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Background Orthodontic appliances hinder proper oral hygiene, leading to plaque retention and biofilm formation which can cause dental diseases. This study investigates the initial impact of orthodontic treatment with fixed appliances versus clear aligners on microbial profiles and gingival/plaque indices.Methods Nine patients were included in this small-scale prospective longitudinal pilot study – five in treatment with fixed appliances and four with clear aligners. Plaque biofilm samples were collected subgingivally, supragingivally and from clear aligner trays, and gingival and plaque indices were recorded at six timepoints over 3 months. DNA was extracted and next-generation 16S rRNA sequencing of the V4 region was performed.Results Fixed appliance patients showed higher plaque and gingival index scores early on (p ≤ 0.05) and higher levels of oral pathogens (p ≤ 0.05). Microbiological analysis revealed distinct microbial communities on clear aligner trays, with health- and disease-associated microbes present at significantly different levels (p ≤ 0.05) compared to intraoral plaque samples.Conclusions Patients undergoing orthodontic therapy with fixed appliances had higher levels of plaque retention and oral pathogens. Clear aligner patients’ had microbial profiles with more oral health-associated species and lower gingival/plaque indices. However, the biofilm on clear aligner trays requires further study, and the small sample size limits the conclusiveness of all findings.Implications Dentists may consider clear aligners as an alternative to fixed appliances, with potential benefits in plaque accumulation and oral microbial composition. All patients undergoing orthodontic therapy should practice good oral hygiene to prevent plaque retention and reduce oral pathogen colonization.Continuing Education Credit Available: A CDA Continuing Education quiz is online for this article: https://www.cdapresents360.com/learn/catalog/view/20The practice worksheet is available online: https://doi.org/10.1080/19424396.2024.2423594

The Royal Institute of British Architects possesses one of the finest collections of architectural drawings and one of the jewels in its crown are over three hundred drawings by the celebrated Renaissance architect Andrea Palladio. Although cataloguing these drawings began in the 1960s as of late 2023 no printed catalogue has been published. This article examines the historiography of Palladio drawings in Britain: half a century of research, in order to set out what many of the issues regarding the project have been in the last fifty years.

Abstract Medicago truncatula is a model legume that has been extensively investigated in diverse subdisciplines of plant science. Medicago littoralis can interbreed with M. truncatula and M. italica; these three closely related species form a clade, i.e. TLI clade. Genetic studies have indicated that M. truncatula accessions are heterogeneous but their taxonomic identities have not been verified. To elucidate the phylogenetic position of diverse M. truncatula accessions within the genus, we assembled 54 plastid genomes (plastomes) using publicly available next-generation sequencing data and conducted phylogenetic analyses using maximum likelihood. Five accessions showed high levels of plastid DNA polymorphism. Three of these highly polymorphic accessions contained sequences from both M. truncatula and M. littoralis. Phylogenetic analyses of sequences placed some accessions closer to distantly related species suggesting misidentification of source material. Most accessions were placed within the TLI clade and maximally supported the interrelationships of three subclades. Two Medicago accessions were placed within a M. italica subclade of the TLI clade. Plastomes with a 45-kb (rpl20-ycf1) inversion were placed within the M. littoralis subclade. Our results suggest that the M. truncatula accession genome pool represents more than one species due to possible mistaken identities and gene flow among closely related species.

Abstract This letter presents a circuit for powering cascode gate driver circuits with super‐junction (SJ) MOSFETs in a bridge‐leg. The cascode gate driver gives inherent MOSFET intrinsic diode deactivation, and the bridge‐leg incorporates an inductive turn‐on snubber which controls the charging currents into the MOSFETs’ output capacitances. The cascode driver circuits and snubber inductor are configured such that the gate‐drive energy is derived from the power stage.

Cardiac arrhythmias are common and often benign in pregnancy. However, haemodynamic instability can occur when tachyarrhythmias are accompanied by aortocaval compression, which can lead to loss of cardiac output. We present an atypical case of a pregnant woman with a supraventricular tachyarrhythmia, which degenerated into ventricular fibrillation arrest while supine due to aortocaval compression. Inducible atypical atrioventricular nodal re-entry tachycardia was subsequently detected on electrophysiological study and presumed to be the most likely initial supraventricular tachyarrhythmia.

Objectives: We sought to investigate the rates of same-day discharge (SDD) post elective percutaneous coronary intervention (PCI) at our institution and review its safety by examining clinical outcomes. We also performed an economic analysis evaluating our hospital’s cost data for SDD following PCI. Methods: Patients undergoing elective PCI at St George Hospital, Australia, from January 2017 to December 2019 were evaluated. Primary outcomes included 7-day major adverse cardiovascular endpoints (MACEs) and readmission to hospital within 30 days. Results: Among 502 patients who underwent elective PCI, 421 patients (83.8%) were managed with SDD. There was one case of acute stent thrombosis and one case of coronary wire-induced perforation requiring a pericardial drain that occurred following elective PCI with SDD (0.54%). Unplanned cardiac re-hospitalisation at 30 days following elective PCI was 5.2%. SDD after elective PCI was associated with a healthcare cost saving of AUD 4817 per case. Conclusion: SDD following elective PCI was demonstrated to be a safe and effective strategy that was also associated with significant cost savings. SDD following elective PCI warrants more widespread use as it lowers healthcare costs, has equivalent patient outcomes and improves patient satisfaction.

Once, when questioned about the originality of Umberto Eco’s Il nome della rosa (1980), Richard Krautheimer gave one of his rare and atypically acerbic replies: “you obviously haven’t read much Sherlock Holmes”. In many ways the volume discussed here provoked in the reviewer a similar response because, when reading through a number of the ten papers presented in these conference proceedings, he kept thinking: “but what about Argan?”. In this case Giulio Carlo Argan playing Canon Doyle, to Gilles Deleuze’s Eco, the latter’s Le Pli of 1988 to Argan’s brilliant but overlooked essay “La retorica e l’arte barocca” of 1955 which is not cited a single time in this book nor present in the bibliography. Acknowledging the importance of Argan (mentioned only in passing on p. 22) would not make Deleuze’s work appear any less innovative, but it certainly would have helped explain more persuasively the significant shifts in post-war perception and reception of the Baroque that were part of the historical preamble to the appearance of Leibniz et le baroque.

A glossary of acronyms used in Digital Humanities compiled by the organizer and contributors to a conference on the subject held at the Università degli studi dell’Aquila in the summer of 2022.

The Summer School dedicated to Digital Humanities (DH) for Art History, held between 27 June and 1 July 2022 at the Department of Excellence of the Department of Human Sciences (DSU Scienze Umane) of the University of L’Aquila, represented a singular opportunity for an in-depth snapshot of a discipline that, de facto, has only recently managed to carve out a defined space within academia. This, all the while for decades having been a fundamental component of many research institutes’ activities and at the same time acquiring a predominant role in the vast world of protection, enhancement and dissemination of cultural patrimony.

The coincidence, in the years 2020–2022, of COVID and increasing worldwide concern about carbon footprints, would appear to culminate in an obvious direction for the future of the DH: the necessity to push forward with mass digitisation so that scholars do not need to fly around the world to study unique objects such as manuscripts, and the move to exclusively digital publishing in order to avoid generating carbon footprints for books and journals which, now in 2022, appear to be entirely unnecessary to publish in print and more akin to driving around in a 1950s Cadillac. The proposition to stop most or all print publishing in academia may have been unthinkable even a decade ago, but in the current situation, and the forseeable future, it would be the most effective way for scholarship and scholars to make their contribution to de-carbonisation.

Future generations of Humanists will likely have no clue of how the Digital Humanities developed so key recollections are here set out in a personal timeline that perhaps can serve as a reference in the future for historiography as experienced by an art and architectural historian.

Objectives: The primary aim was to investigate the efficacy and safety of dual antiplatelet therapy (DAPT) using ticagrelor (T-DAPT) versus clopidogrel (C-DAPT) in a real-world ST-elevation myocardial infarction (STEMI) population. Methods: We retrospectively analyzed 655 consecutive patients having primary percutaneous coronary intervention (PCI) for STEMI at Liverpool Hospital, Sydney, Australia (from January 2013 to April 2016). Medical and procedural therapies were at clinician discretion. Patient data were retrieved from hospital records and primary clinicians. Results: T-DAPT (65%) was used more frequently, and in patients with lower mean CRUSADE (Can Rapid Risk Stratification of Unstable Angina Patients Suppress Adverse Outcomes with Early Implementation of the American College of Cardiology/American Heart Association Guidelines) score, than C-DAPT (24.6 vs. 32.2; p

This paper presents a low-cost approach to experimentally characterise the effect of the inverter supply on the losses in an electrical machine. The set-up comprises of representative subassemblies of the stator and winding, referred to as ‘motorettes’, and a single phase, fast switching silicon-carbide-based converter. The sub-assembly-based thermal analysis has been shown to be a time and cost-effective method to examine winding loss, over a range of operating conditions. In addition to the slot to stator thermal resistance and capacitance. This provides critical information for thermal model calibration and produces more accurate thermal and loss predictions of the full machine behaviour. A test arrangement has been devised to replicate the magnetic excitation of the electrical machine stator during normal operation. This builds upon previous techniques, to allow iron loss to be investigated. Through multiple tests using DC, AC, and PWM-AC supplies, a 20% increase in iron loss is observed due to PWM effects.

A surprising number of art historical texts written before, during and just after the first world war were republished in the 1920s. The fourth edition of Renaissance und Barock: eine Untersuchung über Wesen und Entstehung des Barockstils in Italien by Heinrich Wölfflin was published by Bruckmann in Munich in 1926 with an additional hundred and fifty pages of new text added to it by Hans Rose. This article explores why Rose had been chosen by Wölfflin to undertake the task and why Rose’s approach to the new edition differed quite markedly from the majority of reprints that appeared in this decade.

The Commentary by Hans Rose was published as an appendix to the fourth edition of Heinrich Wölfflin’s Renaissance und Barock - eine Untersuchung über Wesen und Entstehung des Barockstils in Italien, published in 1926. It was almost as long as the original text by Wölfflin and intended to discuss the new approaches on the Italian Baroque developed in German academia in the 1920s. It was divided up into four sections of unequal length, which discuss Subjectivism, Material and Colour, Urban Planning, and St Peter’s respectively.

Temperate grasses belonging to the complex are important throughout the world in pasture and grassland agriculture. Tall fescue ( Schreb.) is the predominant species in the United States, covering approximately 15 million ha. Tall fescue has distinctive morphotypes, two of which are Continental (summer active) and Mediterranean (summer semidormant). This is the first report of a linkage map created for Mediterranean tall fescue, while updating the Continental map with additional simple sequence repeat and sequence-tagged site markers. Additionally, this is the first time that diversity arrays technology (DArT) markers were used in the construction of a tall fescue map. The male parent (Continental), R43-64, map consisted of 594 markers arranged in 22 linkage groups (LGs) and covered a total of 1577 cM. The female parent (Mediterranean), 103-2, map was shorter (1258 cM) and consisted of only 208 markers arranged in 29 LGs. Marker densities for R43-64 and 103-2 were 2.65 and 6.08 cM per marker, respectively. When compared with the other Poaceae species, meadow fescue ( Huds.), annual ryegrass ( Lam.), perennial ryegrass ( L.), (L.) Beauv., and barley ( L.), a total of 171 and 98 orthologous or homologous sequences, identified by DArT analysis, were identified in R43-64 and 103-2, respectively. By using genomic in situ hybridization, we aimed to identify potential progenitors of both morphotypes. However, no clear conclusion on genomic constitution was reached. These maps will aid in the search for quantitative trait loci of various traits as well as help define and distinguish genetic differences between the two morphotypes.

The architecture treatise ‘L'ldea della Architettura Universale’ first appeared in a Dutch translation in 1640, published by Cornelis Danckerts from Amsterdam. This first edition in folio format, abundantly supplied with illustrations and text, only comprised the sixth Book of the treatise, on the subject of the orders of columns. His plan of publishing the whole treatise in a complete translation appears from the journey his son Dancker Danckerts made to Venice in 1655 to buy the original woodcuts from the heirs. However, these plans were thwarted by Symon Bosboom, stonemason by trade and as such involved in the building of the Amsterdam Town Hall, who published a simplified version of the treatise in 1657. This consisted of a short introductory text and chiefly illustrations that could easily be imitated by pupils and amateurs of architecture. In 1662 and 1664 rival editions were published by Joachem Schuym and Joost Vermaarsch respectively, both intended to explain the material of Scamozzi - in the case of Vermaarsch combined with theories of Palladio and Serlio - as simply and clearly as possible. Attempts by Danckerts to retain the market for the luxury edition by means of some cosmetic adaptations and the addition of all the original illustrations, turned out to be useless, however. Consequently, Dancker Danckerts and his successors in the family business opted for taking over the simplified editions. Until in the eighteenth century these adaptations, among which Bosboom's edition proved to have greatest commercial success, were published in a practically unchanged form. Until the middle of the nineteenth century these editions continued to be used as practical sample books for the education of artists and architects. The last editions, from 1784 onwards, introduce some innovations in the text, playing along with the new developments: the rules of architecture appear no longer to be determined from on high, but by the artist himself. This development marks the end of a successful publication history of Bosboom's treatise. The last edition appeared in 1854.

The widely held constructivist view of learning advocates student engagement via interactivity. Within the physics education research community, several specific interactive strategies have been developed to enhance conceptual understanding. One such strategy, the Interactive Lecture Demonstration (ILD) is designed for large lecture classes and, if measured using specific conceptual surveys, is purported to provide learning gains of up to 80%. This paper reports on learning gains for two different Projects over ten years. In Project 1, the ILDs were implemented from 1999 to 2001 with students who had successfully completed senior high school physics. The learning gains for students not exposed to the ILDs were in the range 13% to 16% while those for students exposed to the ILDs was 31% to 50%. In Project 2, the ILDs were implemented from 2007 to 2009 with students who had not studied senior high school physics. Since the use of ILDs in Project 1 had produced positive results, ethical considerations dictated that all students be exposed to ILDs. The learning gains were from 28% to 42%. On the one hand it is pleasing to note that there is an increase in learning gains, yet on the other, we note that the gains are nowhere near the claimed 80%. This paper also reports on teacher experiences of using the ILDs, in Project 2.

Abstract Sleep posture and movements offer insights into neurophysiological health and correlate with overall well-being and quality of life. Clinical practices utilise polysomnography for sleep assessment, which is intrusive, performed in unfamiliar environments, and requires trained personnel. While sensor technologies such as actigraphy are less invasive alternatives, concerns about their reliability and precision in clinical practice persist. Moreover, the field lacks a universally accepted algorithm, with methods ranging from raw signal thresholding to data-intensive classification models that may be unfamiliar to medical staff. This paper proposes a comprehensive framework for objectively detecting sleep posture changes and temporally segmenting postural inactivity using clinically relevant joint kinematics, measured by a custom-made wearable sensor. The framework was evaluated on wrist kinematic data from five healthy participants during simulated sleep. Intuitive three-dimensional visualisations of kinematic time series were achieved through dimension reduction-based preprocessing, providing an out-of-the-box framework explainability that may be useful for clinical monitoring and diagnosis. The proposed framework achieved up to 99.2% F1-score and 0.96 Pearson’s correlation coefficient for posture detection and inactivity segmentation respectively. This work paves the way for reliable home-based sleep movement analysis, serving patient-centred longitudinal care.

There is a huge unmet need for new treatment modalities for ocular surface inflammatory disorders (OSIDs) such as dry eye disease and meibomian gland dysfunction. Mesenchymal stem cell therapies may hold the answer due to their potent immunomodulatory properties, low immunogenicity, and ability to modulate both the innate and adaptive immune response. MSC-like cells that can be isolated from the corneal stroma (C-MSCs) offer a potential new treatment strategy; however, an optimized culture medium needs to be developed to produce the ideal phenotype for use in a cell therapy to treat OSIDs. The effects of in vitro expansion of human C-MSC in a medium of M199 containing fetal bovine serum (FBS) was compared to a stem cell medium (SCM) containing knockout serum replacement (KSR) with basic fibroblast growth factor (bFGF) and human leukemia inhibitory factor (LIF), investigating viability, protein, and gene expression. Isolating populations expressing CD34 or using siRNA knockdown of CD34 were investigated. Finally, the potential of C-MSC as a cell therapy was assessed using co-culture with an in vitro corneal epithelial cell injury model and the angiogenic effects of C-MSC conditioned medium were evaluated with blood and lymph endothelial cells. Both media supported proliferation of C-MSC, with SCM increasing expression of CD34 , ABCG2 , PAX6, NANOG, REX1, SOX2 , and THY1 , supported by increased associated protein expression. Isolating cell populations expressing CD34 protein made little difference to gene expression, however, knockdown of the CD34 gene led to decreased expression of progenitor genes. C-MSC increased viability of injured corneal epithelial cells whilst decreasing levels of cytotoxicity and interleukins-6 and -8. No pro-angiogenic effect of C-MSC was seen. Culture medium can significantly influence C-MSC phenotype and culture in SCM produced a cell phenotype more suitable for further consideration as an anti-inflammatory cell therapy. C-MSC show considerable potential for development as therapies for OSIDs, acting through anti-inflammatory action.

IntroductionTexture changes occur frequently during real-world haptic explorations, but the neural processes that encode perceptual texture change remain relatively unknown. The present study examines cortical oscillatory changes during transitions between different surface textures during active touch.MethodsParticipants explored two differing textures whilst oscillatory brain activity and finger position data were recorded using 129-channel electroencephalography and a purpose-built touch sensor. These data streams were fused to calculate epochs relative to the time when the moving finger crossed the textural boundary on a 3D-printed sample. Changes in oscillatory band power in alpha (8–12 Hz), beta (16–24 Hz) and theta (4–7 Hz) frequency bands were investigated.ResultsAlpha-band power reduced over bilateral sensorimotor areas during the transition period relative to ongoing texture processing, indicating that alpha-band activity is modulated by perceptual texture change during complex ongoing tactile exploration. Further, reduced beta-band power was observed in central sensorimotor areas when participants transitioned from rough to smooth relative to transitioning from smooth to rough textures, supporting previous research that beta-band activity is mediated by high-frequency vibrotactile cues.DiscussionThe present findings suggest that perceptual texture change is encoded in the brain in alpha-band oscillatory activity whilst completing continuous naturalistic movements across textures.

Abstract RT-PCR is the foremost clinical test for diagnosis of COVID-19. Unfortunately, PCR-based testing has limitations and may not result in a positive test early in the course of infection before symptoms develop. Enveloped RNA viruses, such as coronaviruses, alter peripheral blood methylation and DNA methylation signatures may characterize asymptomatic versus symptomatic infection. We used Illumina’s Infinium MethylationEPIC BeadChip array to profile peripheral blood samples from 164 patients who tested positive for SARS-CoV-2 by RT-PCR, of whom 8 had no symptoms. Epigenome-wide association analysis identified 10 methylation sites associated with infection and a quantile–quantile plot showed little inflation. These preliminary results suggest that differences in methylation patterns may distinguish asymptomatic from symptomatic infection.

Highlights Although most COVID-19 patients develop only mild disease, some mount a pronounced inflammatory response that results in severe pneumonia, respiratory failure, and death. Male sex is a known risk factor for severe outcomes in COVID-19. Biological sex-based differences in COVID-19 severity and mortality are influenced by genetic and hormonal differences at the cellular and immunologic level. Biological sex-based differences are further complicated by environmental factors, namely social and cultural norms, that define gender roles which impact risk of infection and severe outcomes. Understanding sex-based differences in COVID-19 outcomes will help inform clinical decision algorithms and treatment strategies to optimize the care of COVID-19 patients.

Konigsberg et al. profile DNA methylation in blood samples from SARS-CoV-2 cases and controls. The authors use machine learning to classify infected vs. non-infected individuals and predict clinical outcomes related to disease severity.

Abstract   Lolium temulentum L. (Darnel ryegrass) has been proposed to be used as a model species for functional genomics studies in forage and turf grasses, because it is a self-fertile, diploid species with a short life cycle and is closely related to other grasses. Embryogenic calluses were induced from mature embryos of a double haploid line developed through anther culture. The calluses were broken up into small pieces and used for Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105 harboring pCAMBIA1301 and pCAMBIA1305.2 vectors were used to infect embryogenic callus pieces. Hygromycin was used as a selection agent in stable transformation experiments. Hygromycin resistant calluses were obtained after 4–6 weeks of selection and transgenic plants were produced in 10–13 weeks after Agrobacterium-mediated transformation. Fertile plants were readily obtained after transferring the transgenics to the greenhouse. Transgenic nature of the regenerated plants was demonstrated by Polymerase chain reaction (PCR), Southern hybridization analysis, and GUS staining. Progeny analysis showed Mendelian inheritance of the transgenes. The transformation system provides a valuable tool for functionality tests of candidate genes in forage and turf grasses. [ABSTRACT FROM AUTHOR]

Many Australian educators believe that it is common in the USA for teachers to be sued by their students for careless teaching. Contends that this is a misconception, as the reported cases indicate that the US courts have been reluctant so far to f ind teachers liable at common law for careless teaching, or "educational malpractice". The courts have justified this reluctance on "policy" grounds, arguing that to impose such a liability on teachers would not be in the public interest. However, in a r ecent decision in Britain, the House of Lords held that educators are under a duty of care when advising on the educational needs of students. Examines the following issues: the extent to which at present Australian teachers could be liable for loss suff ered by students as a result of poor teaching; the House of Lords' decision on educational negligence, the US courts' policy reasons for refusing to hold teachers liable for poor teaching; whether the Australian courts should follow the US courts or the House of Lords; and the likelihood of Australian teachers being sued for educational malpractice in the near future. [ABSTRACT FROM AUTHOR]

The worldwide limited availability of suitable corneal donor tissue has led to the development of alternatives, including keratoprostheses (Kpros) and tissue engineered (TE) constructs. Despite advances in bioscaffold design, there is yet to be a corneal equivalent that effectively mimics both the native tissue ultrastructure and biomechanical properties. Human decellularized corneas (DCs) could offer a safe, sustainable source of corneal tissue, increasing the donor pool and potentially reducing the risk of immune rejection after corneal graft surgery. Appropriate, human-specific, decellularization techniques and high-resolution, non-destructive analysis systems are required to ensure reproducible outputs can be achieved. If robust treatment and characterization processes can be developed, DCs could offer a supplement to the donor corneal pool, alongside superior cell culture systems for pharmacology, toxicology and drug discovery studies.

PURPOSE:Dried amniotic membrane (AM) can be a useful therapeutic adjunct in ophthalmic surgery and possesses logistical advantages over cryopreserved AM. Differences in preservation techniques can significantly influence the biochemical composition and physical properties of AM, potentially affecting clinical efficacy. This study was established to investigate the biochemical and structural effects of drying AM in the absence and presence of saccharide lyoprotectants and its biocompatibility compared to cryopreserved material. METHODS:AM was cryopreserved or dried with and without pre-treatment with trehalose or raffinose and the antioxidant epigallocatechin (EGCG). Structural and visual comparisons were assessed using electron microscopy. Localisation, expression and release of AM biological factors were determined using immunoassays and immunofluorescence. The biocompatibility of the AM preparations co-cultured with corneal epithelial cell (CEC) or keratocyte monolayers were assessed using cell proliferation, cytotoxicity, apoptosis and migration assays. RESULTS:Drying devitalised AM epithelium, but less than cryopreservation and cellular damage was reduced in dried AM pre-treated with trehalose or raffinose. Dried AM alone, and with trehalose or raffinose showed greater factor retention efficiencies and bioavailability compared to cryopreserved AM and demonstrated a more sustained biochemical factor time release in vitro. Cellular health assays showed that dried AM with trehalose or raffinose are compatible and superior substrates compared to cryopreserved AM for primary CEC expansion, with increased proliferation and reduced LDH and caspase-3 levels. This concept was supported by improved wound healing in an immortalised human CEC line (hiCEC) co-cultured with dried and trehalose or raffinose membranes, compared to cryopreserved and fresh AM. CONCLUSIONS:Our modified preservation process and our resultant optimised dried AM has enhanced structural properties and biochemical stability and is a superior substrate to conventional cryopreserved AM. In addition this product is stable and easily transportable allowing it to be globally wide reaching for use in clinical and military sectors.

Limbal epithelial stem cells may ameliorate limbal stem cell deficiency through secretion of therapeutic proteins, delivered to the cornea in a controlled manner using hydrogels. In the present study the secretome of alginate-encapsulated limbal epithelial stem cells is investigated. Conditioned medium was generated from limbal epithelial stem cells encapsulated in 1.2% (w/v) calcium alginate gels. Conditioned medium proteins separated by 1-D gel electrophoresis were visualized by silver staining. Proteins of interest including secreted protein acidic and rich in cysteine, profilin-1, and galectin-1 were identified by immunoblotting. The effect of conditioned medium (from alginate-encapsulated limbal epithelial stem cells) on corneal epithelial cell proliferation was quantified and shown to significantly inhibit (P≤0.05) their growth. As secreted protein acidic and rich in cysteine was previously reported to attenuate proliferation of epithelial cells, this protein may be responsible, at least in part, for inhibition of corneal epithelial cell proliferation. We conclude that limbal epithelial stem cells encapsulated in alginate gels may regulate corneal epithelialisation through secretion of inhibitory proteins.

PURPOSE: To evaluate a panel of ten known endogenous control genes (ECG) with quantitative reverse transcription PCR (qPCR), for identification of stably expressed endogenous control genes in the ocular surface (OS) epithelial regions including cornea, limbus, limbal epithelial crypt and conjunctiva to normalise the quantitative reverse transcription PCR data of genes of interest expressed in above-mentioned regions. METHOD: The lasermicrodissected (LMD) OS epithelial regions of cryosectioned corneoscleral buttons from the cadaver eyes were processed for RNA extraction and cDNA synthesis to detect genes of interest with qPCR. Gene expression of 10 known ECG--glyceraldehyde-3-phosphate dehydrogenase (GAPDH), beta actin (ACTB), peptidylprolyl isomerase (PPIA), TATA-box binding protein (TBP1), hypoxanthine guanine phosphoribosyl transferase (HPRT1), beta glucuronidase (GUSB), Eucaryotic 18S ribosomal RNA (18S), phosphoglycerate kinase (PGK1), beta-2-microglobulin (B2M), ribosomal protein, large, P0 (RPLP0)--was measured in the OS epithelial regions by qPCR method and the data collected was further analysed using geNorm software. RESULTS: The expression stability of ecgs in the os epithelial regions in increasing order as determined with genorm software is as follows: ACTB