Sex- and age-related shift of relapse phenotypes in a cohort of relapsing multiple sclerosis patients: Post hoc analysis from the OPERA phase 3 trials.

BACKGROUND AND PURPOSE: Relapse presentation in relapsing multiple sclerosis (RMS) differs between sexes, leading to differential outcomes. An influence of age seems likely but is less well investigated separately for women and men. METHODS: Using the large well-defined dataset of the pivotal trials of ocrelizumab in RMS, OPERA I and II, and their open-label extension, we performed a post hoc analysis to investigate relapse phenotypes for sex- and age-related differences in n = 929 relapses in 534 subjects (171 men, 363 women). Frequencies of affected functional systems were analyzed separated by sex and for three age strata (<35, 35-44, ≥45 years). Exact p-values are given for this exploratory analysis. RESULTS: Frequencies of mono- versus polysymptomatic relapse presentations were different neither between sexes nor in different age groups. Cerebellar symptoms were more frequent in relapses in men (female [f]: 23.1%, male [m]: 33.0%, p = 0.002), and women's relapses included more sensory (f: 53.8%, m: 32.3%, p < 0.001) and fatigue symptoms (f: 22.6%, m: 14.7%, p = 0.006). Whereas the sex difference for sensory involvement was present over all age groups (<35 years: f: 58.3%, m: 30.4%, p < 0.001; 35-44 years: f: 53.7%, m: 36.0%, p = 0.003; ≥45 years: f: 47.8%, m: 28.8%, p = 0.009), the difference for cerebellar involvement diminished with age (<35 years: f: 20.1%, m: 33.3%, p = 0.009; 35-44 years: f: 22.7%, m: 34.2%, p = 0.034; ≥45 years: f: 27.8%, m: 30.3%, p = 0.750). Relapse presentation seemed to shift with age in women only. CONCLUSIONS: We describe sex-specific relapse presentations and an influence of age only for women. Underlying causal factors warrant further investigations.

Phylogenetic and kinematic constraints on avian flight signals.

Many birds vocalize in flight. Because wingbeat and respiratory cycles are often linked in flying vertebrates, birds in these cases must satisfy the respiratory demands of vocal production within the physiological limits imposed by flight. Using acoustic triangulation and high-speed video, we found that avian vocal production in flight exhibits a largely phasic and kinematic relationship with the power stroke. However, the sample of species showed considerable flexibility, especially those from lineages known for vocal plasticity (songbirds, parrots and hummingbirds), prompting a broader phylogenetic analysis. We thus collected data from 150 species across 12 avian orders and examined the links between wingbeat period, flight call duration and body mass. Overall, shorter wingbeat periods, controlling for ancestry and body mass, were correlated with shorter flight call durations. However, species from vocal learner lineages produced flight signals that, on average, exceeded multiple phases of their wingbeat cycle, while vocal non-learners had signal periods that were, on average, closer to the duration of their power stroke. These results raise an interesting question: is partial emancipation from respiratory constraints a necessary step in the evolution of vocal learning or an epiphenomenon? Our current study cannot provide the answer, but it does suggest several avenues for future research.

Placental growth factor blunts uterine artery responses to angiotensin II.

OBJECTIVE: Changes in maternal serum concentration of placental growth factor (PlGF) and vascular response to intravascular infusion of Angiotensin II (Ang II) follow a bell-shaped curve pattern during gestation. This study evaluates the effects of PlGF and soluble vascular endothelial growth factor receptor-1 (sFlt-1) on responses of human uterine arteries (UA) to Ang II. DESIGN: Experimental. SETTING: Baylor College of Medicine and Texas Children's Hospital-Pavilion for Women. SAMPLE: Uterine arteries samples (n = 14) were obtained from normotensive women undergoing caesarean hysterectomy at ≥32 weeks. METHODS: Uterine arteries rings were incubated with (1) Krebs solution; (2) PlGF at 1.45, 14.5, and 500 pg/ml; (3) sFlt-1 at 2 ng/ml; and (4) a combination of sFlt-1, and PlGF. Dose-contraction responses to Ang II were determined in UA rings incubated in the above-mentioned conditions. Responses were also measured in presence of L-NAME or inhibitors of endothelium-derived hyperpolarising factor: apamine and charybdotoxin. The t-test was used for comparisons. MAIN OUTCOME MEASURE: Changes in vascular reactivity of UA rings. RESULTS: PlGF blunted (P = 0.03) and sFlt-1 increased (P <0.01) the UA maximum responses to Ang II. A combination of sFlt-1 and PlGF blunted UA responses to Ang II (P < 0.05). l-NAME, apamine, and charybdotoxin reversed the relaxation effects of PlGF on UA responses to Ang II (P < 0.05). CONCLUSIONS: PlGF contributes to the blunted vascular response to Angiotensin II during normotensive pregnancies and sFlt-1 appears to attenuate this effect. PlGF and sFlt-1 may contribute to the regulation of vascular tone during pregnancy by altering the vascular response to Angiotensin II. FUNDING: Baylor College of Medicine. TWEETABLE ABSTRACT: Placental growth factor and soluble vascular endothelial growth factor receptor-1 modulate the uterine artery response to Angiotensin II in normotensive pregnant women.

Factors associated with ever use of mammography in a limited resource setting. A mixed methods study.

OBJECTIVE: To evaluate facilitators and barriers influencing mammography screening participation among women. DESIGN: Mixed methods study. SETTING: Three hospital catchment areas in Hidalgo, Mexico. PARTICIPANTS: Four hundred and fifty-five women aged 40-69 years. INTERVENTION: Three hundred and eighty women completed a survey about knowledge, beliefs and perceptions about breast cancer screening, and 75 women participated in semi-structured, in-person interviews. Survey data were analyzed using logistic regression; semi-structured interviews were transcribed and analyzed using elements of the grounded theory method. MAIN OUTCOME MEASURE: Women were categorized as never having had mammography or having had at least one mammogram in the past. RESULTS: From survey data, having had a Pap in the past year was associated with ever having had breast screening (odds ratio = 2.15; 95% confidence interval 1.30-3.54). Compared with never-screened women, ever-screened women had better knowledge of Mexican recommendations for the frequency of mammography screening (49.5% vs 31.7% P < 0.001). A higher percentage of never-screened women perceived that a mammography was a painful procedure (44.5% vs 33.8%; P < 0.001) and feared receiving bad news (38.4% vs 22.2%; P < 0.001) compared with ever-screened women. Women who participated in semi-structured, in-person interviews expressed a lack of knowledge about Mexican standard mammographic screening recommendations for age for starting mammography and its recommended frequency. Women insured under the 'Opportunities' health insurance program said that they are referred to receive Pap tests and mammography. CONCLUSIONS: Local strategies to reduce mammogram-related pain and fear of bad news should work in tandem with national programs to increase access to screening.

VUV and soft x-ray ionization of a plant volatile: Vanillin (C8H8O3).

Plant volatiles are emitted by plants in response to several forms of stress, including interaction with energetic photons. In the present work, we discuss the interaction of extreme UV and soft X-ray photons with a plant volatile, vanillin. The single and double (multiple) ionization of the vanillin molecule have been studied for the first time using time-of-flight mass spectrometry and VUV and soft X-ray photons (synchrotron radiation, at 12.0 eV, 21.2 eV, 130 eV, 310 eV, 531 eV, and 550 eV). At 12.0 and 21.2 eV, only singly charged species are observed and the parent ion, C8H8O3 (+), is the dominant species. Energy differences for some selected fragments were calculated theoretically in this energy region. At 130 eV, direct double and triple ionization of the valence electrons may occur. The fragmentation increases and CHO(+) becomes one of the main cations in the mass spectrum. The molecular ion is still the dominant species, but other fragments, such as C6H5O(+), begin to present similar intensities. At 310 eV, C 1s electrons may be ionized and Auger processes give rise to dissociative doubly ionized cations. Ionization around the O 1s edge has been studied both at the 531 eV resonance and above the ionization edge. Resonant and normal Auger processes play a significant role in each case and a large fragmentation of the molecule is observed at both photon energies, with intense fragments such as CHO(+) and CH3 (+) being clearly observed. A near edge X-ray absorption fine structure spectrum of the vanillin molecule was obtained around the O 1s ionization threshold. In addition, the fragmentation of vanillin has also been studied using a fast beam of electrons (800 eV), for the sake of comparison.

Outcomes after laparoscopic conversion of failed adjustable gastric banding to sleeve gastrectomy or Roux-en-Y gastric bypass.

BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) has a high incidence of long-term complications and failures. The best procedure to handle these failures and the optimal number of stages in such cases is still controversial. The aim of this retrospective study was to compare the results of conversions of LAGB to either laparoscopic sleeve gastrectomy (LSG) or laparoscopic Roux-en-Y gastric bypass (LRYGB) in failed LAGB using a single-stage approach. METHODS: All patients who underwent conversion from LAGB to either LRYGB or LSG between January 2005 and March 2012 were included in the study. Early and late complications were reviewed. The percentage excess weight loss (%EWL) between the two groups was compared at 3, 6, 12 and 24 months of follow-up. RESULTS: Fifty-nine patients, 11 men and 48 women, were included in the study. The most frequent indication was insufficient weight loss or weight regain (non-responders group), in 44 patients (75 per cent); 15 patients had a revision for complicated LAGB. The early complication rate in the non-responders group was 7 per cent (3 of 44 patients), compared with 13 per cent (2 of 15) in the complicated LAGB group. Mean(s.d.) %EWL in the non-responders group was 55(22) per cent in patients converted to LRYGB and 28(25) in those converted to LSG (P = 0·001). CONCLUSION: LRYGB and LSG are both safe and feasible options for failed or complicated LAGB. In the non-responders group, %EWL was superior for conversion to LRYGB. The surgical morbidity rate was highest in patients having revision for band complications.

Endometrial inflammatory markers of the early immune response in mares susceptible or resistant to persistent breeding-induced endometritis.

Transient endometritis after breeding is necessary for clearance of bacteria and spermatozoa; however, in a subpopulation of mares, the inflammation fails to resolve in a timely fashion. The objective of this study was to describe the uterine inflammatory response in mares susceptible or resistant to persistent breeding-induced endometritis (PBIE) during the first 24 h after induction of uterine inflammation.Twelve mares were classified as susceptible (nZ6) or resistant (nZ6) to PBIE. Mares were inseminated over five estrous cycles and endometrial biopsies were collected at one time point per cycle before (0) and 2, 6, 12, and 24 h after insemination. qPCR analysis for IL1B, IL6, IL8, IFNG, TNF (TNFA), IL10, and IL1RN was performed, and endometrial inflammatory cells were counted for each sample. Relative quantification values reported fold changes in mRNA expression from 0 h values. A general pattern of expression post insemination was observed in both groups of mares. Cytokine mRNA increased at 2 h, peaked between 2 and 12 h, and then decreased.Differences were detected between groups of mares 6 h after challenge; resistant mares had higher mRNA expression of IL6, IL1RN,and IL10 than susceptible mares. Susceptible mares had an increased number of polymorphonuclear neutrophils in the endometrium 2 and 12 h after breeding when compared with resistant mares. These findings describe an inherent difference in the initial immune response to insemination and may help explain the transient nature of inflammation in resistant mares, whereas susceptible mares develop a persistent inflammation.

New immediate release formulation for deterring abuse of methadone.

CONTEXT: Drug abusers are known to take a dosage form containing an opioid analgesic and crush, shear, grind, chew, or dissolve it in water or in alcohol, in order to extract the opioid component. OBJECTIVE: Develop an anti abuse immediate release formulation using methadone as model drug. MATERIALS AND METHODS: Tablets combining methadone and alkalizing agents were manufactured. A methadone assay was used to determine extraction efficiency from tablets in aqueous and alcohol solvents. In vitro dissolution testing was used to determine drug release in different media. RESULTS AND DISCUSSIONS: Meglumine-based formulations prevented extraction of 70 to 100% of methadone from tablets. Addition of this alkalizing agent caused methadone to precipitate out of a solution along with other ingredients and be retained on standard filters. Meglumine-containing and control tablets showed similar dissolution profiles in acidic media, suggesting adequate solubilisation of the drug early in the gastrointestinal tract. Finally, stability upon storage of the formulations for 6 months at 25°C/60%RH and 40°C/75%RH was confirmed. CONCLUSION: Incorporation of an alkalizing agent into methadone tablets significantly reduced the preparation of a methadone solution for intravenous administration and abuse, while allowing the formulation to release methadone in gastric media and provide desired pharmacological effect.

[Development of a nested polymerase chain reaction test for the diagnosis of transmissible gastroenteritis of pigs]. / Desarrollo de una prueba de reacción en cadena de la polimerasa anidada para el diagnóstico de la gastroenteritis transmisible del cerdo.

The aim of this study was to develop a nested polymerase chain reaction (nested PCR) for the rapid detection of transmissible gastroenteritis virus (TGEV) of pigs. The primers were designed on the basis of highly conserved regions of several TGEV sequences included in the analysis. External primers were used to amplify a fragment of the expected size (441 bp) in all the samples evaluated using reverse transcriptase polymerase chain reaction (RT-PCR), but with very low intensity. In the second amplification (nested PCR), internal primers were used to amplify a fragment of the expected size (168 bp), with good concentration. The performance of the test based on virus isolates in tissue culture and in clinical samples was judged good for the virological diagnosis of transmissible gastroenteritis of pigs.

[Prevalence of asthma and associated risk factors in schoolchildren from an area of indigenous population]. / Prevalencia de asma y factores de riesgo asociados en escolares provenientes de una zona de pueblos indígenas.

INTRODUCTION: The prevalence of asthma in Chile ranges from 10.2 to 14.9%. In previous studies, the Araucanía Region has not been included. OBJECTIVE: To determine the prevalence of asthma in the school po pulation of the Araucanía Region. SUBJECTS AND METHOD: The ISAAC questionnaires were used for asthma symptoms in addition to a questionnaire for recording sociodemographic data, belonging to the Mapuche ethnicity, type of heating, exposure to indoor air pollution, and family history. The Binomial Regression Model was used to evaluate the effect of each of the different variables of inter est, adjusting by age groups (6-7 and 13-14 years). The model also evaluated the additive interaction between these variables and age. RESULTS: 823 surveys were applied, where the prevalence of asthma was 14.2% and 23.2% in the 6-7 year old group and the 13-14 year old group, respectively. 43.7% de clare to be Mapuche, 32.9% live in rural areas, and 81.4% use firewood as a heating method. Through binomial regression model analysis, where for the study group, living in the coastal commune was significantly associated with a lower prevalence of asthma (-13.2% [CI: -17.7 to -8.6]. The history of an asthmatic mother was significantly associated with a higher prevalence of asthma (17.9% [CI: 2.7 to 33.1]. Living in a rural area, to be Mapuche or the type of heating used, showed differences statistically significant (p 0.18, p 0.609 and p 0.480, respectively) Conclusion: 13-14 year-olds school children in the Araucanía Region presented a higher prevalence of asthma. There was an association with asthma in the mother where this increase is not associated with living in a rural area, to be Ma puche or type of heating.

Mutational analysis of BRCA1 and BRCA2 genes in Mexican breast cancer patients.

Germline mutations in the BRCA1 and BRCA2 genes predispose to breast and ovarian cancer. A variable incidence of mutations has been reported for these genes. The contribution of BRCA1 and BRCA2 mutations to Mexican women with breast and/or ovarian cancer is not known. Because of the increasing prevalence of breast cancer in this population, it is necessary to study the presence of mutations in both genes. We screened BRCA1 and BRCA2 genes in 40 patients: 29 patients with a history of breast and/or ovarian cancer, and 11 patients with early-onset breast cancer (< 40 years), through denaturing high performance liquid chromatography analyses. We found two frameshift mutations in BRCA1 and one missense mutation in each gene. Additionally we found several intronic variants as well as synonymous mutations. We found 5% of deleterious mutations in the BRCA genes. Larger studies are needed to establish the significance and prevalence of BRCA mutations among Mexican women.

Carotid artery restenosis in a Hispanic population.

BACKGROUND AND PURPOSE: Carotid endarterectomy is one of the main surgical procedures used for carotid stenosis and its recurrence. Besides the setting of a randomized controlled trial for asymptomatic and symptomatic carotid artery stenosis, there is little information about the rate of restenosis after carotid endarterectomy in Hispanics. The purpose of this study is to describe the results of carotid endarterectomy on the basis of restenosis in a Hispanic population. METHOD: A retrospective revision of 47 endarterectomies performed on 43 patients by a single surgeon at the VA Caribbean Health Care System and Pavia Hospital, during an eight year period (1990-1998), was conducted. Information about endarterectomies, restenosis and known risk factors for carotid stenosis were obtained from medical records. RESULTS: Of the 43 patients, 31 were male (72%) and 12 female (28%), with a mean age of 67.9 years. Re-operations for recurrent carotid stenosis were performed in 2 patients (4.7%). Restenosis cases were asymptomatic, hence diagnosed through followup ultrasound Duplex studies and confirmed by angiography after 3 and 4 years of the first surgical procedure. The degree of restenosis (70% to 99%) after the initial endarterectomy was 4.3%. The major risk factors found among patients were hypertension (58%), hypercholesterolemia (50%), smoking (46%), and alcohol (34%). CONCLUSIONS: Carotid endarterectomy with primary closure is safe and durable. Repeated surgery using patch grafts in this Hispanic population was also safe. The concordance of risk factors and incidence of carotid stenosis correlated well with other studies.

Myths and realities about knowledge, attitudes and practices of household contacts of tuberculosis patients.

SETTING: Five Colombian cities: Villavicencio, Pereira, Cucuta, Bucaramanga and Bosa. OBJECTIVE: To describe the knowledge, attitudes and practices related to tuberculosis (TB) in household contacts (HHCs) of TB patients. METHODS: A cross-sectional study was conducted. The estimated sample size required was 855. The World Health Organization guide 'Advocacy, communication and social mobilization for TB control: a guide to developing knowledge, attitude and practice surveys' was translated into Spanish and adapted to the dialect of each city. HHCs were invited to participate in the study and included if they agreed. RESULTS: We interviewed 878 HHCs. Most of them knew that TB was transmitted by airborne droplets; however, 52.2% also said that TB could be transferred from one person to another by sharing plates or clothes or shaking hands. Fifty-five per cent of HHCs acquired TB-related information from health care workers, and 44% from family members and friends. Fear was the main reaction reported by HHCs when they were informed of a TB diagnosis (60%). Eighty-five per cent of HHCs answered that the community rejects or avoids TB cases. CONCLUSION: There are significant gaps in the knowledge about TB and the stigmatisation associated with it. Educational strategies should be designed together with the community to demystify many misconceptions about TB.

Fibrous condylar dysplasia: Resection and reconstruction with a custom-made TMJ prosthesis using virtual surgical planning.

The main objective of this article is to describe the indications for custom-made alloplastic reconstruction of the temporomandibular joint (TMJ) using the new techniques of virtual surgical planning and computer-aided design/computer-aided manufacturing (CAD/CAM) technology through the study of a clinical case and an updated review of the literature. The patient was a 45-year-old male with a fast-growing condylar dysplasia that produced a progressive facial deformity with limited mandibular mobility, which required a radical resection. Predictable, satisfactory, and stable results over time are described from the aesthetic, functional, and symptomatic points of view, through a comprehensive approach to all the aspects of this case by following an appropriate line of development and using the most current techniques.

The impact of measurement of respiratory quotient by indirect calorimetry on the achievement of nitrogen balance in patients with severe traumatic brain injury.

BACKGROUND: This study evaluated the impact of IC on the optimization of nutritional support and the achievement of +NB in patients with TBI. MATERIALS AND METHODS: 27 patients (GCS ≤ 8), treated with a 5-day multimodality monitoring and goal-directed therapy protocol, received enteral nutrition on day 1 followed by IC on days 3 and 5 and assessment of NB on day 7. In the first cohort (n = 11), no adjustment in kcal was made. In the second cohort (n = 16), nutrition was targeted to an RQ of 0.83 by day 3. The first cohort was analyzed with respect to NB status; the second cohort was compared to patients with (-) and +NB of the first cohort. Data (mean ± SD) were analyzed with unpaired t test, and Chi square and Fisher exact tests. RESULTS: 4/11(36 %) patients in the first cohort had +NB. The predicted mortality by TRISS, substrate utilization, and RQ was significantly lower compared to the second cohort. The mortality predicted by the CrasH model did not differ between the two cohorts. A RQ of 0.74 was associated with the preferential use of fat and protein and -NB, whereas a RQ of 0.84 favored utilization of carbohydrates and +NB. All patients whose kcal intake was adjusted based on the RQ on day 3 reached a +NB by day 7. CONCLUSION: An increase in kcal ≥25 % in patients with a RQ < 0.83 on day 3 improves substrate utilization, decreases protein utilization and optimizes the achievement of +NB by day 7.

Long-term function (6 years) of islet allografts in type 1 diabetes.

Eight type 1 diabetic patients, ages 29-41 years, with mean diabetes duration of 23 years (range 18-29 years) received islet transplants from 1 to 5 donors. Seven patients had stable kidney allografts 1-11 years before the islet transplant, and one patient had a simultaneous islet-kidney allograft. Patients' blood glucose control was poor as reflected by the mean +/- SD HbA1c of 9.1 +/- 1.7% before transplant. Of the first three patients, two (1 and 3) achieved insulin independence for 36 and 38 days, respectively. Two recipients rejected their islet grafts within 1 month (2 and 8) and therefore were excluded from analysis. The HbA1c and insulin requirement of the six remaining patients who had persistent islet function for more than 60 days was significantly reduced from 9.3 +/- 1.9 to 6.4 +/- 1.0% (P = 0.002) and from 0.75 +/- 0.15 to 0.35 +/- 0.12 U x kg(-1) x day(-1) (P < 0.001), respectively. The two patients with the longest graft survival (4 and 6) achieved a normalization or near-normalization of their HbA1c levels during 6 years in the absence of severe episodes of hypoglycemia. As demonstrated by a decline in C-peptide response during Sustacal challenge tests over a 6-year period, there was a diminution of islet allograft function over time, despite persistence of normal or near normal HbA1c. We concluded that transplantation of allogeneic islets with an islet mass comparable with whole or segmental pancreas transplants in type 1 diabetic patients can result in long-term islet allograft function; further, we concluded that, in conjunction with small dosages of exogenous insulin, a functioning islet allograft can result in near-normalization of blood glucose levels and significant improvement in HbA1c. The occurrence of severe hypoglycemic episodes observed for patients in the Diabetes Control and Complications Trial was not observed in recipients with functioning islet transplants, despite the continuous need for exogenous insulin therapy to sustain normal HbA1c over the 6-year follow-up. The significant improvement in metabolic control observed for the patients described in this study, and the potential to significantly decrease or halt the progression of diabetic complications, support the continued application of islet allotransplantation as a treatment modality for type 1 diabetic patients.

Haldane's sieve and adaptation from the standing genetic variation.

We consider populations that adapt to a sudden environmental change by fixing alleles found at mutation-selection balance. In particular, we calculate probabilities of fixation for previously deleterious alleles, ignoring the input of new mutations. We find that "Haldane's sieve"--the bias against the establishment of recessive beneficial mutations--does not hold under these conditions. Instead probabilities of fixation are generally independent of dominance. We show that this result is robust to patterns of sex expression for both X-linked and autosomal loci. We further show that adaptive evolution is invariably slower at X-linked than autosomal loci when evolution begins from mutation-selection balance. This result differs from that obtained when adaptation uses new mutations, a finding that may have some bearing on recent attempts to distinguish between hitchhiking and background selection by contrasting the molecular population genetics of X-linked vs. autosomal loci. Last, we suggest a test to determine whether adaptation used new mutations or previously deleterious alleles from the standing genetic variation.

Characterisation of the inflammatory cytokine response to anthelmintic treatment in ponies.

REASONS FOR PERFORMING STUDY: Anthelmintic treatments have been associated with local inflammatory reactions. Since each class of anthelmintic has unique mechanisms of action affecting different subpopulations of parasites, we hypothesised that they will also induce characteristic proinflammatory responses. OBJECTIVES: To determine the effect of anthelmintic class on the proinflammatory response post treatment. STUDY DESIGN: Ponies naturally infected with cyathostomins and other parasites after pasture grazing were left untreated or treated with representatives of 3 different classes of anthelmintics: fenbendazole (benzimidazole); pyrantel tartrate (pyrimidine); and moxidectin (macrocyclic lactone). All were monitored for the expression of proinflammatory genes in the peripheral blood using real-time PCR. METHODS: The ponies were divided into 4 treatment groups: Group 1 (n = 4) were untreated controls; Group 2 (n = 5) received 5 daily doses of fenbendazole (10 mg/kg bwt); Group 3 (n = 4) received daily treatment of pyrantel tartrate 2× (2.65 mg/kg bwt); and Group 4 (n = 5) received a single dose of moxidectin (400 µg/kg bwt). Blood samples were collected daily for 2 weeks to determine the effect of deworming on proinflammatory gene expression. Faecal egg counts were used to evaluate the efficacy of each drug. RESULTS: While treatment with the benzimidazole significantly reduced egg counts up to 14 days post treatment, it also stimulated proinflammatory gene expression. Treatment with pyrantel salt also reduced faecal egg counts with less of a proinflammatory response. Treatment with the macrocyclic lactone was the most successful in reducing faecal egg counts and produced no signs of increased proinflammatory cytokine expression. CONCLUSIONS: This study revealed pronounced differences in the cytokine responses to anthelmintic treatment. This inflammatory reaction may play a role in the development of parasitic disease post anthelmintic treatment.

Characterization of the in situ immunological responses to vaccine adjuvants.

Adjuvants are included with many inactivated and some modified live vaccines to enhance immune responses to specific antigens. While early vaccines relied exclusively upon aluminum salts, still the major adjuvant used in human vaccines, other adjuvant products are used in veterinary medicine. In addition to enhancing antigen presentation, adjuvants can also enhance the development of specific immune responses. Thus, alum adjuvants often preferentially stimulate humoral immune responses. By contrast, lipid-based adjuvants are often more effective at stimulating cell-mediated immune responses. Metastim(®) is a lipid-based adjuvant reported to elicit both humoral and cellular immune responses, though the mechanism responsible for this activity remains unclear. In this study, we compared the ability of equine influenza virus vaccines containing either saline or Metastim(®) or an aluminum phosphate adjuvant to stimulate antigen presenting cell function in vivo. Six ponies were intradermally inoculated with inactivated equine influenza (KY97) mixed with either adjuvant or saline. Multiple sites were injected so that biopsies could be collected at different times post injection. The 4mm punch biopsies were formalin-fixed, paraffin-embedded, and stained with hematoxylin and eosin (H&E). Total RNA was isolated from 2mm punch biopsies for the determination of gene expression by real-time PCR. H&E staining revealed a variety of cells recruited to the injection sites, including lymphocytes, neutrophils, eosinophils and macrophages. Real-time PCR analysis of the injection site confirmed this cellular infiltration and identified increased expression of activation markers. Both vaccines also stimulated gene expressions of pro-inflammatory cytokines. The vaccine containing Metastim(®) elicited significantly higher gene expression of interferon-γ, IL-12, CD4 and CD83 compared to alum (p<0.05). While the greater induction of IFNγ-related gene expression indicates that Metastim(®) can elicit Th-1 immune responses more effectively than the aluminum salt, there was also evidence of Th2 cytokine induction.