The use of a rein tension device to compare different training methods for neck flexion in base-level trained Warmblood horses at the walk.

BACKGROUND: Debate surrounds the use of high rein tension for obtaining different head and neck positions in the training of sport horses on account of possible welfare issues. OBJECTIVES: To compare auxiliary rein tension in two methods (Draw Reins and Concord Leader) for obtaining a standardised head and neck position on a hard and a soft surface. STUDY DESIGN: Intervention study. METHODS: Left and right rein tensions were measured in 11 base-level trained client-owned sport horses (mean age ± s.d.; 10 ± 3.2 years) exercised in-hand with, in a random order, conventional draw reins or the newly developed Concord Leader in a standardised head and neck position. Rein tension was measured using a calibrated device operating at 10 Hz during six runs of 15 s in a straight line for each training method on both a hard and a soft surface. A linear mixed model and grouped logistic regression analysis were applied to compare the two methods (P<0.05). RESULTS: The odds of a tension of 0 N were lower with draw reins than with the Concord Leader. The rein tension (mean sum of the force applied, in N) of the draw reins was 13.8 times higher than that of the Concord Leader. MAIN LIMITATIONS: This study was performed on horses exercised in-hand; however, these auxiliary aids are normally used when lungeing. Possible redirection of rein tension towards the poll was not measured. CONCLUSIONS: We showed that when using the Concord Leader a similar head and neck position is achieved with a much lower rein tension than with the draw reins and, more importantly, with a much greater likelihood of 0 N. It is unnecessary to use high auxiliary rein tension to obtain a standard, flexed head and neck position.

Influence of thyroid hormone disruption on the incidence of shingles.

The reactivation of dormant alpha-human herpesvirus (αHHV) has been attributed to various causes often referred to as stressors. However, no clinical study investigating the relationship between stressors and reactivation exists in humans at this time. Herpes simplex virus type-1 (HSV-1), an important αHHV, was shown to have its gene expression and replication regulated by thyroid hormone (TH) using molecular biology approaches. Varicella zoster virus (VZV) is categorized in αHHV superfamily and shares similar homology with HSV-1. We hypothesize that a history of TH imbalance may be associated with the incidence of shingles (VZV reactivation). This current pilot study, based on a hospital medical claims database, was conducted as a retrospective case-controlled investigation to determine if a putative link between TH imbalance and incidence of shingles is present. An odds ratio of 2·95 with a χ 2 value of 51·74 was calculated for the total population diagnosed with TH disruption and shingles. Further analyses indicated that African American males exhibited a much higher chance of simultaneous diagnoses. These results show that a TH imbalance history may affect VZV reactivation at different incidence rates in different races and age groups.

Rare mandibular surgical ciliated cysts: report of two new cases.

The surgical ciliated cyst, also known as the "postoperative maxillary cyst" or "implantation cyst," occurs as a result of iatrogenic implantation of respiratory epithelium into a noncontiguous surgical site after sinus surgery. It typically presents as a well-defined radiolucency in the maxilla in young adults. Histopathologically, the cyst is lined by ciliated columnar, cuboidal, or pseudostratified squamous epithelium with mucous cells. We report two rare cases of a surgical ciliated cyst located in the mandible.

Generation of recombinant antibody fragments that target canine dendritic cells by phage display technology.

One of the main goals in cancer immunotherapy is the efficient activation of the host immune system against tumour cells. Dendritic cells (DCs) can induce specific anti-tumour immune responses in both experimental animal models and humans. However, most preclinical studies using small animal models show only limited correlation with studies carried out in clinical settings, whereas laboratory dogs naturally develop tumours that are biologically and histopathologically similar to their human counterparts. Here, we describe the generation and characterization of recombinant antibodies against canine DCs, isolated using the Tomlinson phage display system. We successfully isolated highly specific single-chain variable fragment (scFv) antibodies in a sequential three-step panning strategy involving depletion on canine peripheral blood mononuclear cells followed by positive selection on native canine DCs. This provides the basis for an antibody-based method for the immunological detection and manipulation of DCs and for monitoring antigen-specific immune responses.

Predominant functional activity of the large, neutral amino acid transporter (LAT1) isoform at the cerebrovasculature.

In this study, we identify the predominant functional expression of the large, neutral amino acid transporter (LAT1) isoform at the blood-brain barrier (BBB). An in situ rat brain perfusion technique allowed perfusion of the radiotracer [(14)C]-L-Leu (a ligand for both LAT1 and LAT2) alone or competed with excess concentration of either LAT1 or LAT2 specific amino acids. The LAT2 specific amino acid, [(14)C]-L-Asn, was perfused alone or with excess concentration of various amino acids. The brain uptake of [(14)C]-L-Leu was not significantly inhibited by LAT2 specific amino acids, but was inhibited significantly (up to 90%) by the LAT1 specific amino acid, D-Met. L-Asn did not demonstrate saturable brain uptake. These data clearly demonstrate that LAT1 is the functionally predominant isoform expressed at the BBB which is responsible for brain uptake of large, neutral amino acids. In addition, the functional activity of cerebrovascular LAT2 is insignificant, or absent.

A bioreversible prodrug approach designed to shift mechanism of brain uptake for amino-acid-containing anticancer agents.

By derivatization at the N-terminus of amino acid-based anticancer agents (e.g. melphalan and acivicin) to form a drug delivery system (TDDS), we demonstrate a change in the mechanism of brain uptake from the large neutral amino acid transporter (LAT) pathway to passive. An in situ rat brain perfusion technique was used to determine the brain capillary permeability-surface area (PA) product for [(14)C]L-Leu as control (5.18 +/- 0.32 x 10(-2) mL/s/g), which was inhibited competitively (to 7-18% of control) by an excess concentration of the amino-acid-containing anticancer agents, acivicin and melphalan. However, TDDS did not compete for LAT-mediated brain uptake of the radiotracer [(14)C]L-Leu. Brain uptake of TDDS was determined after in situ brain perfusion followed by RP-HPLC along with LC-MS/MS detection of the analytes in brain samples. The PA product for CH(3)-TDDS containing melphalan (5.09 +/- 2.0 x 10(-2) mL/s/g) shows that these agents rapidly cross the blood-brain barrier. Furthermore, competition studies of CH(3)-TDDS with [(3)H]verapamil suggest that the TDDS interacts significantly with the multidrug resistant efflux system (P-glycoprotein) at the blood-brain barrier. Therefore, TDDS were shown to lack LAT-mediated brain uptake. The drug delivery systems, however, showed uptake predominantly via the passive route along with recognition by the multidrug resistant efflux protein at the cerebrovasculature.

High bicoid levels render the terminal system dispensable for Drosophila head development.

In Drosophila, the gradient of the Bicoid (Bcd) morphogen organizes the anteroposterior axis while the ends of the embryo are patterned by the maternal terminal system. At the posterior pole, expression of terminal gap genes is mediated by the local activation of the Torso receptor tyrosine kinase (Tor). At the anterior, terminal gap genes are also activated by the Tor pathway but Bcd contributes to their activation. Here we present evidence that Tor and Bcd act independently on common target genes in an additive manner. Furthermore, we show that the terminal maternal system is not required for proper head development, since high levels of Bcd activity can functionally rescue the lack of terminal system activity at the anterior pole. This observation is consistent with a recent evolution of an anterior morphogenetic center consisting of Bcd and anterior Tor function.

Modulating blood-brain barrier interactions of amino acid-based anticancer agents.

The large neutral amino acid (LNAA) transporter at the blood-brain barrier (BBB) mediates brain uptake of amino acid-based anticancer agents (e.g., melphalan and acivicin). In this study, we blocked the amino acid terminus of the anticancer agents using a bioreductive drug delivery system (TDDS). This molecular modification of the anticancer agents is expected to prevent LNAA carrier-mediated transport across the BBB. In this study, we demonstrate that the parent amino acid containing anticancer agents are substrates for the LNAA transporter at the BBB, whereas the TDDS is not recognized by the LNAA transporter. An in situ rat brain perfusion technique was used to determine competition for LNAA carrier-mediated transport at the BBB using [14C]L-leucine. The BBB capillary permeability-surface area (PA) product for the radiotracer [14C]L-leucine (control) was determined to be 5.18 +/- 0.32 x 10(-2) ml/s/g (100%). The control PA value for [14C]L-leucine was competitively inhibited (down to 7-18% of control) by excess L-phenylalanine as well as by excess concentration of the anticancer amino acids, melphalan and acivicin, showing competition for the LNAA transporter at the BBB. In contrast, brain perfusion of [14C]L-leucine in presence of excess TDDS resulted in no competition for brain uptake of [14C]L-leucine via the LNAA transporter. Thus, bioreversible derivatization of the parent anticancer amino acids resulted in blocking the amino acid functional group, thereby leading to loss of recognition for the cerebrovascular LNAA transporter at the BBB.

A green fluorescent protein enhancer trap screen in Drosophila photoreceptor cells.

The Drosophila ommatidia contain two classes of photoreceptor cells (PR's), the outer and the inner PR's. We performed an enhancer trap screen in order to target genes specifically expressed in PR's. Using the UAS/GAL4 method with enhanced green fluorescent protein (eGFP) as a vital marker, we screened 180000 flies. Out of 2730 lines exhibiting new eGFP patterns, we focused on 16 lines expressing eGFP in particular subsets of PR's. In particular, we describe three lines inserted near the spalt major, m-spondin and furrowed genes, whose respective expression patterns resemble those genes. These genes had not been reported to be expressed in the adult eye. These examples clearly show the ability of our screen to target genes expressed in the adult Drosophila eye.

(+/-)-trans-2-[3-methoxy-4-(4-chlorophenylthioethoxy)-5-(N-methyl-N- hydroxyureidyl)methylphenyl]-5-(3,4, 5-trimethoxyphenyl)tetrahydrofuran (CMI-392), a potent dual 5-lipoxygenase inhibitor and platelet-activating factor receptor antagonist.

By incorporating an N-hydroxyurea functionality onto diaryltetrahydrofurans, a novel series of compounds was investigated as dual 5-lipoxygenese (5-LO) inhibitor and platelet-activating factor (PAF) receptor antagonist. These dual functional compounds were evaluated in vitro for 5-LO inhibition in RBL cell extracts and human whole blood, and PAF receptor antagonism in a receptor binding assay. PAF-induced hemoconcentration and arachidonic acid- and TPA-induced ear edema in mice were used to determine in vivo activities. The structure-activity relationship analysis to define a preclinical lead is presented. (+/-)-trans-2-[3-methoxy-4-(4-chlorophenylthioethoxy)-5-(N-methyl- N-h ydroxyureidyl)methylphenyl]-5-(3,4, 5-trimethoxyphenyl)tetrahydrofuran (40, CMI-392) was selected for further study. In the arachidonic acid-induced mouse ear edema model, 40 was more potent than either zileuton (a 5-LO inhibitor) or BN 50739 (a PAF receptor antagonist), and it demonstrated the same inhibitory effect as a physical combination of the latter two agents. These results suggest that a single compound which both inhibits leukotriene synthesis and blocks PAF receptor binding may provide therapeutic advantages over single-acting agents. The clinical development of compound 40 is in progress.

Validation and assessment of a conversational interaction intervention.

Multiple measures of social behavior were obtained in order to (a) evaluate the effects of a systematic replication of a conversational interaction intervention, (b) identify critical target behaviors, (c) select participants likely to benefit from intervention, and (d) assess the social and experimental validity of the intervention. Combined measures (e.g., performance screening, assessing students' goals, behavioral ratings) corroborated the decision to introduce multiple-exemplar, self-instructional training to 4 students with moderate mental retardation who were observed to have limited social interaction with their peers at school. Findings indicated that the procedure was effective at increasing empirical indicators of participants' generalized conversational interaction and that behavioral change was validated socially by ratings of peers and important others.

The effects of multiple-exemplar self-instructional training on high school students' generalized conversational interactions.

A multiple-baseline-across-students design was used to investigate the effects of multiple-exemplar self-instructional training on the acquisition and generalization of conversational interaction of 4 high school students with mental retardation. The multiple-exemplar component of the model consisted of (a) several peers without disabilities teaching the use of a self-instructional social skills strategy across diverse examples of conversational interactions and across two settings and (b) assessing the generalized effects of training across additional peers and one setting. Findings indicated that peers were effective in teaching the multiple-exemplar strategy and that peer training was associated with systematic increases in generalized conversational interactions with familiar and unfamiliar peers with and without disabilities in an additional setting. Social validation data indicated that following multiple-exemplar training, all participants' performances approximated those of general education students and was judged by others to have improved.

Correction of bone marrow nucleated cell counts for the presence of fat particles.

Although the use of bone marrow transplantation has increased greatly in recent years, the quality control procedures used in bone marrow processing laboratories remain less than ideal. Accurate marrow total nucleated cell (TNC) counts are essential for effective monitoring of bone marrow collection and processing. Aspirated marrow is variably contaminated by fat particles, resulting in overestimation of marrow TNC by automated analyzers. A recently-marketed hematological analyzer (Cobas-Helios; Roche Diagnostic Systems, Branchburg, NJ) offers the potential to correct marrow TNC counts for fat particles using available software. The authors investigated the accuracy of corrected TNC counts on 21 marrow samples, using a visual chamber count as the reference method. The correction methods studied were software correction, using the Cobas-Helios differential system, and replacement of the sample plasma with saline. Uncorrected automated marrow TNC counts (mean, 28.4 x 10(9)/L) were significantly higher than the visual reference counts (mean, 23.1 x 10(9)/L). Neither the mean corrected automated count (24.3 x 10(9)/L) nor the mean saline replaced count (24.6 x 10(9)/L) differed significantly from the mean visual reference count. For both the corrected automated and saline replaced counts, 20 of the 21 data points (95%) fell within a 95% confidence interval computed for the reference method. The authors conclude that both the corrected automated method, using the Cobas-Helios, and the saline replacement method are acceptable alternatives to the visual chamber count.

Quality of life in applied research: a review and analysis of empirical measures.

A consensus list of 15 dimensions and corresponding components of quality of life was derived from the literature and proposed as a conceptual model. Eighty-seven empirical studies found in the applied research then were analyzed, and the proposed model was found to be supported by 1,243 empirical measures. In addition, studies were analyzed with respect to (a) demographic variables, (b) type of investigation, (c) method of assessment, (d) psychometric properties of assessments used, and (e) common dimensions of measures investigated and their frequency of measurement. Based upon findings, issues regarding methodology, implications for policy and practice in the field, and recommendations for future research were discussed.

Acute procedural results in the treatment of 30 coronary artery bifurcation lesions with a double-wire atherectomy technique for side-branch protection.

Percutaneous treatment of bifurcation lesions has been consistently shown to be associated with lower acute success rates, higher initial complication rates, and an increased rate of restenosis when compared with findings in nonbifurcation lesions. Recent analysis of data from a CAVEAT subgroup suggests that directional atherectomy of bifurcation lesions can improve initial success rates and lower restenosis rates but at the cost of high complication rates. Reports from several angioplasty series document improved success rates and lower complication rates with the use of a two-wire technique to protect side branches when treating bifurcation lesions. Our experience with a two-wire atherectomy technique that uses a nitinol wire to protect important side branches is presented.

A procedure for successful nonsurgical embryo transfer in swine.

The current study was undertaken to develop a successful procedure for the nonsurgical transfer of pig embryos. A total of 663 embryos were surgically collected on Day 4 or 5 from 55 donors, of which 542 embryos of acceptable quality were nonsurgically transferred to 46 recipients. Nonsurgical recipient gilts were sedated 15 min prior to transfer with 20 mg im acepromazine maleate. A disposable insemination spirette with an attached 3-way stopcock was manipulated into the cervix of each gilt. Embryos were expelled from a tomcat catheter into the spirette, and 10 to 12 ml of Whitten's medium were used to flush embryos through the spirette into the reproductive tract. Sixteen (34.8%) recipient gilts did not return to estrus before Day 36, and 10 (21.7%) gilts farrowed with an average litter size of 4.3 +/- 0.7. Embryos were collected from an additional 20 donors and were surgically transferred to an additional 19 recipients. Surgical transfers conducted at the same time as the nonsurgical transfers resulted in 12 (63.2%) gilts farrowing and 7.1 +/- 0.6 pigs were born per litter. In conclusion, a procedure has been developed for nonsurgical transfer of swine embryos which simplifies the process of embryo transfer and which may increase the potential for utilization of embryo transfer technologies by swine producers.

Examination of common PTCA guide wires used for side branch protection during directional coronary atherectomy of bifurcation lesions performed in vivo and in vitro.

Side branch protection of bifurcation lesions with guide wires treated with directional coronary atherectomy has been performed safely in a small number of patients using a nickel/titanium (Nitinol) alloy guide wire. There is no information about the safety of using other guide wires in this scenario. The purpose of this study was to microscopically examine a variety of guide wires exposed to the directional coronary atherectomy (DCA) device in vitro. Eight common guide wires used in percutaneous coronary angioplasty were positioned in a side branch of a simulated coronary bifurcation and exposed to the operating DCA device. The guide wires were examined grossly and under scanning electron microscopy. The Nitinol guidewire suffered no discernible damage in comparison to varying degrees of damage seen on the other guide wires, and this guidewire appears to be best suited for side branch protection. Examination of the protecting guide wire and cutting cylinder used in the initial patients also showed no damage. Further clinical trials are necessary to assess the efficacy of DCA under such conditions.

Modified "kissing" atherectomy procedure with dependable protection of side branches by two-wire technique.

We report a new two-wire atherectomy technique for side branch protection. Newer, more resilient wire designs are capable of withstanding cutting forces of the Simpson atherocath device. This technique expands atherectomy application to lesions previously excluded from atherectomy as high risk lesions.

Constrictive pericarditis after cardiac surgery.

Forty-five patients were identified as having constrictive pericarditis after cardiac surgery. The mean patient age was 61 years (range, 40 to 77 years). Twenty-three of 37 patients with adequate clinical information were reported to have had a diagnosis of postpericardiotomy syndrome after the original surgery. The mean interval from original surgery to presentation with constriction was 23.4 months (range, 1 to 204 months). Computerized tomography was helpful in establishing a diagnosis of constriction in 23 of 29 patients (79%). Bypass graft patency was 93% (85 of 91 grafts). Severe pulmonary hypertension (pulmonary artery systolic pressure greater than or equal to 60 mm Hg) was present in nine patients; 8 had coexistent valvular disease (seven cases of mitral valve disease, and aortic valve disease in one). Thirty-seven of the 45 patients underwent pericardial stripping, 28 of whom experienced marked symptomatic improvement. One patient had persistent right heart failure, which was not documented to be secondary to constriction. Four patients had persistent constrictive physiologic conditions. Three of these patients had more extensive pericardial stripping and showed clinical improvement. Four patients (11%) died within 30 days of stripping. Eight patients received medical therapy alone. The decision to treat patients medically was based either on favorable response to medical therapy (five patients), or poor general clinical status.