Trimers Conjugated to Fibrin Hydrogels Promote Salivary Gland Function.

New strategies for tissue engineering have great potential for restoring and revitalizing impaired tissues and organs, including the use of smart hydrogels that can be modified to enhance organization and functionality of the salivary glands. For instance, monomers of laminin-111 peptides chemically conjugated to fibrin hydrogel (L1pM-FH) promote cell cluster formation in vitro and salivary gland regeneration in vivo when compared with fibrin hydrogel (FH) alone; however, L1pM-FH produce only weak expression of acinar differentiation markers in vivo (e.g., aquaporin-5 and transmembrane protein 16). Since previous studies demonstrated that a greater impact can be achieved when trimeric forms were used as compared with monomeric or dimeric forms, we investigated the extent to which trimers of laminin-111 chemically conjugated to FH (L1pT-FH) can increase the expression of acinar differentiation markers and elevate saliva secretion. In vitro studies using Par-C10 acinar cells demonstrated that when compared with L1pM-FH, L1pT-FH induced similar levels of acinar-like cell clustering, polarization, lumen formation, and calcium signaling. To assess the performance of the trimeric complex in vivo, we compared the ability of L1pM-FH and L1pT-FH to increase acinar differentiation markers and restore saliva flow rate in a salivary gland wound model of C57BL/6 mice. Our results show that L1pT-FH applied to wounded mice significantly improved the expression of the acinar differentiation markers and saliva secretion when compared with the monomeric form. Together, these positive effects of L1pT-FH warrant its future testing in additional models of hyposalivation with the ultimate goal of applying this technology in humans.

Risk of re-operation after outpatient distal hypospadias repair in a large, multistate cohort.

BACKGROUND: Repair of distal hypospadias is one of the most common pediatric urology procedures in the US. However, the risk of postsurgical complications has been reported primarily from single-institution and tertiary center studies, with short duration of patient follow-up. OBJECTIVE: The aim of the study was to examine the incidence of re-operation and risk factors for re-operation following outpatient distal hypospadias repair in a large, representative sample of US children. METHODS: A retrospective cohort study of patients aged 0-18 years undergoing single-stage distal hypospadias repair was conducted. Data were obtained from the State Ambulatory Surgery and Services Databases of 9 participating states. Patients who underwent outpatient surgery in 2008-2013 were identified using Current Procedural Terminology (CPT) codes. Patients with records suggesting prior surgery for hypospadias (CPT) were excluded, as were patients who underwent the initial repair <2 years before the end of state data availability. Return outpatient surgery visits across institutions within each of the 9 states were tracked to identify re-operations after the single-stage repair, using CPT codes for surgical treatment of hypospadias complications in 2008-2015. Time-to-event analyses were used to estimate the probability (risk) of re-operation over time and to examine whether patient and institutional characteristics were predictive of re-operation (age, race/ethnicity, health insurance, facility ownership, and institutional volume of hypospadias repair). RESULTS: A total of 4673 children treated across 148 institutions were included. The median follow-up time was 4.1 years (range: 2-7.9). Most patients were <1 year of age at the time of initial repair (53%). The risk of re-operation was 2.6% (95% confidence interval [CI]: 2.1-3.0%) at 1 year and 6.7% (95% CI: 6.0-7.5%) at 5 years after initial repair (Figure). Approximately 13% of re-operation patients had the re-operation at a different institution. None of the patient or institutional factors examined was a significant predictor of the risk of re-operation. DISCUSSION: In this population-based cohort, the estimated 5-year risk of re-operation following single-stage distal hypospadias repair was 6.7% (95% CI: 6.0-7.5). Most re-operations occurred after the first year, informing long-term expectations about postoperative complications. This study was limited by a lack of data on severity of hypospadias and surgeon characteristics and the inability to track re-operations outside of the state in which the original repair was performed. CONCLUSION: Approximately 7% of children undergoing distal hypospadias repair undergo a re-operation within 5 years. None of the factors studied were predictive of re-operations.

Broadly tunable femtosecond pulses around 2.06 µm from a diode-pumped Tm3+-doped solid-state laser source.

We report on a broadly tunable diode-pumped femtosecond Tm:LuScO3 laser source around 2.06 µm. Tuning was obtained through the use of a steeply diving birefringent filter, maintaining sub-600 fs pulses over a tuning range of 2019-2110 nm. The minimum pulse duration of 240 fs was recorded at a central wavelength of 2080 nm with an average output power of 93 mW. Higher output coupling of 2% resulted in a narrower tuning range of 2070-2102 nm with generated pulses as short as 435 fs and an average output power of 119 mW at 2090 nm.

Trends in surgical management of multicystic dysplastic kidney at USA children's hospitals.

INTRODUCTION AND OBJECTIVE: Multicystic dysplastic kidney (MCDK) is a congenital renal cystic disease often incidentally diagnosed in children. Historically, children with MCDK underwent early nephrectomy because of concerns for the development of hypertension or malignancy. Over the last decade, management recommendations have not supported routine early surgical removal of MCDK. The study authors sought to determine the current trends in the use of nephrectomy for MCDK in US children's hospitals because national practice patterns have not been investigated. METHODS: A population-based retrospective cohort study using the Pediatric Health Information System (PHIS) was conducted. The study population was comprised of patients aged 0-18 years with a diagnosis of MCDK (International Classification of Diseases-9th revision, code 753.19) admitted to the inpatient department of the study hospital between January 2006 and September 2015. Patients with additional renal anomalies including polycystic kidney, medullary cystic kidney, and medullary sponge kidney were excluded, as were patients treated in a hospital that did not contribute data to the PHIS continuously throughout the study period. Trends in the annual proportion of nephrectomies performed were analyzed among admissions in the study population, along with patient clinical and demographic information. RESULTS: A total of 3792 MCDK admissions, in 34 hospitals, were included in the study. Overall, 569 nephrectomies were performed during the study period. The proportion of nephrectomy decreased annually by 9.2% on average, from 22.1% in 2006 to 7.3% in the first 3 quarters of 2015. No significant trends were observed in the annual number of overall MCDK admissions or patient age at procedure among patients who had a nephrectomy. Among nephrectomies, 84.2% were open and 15.8% were minimally invasive procedures (laparoscopic non-robotic, 10% and robotic, 5.8%). The proportion of minimally invasive nephrectomies increased annually by 13.7%, from 8% in 2006 to 29% in 2015. DISCUSSION: Trends in the use of nephrectomy for MCDK at a national level have not been previously reported. This study is limited by the use of inpatient discharge data, which did not allow estimating the true rate of nephrectomy in patients born with MCDK. CONCLUSIONS: During the study period, there has been a decrease in the use of nephrectomy for MCDK in pediatric hospitals, along with a concurrent increase in utilization of minimally invasive techniques to perform nephrectomies. These results suggest that in general, urologists at freestanding children's hospitals are heeding recommendations for observation and against routine early surgical removal of these kidneys; although trends in the use of nephrectomy varied between hospitals, there is room for continued improvement in following these recommendations.

Diode-pumped femtosecond Tm3+-doped LuScO3 laser near 2.1 µm.

We report on the first demonstration, to the best of our knowledge, of a diode-pumped Tm:LuScO3 laser. Efficient and broadly tunable continuous wave operation in the 1973-2141 nm region and femtosecond mode-locking through the use of an ion-implanted InGaAsSb quantum-well-based semiconductor saturable absorber mirror are realized. When mode-locked, near-transform-limited pulses as short as 170 fs were generated at 2093 nm with an average output power of 113 mW and a pulse repetition frequency of 115.2 MHz. Tunable picosecond pulse generation was demonstrated in the 2074-2104 nm spectral range.

Exogenous mineralization of hard tissues using photo-absorptive minerals and femto-second lasers; the case of dental enamel.

A radical new methodology for the exogenous mineralization of hard tissues is demonstrated in the context of laser-biomaterials interaction. The proposed approach is based on the use of femtosecond pulsed lasers (fs) and Fe3+-doped calcium phosphate minerals (specifically in this work fluorapatite powder containing Fe2O3 nanoparticles (NP)). A layer of the synthetic powder is applied to the surface of eroded bovine enamel and is irradiated with a fs laser (1040 nm wavelength, 1 GHz repetition rate, 150 fs pulse duration and 0.4 W average power). The Fe2O3 NPs absorb the light and may act as thermal antennae, dissipating energy to the vicinal mineral phase. Such a photothermal process triggers the sintering and densification of the surrounding calcium phosphate crystals thereby forming a new, dense layer of typically ∼20 µm in thickness, which is bonded to the underlying surface of the natural enamel. The dispersed iron oxide NPs, ensure the localization of temperature excursion, minimizing collateral thermal damage to the surrounding natural tissue during laser irradiation. Simulated brushing trials (pH cycle and mechanical force) on the synthetic layer show that the sintered material is more acid resistant than the natural mineral of enamel. Furthermore, nano-indentation confirms that the hardness and Young's modulus of the new layers are significantly more closely matched to enamel than current restorative materials used in clinical dentistry. Although the results presented herein are exemplified in the context of bovine enamel restoration, the methodology may be more widely applicable to human enamel and other hard-tissue regenerative engineering. STATEMENT OF SIGNIFICANCE: In this work we provide a new methodology for the mineralisation of dental hard tissues using femtosecond lasers and iron doped biomaterials. In particular, we demonstrate selective laser sintering of an iron doped fluorapatite on the surface of eroded enamel under low average power and mid-IR wavelength and the formation of a new layer to substitute the removed material. The new layer is evaluated through simulated brushing trials and nano-indentation. From the results we can conclude that is more acid resistant than natural enamel while, its mechanical properties are superior to that of current restorative materials. To the best of our knowledge this is the first time that someone demonstrated, laser sintering and bonding of calcium phosphate biomaterials on hard tissues. Although we here we discuss the case of dental enamel, similar approach can be adopted for other hard tissues, leading to new strategies for the fixation of bone/tooth defects.

1.9 µm waveguide laser fabricated by ultrafast laser inscription in Tm:Lu2O3 ceramic.

The ultrafast laser inscription technique has been used to fabricate channel waveguides in Tm3+-doped Lu2O3 ceramic gain medium for the first time to our knowledge. Laser operation has been demonstrated using a monolithic microchip cavity with a continuous-wave Ti:sapphire pump source at 796 nm. The maximum output power achieved from the Tm:Lu2O3 waveguide laser was 81 mW at 1942 nm. A maximum slope efficiency of 9.5% was measured with the laser thresholds observed to be in the range of 50-200 mW of absorbed pump power. Propagation losses for this waveguide structure are calculated to be 0.7 dB⋅cm-1 ± 0.3 dB⋅cm-1 at the lasing wavelength.

Identification of long non-coding RNA in the horse transcriptome.

BACKGROUND: Efforts to resolve the transcribed sequences in the equine genome have focused on protein-coding RNA. The transcription of the intergenic regions, although detected via total RNA sequencing (RNA-seq), has yet to be characterized in the horse. The most recent equine transcriptome based on RNA-seq from several tissues was a prime opportunity to obtain a concurrent long non-coding RNA (lncRNA) database. RESULTS: This lncRNA database has a breadth of eight tissues and a depth of over 20 million reads for select tissues, providing the deepest and most expansive equine lncRNA database. Utilizing the intergenic reads and three categories of novel genes from a previously published equine transcriptome pipeline, we better describe these groups by annotating the lncRNA candidates. These lncRNA candidates were filtered using an approach adapted from human lncRNA annotation, which removes transcripts based on size, expression, protein-coding capability and distance to the start or stop of annotated protein-coding transcripts. CONCLUSION: Our equine lncRNA database has 20,800 transcripts that demonstrate characteristics unique to lncRNA including low expression, low exon diversity and low levels of sequence conservation. These candidate lncRNA will serve as a baseline lncRNA annotation and begin to describe the RNA-seq reads assigned to the intergenic space in the horse.

ß-pyrophosphate: A potential biomaterial for dental applications.

Tooth hypersensitivity is a growing problem affecting both the young and ageing population worldwide. Since an effective and permanent solution is not yet available, we propose a new methodology for the restoration of dental enamel using femtosecond lasers and novel calcium phosphate biomaterials. During this procedure the irradiated mineral transforms into a densified layer of acid resistant iron doped ß-pyrophosphate, bonded with the surface of eroded enamel. Our aim therefore is to evaluate this densified mineral as a potential replacement material for dental hard tissue. To this end, we have tested the hardness of ß-pyrophosphate pellets (sintered at 1000°C) and its mineral precursor (brushite), the wear rate during simulated tooth-brushing trials and the cytocompatibility of these minerals in powder form. It was found that the hardness of the ß-pyrophosphate pellets is comparable with that of dental enamel and significantly higher than dentine while, the brushing trials prove that the wear rate of ß-pyrophosphate is much slower than that of natural enamel. Finally, cytotoxicity and genotoxicity tests suggest that iron doped ß-pyrophosphate is cytocompatible and therefore could be used in dental applications. Taken together and with the previously reported results on laser irradiation of these materials we conclude that iron doped ß-pyrophosphate may be a promising material for restoring acid eroded and worn enamel.

Tissue resolved, gene structure refined equine transcriptome.

BACKGROUND: Transcriptome interpretation relies on a good-quality reference transcriptome for accurate quantification of gene expression as well as functional analysis of genetic variants. The current annotation of the horse genome lacks the specificity and sensitivity necessary to assess gene expression especially at the isoform level, and suffers from insufficient annotation of untranslated regions (UTR) usage. We built an annotation pipeline for horse and used it to integrate 1.9 billion reads from multiple RNA-seq data sets into a new refined transcriptome. RESULTS: This equine transcriptome integrates eight different tissues from 59 individuals and improves gene structure and isoform resolution, while providing considerable tissue-specific information. We utilized four levels of transcript filtration in our pipeline, aimed at producing several transcriptome versions that are suitable for different downstream analyses. Our most refined transcriptome includes 36,876 genes and 76,125 isoforms, with 6474 candidate transcriptional loci novel to the equine transcriptome. CONCLUSIONS: We have employed a variety of descriptive statistics and figures that demonstrate the quality and content of the transcriptome. The equine transcriptomes that are provided by this pipeline show the best tissue-specific resolution of any equine transcriptome to date and are flexible for several downstream analyses. We encourage the integration of further equine transcriptomes with our annotation pipeline to continue and improve the equine transcriptome.

Modelling topical photodynamic therapy treatment including the continuous production of Protoporphyrin IX.

Most existing theoretical models of photodynamic therapy (PDT) assume a uniform initial distribution of the photosensitive molecule, Protoporphyrin IX (PpIX). This is an adequate assumption when the prodrug is systematically administered; however for topical PDT this is no longer a valid assumption. Topical application and subsequent diffusion of the prodrug results in an inhomogeneous distribution of PpIX, especially after short incubation times, prior to light illumination. In this work a theoretical simulation of PDT where the PpIX distribution depends on the incubation time and the treatment modality is described. Three steps of the PpIX production are considered. The first is the distribution of the topically applied prodrug, the second in the conversion from the prodrug to PpIX and the third is the light distribution which affects the PpIX distribution through photobleaching. The light distribution is modelled using a Monte Carlo radiation transfer model and indicates treatment depths of around 2 mm during daylight PDT and approximately 3 mm during conventional PDT. The results suggest that treatment depths are not only limited by the light penetration but also by the PpIX distribution.

Monte Carlo modelling of photodynamic therapy treatments comparing clustered three dimensional tumour structures with homogeneous tissue structures.

We explore the effects of three dimensional (3D) tumour structures on depth dependent fluence rates, photodynamic doses (PDD) and fluorescence images through Monte Carlo radiation transfer modelling of photodynamic therapy. The aim with this work was to compare the commonly used uniform tumour densities with non-uniform densities to determine the importance of including 3D models in theoretical investigations. It was found that fractal 3D models resulted in deeper penetration on average of therapeutic radiation and higher PDD. An increase in effective treatment depth of 1 mm was observed for one of the investigated fractal structures, when comparing to the equivalent smooth model. Wide field fluorescence images were simulated, revealing information about the relationship between tumour structure and the appearance of the fluorescence intensity. Our models indicate that the 3D tumour structure strongly affects the spatial distribution of therapeutic light, the PDD and the wide field appearance of surface fluorescence images.

Monte Carlo modelling of daylight activated photodynamic therapy.

The treatment of superficial skin lesions via daylight activated photodynamic therapy (PDT) has been explored theoretically with three dimensional (3D) Monte Carlo radiation transfer simulations. For similar parameters and conditions, daylight activated PDT was compared to conventional PDT using a commercially available light source. Under reasonable assumptions for the optical properties of the tissue, protoporphyrin IX (PpIX) concentration and a treatment dose of 75 J cm(-2), it was found that during a clear summer day an effective treatment depth of over 2 mm can be achieved after 30 min of daylight illumination at a latitude of 56 degrees North. The same light dose would require 2.5 h of daylight illumination during an overcast summer day where a treatment depth of about 2 mm can be achieved. For conventional PDT the developed model suggests that 15 min of illumination is required to deliver a light dose of 75 J cm(-2), which would result in an effective treatment depth of about 3 mm. The model developed here allows for the determination of photo-toxicity in skin tissue as a function of depth for different weather conditions as well as for conventional light sources. Our theoretical investigation supports clinical studies and shows that daylight activated PDT has the potential for treating superficial skin lesions during different weather conditions.

Diode-pumped femtosecond solid-state waveguide laser with a 4.9 GHz pulse repetition rate.

We report on the first demonstration of a passively mode-locked, diode-pumped, monolithic Yb:glass channel waveguide laser that incorporates a semiconductor saturable absorber mirror. Stable and self-starting mode-locking is achieved in a Fabry-Perot cavity configuration producing a pulse repetition rate up to 4.9 GHz. The shortest pulse duration of 740 fs is generated with 30 mW of average output power at a center wavelength of 1058 nm. A maximum output power of 81 mW is produced during mode-locking with corresponding pulse duration of 800 fs.

Electrically-controlled rapid femtosecond pulse duration switching and continuous picosecond pulse duration tuning in an ultrafast Cr4+:forsterite laser.

We demonstrate rapid switching between picosecond and femtosecond operational regimes in a Cr(4+):forsterite laser, using an electrically-contacted GaInNAs SESAM with saturable absorption characteristics controlled via the quantum-confined Stark effect. Additionally, continuous picosecond pulse duration tuning by over a factor 3 is reported.

Monte Carlo simulations for optimal light delivery in photodynamic therapy of non-melanoma skin cancer.

The choice of light source is important for the efficacy of photodynamic therapy (PDT) of non-melanoma skin cancer. We simulated the photodynamic dose (PDD) delivered to a tumour during PDT using theoretical radiation transfer simulations performed via our 3D Monte Carlo radiation transfer (MCRT) model for a range of light sources with light doses up to 75 J cm(-2). The PDD delivered following superficial irradiation from (A) non-laser light sources, (B) monochromatic light, (C) alternate beam diameters and (D) re-positioning of the tumour within the tissue was computed. (A) The final PDD deposited to the tumour at a depth of 2 mm by the Paterson light source was 2.75, 2.50 and 1.04 times greater than the Waldmann 1200, Photocure and Aktilite, respectively. (B) Tumour necrosis occurred at a depth of 2.23 mm and increased to 3.81 mm for wavelengths 405 and 630 nm, respectively. (C) Increasing the beam diameter from 10 to 50 mm had very little effect on depth of necrosis. (D) As expected, necrosis depths were reduced when the tumour was re-positioned deeper into the tissue. These MCRT simulations show clearly the importance of choosing the correct light source to ensure optimal light delivery to achieve tumour necrosis.

The development and application of femtosecond laser systems.

Some background as well as recent progress in the development of femtosecond lasers are discussed together with a brief outline of a few representative emergent applications in biology and medicine that are underpinned by access to such sources. We also provide a short summary of other contributions in this focus issue.

Algorithm-based continuous pulse duration tuning and performance control of a mode-locked laser diode.

A control algorithm is presented that addresses the stability issues inherent to the operation of monolithic mode-locked laser diodes. It enables a continuous pulse duration tuning without any onset of Q-switching instabilities. A demonstration of the algorithm performance is presented for two radically different laser diode geometries and continuous pulse duration tuning between 0.5 ps to 2.2 ps and 1.2 ps to 10.2 ps is achieved. With practical applications in mind, this algorithm also facilitates control over performance parameters such as output power and wavelength during pulse duration tuning. The developed algorithm enables the user to harness the operational flexibility from such a laser with 'push-button' simplicity.

Optical trapping with "on-demand" two-photon luminescence using Cr:LiSAF laser with optically addressed saturable Bragg reflector.

We demonstrate a diode-pumped Cr:LiSAF laser with controllable and reliable fast switching between its continuous-wave and mode-locked states of operation using an optically-addressed semiconductor Bragg reflector, permitting dyed microspheres to be continuously trapped and monitored using a standard microscope imaging and on-demand two-photon-excited luminescence techniques.

Passively mode locked femtosecond Tm:Sc2O3 laser at 2.1 µm.

We report on the passive mode locking of a Tm3+:Sc2O3 laser at 2.1 µm using a semiconductor saturable absorber mirror based on InGaAsSb quantum wells. Transform-limited 218 fs pulses are generated with an average power of 210 mW. A maximum output power of 325 mW is produced during mode locking with the corresponding pulse duration of 246 fs at a pulse repetition frequency of 124.3 MHz. A Ti:sapphire laser is used as the pump source operating at 796 nm.