TAT Nanobody Exerts Antiviral Effect against PRRSV In Vitro by Targeting Viral Nucleocapsid Protein.

Porcine reproductive and respiratory syndrome (PRRS) is caused by the PRRS virus (PRRSV), which has brought huge economic losses to the pork industry worldwide since its first discovery in the late 1980s in North America. To date, there are no effective commercial vaccines or therapeutic drugs available for controlling the spread of PRRSV. Due to their unique advantages of high affinity and high specificity, nanobodies (Nbs) have received increasing attention in the process of disease diagnosis and treatment. Trans-activator transcription (TAT) can serve as a vector to carry specific proteins into cells by passing through cell membranes. In our previous study, a specific Nb against the PRRSV nucleocapsid (N) protein was screened using phage display technology. For this study, we developed a novel recombinant protein constituting a TAT-conjugated Nb, which we call TAT-Nb1. The target cell entry efficiency of TAT-Nb1 and its effect on PRRSV infection and replication were then investigated. Our results indicate that TAT delivered Nb1 into Marc-145 cells and porcine alveolar macrophages (PAMs) in a dose- and time-dependent manner. Furthermore, TAT-Nb1 dose-dependently suppressed PRRSV infection and replication, where this antiviral effect was independent of PRRSV strain. Co-immunoprecipitation results revealed that Nb1 efficiently interacted with the N protein of PRRSV. Taken together, the presented results suggest that TAT-Nb1 can effectively suppress PRRSV replication, and it may be considered as a new anti-PRRSV candidate drug.

Porcine reproductive and respiratory syndrome virus non-structural protein 4 cleaves guanylate-binding protein 1 via its cysteine proteinase activity to antagonize GBP1 antiviral effect.

Porcine reproductive and respiratory syndrome (PRRS) is a highly infectious disease caused by PRRS virus (PRRSV) that causes great economic losses to the swine industry worldwide. PRRSV has been recognized to modulate the host antiviral interferon (IFN) response and downstream interferon-stimulated gene expression to intercept the antiviral effect of host cells. Guanylate-binding proteins (GBPs) are IFN-inducible GTPases that exert broad antiviral activity against several DNA and RNA viruses, of which GBP1 is considered to play a pivotal role. However, the role of GBP1 in PRRSV replication remains unknown. The present study showed that overexpression of GBP1 notably inhibited PRRSV infection, while the knockdown of endogenous GBP1 promoted PRRSV infection. The K51 and R48 residues of GBP1 were essential for the suppression of PRRSV replication. Furthermore, GBP1 abrogated PRRSV replication by disrupting normal fibrous actin structures, which was indispensable for effective PRRSV replication. By using a co-immunoprecipitation assay, we found that GBP1 interacted with the non-structural protein 4 (nsp4) protein of PRRSV, and this interaction was mapped to the N-terminal globular GTPase domain of GBP1 and amino acids 1-69 of nsp4. PRRSV infection significantly downregulated GBP1 protein expression in Marc-145 cells, and nsp4, a 3C-like serine proteinase, was responsible for GBP1 cleavage, and the cleaved site was located at glutamic acid 338 of GBP1. Additionally, the anti-PRRSV activity of GBP1 was antagonized by nsp4. Taken together, these findings expand our understanding of the sophisticated interaction between PRRSV and host cells, PRRSV pathogenesis and its mechanisms of evading the host immune response.

HRP-conjugated-nanobody-based cELISA for rapid and sensitive clinical detection of ASFV antibodies.

African swine fever (ASF), which is caused by the ASF virus (ASFV), is a highly contagious hemorrhagic disease that causes high mortality to domestic porcine and wild boars and brings huge economic losses to world swine industry. Due to the lack of an effective vaccine, the control of ASF must depend on early, efficient, and cost-effective detection and strict control and elimination strategies. Traditional serological testing methods are generally associated with high testing costs, complex operations, and high technical requirements. As a promising alternative diagnostic tool to traditional antibodies, nanobodies (Nb) have the advantages of simpler and faster generation, good stability and solubility, and high affinity and specificity, although the system is dependent on the immunization of Bactrian camels to obtain the specific VHH library of the target protein. The application of Nbs in the detection of ASFV antibodies has not yet been reported yet. Using a phage display technology, one Nb against the ASFV p54 protein that exhibited high specificity and affinity, Nb8, was successfully screened. A HEK293T cell line stably expressing Nb8-horseradish peroxidase (HRP) fusion protein was established using the lentiviral expression system. Following the optimization of the reaction conditions, the Nb8-HRP fusion protein was successfully used to establish a competitive enzyme-linked immunosorbent assay (cELISA) to detect ASFV-specific antibodies in pig serum, for the first time. There was no cross-reaction with healthy pig serum, porcine pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), porcine epidemic diarrhea virus (PEDV), and classical swine fever virus (CSFV) positive sera. The optimal cut-off value for the cELISA by ROC analysis was 52.5%. A total of 209 serum samples were tested using the developed cELISA and a commercial ELISA kit. The results showed that the relative specificity of the cELISA was 98.97%, and the relative sensitivity of the cELISA was 93.3%, with the percent agreement between the two ELISA methods being 98.56%. In conclusion, a specific, sensitive, and repeatable cELISA was successfully developed based on the Nb8 as a probe, providing a promising method for the detection of anti-ASFV antibodies in clinical pig serum. KEY POINTS: • We successfully screened a specific, high affinity nanobody against ASFV p54 protein. • We establish a method for continuous and stable expression of Nb-HRP fusion protein using a lentiviral packaging system. • We establish a nanobody cELISA detection method that can monitor an ASF infection.

K205R specific nanobody-horseradish peroxidase fusions as reagents of competitive ELISA to detect African swine fever virus serum antibodies.

BACKGROUND: African swine fever virus (ASFV) is a highly contagious hemorrhagic disease and often lethal, which has significant economic consequences for the swine industry. Due to lacking of commercial vaccine, the prevention and control of ASF largely depend on early large-scale detection and screening. So far, the commercial ELISA kits have a long operation time and are expensive, making it difficult to achieve large-scale clinical applications. Nanobodies are single-domain antibodies produced by camelid animals, and have unique advantages such as smaller molecular weight, easy genetic engineering modification and low-costing of mass production, thus exhibiting good application prospects. RESULTS: The present study developed a new method for detection of ASFV specific antibodies using nanobody-horseradish peroxidase (Nb-HRP) fusion proteins as probe. By using camel immunization, phage library construction and phage display technology, five nanobodies against K205R protein were screened. Then, Nb-HRP fusion proteins were produced using genetic modification technology. Based on the Nb-HRP fusion protein as specific antibodies against K205R protein, a new type of cELISA was established to detect ASFV antibodies in pig serum. The cut-off value of the cELISA was 34.8%, and its sensitivity, specificity, and reproducibility were good. Furthermore, the developed cELISA exhibited 99.3% agreement rate with the commercial available ELISA kit (kappa value = 0.98). CONCLUSIONS: The developed cELISA method has the advantages of simple operation, rapid and low-costing, and can be used for monitoring of ASFV infection in pigs, thus providing a new method for the prevention and control of ASF.

A Network Model for Detecting Marine Floating Weak Targets Based on Multimodal Data Fusion of Radar Echoes.

Due to the interaction between floating weak targets and sea clutter in complex marine environments, it is necessary to distinguish targets and sea clutter from different dimensions by designing universal deep learning models. Therefore, in this paper, we introduce the concept of multimodal data fusion from the field of artificial intelligence (AI) to the marine target detection task. Using deep learning methods, a target detection network model based on the multimodal data fusion of radar echoes is proposed. In the paper, according to the characteristics of different modalities data, the temporal LeNet (T-LeNet) network module and time-frequency feature extraction network module are constructed to extract the time domain features, frequency domain features, and time-frequency features from radar sea surface echo signals. To avoid the impact of redundant features between different modalities data on detection performance, a Self-Attention mechanism is introduced to fuse and optimize the features of different dimensions. The experimental results based on the publicly available IPIX radar and CSIR datasets show that the multimodal data fusion of radar echoes can effectively improve the detection performance of marine floating weak targets. The proposed model has a target detection probability of 0.97 when the false alarm probability is 10-3 under the lower signal-to-clutter ratio (SCR) sea state. Compared with the feature-based detector and the detection model based on single-modality data, the new model proposed by us has stronger detection performance and universality under various marine detection environments. Moreover, the transfer learning method is used to train the new model in this paper, which effectively reduces the model training time. This provides the possibility of applying deep learning methods to real-time target detection at sea.

Debulking Surgery Combined with Low-Dose Oral Prednisolone and Azathioprine for Intractable IgG4-Related Orbital Disease: A Case Report.

A case of intractable IgG4-related orbital disease (IgG4-ROD) was successfully treated by debulking surgery combined with low-dose prednisolone and azathioprine as maintenance therapy. A 64-year-old man visited our clinic with progressive bilateral upper eyelid swelling and right eye fullness of a year's duration. He was previously treated with systemic corticosteroids for the IgG4-ROD and experienced a partial clinical response but relapsed upon prednisolone cessation. The patient underwent debulking surgery of the right lacrimal gland and right upper eyelid. His post-operative medication was oral prednisolone (5 mg) every other day and 50 mg azathioprine per day. The patient's right eye remained asymptomatic during the 18 months of follow-up. Debulking surgery combined with low-dose prednisolone and azathioprine, as a maintenance therapy, is an effective and alternative treatment for the long-term control of intractable IgG4-ROD.

Broad-band, reversible nonreciprocal light transmission based on a single nanocavity.

We present a general yet simple method for achieving broad-band, high-contrast-ratio, and reversible all-optical diodes. The mechanism is based on the controllable photonic transitions inside a nonlinear nanocavity, triggered by only one pulse. We demonstrate that the interaction between the signal, pump pulse, and the cavity mode plays a crucial role in dynamically controlling the nonreciprocal light transmissions. Using this mechanism, we show that the nonreciprocal light transmission can be controllably reversed without changing the signal light's wavelength or power, with a broad operation bandwidth over 7.5nm and a transmission contrast rate over 20dB. This approach provides a promising avenue in flexible manipulation of on-chip all-optical signal processing.

Efficient capture of aqueous humic acid using a functionalized stereoscopic porous activated carbon based on poly(acrylic acid)/food-waste hydrogel.

Stereoscopic porous carbons have shown good potential in humic acid (HA) removal. In this work, a novel stereoscopic porous activated carbon (SPAC) was designed and synthesized via the self-assembly of a hydrogel based on food waste during in-situ polymerization, vacuum drying, carbonization, and activation. Then, the SPAC was functionalized with 3-aminopropyltriethoxysilane (APTES) and the adsorption behavior of the modified SPAC (SPAC-NH2) was studied systematically. The effects of pH, contact time, initial concentration of HA, and adsorbent dose were investigated, showing that optimal HA removal efficiency (>98.0%) could be achieved at an initial HA concentration of 100 mg/L. The experimental adsorption isotherm data was fitted to the Langmuir model with a maximum adsorption capacity of 156.0 mg HA/g SPAC-NH2. Analysis of the mechanism indicated that the removal of HA was mainly realized through the amidization reaction between the COOH groups of HA and the NH2 groups of APTES. All of the above results showed that SPAC-NH2 powder is an efficient, eco-friendly, and reusable adsorbent which is suitable for the removal of HA from wastewater.

The healing effect of the collagen-glycosaminoglycan copolymer on corneal thinning.

BACKGROUND: To study the healing processes of partial thickness wounds in the adult rabbit cornea after grafting a porous collagen-glycosaminoglycan copolymer matrix (CG). METHODS: In this study, the regeneration of surgically-induced rabbit corneal defect implanted with CG was investigated. The corneal partial thickness wound was created by 7.5 mm trephine. The wound was implanted with CG. Effects on wound healing was analyzed using clinical data on epithelial migration and corneal thickness, and histological data on collagen and alpha smooth muscle actin distribution. RESULTS: Compared with control group, CG induced a relatively severe inflammatory reaction in grafted cornea until the CG matrix was completely degraded. The new vessel ingrowth and stromal regeneration maintained the corneal thickness. The grafted cornea was significantly thicker (P < 0.001) than the control group. On day 90, the corneal opacity score of the control group was one and the grafted cornea was two. CONCLUSION: CG copolymer matrix can successfully repair the damaged corneal stroma by injury, and regain its thickness. However, CG matrix induced inflammatory healing process thus causing mild corneal haziness and neovascularization.

Correction of involutional entropion with retractor redirection.

The study aims to report the surgical outcome of a retractor redirection procedure for involutional entropion repair for Asians. The study included all cases diagnosed with involutional entropion and significant ocular irritation who presented from 2008 to 2012. Sixty-seven eyelids in 54 patients were included in this study. All cases were operated on by one surgeon and had a minimum of 12-months follow-up. Success was defined as cases showing no recurrence of entropion with forceful eyelid squeezing postoperatively. A retrospective chart review was performed to assess the success rate, recurrences and complications of the procedure. During a mean follow-up period of 26.2 months (range, 12-53 months), 5 patients died during the study period. Two eyelids (3%) of one patient had a recurrence at 34 months postoperatively. One eyelid (1.5%) with a significant horizontal laxity developed postoperative ectropion and required a secondary horizontal shortening procedure. No other postoperative complications or dissatisfaction were reported. The retractor redirection procedure aims to repair the retractors and prevent orbicularis muscle overriding via inserting the retractors to the anterior lamellae. It yields a long-term success rate of 95.5% and is an effective technique for correcting involutional entropion.

Expression of retinoic acid-binding proteins and retinoic acid receptors in sebaceous cell carcinoma of the eyelids.

BACKGROUND: Sebaceous cell carcinoma of the eyelid is a malignant tumor. However, the pathoetiology of sebaceous cell carcinoma is not clear. Retinoic acid (RA) signaling is essential for skin epidermal differentiation including the eyelids. In this study, we investigate the expression of ß-catenin, RA-binding proteins and RA receptors in sebaceous cell carcinoma of the eyelid and try to estimate their influence on its pathoetiology. METHODS: Retrospective, noncomparative, consecutive interventional case series. Sixteen cases of eyelid sebaceous gland carcinoma who received tumor excision at our hospital between 2001 and 2011 were included. Immunohistochemical staining for ß-catenin, cellular retinoic acid binding protein 1 (CRABP1), cellular retinoic acid binding protein 2 (CRABP2), fatty acid-binding protein 5 (FABP5), retinoic acid receptors (RAR-α, -ß, -γ), and retinoid X receptors (RXR-α, -ß, -γ) was performed on tissue samples obtained from tumor excision. RESULTS: Of the 16 sebaceous cell carcinoma cases reviewed, six were male and 10 female. The mean follow-up period was 6.7 ± 3.66 years (range, 0.3-13 years). Of these 16 cases, the expression of ß-catenin was significantly increased in sebaceous cell carcinoma cases. CRABP1 was similarly expressed in the sebaceous cell carcinoma and control groups. CRABP2 and FABP5 were expressed in hair follicles of lid skin in both groups, whereas the CRABP2 and FABP5 were aberrantly expressed in the tumor cells of the sebaceous glands. Notably, the expression of retinoic acid receptor (RAR-ß) and retinoid X receptors (RXR-ß, -γ) was significantly upregulated in sebaceous cell carcinoma of the eyelids. CONCLUSIONS: Our findings indicate that retinoic acid signaling is related to the pathogenesis of sebaceous cell carcinoma of the eyelids.

To explore the neonatal nurses' beliefs and attitudes towards caring for dying neonates in Taiwan.

(1) To explore attitudes and beliefs of neonatal nurses toward nursing care for dying neonates; (2) to estimate the influence of neonatal nurses' personal and professional characteristics on their attitudes towards end-of life care for dying infants. A cross-sectional design was used. A questionnaire was used to collect data from 80 neonatal nurses. Research setting was four level III NICUs at four medical centers around the central region of Taiwan. Research participants were neonatal nurses who had worked for at least 1 year in one of level III NICUs and had been directly involved with the care of dying infants. Research participants were 80 neonatal nurses (response rate 100 %). Research findings identified eight barriers hindering neonatal palliative care practice. These barriers were insufficient communication due to the lack of an in-service educational program; the lack of available counseling help for neonatal clinicians; inability to express personal opinions, values and beliefs towards neonatal palliative care; insufficient staffing; the lack of unit policies/guidelines for supporting palliative care; the technological imperative; parental demands and personal beliefs about death and previous experience caring for dying infants. Further studies are needed to explore each barrier and to provide in-service neonatal palliative care educational programs that are needed to decrease these barriers.

Identification of a Linear B Cell Epitope on p54 of African Swine Fever Virus Using Nanobodies as a Novel Tool.

African swine fever (ASF) has received great attention from the swine industry due to the pandemic and the lack of vaccines or effective treatments. In the present study, 13 African swine fever virus (ASFV) p54-specific nanobodies (Nbs) were successfully screened based on Bactrian camel immunization of p54 protein and phage display technology, and their reactivity with the p54 C-terminal domain (p54-CTD) was determined; however, only Nb8-horseradish peroxidase (Nb8-HRP) exhibited the best reactivity. Immunoperoxidase monolayer assay (IPMA) and immunofluorescence assay (IFA) results indicated that Nb8-HRP specifically reacted with ASFV-infected cells. Then, the possible epitopes of p54 were identified using Nb8-HRP. The results showed that Nb8-HRP could recognize p54-CTD truncated mutant p54-T1. Then, 6 overlapping peptides covering p54-T1 were synthesized to determine the possible epitopes. Dot blot and peptide-based enzyme-linked immunosorbent assay (ELISA) results suggested that one novel minimal linear B cell epitope, 76QQWVEV81, which had never been reported before, was identified. Alanine-scanning mutagenesis revealed that 76QQWV79 was the core binding site for Nb8. Epitope 76QQWVEV81 was highly conserved among genotype II ASFV strains and could react with inactivated ASFV antibody-positive serum from naturally infected pigs, indicating that it was a natural linear B cell epitope. These findings provide valuable insights for vaccine design and p54 as an effective diagnostic tool. IMPORTANCE The ASFV p54 protein plays an important role in inducing neutralization antibodies in vivo after viral infection and is often used as a candidate protein for subunit vaccine development. The full understanding of the p54 protein epitope provides a sufficient theoretical basis for p54 as a vaccine candidate protein. The present study uses a p54-specific nanobody as a probe to identify a highly conserved antigenic epitope, 76QQWVEV81, among different ASFV strains, and it can induce humoral immune responses in pigs. This is the first report using virus-specific nanobodies as a tool to identify some special epitopes that cannot be recognized by conventional monoclonal antibodies. This study opens up nanobodies as a new tool for identifying epitopes and also provides a theoretical basis for understanding p54-induced neutralizing antibodies.

Goldenhar syndrome: a cause of secondary immunodeficiency?

Goldenhar syndrome (GS) results from an aberrant development of the 1st and 2nd branchial arches. There is a wide range of clinical manifestations, the most common being microtia, hemifacial microsomia, epibulbar dermoids and vertebral malformations. We present two cases of GS and secondary immunodeficiency due to anatomical defects characteristic of this disorder. Case 1 (3-year-old female) averaged 6 episodes of sinusitis and otitis media per year. Case 2 (7-year-old female) also had recurrent otitis media, an episode of bacterial pneumonia, and 2 episodes of bacterial meningitis. Their immune evaluation included a complete blood count with differential, serum immunoglobulin levels and specific antibody concentrations, lymphocyte phenotyping, and mitogen and antigen responses, the results of which were all within normal ranges. Both children demonstrated major structural abnormalities of the inner and middle ear structures, retention of fluid in mastoid air cells, and chronic sinusitis by computed tomography. These two cases illustrate how a genetically-associated deviation of the middle ear cleft can cause recurrent infections and chronic inflammation of the middle ear and adjacent sinuses, even meninges, leading to a greatly reduced quality of life for the child and parents.

Fabrication of a facial prosthesis for a 13-year-old child by using a point-and-shoot three-dimensional scanner and CAD/CAM technology.

Patients cannot wear ocular prostheses after undergoing orbital exenteration. They require a facial prosthesis to obtain a more favorable appearance, which greatly affects their social life and psychological health. In addition, conventional prosthesis-making processes require substantial time and expense. The economic burden is particularly heavy on children, who may require many prosthesis replacements as they mature. We report a method of fabricating a facial prosthesis by three-dimensional (3D) facial scanning and 3D printed for a 13-year-old girl who underwent partial orbital exenteration for malignant ciliary body medulloepithelioma 2 years ago. The patient's facial contour was captured with a hand-held, point-and-shoot 3D scanner. A facial prosthesis was designed using a mirror image technique with 3D modeling software and 3D printed. The prosthesis was then postprocessed and cast in silicone rubber. An ocular prosthesis was integrated into the facial prosthesis. The prosthesis was retained by prosthetic adhesives. This digitally assisted, impression-free method may lower the cost and effort of making facial prostheses and improve patient comfort, especially for children.

Equivalent legibility font size for traditional Chinese character compared to early treatment diabetic retinopathy study near visual acuity.

PURPOSES: To investigate the legibility of a standardized logarithmic print size of traditional Chinese (TC) characters and compare it with Early Treatment Diabetic Retinopathy Study (ETDRS) near chart. MATERIALS AND METHODS: A total of 1243 commonly used TC characters were chosen and divided into three groups according to its stroke complexity: Group A with 2-9 strokes, Group B with 10-17 strokes, and Group C with 18-25 strokes. For each group of characters, near charts were created using randomly chosen characters arranged in decreasing logarithmic size. In a well-illuminated room, healthy controls were fully corrected to test both ETDRS near chart and our set of TC near charts. The smallest legible font sizes (SLFS) in TC near charts were recorded and analyzed. RESULTS: Forty-two healthy eyes (21 participants) (age 29 ± 8.9 years old) were included. The mean near best-corrected visual acuity (nBCVA) in ETDRS chart was 0.06 ± 0.05 logMAR. We found that the mean SLFS in TC charts (0.33 ± 0.09 logMAR) was significantly larger than the nBCVA in ETDRS chart (P < 0.001). The SLFS of Group B and the SLFS of Group C was significantly larger than that of Group A (P < 0.001). CONCLUSION: According to our results, to recognize TC characters, normal-sight readers need a 0.22-0.30 logMAR (1.7-2.0 fold) enlargement of the acuity size measured by ETDRS near chart. The low-stroke TC charts may provide a new method to assess the postsurgical outcomes for comparable functional visual acuity in reading TC characters.

Comparing pericanalicular sutures with direct canalicular wall sutures for canalicular laceration.

PURPOSE: To determine whether a statistically significant difference exists between direct canalicular wall sutures (DCs) and pericanalicular sutures (PCs) in the repair of traumatic canalicular lacerations. METHODS: The medical records of 63 patients who underwent primary repairs for traumatic canalicular lacerations were retrospectively reviewed. Patients were divided in 2 groups according to the suturing techniques used: the DC group (n = 41) and the PC group (n = 22). Anatomic results were compared between these 2 groups. A successful result was determined by attempted irrigation and probing of the injured canaliculus at the last follow-up visit. RESULTS: There were 6 failed procedures among the patients who underwent pericanalicular repair and 1 failed procedure among the patients who underwent direct canalicular wall repair (p = 0.024). CONCLUSIONS: These data support the higher success rates in patients treated with direct canalicular repair compared with pericanalicular repair.

An Association Rule Analysis of Combined Acupoints for the Treatment of Patients with Dry Eye Disease.

BACKGROUND: Dry eye disease (DED) has a higher incidence in old age and is seen predominantly in females worldwide. Neurosensory abnormalities, ocular surface inflammation and damage, film instability, and hyperosmolarity are major and proven pathologies responsible for a poor quality of life. Tear breakup time and Schirmer's I test are predominantly used for the evaluation of primary outcomes in patients undergoing conventional treatment. A previous meta-analysis of some relevant studies proved that combination of acupoints could be more effective than single acupoint treatment. OBJECTIVES: The present study aimed to undertake association rule mining and examined the potential kernel acupoint combination in DED treatment constructed from the extracted randomized controlled trials (RCTs) based on a previous meta-analysis. METHODS: We summarized 32 acupoints as binary data from the 12 eligible RCTs and analyzed them based on the Apriori algorithm. RESULTS: TE23, BL2, ST2, ST1, EX-HN5, BL1, LI4, ST36, SP6, and KI3 were the 10 most frequently selected acupoints. The major associated rules in combination of acupoints were {TE23, LI4} ≥ {ST1} and {TE23, ST1} ≥ {LI4}, as inferred from 23 association rules. CONCLUSIONS: For acupuncture treatment of DED, combined TE23, LI4, and ST1 acupoints could be settled as the kernel of acupoint combination.

Sec15 interacts with Rab11 via a novel domain and affects Rab11 localization in vivo.

Sec15, a component of the exocyst, recognizes vesicle-associated Rab GTPases, helps target transport vesicles to the budding sites in yeast and is thought to recruit other exocyst proteins. Here we report the characterization of a 35-kDa fragment that comprises most of the C-terminal half of Drosophila melanogaster Sec15. This C-terminal domain was found to bind a subset of Rab GTPases, especially Rab11, in a GTP-dependent manner. We also provide evidence that in fly photoreceptors Sec15 colocalizes with Rab11 and that loss of Sec15 affects rhabdomere morphology. Determination of the 2.5-A crystal structure of the C-terminal domain revealed a novel fold consisting of ten alpha-helices equally distributed between two subdomains (N and C subdomains). We show that the C subdomain, mainly via a single helix, is sufficient for Rab binding.

Long-Term Results of Autologous Auricular Cartilage Graft Applied in Anophthalmic Orbits Unable to Wear Prosthesis.

In anophthalmic patients, shallow lower fornices make wearing ocular prostheses impossible and maintaining normal social activities difficult. This study retrospectively investigated the long-term surgical outcomes of autologous auricular cartilage grafting for contracted orbits. From 1995 to 2013, 29 anophthalmic contracture sockets with inadequate lower fornices and poor prosthesis retention presented to Chang Gung Memorial Hospital in Linkou, Taiwan, were treated using this surgical method. The success rate, aesthetic outcome, recurrence, and complications were analyzed. Among the 29 patients, 15 were women, 14 were men, their mean age was 45 years, and the mean follow-up time was 52 months (range = 6-159 months). Satisfactory lid position was achieved in 25 cases (86%), and lower fornix retraction recurred in four cases (14%). Neither donor site morbidity nor auricular deformity was noted during the follow-up period. Therefore, an auricular cartilage graft can be used successfully as a compatible spacer for anophthalmic patients with shallow lower fornices and prosthesis-fitting problems in long-term follow-up.