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Abstract. Rationale:. The most common upper limb amputations are finger amputations, resulting in functional limitations that lead to problems with activities of daily living or job loss. For many years, prosthetic options for finger amputations have been limited to passive prostheses. In many countries including South Korea, body-powered finger prostheses have rarely been prescribed due to high cost, lack of experience of physicians and prosthetists, low interest and no coverage by insurance benefits. We report 2 cases of work-related finger amputations in patients who received body-powered 3D-printed finger prostheses. Patient concerns and diagnosis:. Patient 1 was a 25-year-old woman with second and third finger amputations at the proximal interphalangeal level. Patient 2 was a 26-year-old man who sustained a second finger amputation at proximal interphalangeal level. Interventions:. We created body-powered 3D-printed finger prostheses that mimicked distal interphalangeal joint motion through patient-driven metacarpophalangeal joint motion using a string connected to a wrist strap and a linkage system. The source code “Knick Finger” was downloaded from e-NABLE. Outcomes:. After 1 month of prosthesis training, both patients were satisfied with the prostheses and showed improved performance in patient-derived goals of cooking (patient 1) and typing on a computer (patient 2). Lessons:. Over the past decade, significant advances have been made in 3D-printed prosthetics owing to their light weight, low cost, on-site fabrication, and easy customization. Although there are still several limitations in the general application of 3D-printed finger prostheses, our study suggests that for patients with finger amputations, body-powered 3D-printed finger prostheses have high potential as an additional prosthetic option to the existing passive cosmetic prostheses.

Psychological readiness following anterior cruciate ligament reconstruction (ACLR) correlates with different return to sport outcomes. However, the relationship between strength and power and psychological readiness remains unexplored. The aim of this study was to investigate the relationship between anterior cruciate ligament return to sport after injury (ACL-RSI) scores and various hamstrings and quadriceps strength and power variables. Twelve participants (20.7 ± 2.5 years old; 174.2 ± 7.5 cm; 70.2 ± 8.5 kg; 18.2 ± 8.3% of body fat) who had an ACLR nine months or more before the study completed the ACL-RSI questionnaire and isokinetic strength testing of the hamstrings and quadriceps (60°·s -1 and 180°·s -1 ). Based on ACL-RSI scores, they were divided into "cases" and "controls", deemed not psychologically ready and psychologically ready to return to previous sport performance (PILOS), respectively. The main findings are that quadriceps' and hamstrings' rate of torque development (RTD) and time since surgery were determinants of psychological readiness following ACLR. Furthermore, compared to controls, cases showed significantly lower quadriceps torque at angles close to full knee extension (40 deg and 30 deg from extension). They also showed lower RTD than controls, but no difference in peak torque. These results suggest that physiotherapists should facilitate athletes' return to sport (RTS) by focusing on the restoration of RTD and strength at angles close to full knee extension.

Clinical management of many chronic ophthalmological disorders requires direct delivery of drugs into the vitreous. There is an important need to investigate novel needle-less alternatives to deliver drugs to the vitreous. The purpose of this study is to assess the effects of a needle-less system using ultrasound to enhance vitreal delivery of small molecules through the sclera in an ex vivo model and to evaluate whether changes in permeability are mainly due to the heat generated by sonication. An eye cup containing 1 mL of sodium fluorescein 0.1% was placed on top of the sclera of cadaveric rabbit eyes. Treated eyes were sonicated for 10 minutes, and left in contact with the fluorescein solution for an additional 50 minutes. Control eyes received the same exposure to fluorescein solution (60 minutes) in the eye cup without ultrasound treatment. Vitreous humor was collected and analyzed using a fluorescence spectrophotometer to calculate the concentration of fluorescein that diffused into the vitreous humor. An additional set of eyes was treated using a heating probe to evaluate whether changes in permeability were mainly due to heat. Vitreous samples from ultrasound-treated eyes showed a 44.6% higher concentration of fluorescein compared to control eyes. The concentration of fluorescein in the vitreous of heat-treated eyes did not show a significant difference when compared to control eyes. Thus, phonophoresis is a promising needle-less method for vitreal drug delivery, and local heating conducted to the surface of the sclera should be mitigated because it does not enhance the efficacy of the method., Competing Interests: None.

Purpose: This study investigated nonadjacent dependency learning in Cantonese-speaking children with and without a history of specific language impairment (SLI) in an artificial linguistic context., Method: Sixteen Cantonese-speaking children with a history of SLI and 16 Cantonese-speaking children with typical language development (TLD) were tested with a nonadjacent dependency learning task using artificial languages that mimic Cantonese., Results: Children with TLD performed above chance and were able to discriminate between trained and untrained nonadjacent dependencies. However, children with a history of SLI performed at chance and were not able to differentiate trained versus untrained nonadjacent dependencies., Conclusions: These findings, together with previous findings from English-speaking adults and adolescents with language impairments, suggest that individuals with atypical language development, regardless of age, diagnostic status, language, and culture, show difficulties in learning nonadjacent dependencies. This study provides evidence for early impairments to statistical learning in individuals with atypical language development.

Purpose: To evaluate the effect of nonenzymatic cross-linking (glycation) upon the permeability of the vitreous to small- and large-solute diffusion., Methods: Vitreous from freshly excised porcine eyes was treated for 30 minutes with control or 0.01%, 0.1%, or 1% methylglyoxal (MG) solution. The efficacy of the glycation regimen was verified by measuring nonenzymatic cross-link density by fluorescence in the vitreous samples. Resistance to collagenase digestion as well as N(ε)-(carboxyethyl) lysine (CEL) content were also measured. The permeability coefficient for fluorescein and fluorescein isothiocyanate (FITC)-IgG diffusion through 3 mL of the vitreous samples was determined by using a custom permeability tester., Results: Vitreous cross-linking with MG treatment was confirmed by increased fluorescence, increased CEL concentration, and increased resistance to collagenase digestion. Vitreous glycation resulted in a statistically significant decrease in the permeability coefficient for fluorescein diffusion when either 0.1% or 1% MG solution was used (5.36 ± 5.24 × 10(-5) cm s(-1), P = 0.04; and 4.03 ± 2.1 × 10(-5) cm s(-1), P = 0.001; respectively, compared with control, 9.77 ± 5.45 × 10(-5) cm s(-1)). The permeability coefficient for diffusion of FITC-IgG between control (9.9 ± 6.37 × 10(-5) cm s(-1)) and treatment groups was statistically significant at all MG concentrations (0.01% MG: 3.95 ± 3.44 × 10(-5) cm s(-1), P = 0.003; 0.1% MG: 4.27 ± 1.32 × 10(-5) cm s(-1), P = 0.004; and 0.1% MG: 3.72 ± 2.49 × 10(-5) cm s(-1), P = 0.001)., Conclusions: Advanced glycation end-product (AGE) accumulation reduces vitreous permeability when glycation is performed in ex vivo porcine vitreous. The permeability change was more pronounced for the larger solute, suggesting a lower threshold for AGE-induced permeability changes to impact the movement of proteins through the vitreous when compared with smaller molecules.

Background: Known factors affecting the management of vesicoureteral reflux (VUR) include reflux grade, infection frequency, age and gender. We hypothesized that provider preference is highly associated with management., Methods: Utilizing the national billing database, Faculty Practice Solutions Center, a multivariable logistic regression model, was applied to analyze the association of pediatric urologist treatment patterns, patient age, gender, uni- or bilateral disease, insurance type, presence of nephropathy and race with the type of VUR treatment a patient would receive., Results: We identified 59 pediatric urologists who managed 7,882 new reflux patients from 2009 to 2011. Over this 3-year period there was wide variation in surgical utilization between surgeons (mean 50 %) but minimal change for each surgeon (5 %). For every 100 new reflux patients, median utilization of reimplantation surgery and injection of dextranomer/hyaluronic acid copolymer (Deflux) was 26 and 20 %, respectively. Age ranked highest in predicting surgical versus non-surgical management, while a surgeon's historic Deflux utilization rate ranked highest in predicting surgery type. Older age, female gender and white race also increased the odds of Deflux utilization over reimplantation., Conclusions: A surgeon's historic Deflux utilization was the most important predictor of VUR surgery type. Although data on reflux grade were not available, analysis of patient and surgeon characteristics suggests that surgeon preference is the first or second most critical factor in determining a patient's treatment., Competing Interests: The authors have no conflicts of interest to disclose.

Objectives: Qualitative research aimed at identifying patient acceptance of active surveillance (AS) has been identified as a public health research priority. The primary objective of this study was to determine if analysis of a large-sample of anonymous internet conversations (ICs) could be utilized to identify unmet public needs regarding AS., Methods: English-language ICs regarding prostate cancer (PC) treatment with AS from 2002-12 were identified using a novel internet search methodology. Web spiders were developed to mine, aggregate, and analyze content from the world-wide-web for ICs centered on AS. Collection of ICs was not restricted to any specific geographic region of origin. NLP was used to evaluate content and perform a sentiment analysis. Conversations were scored as positive, negative, or neutral. A sentiment index (SI) was subsequently calculated according to the following formula to compare temporal trends in public sentiment towards AS: [(# Positive IC/#Total IC)-(#Negative IC/#Total IC) x 100]., Results: A total of 464 ICs were identified. Sentiment increased from -13 to +2 over the study period. The increase sentiment has been driven by increased patient emphasis on quality-of-life factors and endorsement of AS by national medical organizations. Unmet needs identified in these ICs include: a gap between quantitative data regarding long-term outcomes with AS vs. conventional treatments, desire for treatment information from an unbiased specialist, and absence of public role models managed with AS., Conclusions: This study demonstrates the potential utility of online patient communications to provide insight into patient preferences and decision-making. Based on our findings, we recommend that multidisciplinary clinics consider including an unbiased specialist to present treatment options and that future decision tools for AS include quantitative data regarding outcomes after AS.

Purpose: The literature on secondary surgery after hypospadias repair is limited. We determined risk factors for secondary surgery via a population based approach., Materials and Methods: We used a hospital consortium database to identify a population that underwent hypospadias repair in 2009 and 2010. Specifically meatal advancement and glanuloplasty, distal, proximal and perineal hypospadias repairs were evaluated. Secondary surgeries performed between 2009 and 2011 were captured and the variables of age at primary surgery, insurance, region and surgeon volume were measured. Mixed effects logistic regression analysis was used to analyze independent variables associated with secondary surgery., Results: We identified 5,326 subjects who underwent primary hypospadias repair by 114 surgeons at 47 hospitals in 2009 and 2010. Distal hypospadias repair is associated with a 9% secondary surgery rate. After adjusting for other factors every additional 10 distal repairs that a surgeon performed yearly was associated with a 29% decreased risk of requiring fistula, stricture or diverticulum repair. There were regional differences for secondary surgery following meatal advancement and glanuloplasty and distal repairs. Finally, each additional year of patient age at distal repair was associated with a 15% increased risk of requiring secondary cystoscopy and a 21% increased risk of requiring urethral dilation/incision., Conclusions: This population based study produced significantly different results than small studies of select patients and techniques. Distal hypospadias repair is associated with a 9% secondary surgery rate. Low surgeon volume independently increases the risk of fistula, stricture or diverticulum repair. Increased patient age at primary distal hypospadias repair increases the risk of cystoscopy and urethral dilation/incision., (Copyright © 2013 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.)

Purpose: To determine the flow characteristics of the 3 different models of the Ex-PRESS miniature glaucoma device in a controlled laboratory study., Materials and Methods: The 3 different Ex-PRESS models (P-50, R-50, and P-200; Optonol Ltd; now Alcon Lab) were tested using a gravity-driven flow test. Three samples of each of the 3 Ex-PRESS models were subjected to a constant gravitational force of fluid at 5 different pressure levels (5 to 25 mm Hg). Four measurements per sample were taken at each pressure level. The main outcome measure was flow rate (Q) (µL/min). Resistance (R) was calculated by dividing pressure (P) by the measured flow (Q)., Results: The flow rate was primarily pressure dependent. The P-200 model (internal diameter 200 µm) showed a statistically significant higher flow rate and lower resistance compared with both the P-50 and R-50 models (internal diameter 50 µm) (P<0.0001). The P-50 and R-50 models demonstrated similar flow rates (P=0.08) despite their difference in tube length (2.64 vs. 2.94 mm)., Conclusions: The 3 models of the Ex-PRESS mini shunt behaved in vitro as simple flow resistors by creating a relatively constant resistance to flow. Tube diameter was the only parameter with significant impact on flow and resistance. All models demonstrated flow rates per unit of pressure much higher than the outflow facility of a healthy human eye.

The identification of biomaterials that are well tolerated in the eye is important for the development of new ocular drug delivery devices and implants, and the application of micro- and nanoengineered devices to biomedical treatments is predicated on the long-term preservation within the target organ or tissue of the very small functional design elements. This study assesses the ocular tolerance and durability of micro- and nanostructured biopolymer thin films injected or implanted into the rabbit eye. Structured poly(caprolactone) (PCL) thin films were placed in adult rabbit eyes for survival studies, with serial ophthalmic examinations over 6 months. Morphologic abnormalities and device/tissue reactions were evaluated by histologic studies, and scanning electron microscopy (SEM) of films was used to determine the structural integrity. Structured PCL thin films (20- to 40-μm thick) were constructed to design specifications with 50-μm linear microgrooves or arrays of nanopores with ~30-nm diameters. After up to 9 months of ocular residency, SEM on devices retrieved from the eye showed preservation of micro- and nanostructural features. In ocular safety evaluations carried out over 6 months, serial examinations in 18 implanted eyes showed no evidence of chronic inflammation, cataractogenesis, or retinal toxicity. Postoperative ocular inflammation was seen in 67% of eyes for 1 week, and persistent corneal edema occurred in 1 eye. Histology revealed no ocular inflammation or morphologic abnormalities of ocular tissues. Thin-film/tissue responses such as cellular reaction, fibrosis, or surface biodeposits were not seen. Micro- and nanostructured PCL thin films exhibited acceptable ocular tolerance and maintained the structural integrity of design features while residing in the eye. Thin-film micro- and nanostructured PCL appears to be a feasible biomaterial for intraocular therapeutic applications.

This study investigated the hemodynamic mechanisms underlying the exaggerated blood pressure response to muscle contraction in prehypertensive humans and the potential role of skeletal muscle metabo- and mechanoreceptors in this response. To accomplish this, changes in peak mean arterial blood pressure (ΔMAP), cardiac output, and total peripheral resistance (ΔTPR) were compared between prehypertensive (n = 23) and normotensive (n = 19) male subjects during 2 min of static contraction (at 50% of maximal tension), 2 min of postexercise muscle ischemia (metaboreflex), and 1 min of passive dorsiflexion of the foot (tendon stretch, mechanoreceptor reflex). These variables were assessed before and during the interventions. Percentage increases from baseline in MAP and TPR in response to the exercise pressor reflex were augmented in the prehypertensives, compared with the normotensives (44% ± 5% vs. 33% ± 4% and 34% ± 15% vs. 2% ± 8%, respectively) (p < 0.05). Metaboreflex-induced increases in MAP and TPR were also augmented in the prehypertensives (28% ± 5% vs. 14% ± 4% and 36% ± 12% vs. 14% ± 9%, respectively) (p < 0.05). In response to the mechanoreflex, no differences in the percentage increase in MAP or TPR were seen between groups. The results indicate that the reflex pressor response to static contraction is augmented in prehypertension and suggest that this phenomenon is due, at least in part, to enhanced activation of metaboreceptors.

The purpose of this study was to assess fundamental differences between the mechanics of the posterior sclera in paired eyes using uniaxial and whole globe inflation testing, with an emphasis on the relationship between testing conditions and observed tissue behavior. Twenty porcine eyes, consisting of matched pairs from 10 pigs, were used in this study. Within pairs, one eye was tested with 10 cycles of globe pressurization to 150 mmHg (∼10× normal IOP) while biaxial strains were tracked via an optical system at the posterior sclera. An excised posterior strip from the second eye was subjected to traditional uniaxial testing in which mechanical hysteresis was recorded from 10 cycles to a peak stress of 0.13 MPa (roughly equivalent to the circumferential wall stress produced by an IOP of 150 mmHg under the thin-walled pressure vessel assumption). For approximately equivalent loads, peak strains were more than twice as high in uniaxial tests than in inflation tests. Different trends in the load-deformation plots were seen between the tests, including an extended "toe" region in the uniaxial test, a generally steeper curve in the inflation tests, and reduced variability in the inflation tests. The unique opportunity of being able to mechanically load a whole globe under near physiologic conditions alongside a standard uniaxially tested specimen reveals the effects of testing artifacts relevant to most uniaxially tested soft tissues. Whole globe inflation offers testing conditions that significantly alter load-deformation behavior relative to uniaxial testing; consequently, laboratory studies of interventions or conditions that alter scleral mechanics may greatly benefit from these findings., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Purpose: To investigate the effect of cross-linking treatment on corneal permeability in a live animal model., Methods: Rabbit eyes were selected at random to be left unoperated or to undergo epithelial debridement with or without treatment consisting of cross-linking (CXL) with riboflavin and ultraviolet-A. Nine eyes received a total dose of 3.6 J/cm² and after epithelial healing the corneas were placed in a two-chamber system for quantification of the diffusion of fluorescein compared with controls. Thirty eyes received a total dose of 5.4 J/cm² and, after epithelial healing, in vivo corneal permeability was quantified as the pupillary response over a 30-minute period to a dose of topical pilocarpine compared with controls., Results: In the ex vivo assay, the mean permeability coefficient in the CXL group (2.42 × 10⁻⁷) was reduced when compared with the unoperated controls (3.73 × 10⁻⁷; P = 0.007) and to the eyes that received epithelial debridement alone (3.74 × 10⁻⁷; P = 0.01). In the in vivo permeability assay, the change in pupillary diameter at 30 minutes after pilocarpine administration was smaller in the CXL group (-1.9 mm), compared with the epithelial debridement group (-2.6 mm; P < 0.001) and with the unoperated controls (-2.7 mm; P = 0.003)., Conclusions: Corneal cross-linking with ultraviolet-A and riboflavin results in a statistically significant reduction in corneal permeability. These findings suggest that dosing of topical medications may need to be increased in eyes with a history of CXL to achieve expected therapeutic effects, and they may have implications for the long-term health of the cornea.

Purpose: To investigate the relationship between corneal permeability and nonenzymatic cross-link density., Methods: Corneas were dissected from 90 cadaveric porcine eyes. Samples were incubated for 24 hours with control solution or methylglyoxal at concentrations of 0.01%, 0.10%, and 1.00%. Nonenzymatic cross-link density in treated and control groups was quantified by papain digest and fluorescence spectrophotometry. Control and treated corneas were mounted in a customized Ussing-type chamber connected to vertical tubing, and specific hydraulic conductivity was determined according to the descent of a column of degassed saline at room temperature. Permeability to diffusion of fluorescein in a static chamber was determined for a similar set of corneal samples., Results: Methylglyoxal treatment effectively increased nonenzymatic cross-link content, as indicated by the average fluorescence for each group. Specific hydraulic conductivity (m(2)) was reduced with increasing cross-link density. Similarly, the permeability coefficient for the fluorescein solute consistently decreased with increasing methylglyoxal concentration, indicating diffusion impedance resulting from the treatment., Conclusions: Nonenzymatic cross-link density in the cornea can be significantly increased by treatment with methylglyoxal. Porcine cornea showed a nonlinear reduction in solute permeability and specific hydraulic conductivity with increasing cross-link density. This model suggests that age-related nonenzymatic cross-link accumulation can have a substantial impact on corneal permeability.

Purpose: To investigate the relationship between scleral permeability and nonenzymatic cross-link density., Methods: Scleral discs 18 mm in diameter were dissected from the medial and lateral equatorial regions of 60 cadaveric porcine eyes. Samples were incubated for 24 hours with control solution or methylglyoxal at concentrations of 0.001%, 0.01%, 0.10%, and 1.00%. Nonenzymatic cross-link density in treated and control groups was quantified with the use of papain digest and fluorescence spectrophotometry. Treated scleral discs were mounted in a customized Ussing-type chamber connected to vertical tubing, and specific hydraulic conductivity was determined according to the descent of a column of degassed saline at room temperature. Permeability to diffusion of fluorescein in a static chamber was determined for another set of treated scleral samples., Results: Methylglyoxal treatment effectively increased nonenzymatic cross-link content, as indicated by the average fluorescence for each group. Specific hydraulic conductivity (m(2)) was reduced with increasing cross-link density. Similarly, the permeability coefficient for the fluorescein solute consistently decreased with increasing methylglyoxal concentration, indicating diffusion impedance from the treatment., Conclusions: Nonenzymatic cross-link density can be significantly increased by treatment with methylglyoxal. Porcine sclera showed a nonlinear reduction in solute permeability and specific hydraulic conductivity with increasing cross-link density. This model indicates that age-related nonenzymatic cross-link accumulation can have a substantial impact on scleral permeability.

Purpose: To assess the ultrastructural and fluid flow characteristics of cultured trabecular meshwork (TM) cells derived from fetal sources., Methods: Fetal eyes were carefully dissected to isolate the developing TM tissue for culture. Immunostaining was used to assess the expression of the junction-associated proteins zonula occludens-1 (ZO-1) and occludin. Fetal and adult TM cells were grown to confluence on permeable membranes for both flow and ultrastructural studies. Fluid flow resistance was measured by permeation of horseradish peroxidase (hrp) activity and hydraulic conductivity (HC) experiments. The effects of dexamethasone (Dex) on permeability and HC were also evaluated., Results: ZO-1 and occludin are expressed in the TM region of tissue sections and at cell borders in cultured fetal and adult TM cells. Transmission electron microscopy demonstrated that cultured TM cells possessed numerous mitochondria, electron-dense bodies, surface microvilli, and adherens and gap junctions. The permeation of hrp across fetal TM cell monolayers (0.030 +/- 0.010) and of adult TM cells (0.031 +/- 0.010) had similar values for absorbance at 470 nm (p = 0.83, 95% CI: -0.004, 0.005). Dex treatment significantly reduced the permeability to 0.022 +/- 0.008 (p = 0.002) and 0.018 +/- 0.009 (p = 0.004) for fetal and adult TM cells, respectively. The average HC (microl/min/mmHg/cm(2)) of fetal cells (2.78 +/- 1.03) and of the adult cells (2.15 +/- 1.31) was not significantly different (p = 0.24, 95% CI: -1.01, 0.26). Dex treatment significantly reduced HC in both fetal (1.24 +/- 0.72, p = 0.0004) and adult (1.29 +/- 0.29, p = 0.00001) TM., Conclusions: Cultured fetal TM cells exhibited similar expression of junctional proteins and ultrastructural features as their adult counterparts. The permeability and HC of the fetal cells were similar to their older adult counterparts. Dex treatment induced increased fluid flow resistance in both cell types. These cells may serve as a source for in vitro studies of meshwork physiology.

Purpose: To study photoreceptor apoptosis and iron migration as mechanisms of retinotoxicity in a rabbit model of subretinal hemorrhage (SRH) and to assess intravitreal triamcinolone acetonide (IVTA) for anti-apoptotic and neuroprotective effects., Methods: In adult rabbits, eyes were studied histologically after subretinal injection of autologous blood. For comparisons of control eyes with eyes injected with 2 mg IVTA, morphometric analysis was performed with light microscopy, whereas apoptosis was quantified with terminal dUTP nick end labeling (TUNEL) and fluorescence microscopy. Localization of retinal iron was assessed with Perls' stain., Results: Photoreceptor degeneration was initiated 48 hours after exposure to subretinal blood and progressed over 7 days. Increased TUNEL positivity demonstrating apoptotic cell death was associated with SRH and photoreceptor loss. VIP-Perls staining demonstrated iron in the photoreceptor layer and retinal pigment epithelium that correlated with photoreceptor degeneration. Treatment with IVTA enhanced photoreceptor cell survival by 11% at 48 hours and by 45% at 72 hours (P = 0.01) and reduced photoreceptor apoptosis ratios by 25% at 48 hours (P = 0.006)., Conclusions: Photoreceptor toxicity caused by SRH occurs at least in part by apoptosis and is associated with iron migration to the photoreceptor layer. Treatment with IVTA reduced photoreceptor loss and apoptosis, indicating a neuroprotective action. Therapies to target SRH may augment anti-VEGF treatments in exudative age-related macular degeneration and other diseases of choroidal neovascularization.

Purpose: To isolate and characterize fetal trabecular meshwork (FTM) cells for study in culture. Cultured adult trabecular meshwork (TM) cells often possess a slower rate of growth and restricted number of population doublings, limiting the ability to perform expanded testing., Methods: Fetal eyes from 24-week gestation abortions were delicately dissected to isolate the developing trabecular meshwork tissue. Three primary cultures were achieved and passaged. Light microscopy was used to compare the FTM cells to two cultured adult TM cell lines. Immunocytochemistry and Western blot analysis were utilized to identify specific protein expression., Results: The FTM cells demonstrated similar microscopic characteristics to adult TM cells, including monolayer formation, cobblestone pattern, and comparable size. FTM cells exhibited faster, more consistent doubling times when compared with adult TM cells. They grew rapidly even after passage 8, whereas their adult counterparts slowed significantly with each successive passage and failed to reach confluence at passages 4 to 5. Immunofluorescent staining was positive for actin, vimentin, fibronectin, laminin, aquaporin-1, CD-44, and myocilin in both FTM and adult TM cells. In both fetal and adult cells, Western blots showed substantial increase in myocilin after exposure to dexamethasone., Conclusions: Characterization by microscopy and immunocytochemistry suggest that FTM cells have properties similar to adult TM cells. Fetal tissues may be a useful source of abundant, rapidly dividing FTM cells for in vitro investigation. The ability to do expanded research in this field may contribute to a better understanding of the molecular mechanisms in glaucoma development.

Background & Aims: Early events in the pancreatic acinar cell critical for development of pancreatitis include activation of the transcription factor nuclear factor kappa B (NF-kappa B), abnormal Ca(2+) responses, and trypsinogen activation. Mechanisms underlying these responses, which can be studied in isolated pancreatic acini stimulated with supraphysiologic doses of cholecystokinin (CCK-8), remain poorly understood. We here report that these responses are regulated by phosphatidylinositide 3-kinase (PI3K) gamma., Methods: To inactivate PI3K, we used mice deficient in the catalytic PI3K gamma subunit p110 gamma as well as the PI3K inhibitors LY294002 and wortmannin. We measured Ca(2+) responses by using Fura-2, NF-kappa B-binding activity by electromobility shift assay, I kappa B degradation by Western blotting, and trypsinogen activation by fluorogenic assay., Results: CCK-induced intracellular Ca(2+) mobilization, Ca(2+) influx, trypsinogen, and NF-kappa B activation were all diminished in pancreatic acini isolated from p110 gamma(-/-) mice. Both in mouse and rat acini, these responses were inhibited by the PI3K inhibitors. The Ca(2+) signal and trypsinogen activation were similarly reduced in acini isolated from p110 gamma(-/-) and p110 gamma(+/-) mice compared with wild-type mice. By contrast, NF-kappa B activation was inhibited in p110 gamma(-/-) acini but not in p110 gamma(+/-) acini. These differences indicate that the mechanism of NF-kappa B regulation by PI3K gamma differs from those for the Ca(2+) and trypsinogen responses. CCK-induced responses in p110 gamma(-/-) acini were all further inhibited by LY294002, indicating the involvement of other PI3K isoform(s), in addition to PI3K gamma., Conclusions: The results show that key pathologic responses of the pancreatic acinar cell are regulated by PI3K gamma and suggest an important role for this PI3K isoform in pancreatitis.