Progressive ulcerative keratitis in dogs in the United Kingdom: Microbial isolates, antimicrobial sensitivity, and resistance patterns.

OBJECTIVES: The objective of the study was to identify bacterial pathogens and their antimicrobial sensitivity profile associated with cases of canine progressive ulcerative keratitis. MATERIALS AND METHODS: Analysis of microbial culture and sensitivity results from dogs with progressive ulcerative keratitis presenting to a UK referral practice between December 2018 and August 2020. RESULTS: Positive bacterial cultures were obtained from 80/148 (54%) of the canine ulcers sampled with 99 bacterial isolates cultured. Streptococcus canis (n = 29), Pseudomonas aeruginosa (n = 19), and Staphylococcus pseudintermedius (n = 16) were the most common isolates. Pseudomonas aeruginosa was more likely to be isolated whether the ulcer was clinically malacic at the time of sampling (OR = 10.1, p < .001). Ulcers treated prior to culture with fusidic acid were 7.6 times more likely to be positive than those treated with any other antimicrobial(s). Bacterial isolates demonstrated resistance against neomycin (85%), fusidic acid (78%), and tetracycline (68%). Conversely, isolates were most likely to be sensitive to gentamicin (88%), ofloxacin (77%), ciprofloxacin (73%), and chloramphenicol (64%). Antimicrobial combinations of chloramphenicol or gentamicin with a fluoroquinolone (ofloxacin or ciprofloxacin) or chloramphenicol combined with gentamicin were the most effective on in vitro analysis (over 90% susceptibility of all isolates). CONCLUSION: The most common bacterial species associated with canine progressive ulcerative keratitis in a UK referral population were S. canis, P. aeruginosa, and S. pseudintermedius. Combination antimicrobial therapy is recommended pending culture and sensitivity results given the varied antimicrobial susceptibility profiles and significant bacterial in vitro resistance to antimicrobial monotherapy.

Estimation of the intraoperative blood loss in dogs undergoing enucleation.

PURPOSE: To quantify the surgical blood loss during canine enucleation and to investigate the relationship between this and any patient, surgical, and anesthetic factors. METHODS: A prospective observational analysis was conducted on 121 client-owned dogs (130 eyes) undergoing enucleation at a referral ophthalmology clinic. Blood loss was estimated by the gravimetric method (weight difference between dry and blood-containing surgical materials) to provide absolute blood loss (ABL) in milliliters, expressed as a percentage of circulating blood volume, to establish relative blood loss (RBL). RESULTS: Median ABL was 12 ml (1.6-116 ml), and median RBL was 1.3% (0.1%-6.7%). A higher RBL was associated with the following: use of a bupivacaine splash block versus retrobulbar nerve block (1.9 vs. 1%; p < .001), transpalpebral versus subconjunctival approach (2.2 vs. 1.3%; p = .003), and small versus large breed dogs (1.7% vs. 1.1%; p = .001). Both ABL and RBL differed significantly between surgeons. There was no significant difference in hemorrhage associated with the presence of ocular hypertension, systemic illness, surgical time, administration of meloxicam or choice of pre-medicant (acepromazine vs medetomidine). No dog required supportive intervention in response to surgical hemorrhage. CONCLUSIONS: This study has established a surgical blood loss estimate for dogs undergoing enucleation at an ophthalmology referral centre. Subconjunctival enucleation may be preferred for patients at greater risk of haemodynamic complications.

Bilateral third eyelid ectopic cilia in an adult horse.

A 15-year-old Cob mare presented with a 4-month history of chronic epiphora and intermittent blepharospasm in the right eye. On ophthalmic examination, two translucent aberrant hairs were identified at the third eyelid margin corresponding to an area of corneal fibrosis and neovascularization. Partial excision of the third eyelid was performed, and histopathology confirmed ectopic hair follicles. Two weeks later, clinical signs recurred in the same eye. Examination revealed another pair of aberrant hairs on the bulbar surface of the third eyelid near its leading edge. This portion of the third eyelid was also excised, and histopathology confirmed two additional ectopic hair follicles. Eight months later, the horse developed similar clinical signs in the left eye. Ophthalmic examination showed a single aberrant translucent hair at the third eyelid margin associated with focal fibrosis and neovascularization of the ventromedial cornea. Partial excision of the third eyelid was performed, and histopathology confirmed an ectopic hair follicle within the third eyelid conjunctiva. Excision was curative at 4 years postoperatively with no further recurrence in either eye.

Clinical and pathological findings in three dogs with a corneocentric presentation of nodular granulomatous episcleritis.

PURPOSE: To describe the clinical signs, management, histopathologic findings, and outcome of three dogs with a corneocentric presentation of nodular granulomatous episcleritis (NGE). METHODS: Three dogs of varying breeds were presented for a unilateral, nonpainful, and infiltrative corneal lesion in the dorsal aspect of the eye. Clinical response to symptomatic topical treatment directed at a presumed inflammatory or immune-mediated cause was poor. Due to this, and concerns of neoplasia, ultrasonography (n = 1), incisional biopsy (n = 2), and/or enucleation (n = 2) were performed. RESULTS: The inflammatory infiltrate observed on histopathology was identical to that seen in nodular granulomatous episcleritis in all three cases. However, atypically the inflammation was confined to the cornea and limbus, without episcleral or conjunctival involvement. Inflammation of the cornea was full thickness to Descemet's membrane. Following enucleation (n = 2), there were no postoperative complications, and no reported ophthalmic disease in the remaining eye. Currently, the single non-enucleated case remains controlled with systemic and topical immunosuppression. CONCLUSION: To the best of the authors' knowledge, this is the first report of an NGE condition purely affecting the full thickness of the cornea, without episcleral or conjunctival involvement. The authors propose this to represent an atypical corneocentric variant of NGE. This clinical presentation can resemble neoplasia; incisional biopsy is recommended for a definitive diagnosis. Further research into the optimal treatment strategies for this variant of NGE is required.

An investigation into the bacterial contamination of goniolenses in use in clinical practice.

Objectives To report the incidence and evaluate the clinical significance of goniolens bacterial contamination in clinical use in dogs with three different usage protocols and one with an added cleaning protocol. Animals Studied and Methods Three groups of twenty dogs undergoing gonioscopy at a private practice in the UK had the goniolenses swabbed for bacteriology culture and identification prior to placement on the cornea. Three protocols of lens use, with 2 different types of goniolens, were studied. One protocol was then repeated with 21 dogs with a lens cleaning protocol prior to storage. Results Low levels of bacterial contamination were found in all 3 initial groups (10-15%). No correlation was found between usage protocol used and rate of contamination and no correlation was found between length of storage between use and contamination. All bacteria cultured were considered naturally occurring commensals for the canine eye and environment. The group with a cleaning protocol had a 4.7% contamination rate. This was not statistically different from the non-cleaning groups. Conclusions The rate of bacterial contamination of goniolenses in clinical practice is low and the bacterial contaminants consist of commensal bacteria, unlikely to be of detriment to the eye. Minimal contamination of the goniolenses was found and this did not appear to be of clinical significance. The introduction of a simple cleaning protocol did not produce a statistically significant reduction in bacterial contamination.

Ocular biometry by computed tomography in different dog breeds.

OBJECTIVE: To (i) correlate B-mode ocular ultrasound (US) and computed tomography (CT) (prospective pilot study), (ii) establish a reliable method to measure the normal canine eye using CT, (iii) establish a reference guide for some dog breeds, (iv) compare eye size between different breeds and breed groups, and (v) investigate the correlation between eye dimensions and body weight, gender, and skull type (retrospective study). PROCEDURE: B-mode US and CT were performed on ten sheep cadaveric eyes. CT biometry involved 100 adult pure-bred dogs with nonocular and nonorbital disease, representing eleven breeds. Eye length, width, and height were each measured in two of three planes (horizontal, sagittal, and equatorial). RESULTS: B-mode US and CT measurements of sheep cadaveric eyes correlated well (0.70-0.71). The shape of the canine eye was found to be akin to an oblate spheroid (a flattened sphere). A reference guide was established for eleven breeds. Eyes of large breed dogs were significantly larger than those of medium and small breed dogs (P < 0.01), and eyes of medium breed dogs were significantly larger than those of small breed dogs (P < 0.01). Eye size correlated with body weight (0.74-0.82) but not gender or skull type. CONCLUSIONS: Computed tomography is a suitable method for biometry of the canine eye, and a reference guide was established for eleven breeds. Eye size correlated with breed size and body weight. Because correlation between B-mode US and CT was shown, the obtained values can be applied in the clinical setting, for example, for the diagnosis of microphthalmos and buphthalmos.

Outcome of phacoemulsification following corneal and lens laceration in cats and dogs (2000-2010).

OBJECTIVE: To investigate the success rate of phacoemulsification following corneal and lens laceration in dogs and cats. PROCEDURE: Retrospective review of cats and dogs presenting with corneal and lens laceration and treated with phacoemulsification. RESULTS: The records of 33 patients (33 eyes: six feline, 27 canine) presenting to a private referral center were reviewed. Affected dogs were younger (median 18 months) than affected cats (median 30 months). The lacerations were caused by cat scratch trauma (9/33), thorn injury (6/33), and glass shards (1/33); the cause was unknown in 17/33 cases. All cats and 85.2% of all dogs were visual at the last examination. The median follow-up was 4 and 8 months for cats and dogs, respectively. In all canine cases that developed vision loss, this occurred within the first 14 weeks postoperatively. The ultimate cause for vision loss in dogs was secondary glaucoma (4/4) and retinal detachment (1/4). CONCLUSION: Cats have an excellent outcome and dogs a very good outcome following surgery for corneal and lens laceration. The cause of the trauma, the size of the lesion, the time interval between the ocular trauma and surgery, and the type of surgery were not found to have an influence on the outcome of patients in this study. We postulate that vision loss might develop more often in cases with complications associated with the initial corneal laceration wound.

Study of Preoperative Radiotherapy for Sarcomas of the Extremities with Intensity-Modulation, Image-Guidance and Small Safety-margins (PREMISS).

BACKGROUND: The aim of the trial is to demonstrate that with the use of modern IMRT/IGRT and reduction of safety margins postoperative wound complications can be reduced. METHODS/ DESIGN: The trial is designed as a prospective, monocentric clinical phase II trial. The treatment is performed with helical IMRT on the Tomotherapy HiArt System© or with RapidArc© IMRT as available. All treatments are performed with 6 MV photons and daily online CT-based IGRT. A dose of 50 Gy in 2 Gy single fractions (5 fractions per week) is prescribed. Restaging including MRI of the primary tumor site as well as CT of the thorax/abdomen is planned 4 weeks after RT. PET-examinations or any other imaging can be performed as required clinically. In cases of R1 resection, brachytherapy is anticipated in the 2nd postoperative week. Brachytherapy catheters are implanted into the tumor bed depending on the size and location of the lesion. Surgery is planned 5-6 weeks after completion of neoadjuvant RT. All patients are seen for a first follow-up visit 2 weeks after wound healing is completed, thereafter every 3 months during the first 2 years. The endpoints of the study are evaluated in detail during the first (2 weeks) and second (3 months) follow-up. Functional outcome and QOL are documented prior to treatment and at year 1 and 2. Treatment response and efficacy will be scored according to the RECIST 1.1 criteria. A total patient number of 50 with an expected 20% rate of wound complications were calculated for the study, which translates into a 95% confidence interval of 10.0-33.7% for wound complication rate in a binomial distribution. DISCUSSION: The present study protocol prospectively evaluates the use of IMRT/IGRT for neoadjuvant RT in patients with soft tissue sarcomas of the extremity with the primary endpoint wound complications, which is the major concern with this treatment sequence. Besides complications rates, local control rates and survival rates, as well as QOL, functional outcome and treatment response parameters (imaging and pathology) are part of the protocol. The data of the present PREMISS study will enhance the current literature and support the hypothesis that neoadjuvant RT with IMRT/IGRT offers an excellent risk-benefit ratio in this patient population. TRIAL REGISTRATION: NCT01552239.

Risk of very low birth weight based on perinatal periods of risk.

OBJECTIVE: To determine the risk factors associated with having a very low birth weight (VLBW) infant as a follow-up to the first phase of a Perinatal Periods of Risk approach. DESIGN AND SAMPLE: Retrospective cohort analysis of birth certificates. Population-based sample of 53,427 birth certificates for the city under study during the years 1999-2006. MEASURES: The relationship of selected maternal characteristics as predictors of VLBW using multivariate logistic regression analysis. RESULTS: The maternal characteristics associated with VLBW were as follows: no prenatal care (OR = 4.04), inadequate weight gain (OR = 3.97), Black, non-Hispanic race (OR = 1.50), less than 20 years old (OR = 1.42) and more than 35 years old (OR = 1.43). After analyzing age and race/ethnicity together, Black non-Hispanic women less than 20 years of age (OR = 2.70) or over 35 years of age (OR = 2.45) still had an increased odds for having a VLBW infant whereas Black non-Hispanic women between the ages of 20 and 35 did not. CONCLUSIONS: The findings of this study suggest educating women on the importance of preconception care, prenatal care, and adequate pregnancy weight gain to reduce the odds of having a VLBW infant.

Choline and Betaine Levels in Plasma Mirror Choline Intake in Very Preterm Infants.

Choline is essential for cell membrane formation and methyl transfer reactions, impacting parenchymal and neurological development. It is therefore enriched via placental transfer, and fetal plasma concentrations are high. In spite of the greater needs of very low birth weight infants (VLBWI), choline content of breast milk after preterm delivery is lower (median (p25-75): 158 mg/L (61-360 mg/L) compared to term delivery (258 mg/L (142-343 mg/L)). Even preterm formula or fortified breast milk currently provide insufficient choline to achieve physiological plasma concentrations. This secondary analysis of a randomized controlled trial comparing growth of VLBWI with different levels of enteral protein supply aimed to investigate whether increased enteral choline intake results in increased plasma choline, betaine and phosphatidylcholine concentrations. We measured total choline content of breast milk from 33 mothers of 34 VLBWI. Enteral choline intake from administered breast milk, formula and fortifier was related to the respective plasma choline, betaine and phosphatidylcholine concentrations. Plasma choline and betaine levels in VLBWI correlated directly with enteral choline intake, but administered choline was insufficient to achieve physiological (fetus-like) concentrations. Hence, optimizing maternal choline status, and the choline content of milk and fortifiers, is suggested to increase plasma concentrations of choline, ameliorate the choline deficit and improve growth and long-term development of VLBWI.

Parotid duct transposition in dogs: a retrospective review of 92 eyes from 1999 to 2009.

OBJECTIVE: The aim of this study was to perform a retrospective review of parotid duct transposition (PDT) in the dog to determine the rate and nature of complications, the success and failure rates and to evaluate owner satisfaction. METHODS: Medical records of 56 dogs (92 eyes) that underwent PDT and subsequent follow-up by a veterinary ophthalmologist were reviewed. Forty owners (40 dogs/66 eyes) were contacted by telephone and 37 owners (37 dogs/60 eyes) also completed a visual analog scale questionnaire. Statistical evaluation included Wilcoxon Signed Rank tests, one-way analysis of variance and Kaplan-Meier survival analysis with Wilcoxon and Log-rank tests. RESULTS: The mean follow-up was 38.7 months (range 1-109 months). The surgical success rate was 92% (85/92 eyes). Total failures (8%, 7/92 eyes) were because of severe saliva intolerance (n = 5 eyes) and PDT failure (n = 2 eyes). The complication rate was 50% (46/92 eyes) of which 61% (28/46 eyes) were managed medically and 39% (18/46 eyes) required further surgery. Ninety percent (36/40) of owners indicated that they would proceed with surgery again. Statistically significant improvements in owner perception of ocular comfort, number of daily topical treatments, ocular wetness, and postoperative vision were identified. CONCLUSIONS: This study shows that PDT is a successful procedure based on clinical findings and in terms of owner perception. It has also demonstrated that PDT improves ocular comfort and vision in medically refractive cases of keratoconjunctivitis sicca, and that a low level of on-going management is required in 33% of cases.

Cataracts are not associated with retinal detachment in the Bichon Frise in the UK--a retrospective study of preoperative findings and outcomes in 40 eyes.

OBJECTIVE: The aim of this study was to evaluate whether the Bichon Frise population in the UK is at the same risk of developing retinal detachment in association with cataract formation and following phacoemulsification as described in reports from the USA. PROCEDURES: The medical records of Bichon Frises which were presented for cataract assessment and of those which were treated with phacoemulsification at Willows Referral Service between 1997 and 2009 were reviewed. RESULTS: Forty eyes (26 dogs) with unilateral or bilateral cataracts were included in the study. There was no evidence of retinal detachment associated with the cataracts at initial presentation. Phacoemulsification was performed on 34 eyes (20 dogs). Clinically evident lens-induced uveitis was treated preoperatively in 17/34 eyes. Artificial lens implantation was carried out in 30/34 eyes; automated anterior vitrectomy was performed in 7/34 eyes. The mean follow-up time was 16.6 months (range 1.5-73 months). At the last re-examination, 31/34 eyes (91.2%) were visual. Three eyes (8.8%) were blind--two (in the same dog) because of presumptive bilateral optic nerve disease and one because of uveitis and secondary glaucoma. There was no evidence of retinal detachment following phacoemulsification in any of the 34 eyes. CONCLUSION: This study suggests that the Bichon Frise population in the UK does not appear to have a predisposition for retinal detachment in association with cataract formation or following cataract surgery. Prophylactic random transscleral laser retinopexy or transscleral cryopexy cannot therefore be routinely recommended for Bichon Frises with cataracts in the UK.

Lower lid entropion in dogs: A modified technique of the combined Hotz-Celsus and wedge resection procedure.

BACKGROUND: This study aimed to describe a modification of the combined Hotz-Celsus and wedge resection technique for the treatment of lower lid entropion in dogs and evaluate its success rate. METHODS: To reduce tissue trauma, facilitate tissue handling and shorten the surgical time, shortening of the eyelid was performed by excising a rectangular piece of the eyelid margin only, in the central section, extending to but not below the first Hotz-Celsus incision. The eyelid margin surgical wound was then sutured before excision of the Hotz-Celsus crescent which had been surgically scored at the start of surgery. Records of 31 dogs were reviewed for signalment, outcome, need for further surgery, and follow up from 6 to 48 months after surgery. RESULTS: Sixty-one eyes with lower lid entropion were treated. The most common breeds were English bulldog and English cocker spaniel and the median age was 16 months. The success rate for a single entropion correction surgery was 98.4%. One eyelid required a second surgical correction and four eyelids of three large breed dogs had wound breakdown. CONCLUSIONS: This modified technique is successful at correcting lower eyelid entropion associated with overlong eyelid length in dogs while being technically easier to perform.

Longitudinal analysis of quality of life in patients receiving conformal radiation therapy for prostate cancer.

PURPOSE: To prospectively assess quality of life (QoL) in patients receiving conformal radiation therapy (CRT) for prostate cancer. PATIENTS AND METHODS: 78 men with definitive CRT for prostate cancer were entered into the study. Patients were assessed before CRT, at 40 and 60 Gy, and 2, 12 and 24 months after the end of treatment. QoL was assessed using the EORTC Quality of Life Questionnaire C30 and the prostate module PR25. Changes in mean QoL scores with time of >or= 10 points were considered clinically relevant. RESULTS: Global QoL did not change statistically significant during CRT and was slightly above baseline levels during follow-up. CRT had a statistically significant negative short-term impact on role functioning, fatigue, and PR25 urinary symptoms. The scores recovered within 2 months to 1 year after CRT. Emotional functioning and social functioning scores slightly increased during and after CRT. Role functioning decreased by > 10 points at 60 Gy and urinary symptoms decreased by > 10 points at 40 and 60 Gy. All other differences were < 10 points. A high number of concomitant diseases and having no children were negative pretreatment predictors for long-term global QoL. CONCLUSION: Definitive CRT for prostate cancer does not compromise global QoL during therapy and up to 2 years after treatment. It has a limited negative effect on role functioning, urinary symptoms and, to a lesser extent, on fatigue with restitution within 2 months to 1 year after treatment.

Calculating Protein Content of Expressed Breast Milk to Optimize Protein Supplementation in Very Low Birth Weight Infants with Minimal Effort-A Secondary Analysis.

Breast milk does not meet the nutritional needs of preterm infants, necessitating fortification. Breast milk is particularly variable in protein content, hence standardized (fixed dosage) supplementation results in inadequate supply. This was a secondary analysis of 589 breast milk protein content measurements of 51 mothers determined by mid-infrared spectroscopy during a clinical trial of higher versus lower protein supplementation in very low birth weight infants. Mothers (and breast milk samples) were divided into a test (41 mothers) and a validation cohort (10 mothers). In the test cohort, the decrease in protein content by day of lactation was modeled resulting in the breast milk-equation (BME)). In the validation cohort, five supplementation strategies to optimize protein supply were compared: standardized supplementation (adding 1.0 g (S1) or 1.42 g protein/100 mL (S2)) was compared with 'adapted' supplementation, considering variation in protein content (protein content according to Gidrewicz and Fenton (A1), to BME (A2) and to BME with adjustments at days 12 and 26 (A3)). S1 and S2 achieved 5% and 24% of adequate protein supply, while the corresponding values for A1-A3 were 89%, 96% and 95%. Adapted protein supplementation based on calculated breast milk protein content is easy, non-invasive, inexpensive and improves protein supply compared to standardized supplementation.

Efficacy of lamellar resection, cryotherapy, and adjunctive grafting for the treatment of canine limbal melanoma.

OBJECTIVE: To evaluate the efficacy and complication rate of partial lamellar resection followed by cryotherapy for the management of canine limbal melanoma. ANIMALS STUDIED: Fourteen dogs with unilateral canine limbal melanoma which were managed surgically by partial lamellar resection, cryotherapy (1,1,1,2-tetrafluoroethane) and an adjunctive graft procedure. METHODS: The clinical records of dogs treated between June 1998 and June 2008 were reviewed. The signalment, approximate size and location of the melanoma, variation in surgical technique, recurrence rate, short-term (< three months) and long-term (> three months) complications were assessed. Follow-up information was collected by patient re-examination or telephone interview. RESULTS: The mean age at diagnosis was 6.3 years, with a range from 3.2 to 12 years. Seven breeds were affected, including five cross-breed dogs and four Golden Retrievers. The tumour involved the dorsal arc (from the dorsomedial to the ventrolateral quadrant) in 12 eyes and the ventral arc in two eyes. The size of the tumour ranged from 30 to 180 degrees of the limbal circumference. A double freeze-thaw cycle of cryotherapy was performed in 7/14 eyes and a triple freeze-thaw cycle in 7/14 eyes. An adjunctive conjunctival graft was performed in 13/14 eyes (free graft n = 3, posterior nictitans conjunctiva/cartilage n = 4, advancement graft n = 5, small intestinal submucosa/advancement graft n = 1) and a frozen homologous graft in 1/14 eyes. The duration of follow-up ranged from 6 months to 8.5 years with a median of 2.1 years. Recurrence was not clinically detected in any of the 14 eyes. Early complications occurred in 8/14 eyes and included anterior uveitis (7/14), corneal ulceration (5/14), marked corneal granulation tissue at the graft margin (2/14), dyscoria (2/14), corneal lipidosis (1/14) and corneal oedema (1/14). Intra-operative globe perforation had occurred in 5/7 eyes with anterior uveitis and 2/2 eyes with transient dyscoria. Late complications occurred in 3/14 eyes and included corneal lipidosis which was either mild (< 2mm in diameter, 1/14) or marked (>1cm in diameter, 2/14). Marked lipidosis only occurred following the treatment of extensive limbal melanomas which involved approximately 50% of the limbal circumference. CONCLUSIONS: Surgical management comprising partial lamellar resection, cryotherapy and adjunctive graft placement is technically straightforward, minimally invasive, well tolerated and highly effective. Marked corneal lipidosis is most likely to occur as a post-operative complication when the limbal melanoma is extensive.

Neuron-specific enolase antibodies in patients with sudden acquired retinal degeneration syndrome.

Sudden acquired retinal degeneration syndrome (SARDS) is a disease characterised by sudden and bilateral vision loss of dogs. Previous studies failed to identify the underlying cause [Mattson, A., Roberts, S.M., Isherwood, J.M.E., 1992. Clinical features suggesting hyperadrenocorticism associated with sudden acquired retinal degeneration syndrome in a dog. J. Am. Anim. Hosp. Assoc. 28, 199-202; Van der Woerdt, A., Nasisse, M.P., Davidson, M.G., 1991. Sudden acquired retinal degeneration in the dog: clinical and laboratory findings in 36 cases. Prog. Vet. Comp. Ophthamol. 1, 11-18] and earlier investigations about the occurrence of anti-retinal antibodies in SARDS patients showed inconsistent results. To provide a novel approach to those findings we designed a more detailed study. Autoantibodies of SARDS patients and normal controls were tested against the purified autoantigens S-antigen and cellular retinaldehyde binding protein (CRALBP) that play a role in human autoimmune uveitis. Next we tested the autoantibody binding pattern to whole retinal lysate. No difference in the incidence of autoantibodies could be found between SARDS patients and healthy controls while testing the well-known autoantigens S-antigen and CRALBP. Potential novel, yet unknown autoantigens were identified by a screening test using the retinal proteome as an autoantigenic source. In SARDS patients and normal controls, several retinal proteins were bound by IgG antibodies, but one band was strongly marked by SARDS patients. That band was excised, subjected to mass spectrometry (matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF/TOF)) and identified as neuron-specific enolase. Binding of the IgG autoantibodies of SARDS-affected dogs to this protein was verified using purified NSE, revealing 25% of NSE autoantibody-positive SARDS patients and 0% of negative controls. Our findings indicate that at least some dogs with SARDS have autoantibodies against NSE, although it is unclear whether these play a causative role in SARDS or whether they are the result of retinal destruction by another mechanism.

Alloantigen expression on malignant cells and healthy host tissue influences graft-versus-tumor reactions after allogeneic hematopoietic stem cell transplantation.

Durable remissions of hematological malignancies regularly observed following allogeneic hematopoietic stem cell transplantation (aHSCT) are due to the conditioning regimen, as well as an immunological phenomenon called graft-versus-leukemia (GVL) or graft-versus-tumor (GVT) effect. The development of GVL is closely linked to graft-versus-host disease (GVHD), the main side effect associated with aHSCT. Both, GVHD and GVL are mediated by donor T cells that are initially activated by antigen-presenting cells that present recipient-derived alloantigens in the context of either matched or mismatched MHC class I molecules. Using murine models of aHSCT we show that ubiquitously expressed minor histocompatibility alloantigens (mHAg) are no relevant target for GVT effects. Interestingly, certain ubiquitously expressed MHC alloantigens augmented GVT effects early after transplantation, while others did not. The magnitude of GVT effects correlated with tumor infiltration by CD8+ cytotoxic T cells and tumor cell apoptosis. Furthermore, the immune response underlying GVHD and GVT was oligoclonal, highlighting that immunodominance is an important factor during alloimmune responses. These results emphasize that alloantigen expression on non-hematopoietic tissues can influence GVT effects in a previously unrecognized fashion. These findings bear significance for harnessing optimal GVL effects in patients receiving aHSCT.

Differential tumor biological role of the tumor suppressor KAI1 and its splice variant in human breast cancer cells.

The tetraspanin and tumor suppressor KAI1 is downregulated or lost in many cancers which correlates with poor prognosis. KAI1 acts via physical/functional crosstalk with other membrane receptors. Also, a splice variant of KAI1 (KAI1-SP) has been identified indicative of poor prognosis. We here characterized differential effects of the two KAI1 variants on tumor biological events involving integrin (αvß3) and/or epidermal growth factor receptor (EGF-R). In MDA-MB-231 and -435 breast cancer cells, differential effects were documented on the expression levels of the tumor biologically relevant integrin αvß3 which colocalized with KAI1-WT but not with KAI1-SP. Cellular motility was assessed by video image processing, including motion detection and vector analysis for the quantification and visualization of cell motion parameters. In MDA-MB-231 cells, KAI1-SP provoked a quicker wound gap closure and higher closure rates than KAI1-WT, also reflected by different velocities and average motion amplitudes of singular cells. KAI1-SP induced highest cell motion adjacent to the wound gap borders, whereas in MDA-MB-435 cells a comparable induction of both KAI1 variants was noticed. Moreover, while KAI1-WT reduced cell growth, KAI1-SP significantly increased it going along with a pronounced EGF-R upregulation. KAI1-SP-induced cell migration and proliferation was accompanied by the activation of the focal adhesion and Src kinase. Our findings suggest that splicing of KAI1 does not only abrogate its tumor suppressive functions, but even more, promotes tumor biological effects in favor of cancer progression and metastasis.