High-resolution protein structure determination by serial femtosecond crystallography

Structure determination of proteins and other macromolecules has historically required the growth of high-quality crystals sufficiently large to diffract x-rays efficiently while withstanding radiation damage. We applied serial femtosecond crystallography (SFX) using an x-ray free-electron laser (XFEL) to obtain high-resolution structural information from microcrystals (less than 1 micrometer by 1 micrometer by 3 micrometers) of the well-characterized model protein lysozyme. The agreement with synchrotron data demonstrates the immediate relevance of SFX for analyzing the structure of the large group of difficult-to-crystallize molecules.     

Multilocus sequence typing of human and canine C. upsaliensis isolates

Risk of Campylobacter infection in humans has been associated with many sources, including dogs. C. upsaliensis is the most common species found in canines, and has been occasionally isolated from symptomatic humans. This study aimed to investigate the genetic diversity of 41 C. upsaliensis isolates carried by dogs and from nine isolates carried by humans using Multilocus sequence typing (MLST). We identified considerable genetic diversity amongst the C. upsaliensis isolates from both dogs and humans, identifying 45 different sequence types (STs). All STs were new, apart from that of the reference strain. Only three STs were found in more than one isolate: ST-72 (2 isolates), ST-98 (2 isolates) and ST-104 (3 isolates). ST-104 was the only ST to be encountered in both dogs and humans. Thirty-one of the 45 STs were assigned to one of 13 clonal complexes (CCs). Four of these CCs contained STs originating from both humans and dogs. None of the CCs contained exclusively human isolates, and two isolates from dogs within the same kennel belonged to the same CC. The large amount of diversity found in both dog and human isolates of C. upsaliensis, combined with the relatively small database, made it difficult to assign strains to sources of infection. This emphasizes the need to increase the size of the database. Dog and human isolates occasionally grouped together, however there were insufficient human-derived isolates to determine whether or not dogs are a common source of infection. Although C. upsaliensis infection is rare in humans, dogs still remain a potential source, and are therefore a possible zoonotic risk. Further work is needed to investigate the epidemiology of C. upsaliensis infection in humans.     

Open-label trial of a multi-strain synbiotic in cats with chronic diarrhea

This study was designed to test the hypothesis that in cats with chronic diarrhea the daily administration of a proprietary synbiotic (Proviable-DC) would result in an improvement in stool character, as assessed by the owner. Adult cats with chronic diarrhea were recruited for the study and screened for systemic diseases. Fecal flotation, wet mount, immunofluorescence assay (IFA) for Giardia and Cryptosporidium species, and Tritrichomonas species polymerase chain reactions (PCRs) were used to screen for intestinal parasitism. The synbiotic was administered for 21 days; otherwise, no changes were made to ongoing treatment(s) or diet. The severity of the diarrhea was assessed using a standardized fecal scoring system and the owner's subjective perception before, and after, supplementation. The mean fecal score for the 53 cats completing the study decreased from 6.0 to 4.4, representing a significantly (P <0.001) firmer stool character. Seventy-two percent of owners perceived an improvement in their cat's diarrhea following a 21-day course of synbiotic supplementation.     

Diaphragmatic hernia in horses: 44 cases (1986–2006)

Objectives – To present a case series of horses diagnosed with diaphragmatic hernia, and to determine the significance of (1) historical information, examination findings, and laboratory data; and (2) exploratory laparotomy or necropsy findings on short- and long-term outcome.
Setting – University Referral Hospital.
Design – Retrospective study.
Animals – Forty-four horses/foals admitted between 1986 and 2006 with a diagnosis of diaphragmatic hernia made either at exploratory laparotomy or necropsy.
Interventions – None.
Measurements and Main Results – Information from the medical records included history, clinical examination findings at presentation, and findings of exploratory laparotomy or necropsy. Logistic regression or the Fisher exact test was used to determine factors associated with survival. Outcome was defined as survival to discharge (short-term survival), and long-term survival was defined as horses alive at least 1-year post surgery. Of the 44 horses, 18 died or were euthanized before surgery. Twenty-six were taken to surgery, 17 were euthanized. Nine horses recovered from anesthesia, 7 of which survived to hospital discharge. Of these, 5 were alive at long-term follow-up. Survival was significantly associated with the age of the horse (≤2 y old) at presentation, presence of normal peritoneal fluid at presentation, amount of compromised viscera at surgery (<50% small intestine), and the size (<10 cm) and location (ventral) of the diaphragmatic tear.
Conclusion – This study confirms that size and location of the lesion do play a significant role in prognosis. And, although the prognosis for horses with diaphragmatic hernia is poor, if horses have operable lesions there is a fair prognosis for long-term survival.     

Control of urine marking by use of long-term treatment with fluoxetine or clomipramine in cats

OBJECTIVES: To determine whether clomipramine differs from fluoxetine in reducing feline urine marking; whether reduction of marking continues in cats treated >8 weeks; whether recurrence of marking, after abrupt drug withdrawal, is less in cats treated >8 weeks; and whether cats that are successfully treated but resume marking after drug withdrawal can be successfully treated again with the same drug regimen. DESIGN: Positive-controlled, double-masked clinical trial. ANIMALS: 22 neutered cats (2 females, 20 males) > or =1 year old with objectionable urine marking. PROCEDURE: Cats that marked vertically > or =3 times/wk were treated with fluoxetine (1 mg/kg [0.45 mg/lb], q 24 h, PO) or clomipramine (0.5 mg/kg [0.23 mg/lb], q 24 h, PO) for 16 weeks, and efficacy was compared. Recurrence of marking was determined after abrupt withdrawal of fluoxetine at 16 or 32 weeks. Reduction in marking in cats treated with fluoxetine for 8 weeks after returning to marking following drug withdrawal was compared with the initial 8 weeks of successful treatment. RESULTS: Efficacy of fluoxetine and clomipramine was similar. Treatment >8 weeks revealed increasing efficacy in reduction of marking. Return of marking after termination of fluoxetine administration occurred in most cats. Cats successfully treated initially with fluoxetine responded similarly to repeated treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Clomipramine and fluoxetine were equivalent in treating urine marking. Longer treatment increased efficacy. Most cats return to marking after abrupt drug withdrawal. A second course of treatment can be expected to be as effective as the first.     

Effects of moxifloxacin on QT interval in conscious dogs

Moxifloxacin has been shown to induce QT prolongation in both clinical and preclinical models. However, the ability to observe this effect at clinically relevant concentration in normal conscious dogs has not been reported. The purpose of this study was to investigate the effects of moxifloxacin on the QT interval in conscious, healthy dogs. Four male mongrel dogs were chronically instrumented for the measurement of arterial blood pressure, left ventricular blood pressure, cardiac output, electrocardiograms (ECGs), and body temperature. Animals were administered a 1-h i.v. infusion of moxifloxacin once per day via a catheter in the cephalic vein. Each dog received all doses (0, 1, 10, 25 and 50 mg/kg) in an escalating fashion. Moxifloxacin caused a statistically significant increase in arterial blood pressure at 50 mg/kg. A dose-response effect on QT and QTc prolongation was observed. A statistically significant prolongation in the QT interval was observed at 10, 25 and 50 mg/kg and a prolongation of QTc was observed at 25 and 50 mg/kg. These effects occurred at clinically relevant plasma concentrations. This study demonstrate that a study design with four dogs was sensitive enough to measure moxifloxacin-induced QT prolongation at clinically relevant plasma concentrations.     

The effect of grazing forage containing condensed tannins on gastro-intestinal parasite infection and milk composition in Angora does

The objective of this study was to evaluate effects of the condensed tannin (CT)-containing forage sericea lespedeza (sericea lespedeza (SL); Lespedeza cuneata; 15.2% CT), on fecal egg count (FEC), larval development (larvae/10 g of feces), worm burden and immune response compared with a crabgrass (Digitaria ischaemum)/Kentucky 31 tall fescue (Festuca arundinacea; control forage (CTF)) forage low in CT (0.32% CT) in grazing Angora does and their kids. Fifty worm-free mixed-age does were randomly allocated to three treatments. One treatment (10 does; initial liveweight (LW) = 45+/-1.5 kg) entailed grazing of SL forage from April 25 to July 15, 2002 with a second treatment of CTF (20 does; initial LW = 43+/-1.4 kg) grazing during the same period. Does of the third treatment (20 does; initial LW = 44+/-1.4 kg) grazed a sward of SL for 2 weeks and then one of CTF for 2 weeks followed by alternating between the two pastures every 2-week rotational grazing (ROT). To gauge levels of infective larvae on pasture, three worm-free Angora kids (initial LW = 3.6+/-0.2 kg) were randomly selected as tracers. Tracers grazed for final 60 days and were euthanized for determination of worm burden. The immune response of does was measured by skin thickness reaction after the intradermal injection of 250 microg phytohemagglutinin (PHA). Mean FEC for SL and ROT were substantially lower (P < 0.01) than for CTF does (145, 329 and 894 eggs/g, respectively). The FEC for kids was lower (P < 0.05) for SL than for ROT and CTF (550, 2757 and 3600 eggs/g, respectively). Total fecal egg output (3.3, 6.0 and 26.9 x 10(5) eggs/day, respectively) and larval development (242, 263 and 792 larvae/10 g, respectively) were lower (P < 0.05) for SL and ROT than for CTF. Tracers grazing on SL had lower total worm burdens than ROT and CTF (P < 0.01). The immune response was higher (P < 0.01) for SL (4.9 mm) and ROT (6.0 mm) than for CTF (3.0 mm) at 12 h after injection of PHA. The packed cell volume (PCV) in does was higher (P < 0.01) for SL and ROT than for CTF (27, 26 and 23%, respectively). Does that grazed CT-containing forage had considerably lower milk somatic cell counts (SCC) than does grazing non-CT-containing forage. In summary, grazing CT forages reduced FEC, larval development and worm burden, and also appeared to enhance immune response. The CT-containing forage SL reduced gastro-intestinal parasite infections of Angora does and kids.     

Effective bibliographic searching for animal alternatives in veterinary medical education: the UC Davis web site

To prepare students in just four years to enter veterinary practice, veterinary medical educators offer an array of laboratory and clinical experiences coordinated with didactic instruction. Recent curricular changes have reduced the numbers of animals involved in painful or terminal procedures. For each use of animals, veterinary educators are required by the USDA's policies 11 and 12 to complete animal-use protocols that include questions on alternatives to procedures causing more than momentary pain or distress. Veterinary medical educators seeking improved teaching resources and methods or completing animal-use protocols may find it frustrating to locate the relevant information, which is dispersed across many databases. This paper addresses a gap facing veterinary educators by presenting user-friendly searching tools that are targeted toward (a) locating teaching resources and (b) conducting effective bibliographic searches on standard teaching laboratory procedures, as required for animal-use protocols. These tools simplify searching by providing streamlined access to the resources being sought. Facilitating efficient and effective searching by users can improve teaching and simplify compliance with USDA requirements.