Automation of route identification and optimisation based on data-mining and chemical intuition.

Data-mining of Reaxys and network analysis of the combined literature and in-house reactions set were used to generate multiple possible reaction routes to convert a bio-waste feedstock, limonene, into a pharmaceutical API, paracetamol. The network analysis of data provides a rich knowledge-base for generation of the initial reaction screening and development programme. Based on the literature and the in-house data, an overall flowsheet for the conversion of limonene to paracetamol was proposed. Each individual reaction-separation step in the sequence was simulated as a combination of the continuous flow and batch steps. The linear model generation methodology allowed us to identify the reaction steps requiring further chemical optimisation. The generated model can be used for global optimisation and generation of environmental and other performance indicators, such as cost indicators. However, the identified further challenge is to automate model generation to evolve optimal multi-step chemical routes and optimal process configurations.

Aminopiperidine based complexes for lactide polymerisation.

Herein we report the synthesis and characterisation of a series of salalen and salan ligands derived from 2-(aminomethyl)piperidine. Depending on the choice of starting salicylaldehyde, a bicyclic salan type ligand (1-3H2) or imino salalen type ligand (4-6H, 7-9H2) were prepared. The ligands were successfully complexed to group 4 metals and aluminium; with hafnium and zirconium octahedral complexes, M(1-3)2, were realised; whilst with aluminium tetrahedral and trigonal bipyramidal complexes, Al(1-9)Mex (x = 1,2), were isolated. The complexes have been characterised in solution via(1)H and (13)C{(1)H} NMR spectroscopy and in the solid state by X-ray crystallography. The group 4 complexes were observed to have a fac-fac arrangement of ligands and there were two isomers present when 3H2 was ligated. The imino aluminium complexes Al(7-9)Me were isolated as a mixture of diastereoisomers. The resultant complexes were trialed in the ring opening polymerisation of rac-lactide with both heterotactic and isotactic PLA being demonstrated. Tacticity was found to be dependent on the nature of the ligand and metal used; the M(1-3)2 complexes were generally found to have a heterotactic preference (Pr = 0.67-0.76) and the aluminium polymerisation outcome was dictated more by the steric influence of the ligand, particularly for Al(4/6)Me2/Al(7/9)Me.

Preliminary study to assess mycotoxin concentrations in whole corn in the California feed supply.

Mycotoxins are naturally occurring environmental contaminants recognized worldwide in a variety of food and feed products. Produced as secondary metabolites by filamentous fungi, mycotoxins can have acute and chronic effects. Differing seasonal weather patterns and harvesting and storage conditions put corn grain at high risk for mycotoxin contamination. The objective of this study was to assess the risk of mycotoxin exposure posed to California livestock from whole corn. Random samples (n=50) of whole corn were collected and analyzed for 6 different mycotoxins, including aflatoxins, fumonisins, ochratoxins, trichothecenes (deoxynivalenol and T-2 toxin), and zearalenone. The samples represented a cross section of the corn entering California from various corn-growing states (n=43) as well as additional samples from California-grown corn (n=7). The experiment was a randomized sampling design. Over the course of a 6-mo period, 16 trains in California (100-110 railcars) and 5 California grain elevators were randomly sampled. Aflatoxins were detected in 14 samples, with 1 sample containing a concentration of 41.3 µg/kg (as-is basis), which was above the action level of 20 µg/kg for corn fed to dairy cattle. The average concentration of aflatoxins for the 13 samples below the regulatory action level was 8.69 µg/kg (range 4.67 to 13.82 µg/kg). Deoxynivalenol was found in 15 samples and averaged 553 µg/kg (range 340 to 1,072 µg/kg), which was below the federal advisory level of 5,000 µg/kg for grain fed to dairy cattle. Fumonisins were found in 38 samples and averaged 1,687 µg/kg (range 435 to 4,843 µg/kg), which was below the federal guidance level of 30,000 µg/kg in corn for dairy cattle. Ochratoxins, T-2 toxins, and zearalenone were not detected in any samples of whole corn. Fumonisins were the most prevalent mycotoxins found.

Histopathological features of ocular leishmaniosis in the dog.

Canine leishmaniosis (CL) can present with multiple clinical signs and ocular disease is reported to occur in almost 25% of affected dogs. The purpose of the present study was to characterize the nature of inflammation within the eyes of dogs with leishmaniosis and to determine whether parasites were present in these lesions. Eyes from 60 dogs with confirmed leishmaniosis that died or were humanely destroyed over a 4 year period were included in the study. Sections of formalin-fixed globes were stained with haematoxylin and eosin (HE) and subjected to immunohistochemistry using a Leishmania-specific antibody. Clinically evident ocular signs were present in 15 of 60 dogs (13 bilaterally and 2 unilaterally). Thirty-five of 60 dogs received some form of anti-protozoal treatment. In 36 of 120 eyes (30%) a granulomatous inflammatory infiltrate was found and in 32 of 120 eyes (26.6%) the parasite was identified immunohistochemically within the globe. Ocular tissues affected, in order of frequency, were conjunctiva and limbus, ciliary body, iris, cornea, sclera and iridocorneal angle, choroid and the optic nerve sheath. Different microscopical patterns were defined in each of these structures. Leishmania organisms and associated inflammation can be found in different ocular tissues, accounting for some of the ocular clinical signs described for this disease.

Superficial, nonhealing corneal ulcers in horses: 23 cases (1989-2003).

OBJECTIVE: To characterize superficial, nonhealing corneal ulcers in the horse and to assess the affect of age, breed, sex, inciting cause of the ulcer, and treatment on healing time of these ulcers. DESIGN: Retrospective study. ANIMALS: Twenty-three horses with superficial, nonhealing corneal ulcerations. PROCEDURE: Medical records from 1989 to 2003 of horses diagnosed with superficial, nonhealing corneal ulcers were reviewed. Signalment, duration of clinical signs, ophthalmic abnormalities, treatment and response to treatment were recorded. Horses were treated, in addition to medical therapy, using debridement, debridement and grid keratotomy, superficial keratectomy, and superficial keratectomy with conjunctival graft placement. RESULTS: Mean age +/- SD of horses was 13.7 +/- 5.8 years. The mean time of presence of the corneal ulceration was 38.9 +/- 21.3 days. Mean time to complete re-epithelialization of the corneal ulcers after treatment was 20 +/- 14.7 days. Horses treated with debridement alone, grid keratotomy and superficial keratectomy healed in a mean time of 15.3 +/- 14.6 days, 16 +/- 12.6 days, and 22.8 +/- 6.7 days, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Horses treated with a single debridement at initial evaluation healed in a significantly shorter time period than horses treated with grid keratotomy or superficial keratectomy. The latter two procedures may be beneficial in the treatment of nonhealing ulceration in horses, but the results of this study suggest that these procedures should only be performed following failure of the ulcer to heal after epithelial debridement.

Bartonella vinsonii subspecies berkhoffi as a possible cause of anterior uveitis and choroiditis in a dog.

A 2-year old, neutered, female spaniel mixed breed was referred to the North Carolina State University Veterinary Teaching Hospital for evaluation of bilateral anterior uveitis. The dog was febrile and, in addition to anterior uveitis, multifocal hyporeflective lesions were present in the tapetal fundus of both eyes. The antibody titer for Bartonella vinsonii subspecies berkhoffi was positive (1 : 512). Aqueous paracentesis was performed for PCR in an attempt to detect B. vinsonii in the eye but was unsuccessful. The ocular manifestations of Bartonella infection in humans are currently expanding as more sensitive serologic and PCR techniques are being developed to identify Bartonella spp. In addition to optic neuritis and neuroretinitis, retinochoroidal lesions are one of the most common manifestations of B. henselae infection, and are frequently accompanied by vitreous or anterior segment inflammation. Diagnosis of a Bartonella infection in humans can be made on serology alone, in conjunction with ocular examination findings. The ultimate proof of B. vinsonii (berkhoffi) as a direct cause of ocular disease would be detection of the infectious agent in the eye. However, it is unknown at this time whether Bartonella causes ocular disease primarily, secondarily via an autoimmune reaction, or both. Due to the difficulties associated with culture of Bartonella spp. and the limitations of PCR, serology is currently the most useful tool for screening dogs for possible Bartonella spp. infection. In the case presented here, even though the PCR was negative, the clinical signs of anterior uveitis and choroiditis might reasonably be associated with B. vinsonii (berkhoffi) seroreactivity, which was repeatable on three separate occasions. Clinical improvement was also accompanied by a post-treatment decrease in B. vinsonii (berkhoffi) seroreactivity, potentially supporting resolution of Bartonella infection in this dog. This is the first reported case of a possible association between uveitis, choroiditis and Bartonella infection in the dog, without clinical manifestations of other organ or tissue involvement. Future studies based on PCR analysis of intraocular fluids may clarify the involvement of B. vinsonii (berkhoffi) in dogs with intraocular inflammatory disease. Furthermore, performing fluorescein angiography in dogs with elevated Bartonella titers may also prove useful in the identification and characterization of lesions.

Transpupillary diode laser retinopexy in dogs: ophthalmoscopic, fluorescein angiographic and histopathologic study.

OBJECTIVE: To evaluate the ophthalmoscopic, fluorescein angiographic and light microscopic effects of diode laser retinopexy application in the tapetal and nontapetal fundus in the dog, and to ascertain appropriate laser power settings for production of photocoagulative lesions in these two regions. ANIMALS STUDIED: Three adult female Beagle dogs. PROCEDURES: Laser burns were applied to selected areas in the fundus with an indirect headset delivery system using settings varying from 100 to 200 milliWatts (mW) and from 100 to 600 milliSeconds (mS) with total delivered energy ranging between 15 and 100 milliJoules (mJ). The dogs were then monitored by ophthalmoscopic examination and fluorescein angiography at regular intervals for 7-28 days. Histopathologic studies were performed at 7, 14 and 28 days after laser application. RESULTS: The diode laser produced ophthalmoscopically visible lesions in the nontapetal fundus with all laser settings used, and the appearance of these lesions corresponded to the energy levels used, and degree of pigment in the lased region. Gray-white colored lesions with minimal subsensory retinal edema were seen with settings as low as 100 mWatts/150 mSeconds. In the tapetal fundus, laser burns were more difficult to produce, less repeatable, and required higher energy levels. Laser burns appeared as bronze, dark green or black discolorations of the tapetum with varying degrees of subsensory retinal edema. Lesions were more reproducible and were achieved with lower settings in the tapetal area of the tapetal/nontapetal junction. Ophthalmoscopically, depigmentation and repigmentation of the RPE (nontapetal fundus) and degenerative changes in the overlying retina (tapetal fundus) developed in the laser burns over the 28-day study period. Fluorescein angiographic studies showed disruption of the blood-retinal barrier at the level of the RPE and fluorescein leakage into the subsensory retinal space was seen in most lesions at 24 h, was minimal at 3 days, and had resolved by 7 days. Histologically, grayish-white lesions in the nontapetal fundus, and bronze to small black lesions in the tapetal fundus were typically characterized by outer retinal necrosis and RPE migration. Gliosis was considered minimal, was confined to the retina, and no inflammatory cells were seen. Peripheral intense white lesions (nontapetum) and lesions with a black center (tapetal fundus) were characterized by more extensive panretinal and choroidal necrosis. Most of the nontapetal lesions and a few in the tapetal fundus showed the formation of a central retinal detachment. CONCLUSIONS: The diode laser effectively produces lesions suitable for retinopexy in both the nontapetal, pigmented fundus and the tapetal fundus, although variably so in the latter region. Initial laser settings of 100-150 mW/200 mS for the pigmented fundus, and 150 mW/200-300 mS for the peripheral tapetal fundus are recommended, and the clinician should gradually increase time interval settings to achieve a grayish-white lesion in the nontapetum, and a bronze to slightly black lesion in the tapetal fundus. If possible, retinopexy should be applied to the peripheral tapetal area or tapetal/nontapetal junction.

Acute vision loss after general anesthesia in a cat.

A 6-year-old, male castrated, domestic shorthair cat presented with a 2-week history of blindness following general anesthesia. Behavioral changes, lethargy, inappetence and neurological deficits were also noted. Ophthalmic evaluation revealed blindness, normal pupillary light response and dazzle response, but no other abnormalities. Serodiagnostic testing for common infectious agents was negative and a cerebrospinal fluid analysis was normal. History and postmortem examination following euthanasia revealed cerebrocortical necrosis most consistent with anesthesia related hypoxia.

Use of an intravitreal sustained-release cyclosporine delivery device for treatment of equine recurrent uveitis.

OBJECTIVE: To evaluate the use of an intravitreal sustained-release cyclosporine (CsA) delivery device for treatment of horses with naturally occurring recurrent uveitis. ANIMALS: 16 horses with recurrent uveitis. PROCEDURES: Horses with frequent recurrent episodes of uveitis or with disease that was progressing despite appropriate medication were selected for this study. Additional inclusion criteria included adequate retinal function as determined by use of electroretinography, lack of severe cataract formation, and no vision-threatening ocular complications (eg, retinal detachment, severe retinal degeneration, and posterior synechia). Sustained-release CsA delivery devices (4 microg of CsA/d) were implanted into the vitreous through a sclerotomy at the pars plana. Reexaminations were performed 1, 3, 6, and 12 months after implantation, then continued annually. Ophthalmic changes, number of recurrent episodes of uveitis, and vision were recorded. RESULTS: The rate of recurrent episodes after device implantation (0.36 episodes/y) was less than prior to surgery (75 episodes/y). In addition, only 3 horses developed episodes of recurrent uveitis after surgery. Vision was detected in 14 of 16 affected eyes at a mean follow-up time of 13.8 months (range, 6 to 24 months). CONCLUSIONS AND CLINICAL RELEVANCE: This intravitreal sustained-release CsA delivery device may be a safe and important tool for long-term treatment of horses with chronic recurrent uveitis.

Postnatal development of corneal curvature and thickness in the cat.

OBJECTIVE: To evaluate the postnatal development of central corneal curvature and thickness in the domestic cat. Animals studied Six Domestic Short-haired (DSH) kittens starting at 9 weeks of age and 6 adult cats. PROCEDURES: Kittens were evaluated biweekly to monthly for a 12-month period, starting at age 9 weeks. Corneal development was monitored by hand-held keratometry and ultrasound biomicroscopy. Standard regression analysis using a nonlinear least squares method was used to generate a formula that would predict corneal curvature as a function of age. RESULTS: Mean keratometry (K) values for the 9-week-old cats were 54.51 (+/-1.02) diopters (D) and these values steeply declined over the next 3 months to 44.95 (+/-0.90) D. Thereafter, K-values gradually decreased to reach a plateau by 12-15 months of age of 39.90 (+/-0.42) D. Because K-values still appeared to be slightly diminishing at this point, six other > 2-year-old cats were evaluated by keratometry and were found to have K-values of 38.99 (+/-0.81). Two to four diopters of astigmatism was common in young kittens whereas adult cats had a low mean degree of astigmatism (< 1 D). A formula that predicted keratometry values in diopters (K) as a function of age in weeks (w) was established as follows: K = 39.83 + 26.87 exp(-0.074 w). The central cornea increased in thickness primarily during the first 4 months of life with 9 week-old kittens having values of 0.379 (+/-0.012) mm; 16-week-old kittens, 0.548 (+/-0.021) mm and 67 week-old cats, 0.567 (+/-0.012) mm. CONCLUSIONS: The maturation process of the feline cornea proceeds over the first 1-2 years of life to attain an adult status that is characterized by a roughly spherical state of approximately 39 D corneal curvature, substantially flatter than the human cornea, and a central thickness similar to the human cornea. Research studies of the refractive or optical properties of the cornea in which cats are used as experimental animals should be conducted on animals greater than 18 months of age.

Toxoplasmosis.

Toxoplasmosis in dogs and cats can cause chorioretinitis, anterior uveitis, or both. Ocular lesions are a common manifestation of generalized toxoplasmosis. The prevalence of toxoplasmosis as a cause of idiopathic anterior uveitis in cats is not clear, although there is a significant association between exposure to T. gondii and feline anterior uveitis. The pathogenesis of ocular toxoplasmosis may be different in humans and cats, and the anterior uveitis may represent a type of immune-mediated inflammation. A diagnosis is made by observing compatible clinical findings and obtaining supportive findings on serologic tests. Despite improved diagnostic techniques, including determination of IgM class antibodies and PCR testing, definitive diagnosis of ocular toxoplasmosis remains a challenge. Topical anti-inflammatory therapy should be used in cats with anterior uveitis, a positive serum titer, and no concurrent systemic signs. Systemic clindamycin should be given to cats with ocular and systemic signs and to cats with suggestive serology and idiopathic anterior uveitis that fails to respond to topical therapy alone.

Effect of surgical technique on in vitro posterior capsule opacification.

PURPOSE: To compare the effect of different cataract extraction surgical techniques on residual lens epithelial cell (LEC) density and cell regrowth rates using an in vitro model of posterior capsule opacification (PCO). SETTING: Comparative Ophthalmology Research Laboratories, North Carolina State University, Raleigh, North Carolina, USA. METHODS: Lens capsule explants were prepared from freshly enucleated canine globes after extracapsular cataract extraction (ECCE), phacoemulsification, or phacoemulsification followed by capsule vacuuming. Initial cell density on the capsule and cell proliferation were determined by phase contrast microscopy. The effects of the surgical technique on time to confluent growth of the cells across the posterior lens capsule were determined. RESULTS: Residual cell density on the remaining anterior capsule immediately after lens removal was 31.6% +/- 19.3%, 16.1% +/- 8.9%, and 7.7% +/- 5.7% in the ECCE, phacoemulsification, and phacoemulsification/capsule-vacuuming groups, respectively. Time to confluence (range 5.0 to 6.3 days) was not significantly different among the 3 groups when the lens capsules were cultured in serum-supplemented media. The confluence rate was significantly longer (by approximately 5 to 7 days) in the phacoemulsification/capsule-vacuuming group than in the other 2 groups when the capsules were cultured in serum-free media. CONCLUSIONS: Phacoemulsification with and without anterior and equatorial capsular vacuuming led to less initial LEC density in the capsular bag than ECCE. However, because cell proliferation rates among the 3 groups were only marginally affected, near 100% removal of LEC at the time of cataract extraction may be necessary to prevent PCO.

Ex vivo canine lens capsular sac explants.

BACKGROUND: Lens capsular sac explants from human cadaver eyes were used to investigate posterior capsular opacification (PCO). The purpose of this study was to characterize a similar model using canine tissue and to determine whether transferrin (Tf), transforming growth factor beta-2 (TGF-beta2), and insulin-like growth factor-1 (IGF-1) are secreted by lens epithelial cells (LEC) of these ex vivo sacs. METHODS: The lens from canine eyes was removed by extracapsular cataract extraction, the lens sac dissected free, pinned to a petri dish, and cultured in either serum-supplemented or serum-free medium. Morphologic characteristics and growth rate to confluence on the posterior capsule were studied by phase-contrast microscopy. Vimentin, alpha smooth muscle actin, and panTGF-beta expression by LEC were determined by immunohistochemistry. Tf, TGF-beta2, and IGF-1 levels were measured by ELISA in the supernatant of sacs cultured in serum-free medium. RESULTS: The mean time to confluence of LEC onto the posterior capsule was 5.4+/-1.1 days (n=22) and 14.7+/-3.7 days (n=14) for sacs in serum-supplemented and serum-free medium, respectively. Following development of confluence, explants displayed opacification and light scatter from cellular proliferation and capsular contraction. Confluent LEC expressed vimentin, alpha smooth muscle actin, and TGF-beta2, and both Tf and TGF-beta2 were secreted into the culture supernatant. CONCLUSION: Canine lens sac explants have characteristics virtually identical to those of human origin, and appear to be a useful alternative tissue source for this model when human cadaver eyes are unavailable. Tf and TGFbeta-2, but not IGF-1, are secreted by LEC in explanted lens sacs and may influence the proliferation and metaplasia of LEC during the development of PCO.

Ocular lesions associated with systemic hypertension in cats: 69 cases (1985-1998).

OBJECTIVE: To characterize clinical and clinicopathologic findings, response to treatment, and causes of systemic hypertension in cats with hypertensive retinopathy. DESIGN: Retrospective study. ANIMALS: 69 cats with hypertensive retinopathy. PROCEDURE: Medical records from cats with systemic hypertension and hypertensive retinopathy were reviewed. RESULTS: Most cats (68.1%) were referred because of vision loss; retinal detachment, hemorrhage, edema, and degeneration were common findings. Cardiac abnormalities were detected in 37 cats, and neurologic signs were detected in 20 cats. Hypertension was diagnosed concurrently with chronic renal failure (n = 22), hyperthyroidism (5), diabetes mellitus (2), and hyperaldosteronism (1). A clearly identifiable cause for hypertension was not detected in 38 cats; 26 of these cats had mild azotemia, and 12 did not have renal abnormalities. Amlodipine decreased blood pressure in 31 of 32 cats and improved ocular signs in 18 of 26 cats. CONCLUSIONS AND CLINICAL RELEVANCE: Retinal lesions, caused predominantly by choroidal injury, are common in cats with hypertension. Primary hypertension in cats may be more common than currently recognized. Hypertension should be considered in older cats with acute onset of blindness; retinal edema, hemorrhage, or detachment; cardiac disease; or neurologic abnormalities. Cats with hypertension-induced ocular disease should be evaluated for renal failure, hyperthyroidism, diabetes mellitus, and cardiac abnormalities. Blood pressure measurements and funduscopic evaluations should be performed routinely in cats at risk for hypertension (preexisting renal disease, hyperthyroidism, and age > 10 years). Amlodipine is an effective antihypertensive agent in cats.

Histologic and immunohistochemical characterization of lens capsular plaques in dogs with cataracts.

OBJECTIVE: To determine histologic and immunohistochemical characteristics of the multifocal adherent plaques that commonly develop on the internal surfaces of the anterior and posterior lens capsules in dogs with cataracts. SAMPLE POPULATION: 31 anterior and 4 posterior capsular specimens collected during lens extraction surgery in dogs with cataracts. PROCEDURE: Specimens were evaluated, using light and transmission electron microscopy. Immunohistochemical techniques were used to localize cytokeratin, vimentin, alpha-smooth muscle-specific actin, fibronectin, tenascin, and transforming growth factor-beta (TGF-beta) within plaques. RESULTS: Histologically, plaques comprised elongated spindle-shaped cells that formed a placoid mass. Cells were embedded in an extracellular matrix containing collagen fibrils, often with duplicated or split basement membranes. Immunohistochemically, normal lens epithelial cells and cells within plaques stained for vimentin. Most cells and some areas of the extracellular matrix within plaques stained for TGF-beta and alpha-smooth muscle-specific actin. Fibronectin and tenascin were also detected in the extracellular matrix. CONCLUSIONS AND CLINICAL RELEVANCE: Canine lens capsular plaques are histologically and immunohistochemically similar to posterior capsule opacification and subcapsular cataracts in humans, which suggests that the canine condition, like the human conditions, is associated with fibrous metaplasia of lens epithelial cells. Transforming growth factor-beta may play a role in the genesis of capsular plaques. Because severity of plaques was correlated with stage of cataract development, earlier surgical removal of cataracts may be useful to avoid complications associated with plaque formation.

Ocular and periocular manifestations of leishmaniasis in dogs: 105 cases (1993-1998).

The purpose of this retrospective study was to determine the prevalence, type, and prognosis of ocular lesions associated with leishmaniasis in dogs. One hundred and five dogs (24.4% of all cases of leishmaniasis diagnosed during the study period) had ocular or periocular leishmaniasis, and 16 dogs (15.2% of ocular cases) had only ocular lesions and systemic signs were not apparent. Anterior uveitis was the most common manifestation and other prevalent findings included blepharitis and keratoconjunctivitis. Several distinct variations of eyelid lesions were seen including a dry dermatitis with alopecia, diffuse blepharedema, cutaneous ulceration, and discrete nodular granuloma formation. In some cases with keratoconjunctivitis, corneal lesions clinically resembled nodular granulomatous episclerokeratitis. Twenty-seven of the 34 cases with ocular lesions had improvement in signs following systemic antiprotozoal and topical anti-inflammatory therapy, although many cases with anterior uveitis required long-term topical therapy. Response of ocular signs correlated highly with overall, systemic response to therapy. Ophthalmic manifestations of systemic leishmaniasis are common in the dog, and this disease should be considered in the differential diagnosis of most adnexal and anterior segment ocular inflammatory lesions in dogs in endemic areas.