glucose on Ocular Surface Epithelial Cell Barrier and Tight Junction Proteins

PURPOSE. Patients with diabetes mellitus are reported to have ocular surface defects, impaired ocular surface barrier function, and a higher incidence of corneal and conjunctival infections. Tight junctions are critical for ocular surface barrier function. The present study was designed to investigate the effect of high glucose exposure on human corneal and conjunctival epithelial cell barrier function and tight junction proteins. METHODS. Human corneal and conjunctival epithelial cells were exposed to 15 mM and 30 mM glucose for 24 and 72 hours. The barrier function was measured using transepithelial electrical resistance (TEER). The cell migration was quantified using scratch assay. The cells were harvested for protein extraction and mRNA isolation. Gene and protein expression of claudins, zonula occludens (ZOs), and occludin was quantified using realtime PCR and Western blot. RESULTS. Glucose caused a significant decrease in TEER after 72 hours of exposure in both corneal and conjunctival epithelial cells. Glucose did not cause any notable change in migration of either corneal or conjunctival epithelial cells. Glucose exposure did not cause any notable change in protein expression of claudin-1, ZO-1, ZO-2, ZO-3, or occludin. On the other hand, 15 mM glucose caused an increase in gene expression of claudin-1, claudin-3, ZO-2, ZO-3, and occludin, a likely response to osmotic stress since 15 mM mannitol also caused consistently similar increase in gene expression of these proteins. CONCLUSIONS. High glucose exposure causes impairment of corneal and conjunctival epithelial cell barrier function, but this detrimental effect is not caused by a decrease in expression of tight junction proteins: claudin-1, ZO-1, ZO-2, ZO-3, and occludin

The effect of ageing on shear wave elastography muscle stiffness in adults

Background: Skeletal muscle undergoes structural changes with ageing which may alter its biomechanical properties. Shear wave elastography (SWE) may detect these changes by measuring muscle stiffness. Aims: To investigate muscle stiffness in healthy young, middle-aged and elderly cohorts using SWE and correlate it with muscle strength and mass. Methods: Shear wave velocity (SWV) was measured in the quadriceps, hamstrings and biceps brachii of 26 young (range 20–35 years), 21 middle-aged (40–55) and 30 elderly (77–94) volunteers. The participants performed several muscle tests to evaluate their strength. The One-way ANOVA was used to test the muscle stiffness differences between the groups and the Pearson’s correlation coefficient to evaluate the relationship between SWV and muscle strength. Results: The overall resting muscle SWV gradually decreased with age but was only significantly reduced in the elderly group (p < 0.001); with the exception of the vastus lateralis SWV where a significant difference was noted (p < 0.05) between young (1.77 m/s), middle-aged (1.64 m/s) and elderly (1.48 m/s). The elderly group had on average 16.5% lower muscle stiffness compared to the young. SWV significantly correlated with muscle mass (r = 0.316), walking time (r = − 0.560), number of chair stands (r = 0.522), handgrip strength (r = 0.436) and isokinetic knee strength (r = 0.640). Sex and BMI did not explain any significant variation in SWV. Conclusions: Ageing was associated with a decline in skeletal muscle stiffness which positively correlates with muscle weakness. Further research is needed to evaluate the promising role of SWE as a biomarker for sarcopenia assessment and potential falls risk prediction in elderly individuals

Muscle stiffness in rheumatoid arthritis is not altered or associated with muscle weakness: a shear wave elastography study

Objective: To investigate muscle stiffness and strength in rheumatoid arthritis patients compared to healthy controls. Methods: A sample of 80 RA patients from three discrete groups: 1-newly diagnosed treatment-naïve RA (n = 29), 2-active RA for at least 1 year (n = 18) and 3-in remission RA for at least 1 year (n = 33), was compared to 40 healthy controls. Shear wave velocity (SWV) was measured using shear wave elastography as a surrogate for tissue stiffness in multiple muscles. All participants performed isometric grip strength, timed get-up-and-go test, 30-sec chair stand test and isokinetic knee extension/flexion(60°/sec). The difference in SWV amongst the groups was tested using one-way ANOVA, and the correlation between SWV and muscle strength results were calculated using Pearson's coefficients. Results: The mean age ± SD was 61.2 ± 12.8 for RA patients and 61.5 ± 10.5 years for controls. SWV was not significantly different amongst the groups on all muscles (p > 0.05). In comparison to controls, the new and active RA groups showed a significantly lower isokinetic strength by -29%(p = 0.013) and -28%(p = 0.040), fewer chair stands by -28%(p = 0.001) and -44%(p  0.05). Conclusions: Significant muscle weakness was demonstrated in patients with RA disease. However, muscle stiffness was normal and not associated with muscle strength