Efficacy and Safety Outcomes of XEN Implantation and Gonioscopy-assisted Transluminal Trabeculotomy for the Management of Advanced Open-angle Glaucoma.

Aim: Minimally invasive glaucoma surgery (MIGS) is not typically used in patients with advanced-stage glaucoma. This study describes the outcomes and complications of patients with advanced open-angle glaucoma (OAG) who underwent XEN implantation with cataract surgery or gonioscopy-assisted transluminal trabeculotomy (GATT) with cataract surgery. Methods: This retrospective study identified patients who had undergone XEN implantation or GATT for the management of advanced OAG. Outcomes included surgical success, intraocular pressure (IOP) reduction, number of topical IOP-lowering drops, visual field mean deviation (MD), best-corrected visual acuity (BCVA), and complications. Surgical success was defined as an IOP of <14 mm Hg and a 20% reduction at 12 months without topical IOP-lowering drops (complete success) or with topical IOP-lowering drops (qualified success). Results: Exactly 70 eyes were enrolled in this study, including 35 who had undergone XEN implantation and 35 who had undergone GATT. The overall surgical success rate was 74.3% (26 of 35) for eyes that underwent XEN implantation and 71.4% (25 of 35) for eyes that underwent GATT. Percent IOP reduction from baseline to 12 months postoperatively was 48% in the XEN cohort and 32% in the GATT cohort. Significant reduction in the use of topical IOP-lowering drops was demonstrated for both XEN (3.26 ± 1.15-1.23 ± 1.28) (p < 0.001) and GATT (2.46 ± 1.12-0.43 ± 0.78) (p < 0.001) cohorts at 12 months postoperatively. The only complication reported was transient hyphema, which occurred in three patients from the XEN group and four from the GATT group, and resolved spontaneously. Conclusions: Both XEN implantation and GATT may be safe and effective management options when treating patients with advanced OAG. However, larger sample sizes are required to make direct statistical comparisons between these techniques. Clinical significance: In this study, XEN implantation and GATT combined with cataract surgery were each associated with favorable outcomes in patients with advanced OAG. How to cite this article: Ruparelia S, Sharif M, Shoham-Hazon N. Efficacy and Safety Outcomes of XEN Implantation and Gonioscopy-assisted Transluminal Trabeculotomy for the Management of Advanced Open-angle Glaucoma. J Curr Glaucoma Pract 2023;17(2):63-67.

Emergence of Small Interfering RNA-Based Gene Drugs for Various Diseases.

Small molecule, peptide, and protein-based drugs have been developed over decades to treat various diseases. The importance of gene therapy as an alternative to traditional drugs has increased after the discovery of gene-based drugs such as Gendicine for cancer and Neovasculgen for peripheral artery disease. Since then, the pharma sector is focusing on developing gene-based drugs for various diseases. After the discovery of the RNA interference (RNAi) mechanism, the development of siRNA-based gene therapy has been accelerated immensely. siRNA-based treatment for hereditary transthyretin-mediated amyloidosis (hATTR) using Onpattro and acute hepatic porphyria (AHP) by Givlaari and three more FDA-approved siRNA drugs has set up a milestone and further improved the confidence for the development of gene therapeutics for a spectrum of diseases. siRNA-based gene drugs have more advantages over other gene therapies and are under study to treat different types of diseases such as viral infections, cardiovascular diseases, cancer, and many more. However, there are a few bottlenecks to realizing the full potential of siRNA-based gene therapy. They include chemical instability, nontargeted biodistribution, undesirable innate immune responses, and off-target effects. This review provides a comprehensive view of siRNA-based gene drugs: challenges associated with siRNA delivery, their potential, and future prospects.

Biochemical, clinical, demographic and imaging biomarkers for disease progression in knee osteoarthritis.

Aim: To identify prognostic biomarker(s) for knee osteoarthritis (OA) in the Osteoarthritis Initiative (OAI) cohort. Methods: Multilevel regression was used to determine the association between baseline biomarkers and change in biomarkers from baseline to 24 months with clinical and radiographic OA progression over 48 months of follow-up. Results: Higher values of baseline urinary CTXII were consistently associated with an increased risk of OA disease progression outcomes: Kellgren & Lawrence grade (odds ratio [OR]: 1.15, 95% CI: 1.03-1.28); medial joint space narrowing (OR: 1.06, 95% CI: 1.02-1.10); lateral osteophytes (OR: 1.05, 95% CI: 1.01-1.10); joint space width (regression coefficient: -0.005, 95% CI: -0.008-0.001); and Western Ontario and McMaster Universities Arthritis Index pain scores (OR: 1.02, 95% CI: 1.01-1.04). Changes in serum PIIANP and serum COMP over 24 months were associated with clinical disease progression. Conclusion: Urinary CTXII showed stronger associations with radiographic OA and appears to be a reliable prognostic marker, while changes in other biomarkers were found in early symptomatic OA, supporting the phasic nature of OA.

Subchondral Bone Microarchitectural and Mineral Properties and Expression of Key Degradative Proteinases by Chondrocytes in Human Hip Osteoarthritis.

BACKGROUND: The purpose of this study was to investigate the relationship between the expression of key degradative enzymes by chondrocytes and the microarchitectural and mineral properties of subchondral bone across different stages of cartilage degradation in human hip osteoarthritis (OA). METHODS: Osteochondral samples at different stages of cartilage degradation were collected from 16 femoral heads with OA. Osteochondral samples with normal cartilage were collected from seven femoral heads with osteoporosis. Microcomputed tomography was used for the investigation of subchondral bone microarchitecture and mineral densities. Immunohistochemistry was used to study the expression and distribution of MMP13 and ADAMTS4 in cartilage. RESULTS: The microarchitecture and mineral properties of the subchondral plate and trabecular bone in OA varied with the severity of the degradation of the overlying cartilage. Chondrocytes expressing MMP13 and ADAMTS4 are mainly located in the upper zone(s) of cartilage regardless of the histopathological grades. The zonal expression of these enzymes in OA (i.e., the percentage of positive cells in the superficial, middle, and deep zones), rather than their overall expression (the percentage of positive cells in the full thickness of the cartilage), exhibited significant variation in relation to the severity of cartilage degradation. The associations between the subchondral bone properties and zonal and overall expression of these enzymes in the cartilage were generally weak or nonsignificant. CONCLUSIONS: Phenotypic changes in chondrocytes and remodelling of subchondral bone proceed at different rates throughout the process of cartilage degradation. Biological influences are more important for cartilage degradation at early stages, while biomechanical damage to the compromised tissue may outrun the phenotypic change of chondrocytes and is critical in the advanced stages.

Subchondral bone microarchitecture and mineral density in human osteoarthritis and osteoporosis: A regional and compartmental analysis.

Osteoarthritis (OA) and osteoporosis (OP) are historically considered to be inversely correlated but there may be an overlap between the pathophysiology of the two diseases. This study aimed to investigate the subchondral bone microarchitecture and matrix mineralization, and the association between them in OA and OP in relation to the degree of cartilage degeneration. Fifty-six osteochondral plugs were collected from 16 OA femoral heads. They were graded on a regional basis according to the stages of cartilage degeneration, as evaluated by a new macroscopic and a modified microscopic grading system. Twenty-one plugs were collected from seven femoral heads with OP. Plugs were scanned by microcomputed tomography and the microarchitectural and mineral properties were obtained for both subchondral plate and trabecular bone. Microarchitecture and material and apparent densities of subchondral bone in OP were similar to regions with early cartilage degeneration but different from regions with advanced cartilage degradation in OA femoral heads. Subchondral trabecular bone was more mineralized than subchondral plate in both OP and OA, and this compartmental difference varied by severity of cartilage degradation. Furthermore, the relationship among trabecular bone volume fraction, tissue mineral density, and apparent bone density was similar in OP and different stages of OA. Subchondral bone microarchitecture and mineral properties in OP are different from OA in a regionalized manner in relation to stages of cartilage degeneration. Both regional and compartmental differences at structural, material, and cellular levels need to be studied to understand the transition of OA subchondral bone from being osteoporotic to sclerotic.

Pathways to service access for pre-eclampsia and eclampsia in rural Bangladesh: Exploring women's care-seeking.

BACKGROUND: While women in low- and middle-income countries face a range of barriers to accessing care for hypertensive disorders of pregnancy, there is little understanding of the pathways taken to overcome these constraints and reach the services they need. This study explores the perspectives of women and communities on the influences that impact care-seeking decisions and pathways to health services. METHODS: To understand individual perspectives, we conducted 22 in-depth interviews (IDIs) with pre-eclampsia and eclampsia survivors (PE/E) in a tertiary hospital, where they received care after initiating PE/E services in different parts of the country. In four districts, we conducted one male and one female focus group discussion (FGD) to unearth care-seeking pathways and explore normative perspectives and the range of internal and external influences. Careful thematic analysis using Atlas-ti was applied. RESULTS: Prevailing views of women and communities across settings in Bangladesh indicate varied pathways to care throughout their pregnancy, during childbirth, and in the postnatal period influenced by internal and external factors at the individual, familial, social, and health systems levels. Internal influences draw on women's own awareness of hypertension complications and options, and their ability to decide to seek care. External factors include social influences like family and community norms, culturally-accepted alternatives, and community perceptions of the health system's capacity to provide quality care. The interaction of these factors often delay care seeking and can lead to complex pathways to care. CONCLUSION: Women's individual pathways to care were diverse, despite the homogenous community perceptions of the influences on women's care-seeking behaviors. This finding supports the need for improving quality of care in primary healthcare facilities and strengthening gender equity and community-based promotion activities through targeted policy and programming.

Knowledge of pharmacy workers on antihypertensive and anticonvulsant drugs for managing pre-eclampsia and eclampsia in Bangladesh.

BACKGROUND: Pharmacy workers in Bangladesh play an important role in managing pregnancy complications by dispensing, counselling and selling drugs to pregnant women and their families. This study examined pharmacy workers' drug knowledge and practice for pre-eclampsia and eclampsia (PE/E) management, including antihypertensives and anticonvulsants, and determine factors associated with their knowledge. METHODS: A cross-sectional survey with 382 pharmacy workers in public facilities (government) and private pharmacies and drug stores assessed their knowledge of antihypertensive and anticonvulsant drugs. 'Pharmacy workers' include personnel who work at pharmacies, pharmacists, family welfare visitors (FWVs), sub-assistant community medical officers (SACMOs), drug storekeepers. Exploratory and multivariate logistic models were used to describe association between knowledge of medicines used in pregnancy and demographic characteristics of pharmacy workers. RESULTS: Overall, 53% pharmacy workers interviewed were drug store owners in private pharmacies while 27% FWVs/SACMOs, who are government service providers also work as drug prescribers and/or dispensers in public facility pharmacies. Majority of pharmacy workers had poor knowledge compared to correct knowledge on both antihypertensive (77.8% vs 22.3%; p < 0.001) and anticonvulsant drugs (MgSO4) (82.2% vs 17.8%; p < 0.001). Multivariate analysis showed SACMOs and FWVs were greater than 4 times more likely to have correct knowledge on anti-hypertensives (AOR = 4.2, 95% CI:1.3-12.3, P < 0.01) and anticonvulsant drugs (AOR = 4.9, 95% CI:1.3-18.1, P < 0.01) compared to pharmacists. Pharmacy workers who had received training were more likely to have correct knowledge on antihypertensive and anticonvulsant drugs than those who had no training. CONCLUSIONS: Pharmacy workers' knowledge and understanding of antihypertensive and anticonvulsant drugs, particularly for prevention and management of PE/E is limited in Bangladesh. Most pharmacies surveyed are private and staffed with unskilled workers with no formal training on drugs. Expansion of maternal and newborn health programs should consider providing additional skills training to pharmacy workers, as well as regulating these medicines at informal pharmacies to mitigate any harmful practices or adverse outcomes of unauthorized and incorrectly prescribed and used drugs. It is important that correct messaging and medicines are available as drug stores are often the first point of contact for most of the women and their families.

Multivariable logistic and linear regression models for identification of clinically useful biomarkers for osteoarthritis.

Osteoarthritis (OA) is the most common chronic degenerative joint disease which causes substantial joint pain, deformity and loss of activities of daily living. Currently, there are over 500 million OA cases worldwide, and there is an urgent need to identify biomarkers for early detection, and monitoring disease progression in patients without obvious radiographic damage to the joint. We have used regression modelling to describe the association of 19 of the currently available biomarkers (predictors) with key radiographic and clinical features of OA (outcomes) in one of the largest and best characterised OA cohort (NIH Osteoarthritis Initiative). We demonstrate that of the 19 currently available biomarkers only 4 (serum Coll2-1 NO2, CS846, COMP and urinary CTXII) were consistently associated with established radiographic and/or clinical features of OA. These biomarkers are independent of one another and provide additional predictive power over, and above established predictors of OA such as age, gender, BMI and race. We also show that that urinary CTXII had the strongest and consistent associations with clinical symptoms of OA as well as radiographic evidence of joint damage. Accordingly, urinary CTXII may aid in early diagnosis of OA in symptomatic patients without radiographic evidence of OA.

Use of rheumatoid arthritis impact of disease (RAID) in routine care; identification of DAS28 remission and unmet patient-reported outcomes.

OBJECTIVE: The aim was to assess how the patient-reported outcome RA impact of disease (RAID) relates to DAS28 categories in routine care, its utility in identifying patients in DAS28 remission (RDAS) or low disease activity (LDAS) and the burden of unmet patient-reported needs in those achieving RDAS/LDAS. METHODS: DAS28 and RAID scores were collected from patients with established RA attending for routine review. The relationship between RAID and DAS28 was assessed with univariate pairwise correlation and mixed-effects linear regression analyses. RAID <2 was defined as a patient-acceptable state. RESULTS: One hundred and ninety-eight patients were assessed, with 220 observations, using DAS28-CRP categories: 47.5% RDAS, 14.1% LDAS, 31.8% moderate DAS (MDAS) and 6.6% high DAS (HDAS). Both patient visual analog scale score and tender joint count exhibited a high statistical association with RAID using linear regression (P < 0.0001). The mean RAID score per DAS28-CRP category was RDAS 1.84, LDAS 4.78, MDAS 5.60 and HDAS 7.68, with a statistically significant increase in RAID per unit increase in DAS-CRP or DAS28-ESR on linear regression (P < 0.001). Of 66 patients with RAID <2, 64 (97%) were in RDAS and 65 (98.5%) in RDAS/LDAS. Of 134 patients in RDAS/LDAS, RAID was ≥2 in 69 (51.5%), with fatigue and sleep being the worst-scoring domains. CONCLUSION: RAID functions well as a patient-reported outcome in routine care. Patients with RAID <2 have a high likelihood of being in RDAS/LDAS and, if pre-screened, could avoid a clinic visit. Analysis of RAID domains provides individualized targets for holistic care in RA management, with fatigue and sleep problems dominating unmet needs in those in RDAS/LDAS.

A primary health care model for managing pre-eclampsia and eclampsia in low- and middle- income countries.

BACKGROUND: Hypertensive disorders in pregnancy, specifically pre-eclampsia and eclampsia (PE/E), are the second biggest killer of pregnant women globally and remains the least understood and most challenging maternal morbidity to manage. Although great strides were made in reducing maternal and newborn mortality between 1990 and 2015, this was clearly not enough to achieve the global health goals. To reduce maternal deaths: 1) early detection of PE needs to be improved; 2) effective management of PE/E needs to occur at lower health system levels and should encourage timely care-seeking; and 3) prioritizing the scale up of a comprehensive package of services near to where women live. FINDINGS: This commentary describes a pragmatic approach to test scalable and sustainable strategies for expanding access to quality under-utilized maternal health commodities, interventions and services. We present a primary health care (PHC) PE/E Model based on implementation research on identified gaps in care in several countries, accepted global best practice and built on the basic premise that PHC providers can take on additional skills with adequate capacity building, coaching and supervision, and community members desire control over their own health. The PHC PE/E model displays the linkages and opportunities to prevent and treat PE/E in a simplified way; however, there are numerous interlinking factors, angles, and critical points to consider including leadership, policies and protocols; relevant medicines and commodities, ongoing capacity building strategies at lower levels and understanding what women and their communities want for safe pregnancies. CONCLUSION: The PHC model described here uses PE/E as an entry to improve the quality of ANC and by extension the pregnancy continuum. Bringing preventive and treatment services nearer to where pregnant women live makes sense.

Opportunities and challenges for the discovery and validation of proteomic biomarkers for common arthritic diseases.

Osteoarthritis (OA) and rheumatoid arthritis (RA) are most prevalent among all the rheumatic diseases, and currently, there are no reliable biochemical measures for early diagnosis or for predicting who is likely to progress. Early diagnosis is important for making decisions on treatment options and for better management of patients. This narrative review highlights the first-generation biomarkers identified over the last two decades and focuses on the discovery and validation of candidate OA biomarkers from recent mass-spectrometry-based proteomic studies for diagnosis and monitoring disease outcomes in human. It discusses the challenges and opportunities for discovery of novel biomarkers and progress in the development of techniques for measuring biomarkers, and provides directions for future discovery and validation of biomarkers for OA and rheumatoid arthritis.

Fertility differential of women in Bangladesh demographic and health survey 2014.

BACKGROUND: The aim of this study was to examine the fertility differential of women age 15 to 49 using data from Bangladesh Demographic and Health Survey 2014- a survey of women who were born from 1963 to 1999. METHODS: The secondary data analysis was carried out using the BDHS 2014 in order to discuss differences in childbearing practices in Bangladesh. Descriptive statistics were used to analyze the data including education level, geographic location, and religion. A trend test used to assess the inferences. RESULTS: On average, women had 2.3 children in the BDHS 2014; more than 90% of them gave birth to at least one child by age 49 and the average age of first birth was 18 years. Fertility of women strongly differed by education (p < 0.001). The percentage of women with secondary education who had no child was 50.3% and never attended school 8.4%;those with secondary education were six times as likely as those who never attended school to have no child and this pattern was stronger among urban compared with rural women. CONCLUSIONS: Fertility differential becomes robust as education increases. Women's fertility is also related to religion and residence, but these factors were not strongly related as those educational attainments.

Development and validation of novel biomarker assays for osteoarthritis.

BACKGROUND: Osteoarthritis (OA) is the most common chronic joint disease usually diagnosed at relatively advanced stages when there is irreparable damage to the joint(s). Recently, we have identified two novel biomarkers C3f and V65 which appear to be OA-specific and therefore potential markers of early disease. We report the development of immunoassays for quantitative measure of these two novel biomarkers. METHOD: Monoclonal and polyclonal antibodies were generated by immunising mouse and rabbits respectively with peptide-carrier conjugates of C3f and V65. Affinity purified antibodies were used for immunoassays development and assays validated using serum from OA patients and controls. RESULTS: The ELISAs developed showed spiked recovery of up to 96% for C3f and V65 peptides depending on serum dilutions with a coefficient of variation (CV) <10%. The intra- and inter-assay CVs for C3f and V65 were 1.3-10.8% and 4.2-10.3% respectively. Both assays were insensitive for measurements of the peptides in patients and the use of different signal amplification systems did not increase assay sensitivity. CONCLUSION: We have developed two immunoassays for measurements of C3f and V65 peptides biomarkers discovered by our earlier proteomic study. These assays could detect the endogenous peptides in serum samples from patients and controls but lacked sensitivity for accurate measurements of the peptides in patients. Our study highlights the difficulties and challenges of validating biomarker from proteomic studies and demonstrates how to overcome some of the technical challenges associated with developing immunoassays for small peptides.

Validation of a new method by nano-liquid chromatography on chip tandem mass spectrometry for combined quantitation of C3f and the V65 vitronectin fragment as biomarkers of diagnosis and severity of osteoarthritis.

Microfluidic liquid chromatography coupled to a nanoelectrospray source ion trap mass spectrometry was used for the absolute and simultaneous quantitation of C3f and the V65 vitronectin fragment in serum. The method was first carefully optimized and then validated in serum biological matrix. Stable isotopes for the two biomarkers of interest were used as stable isotope labeled peptide standards. A weighted 1/x2 quadratic regression for C3f and a weighted 1/x quadratic regression for the V65 vitronectin peptide were selected for calibration curves. Trueness (with a relative bias <10%), precision (repeatability and intermediate precision <15%) and accuracy (risk <15%) of the method were successfully demonstrated. The linearity of results was validated in the concentration range of 2.5-200ng/mL for C3f and 2.5-100ng/mL for the V65 vitronectin fragment. Serum samples (n=147) classified in 7 groups [(healthy volunteers, OA with 5 grades of severity and rheumatoid arthritis (RA) patients] were analyzed with our new quantitative method. Our data confirm that C3f and the V65 vitronectin fragment are biomarkers of OA severity, but also that C3f fragment is further related to OA severity whereas the V65 vitronectin fragment is more related to early OA detection.

Subchondral Bone Plate Changes More Rapidly than Trabecular Bone in Osteoarthritis.

Osteoarthritis (OA) is the most common joint disorder, characterised by focal loss of cartilage and increased subchondral bone remodelling at early OA stages of the disease. We have investigated the temporal and the spatial relationship between bone remodelling in subchondral bone plate (Sbp) and trabecular bone (Tb) in Dunkin Hartley (DH, develop OA early) and the Bristol Strain 2 (BS2, control which develop OA late) guinea pigs. Right tibias were dissected from six male animals of each strain, at 10, 16, 24 and 30 weeks of age. Micro-computed tomography was used to quantify the growth plate thickness (GpTh), subchondral bone plate thickness (SbpTh) and trabecular bone thickness (TbTh), and bone mineral density (BMD) in both Sbp and Tb. The rate of change was calculated for 10-16 weeks, 16-24 weeks and 24-30 weeks. The rate of changes in Sbp and Tb thickness at the earliest time interval (10-16 weeks) were significantly greater in DH guinea pigs than in the growth-matched control strain (BS2). The magnitude of these differences was greater in the medial side than the lateral side (DH: 22.7 and 14.75 µm/week, BS2: 5.63 and 6.67 µm/week, respectively). Similarly, changes in the BMD at the earliest time interval was greater in the DH strain than the BS2, again more pronounced in the disease prone medial compartment (DH: 0.0698 and 0.0372 g/cm³/week, BS2: 0.00457 and 0.00772 g/cm³/week, respectively). These changes observed preceded microscopic and cellular signs of disease as previously reported. The rapid early changes in SbpTh, TbTh, Sbp BMD and Tb BMD in the disease prone DH guinea pigs compared with the BS2 control strain suggest a link to early OA pathology. This is corroborated by the greater relative changes in subchondral bone in the medial compared with the lateral compartment.

Subchondral bone plate thickening precedes chondrocyte apoptosis and cartilage degradation in spontaneous animal models of osteoarthritis.

Osteoarthritis (OA) is the most common joint disorder characterised by bone remodelling and cartilage degradation and associated with chondrocyte apoptosis. These processes were investigated at 10, 16, 24, and 30 weeks in Dunkin Hartley (DH) and Bristol Strain 2 (BS2) guinea pigs that develop OA spontaneously. Both strains had a more pronounced chondrocyte apoptosis, cartilage degradation, and subchondral bone changes in the medial than the lateral side of the tibia, and between strains, the changes were always greater and faster in DH than BS2. In the medial side, a significant increase of chondrocyte apoptosis and cartilage degradation was observed in DH between 24 and 30 weeks of age preceded by a progressive thickening and stiffening of subchondral bone plate (Sbp). The Sbp thickness consistently increased over the 30-week study period but the bone mineral density (BMD) of the Sbp gradually decreased after 16 weeks. The absence of these changes in the medial side of BS2 may indicate that the Sbp of DH was undergoing remodelling. Chondrocyte apoptosis was largely confined to the deep zone of articular cartilage and correlated with thickness of the subchondral bone plate suggesting that cartilage degradation and chondrocyte apoptosis may be a consequence of continuous bone remodelling during the development of OA in these animal models of OA.

Increased chondrocyte apoptosis is associated with progression of osteoarthritis in spontaneous Guinea pig models of the disease.

Osteoarthritis (OA) is the most common joint disease characterised by degradation of articular cartilage and bone remodelling. For almost a decade chondrocyte apoptosis has been investigated as a possible mechanism of cartilage damage in OA, but its precise role in initiation and/or progression of OA remains to the determined. The aim of this study is to determine the role of chondrocyte apoptosis in spontaneous animal models of OA. Right tibias from six male Dunkin Hartley (DH) and Bristol Strain 2 (BS2) guinea pigs were collected at 10, 16, 24 and 30 weeks of age. Fresh-frozen sections of tibial epiphysis were microscopically scored for OA, and immunostained with caspase-3 and TUNEL for apoptotic chondrocytes. The DH strain had more pronounced cartilage damage than BS2, especially at 30 weeks. At this time point, the apoptotic chondrocytes were largely confined to the deep zone of articular cartilage (AC) with a greater percentage in the medial side of DH than BS2 (DH: 5.7%, 95% CI: 4.2-7.2), BS2: 4.8%, 95% CI: 3.8-5.8), p > 0.05). DH had a significant progression of chondrocyte death between 24 to 30 weeks during which time significant changes were observed in AC fibrillation, proteoglycan depletion and overall microscopic OA score. A strong correlation (p ≤ 0.01) was found between chondrocyte apoptosis and AC fibrillation (r = 0.3), cellularity (r = 0.4) and overall microscopic OA scores (r = 0.4). Overall, the rate of progression in OA and apoptosis over the study period was greater in the DH (versus BS2) and the medial AC (versus lateral). Chondrocyte apoptosis was higher at the later stage of OA development when the cartilage matrix was hypocellular and highly fibrillated, suggesting that chondrocyte apoptosis is a late event in OA.

Quantitative light fluorescence (QLF) and polarized white light (PWL) assessments of dental fluorosis in an epidemiological setting.

BACKGROUND: To determine if a novel dual camera imaging system employing both polarized white light (PWL) and quantitative light induced fluorescence imaging (QLF) is appropriate for measuring enamel fluorosis in an epidemiological setting. The use of remote and objective scoring systems is of importance in fluorosis assessments due to the potential risk of examiner bias using clinical methods. METHODS: Subjects were recruited from a panel previously characterized for fluorosis and caries to ensure a range of fluorosis presentation. A total of 164 children, aged 11 years (±1.3) participated following consent. Each child was examined using the novel imaging system, a traditional digital SLR camera, and clinically using the Dean's and Thylstrup and Fejerskov (TF) Indices on the upper central and lateral incisors. Polarized white light and SLR images were scored for both Dean's and TF indices by raters and fluorescence images were automatically scored using software. RESULTS: Data from 164 children were available with a good distribution of fluorosis severity. The automated software analysis of QLF images demonstrated significant correlations with the clinical examinations for both Dean's and TF index. Agreement (measured by weighted Kappa's) between examiners scoring clinically, from polarized photographs and from SLR images ranged from 0.56 to 0.92. CONCLUSIONS: The study suggests that the use of a digital imaging system to capture images for either automated software analysis, or remote assessment by raters is suitable for epidemiological work. The use of recorded images enables study archiving, assessment by multiple examiners, remote assessment and objectivity due to the blinding of subject status.

Chondrocyte apoptosis determined by caspase-3 expression varies with fibronectin distribution in equine articular cartilage.

AIM: The purpose of this study was to investigate associations between the extent of chondrocyte apoptosis and expression of the articular cartilage (AC) extracellular matrix (ECM) molecules, cartilage oligomeric matrix protein (COMP) and fibronectin. METHOD: Cartilage from four sites (when available) on equine left middle carpal joints (n = 12) were used. Expression of COMP and fibronectin was determined using specific polyclonal antibodies and a biotin-streptavidin/peroxidase method. The intensity of staining for matrix molecules was graded (none, mild, moderate, strong) in each cartilage zone. Apoptosis of chondrocytes in AC sections was assessed by their expression of active caspase-3 using immunohistochemistry. RESULTS: The intensity of fibronectin expression varied significantly according to cartilage depth, with greater expression in the deep zone than in either the superficial or middle layers (P < 0.001). A significant positive association was found overall between intensity of fibronectin expression and chondrocyte apoptosis (r = 0.44, P = 0.0187). The data were also significant for superficial and deep zones (r = 0.44, P = 0.0239 and r = 0.42, P = 0.0279 respectively). Conversely, intensity of COMP expression did not show zonal differences and was un-associated with degree of apoptosis. However, COMP expression was significantly more intense in cartilage than fibronectin (P = 0.0007), and the correlation between overall intensity of COMP and fibronectin was statistically significant (r = 0.56, P = 0.0018). CONCLUSION: The positive correlation between the incidence of apoptosis and expression of fibronectin, a key ECM molecule involved in communication between the chondrocyte and surrounding matrix, suggests that chondrocyte death by apoptosis may alter cartilage metabolism, supporting the role of this process in the pathogenesis of osteoarthritis.