The effect of emulsifier type and oil inclusion on stress-related gene expression of Salmonella typhimurium in oil-in-water emulsion.

Salmonella has been associated with numerous outbreaks from contaminated food products, including emulsions. Emulsions are influenced by emulsifier type and oil presence, which can have varying degrees of stress or protection on bacteria. Although our previous research has shown that emulsifier solutions, rather than emulsions, provide a protective effect on Salmonella typhimurium after thermal treatment, the underlying mechanism remains unclear. This study selected S. typhimurium as the model microorganism and utilized the same emulsifiers (Tween 20, Tween 80, Triton X-100) to create emulsifier solutions and emulsions with the same oil fraction (60% (v/v)) to examine their effect on the expression of nine selected genes (rpoE, rpoH, otsB, proV, fadA, fabA, dnaK, ibpA, ompC) associated with stress response. Specifically, the study observed variations in gene expression under normal and thermal stress at 55°C. After 20-h incubation, Triton X-100 emulsion caused an upregulation of stress-related genes, rpoE, otsB, and fabA, suggesting stressful environment. After thermal treatment, S. typhimurium in Triton X-100 solution showed a longer 5-log reduction time with increased proV and decreased fabA and ompC expression, suggesting enhanced thermal protection compared to its emulsion. Conversely, Tween 80 solution increased fabA and ompC expression, indicating greater membrane fluidity and passive diffusion, potentially reducing thermal resistance. However, according to the upregulation of ibpA, this effect was likely mitigated by the overproduction of heat shock proteins. Notably, Triton X-100 environments exhibited the most significant gene expression changes after heat treatment, whereas Tween 80 without oil was the most inhospitable for bacterial survival. These findings inform bacterial responses under various conditions, aiding food safety strategies.

The effect of emulsifier type and oil fraction on Salmonella Typhimurium growth and thermal inactivation in oil-in-water emulsion.

High water activity oil-in-water emulsions can promote survival and growth of Salmonella Typhimurium. Nevertheless, the precise effect of emulsifier type and oil content on bacterial growth and inactivation is not fully understood. Here, emulsions were prepared using different emulsifiers (Tween 20, Tween 80, and Triton X-100) and different oil fractions (20%, 40%, and 60% (v/v)). TSB (control), emulsifier solutions, and emulsions were inoculated with S. Typhimurium. Bacterial growth rate was measured at 7, 22, and 37°C, whereas thermal inactivation was performed at 55°C. Growth and inactivation data was fitted into Logistic and Weibull models, respectively. At an incubation temperature of 37°C, the presence of high amount of oil (60%) in Tween 20 and Triton X stabilized emulsions extended the lag phase (5.83 ± 2.20 and 9.43 ± 1.07 h, respectively, compared to 2.28 ± 1.54 h for TSB, p < 0.05), whereas individual emulsifiers had no effect on growth behavior compared to TSB. This effect was also prevalent but attenuated at 22°C, whereas no growth was observed at 7°C. In thermal inactivation, we observed protective effect in Tween 80 and Triton X-100 solutions, where time required for five-log reduction was 1914.70 ± 706.35 min and 795.34 ± 420.09 min, respectively, compared to 203.89 ± 10.18 min for TSB (p < 0.05). Interestingly, the presence of high amount of oil did not offer protective effect during thermal inactivation. We hypothesize that oleic acid in Tween 80 and lower hydrophobicity value of Triton X-100 help maintain membrane integrity and improve the resistance of bacteria to heat inactivation.

Latanoprost-associated cystoid macular edema in a patient with phakic eyes.

Here, we report a case of bilateral cystoid macular edema (CME) in a woman with phakic eyes after 22 years of latanoprost use. Optical coherence tomography revealed multiple intraretinal cysts, and fluorescein angiography revealed characteristic petaloid dye leakage from the perifoveal capillaries. A cause-effect relationship was suspected when CME resolved after switching from latanoprost to dorzolamide, as confirmed by positive rechallenge and dechallenge tests. Since prostaglandin analog-induced CME has only been reported in patients with pseudophakic, aphakic, or phakic eyes with retinal conditions predisposing to macular edema, this seems to be the first reported case of latanoprost-induced CME in a patient with phakic eyes without retinal risk factors.

Retinal vasoproliferative tumor regression after intravitreal aflibercept.

Retinal vasoproliferative tumors (RVPTs) are rare benign retinal lesions typically located in the inferotemporal peripheral retina. Several treatment options exist for the management of RVPTs, but no consensus has been proposed. There are only a few reports on the use of anti-vascular endothelial growth factor with bevacizumab to treat exudative or neovascular retinal changes secondary to RVPTs. This report describes a 68-year-old female with a history of systemic hypertension that presented with a 2-week history of gradual loss of visual acuity in the right eye. Fundoscopic examination showed a RVPTs with atypical location that had a favorable response to two-intravitreal aflibercept injections 1 month apart, with resulting subretinal fluid absorption and tumor regression.

A homozygous in-frame duplication within the LRRCT consensus sequence of CFAP410 causes cone-rod dystrophy, macular staphyloma and short stature.

Ciliopathies are a group of genetic dystrophies causing syndromic and non-syndromic retinal degeneration. We identified CFAP410 as the causative gene in a patient with childhood-onset retinal dystrophy without other systemic symptoms at the age of 20. This 20-year-old man presented with cone-rod dystrophy and CFAP410 homozygous in-frame duplication variants (c.340_351dup). His clinical features included early subnormal vision, posterior pole staphyloma, and short stature. Unlike the previously reported features of retinal ciliopathy, our patient showed no obvious retinal pigmentation and only a slight hyper-autofluorescent parafoveal ring at the 16-year follow up. This case report aims to characterize the clinical features in a patient with novel, homozygous and likely pathogenic in-frame duplication variants in the CFAP410 gene. Ultimately, this report will help contribute to the understanding of CFAP410-associated ciliopathies.

Clinical features of pachyvessels associated with polypoidal choroidal vasculopathy in chronic central serous chorioretinopathy.

To investigate the association between clinical features of chronic central serous chorioretinopathy (CSC) and subsequent development of polypoidal choroidal vasculopathy (PCV). Characteristics and treatment response of PCV secondary to CSC were described. This retrospective observational study included 18 patients with chronic CSC (18 eyes) with subsequent PCV and 36 controls (36 eyes) with chronic CSC without PCV development during follow-up. Clinical features were compared between the two groups. A logistic regression model was used to evaluate the risk factor of PCV formation. Treatments for PCV included anti-vascular endothelial growth factor (VEGF) monotherapy, photodynamic therapy (PDT), or PDT and anti-VEGF combination treatment. Subretinal fluid on optical coherence tomography images were assessed after treatments. Significant between-group differences were observed in best-corrected visual acuity after disease resolution and presence of pachyvessels (P = .001 and P = .003, respectively). The presence of pachyvessels in chronic CSC was associated with subsequent PCV (odds ratio = 6.00; 95% CI, 1.74-20.68; P = .005). CSC recurrence and subfoveal choroidal thickness (SFCT) were not significantly associated with subsequent PCV development (P = .393 and P = .911, respectively). The mean age of PCV diagnosis was 51 years, and the mean time from CSC diagnosis to PCV confirmation was 77.8 months. The mean (range) SFCT of PCV was 327.7 (134-599) µm. Nine patients received anti-VEGF monotherapy and 5 had disease remission. Four patients received PDT and anti-VEGF combination treatment and all of the 4 had disease remission. In chronic CSC, pachyvessel characteristics are associated with subsequent PCV development. This result will assist clinicians to evaluate CSC in clinical practice and provide insights into the pathogenesis of PCV.

Curcumin-Loaded Pickering Emulsion Formed by Ultrasound and Stabilized by Metal Organic Framework Optimization.

The ultrasound-assisted preparation of a curcumin-loaded metal organic framework (MOF) UiO-66-NH2 stabilized Pickering emulsion system was carried out in this study. A 3-level-4-factor Box-Behnken design (BBD) and response surface methodology (RSM) analysis were employed to systematically evaluate the effect of different experimental parameters (i.e., ultrasonic power, ultrasonic time, oil content, and MOF content) on curcumin loading capacity (LC) and encapsulation efficiency (EE). The results indicated that ultrasonic power and MOF content significantly affected LC and EE, whereas ultrasonic time and oil content had little effect. A mathematical model for optimizing the preparation of emulsion systems was established. Based on the ridge max analysis, an optimal condition for the newly developed curcumin-loaded MOF-Pickering emulsion was identified, i.e., ultrasonic power 150 W, ultrasonic time 11.17 min, oil content 20.0%, and MOF content 1.10%. At this condition, the LC and EE of curcumin obtained from the experiment reached 7.33% ± 0.54% and 56.18% ± 3.03%, respectively, which were within the prediction range of LC (7.35% ± 0.29%) and EE (54.34% ± 2.45%). The emulsion systems created in this study may find new applications for the delivery of bioactive compounds in food and pharmaceutical areas.

Synthesize of alginate/chitosan bilayer nanocarrier by CCD-RSM guided co-axial electrospray: A novel and versatile approach.

To improve oral bioavailability of bioactive compounds, nano-scopic carrier systems with multiple layers has become the recent popular trend. The preparation of the nano-sized bilayer system usually requires multiple operations and intense energy input. For better efficiency, co-axial electrospraying was developed as the novel method to manufacture dual-layered nanocarrier. To improve the versatility of the production, the co-axial electrospray process was further guided by CCD-RSM (Central Composite Design-Response Surface Methodology) when generating bilayer nanocarrier. Chitosan and alginate were arranged into the core and shell part of the carrier, respectively. Through the RSM analysis, the optimum processing parameters to prepare the chitosan/alginate bilayer carrier with 112.1 ±â€¯35.2 nm diameter were determined. The surface morphology and dual layer structure were characterized microscopically. In summary, this work has successfully developed a novel and versatile mean to efficiently produce bilayer nanocarrier using co-axial electrospray combine with CCD-RSM analysis. These results could serve as a firm foundation to promote electrospray technique for the preparation of oral carrier system containing bioactive components.

Unveiling the role of microRNA-7 in linking TGF-ß-Smad-mediated epithelial-mesenchymal transition with negative regulation of trophoblast invasion.

Several pregnancy complications result from abnormal trophoblast invasion. The dichotomous effect of TGF-ß on epithelial-mesenchymal transition (EMT) between trophoblast invasion and cancer progression remains unknown and a critical concern. We attenuated the expression of TGF-ß type 1 receptor (coding by TGFBR1) with RNA interference in trophoblastic cells and significantly enhanced the trophoblastic invasion. Analysis of microRNA profiles in trophoblasts indicated microRNA-7 as a key molecule linking TGF-ß with the negative regulation of trophoblast invasion. We then attenuated TGFBR1 and miR-7 transcription by transducing either short hairpin RNA targeting TGFBR1 or anti-miR-7-locked nucleonic acid, and we observed an up-regulation of EMT-related transcription factors (TFs) and their downstream effectors, causing a mesenchymal transition of trophoblasts. Conversely, overexpression of TGFBR1 or miR-7 led to the epithelial transition of trophoblasts. Our results showed that TGF-ß-induced miR-7 expression negatively modulated the TGF-ß-SMAD family member 2-mediated EMT pathway via targeting EMT-related TFs and down-regulating their mesenchymal markers. These findings possibly explain, at least in part, why TGF-ß exerts an opposite effect on EMT during trophoblast invasion and cancer progression.-Shih, J.-C., Lin, H.-H., Hsiao, A.-C., Su, Y.-T., Tsai, S., Chien, C.-L., Kung, H.-N. Unveiling the role of microRNA-7 in linking TGF-ß-Smad-mediated epithelial-mesenchymal transition with negative regulation of trophoblast invasion.

PEDF-derived peptide promotes tendon regeneration through its mitogenic effect on tendon stem/progenitor cells.

BACKGROUND: Tendon stem/progenitor cells (TSPC) exhibit a low proliferative response to heal tendon injury, leading to limited regeneration outcomes. Exogenous growth factors that activate TSPC proliferation have emerged as a promising approach for treatment. Here, we evaluated the pigment epithelial-derived factor (PEDF)-derived short peptide (PSP; 29-mer) for treating acute tendon injury and to determine the timing and anatomical features of CD146- and necleostemin-positive TSPC in the tendon healing process. METHODS: Tendon cells were isolated from rabbit Achilles tendons, stimulated by the 29-mer and analyzed for colony-forming capacity. The expression of the TSPC markers CD146, Oct4, and nestin, induced by the 29-mer, was examined by immunostaining and western blotting. Tendo-Achilles injury was induced in rats by full-thickness insertion of an 18-G needle and immediately treated topically with an alginate gel, loaded with 29-mer. The distribution of TSPC in the injured tendon and their proliferation were monitored using immunohistochemistry with antibodies to CD146 and nucleostemin and by BrdU labeling. RESULTS: TSPC markers were enriched among the primary tendon cells when stimulated by the 29-mer. The 29-mer also induced the clonogenicity of CD146+ TSPC, implying TSPC stemness was retained during TSPC expansion in culture. Correspondingly, the expanded TSPC differentiated readily into tenocyte-like cells after removal of the 29-mer from culture. 29-mer/alginate gel treatment caused extensive expansion of CD146+ TSPC in their niche on postoperative day 2, followed by infiltration of CD146+/BrdU- TSPC into the injured tendon on day 7. The nucleostemin+ TSPC were located predominantly in the healing region of the injured tendon in the later phase (day 7) and exhibited proliferative capacity. By 3 weeks, 29-mer-treated tendons showed more organized collagen fiber regeneration and higher tensile strength than control tendons. In culture, the mitogenic effect of the 29-mer was found to be mediated by the phosphorylation of ERK2 and STAT3 in nucleostemin+ TSPC. CONCLUSIONS: The anatomical analysis of TSPC populations in the wound healing process supports the hypothesis that substantial expansion of resident TSPC by exogenous growth factor is beneficial for tendon healing. The study suggests that synthetic 29-mer peptide may be an innovative therapy for acute tendon rupture.

Choroidal metastasis from esophageal squamous cell carcinoma.

We presented a rare case of a sole choroidal metastatic tumor from esophageal squamous cell carcinoma (ESCC) without other organ metastasis in Taiwan. A 43-year-old male with ESCC was referred for a 1-month history of decreased vision in his left eye. A 5.7 mm thick, yellow choroidal tumor occupied posterior pole, featured with pinpoint hyperfluorescence on angiography and subretinal fluid on optical coherence tomography. Positron emission tomography showed a singular hypermetabolic focus in the left eye. The tumor regressed with complete response and the vision preserved after radiation with total 57.60 gray applied by tomotherapy. The gastrointestinal system is the third most common metastatic origin in Taiwan while esophageal cancer metastasizing to choroid is rare. The discrepancy between the high prevalence of primary ESCC and the rareness of choroidal metastasis from ESCC is undetermined.

Pigment epithelium-derived factor short peptides facilitate full-thickness cutaneous wound healing by promoting epithelial basal cell and hair follicle stem cell proliferation.

A previous study by our group showed that a 44-amino-acid fragment of pigment epithelium-derived factor (PEDF) facilitated corneal epithelial wound healing. In the present study this fragment was shortened to obtain peptides of 18, 20 and 29 amino acids in length, and their promoting effects on the healing of full-thickness skin wounds were assessed. Peptides were delivered periodically by topical application to punch wounds of mice. The wound healing speed was evaluated by measuring the reduction of wound areas at 4 and 7 days after injury. Histological analysis with Masson's trichrome staining was used to confirm epithelialization and dermal collagen deposition. Proliferation of epithelial basal cells was documented by 5-bromo-2'-deoxyuridine incorporation. Hair follicle stem cells were identified by immunostaining for leucine-rich repeat-containing G protein-coupled receptor 6. The results indicated that the 20- and 29-amino-acid short peptides significantly reduced the time required for wound healing compared to the vehicle. Histological analysis confirmed faster epithelial cell coverage of open wounds. Treatment with the PEDF peptide fragments also contributed to granulation, tissue formation by increasing the fibroblast population and enhancing collagen deposition in the dermis. Wounds treated with PEDF peptide fragments contained more basal cells proliferated in the epithelium. Moreover, hair follicle stem cells were also stimulated to proliferate by peptide exposure. In conclusion, the present study reported the identification of two short peptides that can enhance the healing of full-thickness skin wounds following topical application. The underlying mechanisms may involve activation of basal cell proliferation and mobilization of hair follicle stem cells.

Real-World Experience with Half-Time Versus Half-Dose Photodynamic Therapy in Chronic Central Serous Chorioretinopathy.

PURPOSE: To evaluate the real-world experience with half-time photodynamic therapy (PDT) versus half-dose PDT for chronic central serous chorioretinopathy (CSC). METHODS: This multicenter retrospective study enrolled patients who received half-time PDT (with irradiation time shortened to 42 s) or half-dose PDT (with the dosage of verteporfin reduced to 3 mg/m2) for chronic CSC and who were followed up for ≧12 months. The success rate, central subfield retinal thickness (CST), and best-corrected visual acuity (BCVA) were documented in each group of patients. RESULTS: A total of 53 eyes from 49 patients were enrolled in this study. Seventeen eyes (15 patients) received half-time PDT and 36 eyes (34 patients) received half-dose PDT. The success rates in both groups were similar at 12 months (94.1% vs. 94.4%; P = 0.543). The mean CST at 1, 6, 12 months decreased significantly when compared with the baseline in both groups (all P < 0.001). The BCVA significantly improved at 6 and 12 months in both groups (all P < 0.05). There were no significant differences in changes of BCVA and changes of CST between the 2 groups at any time point. CONCLUSIONS: Half-time PDT is a feasible treatment for chronic CSC. It has success rates similar to half-dose PDT at 12 months. There were no significant differences in changes of BCVA and changes of CST between the 2 groups at 1, 6, and 12 months after treatment.

Clinical presentations of X-linked retinoschisis in Taiwanese patients confirmed with genetic sequencing.

PURPOSE: To investigate the clinical characteristics of X-linked retinoschisis (XLRS) and identify genetic mutations in Taiwanese patients with XLRS. METHODS: This study included 23 affected males from 16 families with XLRS. Fundus photography, spectral domain optical coherent tomography (SD-OCT), fundus autofluorescence (FAF), and full-field electroretinograms (ERGs) were performed. The coding regions of the RS1 gene that encodes retinoschisin were sequenced. RESULTS: The median age at diagnosis was 18 years (range 4-58 years). The best-corrected visual acuity ranged from no light perception to 20/25. The typical spoke-wheel pattern in the macula was present in 61% of the patients (14/23) while peripheral retinoschisis was present in 43% of the patients (10/23). Four eyes presented with vitreous hemorrhage, and two eyes presented with leukocoria that mimics Coats' disease. Macular schisis was identified with SD-OCT in 82% of the eyes (31/38) while foveal atrophy was present in 18% of the eyes (7/38). Concentric area of high intensity was the most common FAF abnormality observed. Seven out of 12 patients (58%) showed electronegative ERG findings. Sequencing of the RS1 gene identified nine mutations, six of which were novel. The mutations are all located in exons 4-6, including six missense mutations, two nonsense mutations, and one deletion-caused frameshift mutation. CONCLUSIONS: XLRS is a clinically heterogeneous disease with profound phenotypic inter- and intrafamiliar variability. Genetic sequencing is valuable as it allows a definite diagnosis of XLRS to be made without the classical clinical features and ERG findings. This study showed the variety of clinical features of XLRS and reported novel mutations.

Natural history of retinopathy of prematurity: two-year outcomes of a prospective study.

PURPOSE: To investigate the 2-year outcomes of the natural history of retinopathy of prematurity (ROP) in Taiwan. METHODS: A prospective study was conducted at two tertiary medical centers. Premature infants were screened and examined for ROP. The postmenstrual ages of developing each stage of ROP and the associated risk factors were recorded. RESULTS: A total of 698 infants were included. The incidences of ROP and treatment-requiring ROP in all patients with ROP were 29.7% and 37.2%. When only including patients with birth weight of 1,250 g or less, the incidences were 62.2% and 39.1%, respectively. In patients with ROP with birth weight over 1,250 g, 25% of them developed treatment-requiring ROP. The median postmenstrual ages for the development of Stage 1, Stage 2, and Type 1 ROP were 33.0, 34.0, and 34.7 weeks, respectively. Gestational age at birth and birth weight were the most important factors associated with treatment-requiring ROP (hazard ratios of 0.3 and 0.6). CONCLUSION: Our hospital-based study reveals an earlier postmenstrual age of developing ROP in this Asian population than in the Early Treatment for ROP study. Infants with birth weight over 1,250 g could still develop treatment-requiring ROP. Suboptimal oxygen control, different genetic dispositions among different races, inconsistencies in ROP diagnosis, and earlier screening might account for such a phenomenon.

Half-dose verteporfin combined with half-fluence photodynamic therapy for chronic central serous chorioretinopathy.

PURPOSE: This study evaluated the safety and efficacy of half-dose verteporfin combined with half-fluence photodynamic therapy (half-half photodynamic therapy (PDT) for chronic central serous chorioretinopathy (CSC). METHODS: This was a retrospective case series. Fourteen eyes with chronic CSC receiving half-half PDT were included in group 1. Another 28 eyes receiving half-dose verteporfin combined with standard fluence PDT were included in group 2 as a control group. Main outcome measures were the success rates, major complications, best-corrected visual acuity (BCVA), and central subfield foveal thickness (CFT) on optical coherence tomography (OCT) at 6 months in both groups. Success of treatment was defined as complete resolution of subretinal fluid on OCT after treatment without recurrence. RESULTS: There was no significant difference between groups in their age, gender, duration of symptoms, baseline BCVA, baseline CFT, PDT spot size, and follow-up duration. The success rate was 64% (9/14 eyes) in group 1 and 93% (26/28 eyes) in group 2 at 6 months (P=0.031). No major complications were found in either group. Mean CFT showed significant reduction at 6 months in both groups (-115 µm and P<0.001 in group 1; -150 µm and P<0.001 in group 2). The mean BCVA in group 2 improved significantly (P<0.001) at 6 months. The mean BCVA in group 1 showed a trend of improvement but was not statistically significant (P=0.25) at 6 months. CONCLUSIONS: Half-half PDT is a feasible treatment for chronic CSC. However, there was a lower success rate at 6 months compared with the control group.

Choroidal thickness in patients with a history of retinopathy of prematurity.

IMPORTANCE: The cause of reduced vision in patients with a history of retinopathy of prematurity (ROP) is not yet fully understood. The role of the choroid in ROP remains unknown and existing studies of choroidal thickness in patients with a history of ROP are limited. It might be helpful to understand the association of the choroid with ROP by measuring the choroidal thickness in patients with a history of ROP and correlating these findings with the visual outcome of these patients. OBJECTIVE: To examine choroidal thickness by spectral-domain optical coherence tomography in children with a history of ROP and assess the impact of choroidal thickness on visual acuity. DESIGN: A prospective cross-sectional analysis from August 2011 to September 2012. SETTING: Institutional referral centers. PARTICIPANTS: Children aged 6 to 14 years with a history of ROP were classified into the following 2 groups: patients with a history of threshold ROP and treatment with laser or cryotherapy (treated group) and those with regressed ROP who had not received any treatment (nontreated group). All of the patients had a normal-appearing posterior pole. INTERVENTION: Examinations of visual acuity, refractive errors, and optical components and measurement of choroidal thickness. MAIN OUTCOMES AND MEASURES: Best-corrected visual acuity, optical components, and optical coherence tomography findings. RESULTS: In total, 49 patients were enrolled in the study. Patients in the treated group had a significantly thinner choroidal thickness than the patients in the nontreated group after adjusting for age, axial length, and spherical power. Choroidal thickness was found to be positively associated with spherical power and spherical equivalent and negatively associated with axial length and vitreous depth. In addition, a thin choroidal thickness was associated with a worse best-corrected visual acuity. CONCLUSIONS AND RELEVANCE: Choroidal thickness is thinner in patients with threshold ROP compared with the patients with spontaneously regressed ROP. A thinner choroid is associated with worse vision in these patients. This study might imply the association of choroid circulation with ROP.