bluuebunny/crossref_metadata_2025 huggingface dataset:
Efficacy of electrical stimulation to increase muscle strength in people with neurological conditions: a systematic review
Authors: Glinsky Joanne , Harvey Lisa , Van Es Pauline
Published: July 2007

Background and Purpose. Weakness in partially paralysed muscles is a disabling impairment for people with neurological conditions. Strength training programmes are widely administered to address this impairment. There is a common belief that the effectiveness of strength training programmes can be enhanced by the addition of electrical stimulation. The purpose of this systematic review was to assess the efficacy of electrical stimulation for increasing voluntary strength in people with neurological conditions. Method. Eligible randomized trials of electrical stimulation were identified by searches of computerized databases. The search yielded 11 267 abstracts, of which 60 were retrieved. Two assessors independently reviewed full text versions of these articles. Results. Eighteen studies satisfied the inclusion criteria. These studies involved participants with spina bifida (n = 1), cerebral palsy (n = 1), peripheral nerve lesion (n = 1), multiple sclerosis (n = 1), spinal cord injury (n = 3) and stroke (n = 11). The mean (SD) PEDro score for trial quality was 4.9 (1.0) out of 10. Meta‐analyses of studies involving similar patients were not done because of insufficient data or lack of homogeneity. The results of all studies were analysed individually. Conclusion. Several studies suggest a modest beneficial effect of electrical stimulation in patients with stroke. It is not clear whether patients with other types of neurological disabilities benefit from electrical stimulation in the same way. Copyright © 2007 John Wiley & Sons, Ltd.
Investigation of the advanced functionalities of a hybrid quadrupole orthogonal acceleration time‐of‐flight mass spectrometer
Authors: Weaver Paul J. , Laures Alice M.‐F. , Wolff Jean‐Claude
Published: July 2007

Time‐of‐flight mass spectrometry (ToF‐MS) has gained wide acceptance in many fields of chemistry, proteomics, metabolomics and small molecule analysis. ToF‐MS, however, has some inherent advantages and drawbacks. Numerous developments have been made to hybrid ToF instruments to improve their capabilities. We have used a quadrupole orthogonal acceleration ToF (Q‐oa‐ToF) instrument to assess developments made to improve resolution, dynamic range and signal‐to‐noise (S/N) ratios (i.e. sensitivity). Higher mass resolution can improve the analysis of mixtures containing compounds with similar m/z values and improved mass accuracy gives greater confidence for structural elucidation applications. Wide dynamic ranges are necessary for the analysis of unknown samples or samples that vary widely in analyte concentrations. The performance of the advanced functionalities for routine structural elucidation in terms of resolution, dynamic range and S/N ratios was investigated using test compounds. The results presented in this work demonstrate and validate the use of these new enhancements for Q‐ToF instruments and also show their limitations. Copyright © 2007 John Wiley & Sons, Ltd.
A liquid chromatographic/tandem mass spectroscopic method for quantification of the cyclic peptide melanotan‐II. Plasma and brain tissue concentrations following administration in mice
Authors: Hatziieremia Sophia , Kostomitsopoulos Nikolaos , Balafas Vagelis , Tamvakopoulos Constantin
Published: July 2007

Melanotan‐II (MT‐II), a synthetic analogue of the natural melanocortin peptide, α‐melanocyte‐stimulating hormone (α‐MSH), is well known for the anorexic effects it elicits in rodents. These effects are, at least partly, associated with agonistic action on the centrally located melanocortin receptors, MC3R and MC4R. Whether MT‐II exerts this effect via brain penetration still remains unclear. In order to address this question we administered MT‐II in rodents at efficacious doses and then employed a sensitive methodology for the determination of MT‐II in plasma and brain samples. MT‐II was extracted from mouse plasma and brain tissue by acetonitrile precipitation followed by liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis. The described assay improved significantly previously reported MT‐II levels of quantification in rat plasma and brain. The lower limits of quantification (LLOQs) of 0.5 ng/mL and 2.5 ng/g were obtained in 50 µL plasma and 100 µL brain homogenate, respectively. The calibration curve was linear over the concentration range of 0.5–500 ng/mL for plasma and 2.5–250 ng/g for brain tissue. The method was successfully applied in measuring levels of MT‐II in plasma and brain tissue following intraperitoneal (ip) administration of 1 mg/kg of peptide in mice. Following administration of MT‐II, clearance from plasma was rapid. The sensitivity of the assay allowed the determination of low concentrations of MT‐II (11.4 ± 5.5 ng/g) in brain homogenate at 30 min after dosing. However, the brain concentrations when compared with the high plasma levels of MT‐II at the same time point confirmed the low penetrability of the peptide in mouse brain. Copyright © 2007 John Wiley & Sons, Ltd.
Fast and simultaneous determination of darunavir and eleven other antiretroviral drugs for therapeutic drug monitoring: method development and validation for the determination of all currently approved HIV protease inhibitors and non‐nucleoside reverse transcriptase inhibitors in human plasma by liquid chromatography coupled with electrospray ionization tandem mass spectrometry
Authors: ter Heine Rob , Alderden‐Los Carolien G. , Rosing Hilde , Hillebrand Michel J. X. , van Gorp Eric C. M. , Huitema Alwin D. R. , Beijnen Jos H.
Published: July 2007

For the quantification of all currently approved non‐nucleoside reverse transcriptase inhibitors and protease inhibitors, including the new protease inhibitor darunavir and the active nelfinavir metabolite M8, an assay was developed, using liquid chromatography coupled with tandem mass spectrometry. The sample pretreatment consisted of a protein precipitation with a mixture of methanol and acetonitrile using only 100 µL plasma. Chromatographic separation was performed on a reversed‐phase C18 column (150 × 2.0 mm, particle size 5 µm) with a quick stepwise gradient using an acetate buffer (pH 5) and methanol, at a flow rate of 0.25 mL/min. The analytical run time was only 10 min. The triple quadrupole mass spectrometer was operated in the positive ion mode and multiple reaction monitoring was used for drug quantification. The method was validated over a range of 0.1 to 20 µg/mL for amprenavir, atazanavir, efavirenz, indinavir, lopinavir, nelfinavir, the active nelfinavir metabolite M8, nevirapine and ritonavir, a range of 0.05 to 10 µg/mL for saquinavir and darunavir and a range of 0.5 to 100 µg/mL for tipranavir, based on observed concentration ranges in patients treated with these drugs. D5‐squinavir, D6‐indinavir, 13C6‐efavirenz and dibenzepine were used as internal standards. The method was proven to be specific, accurate, precise and robust. Accuracies ranged from 88.5% to 102.2% and all precisions were less than 9.5%. Furthermore, the assay demonstrates a high sensitivity for all analytes and the stepwise gradient allows addition of new analytes into the same method. The method is now successfully applied for routine therapeutic drug monitoring and pharmacokinetic studies in HIV‐infected patients. Copyright © 2007 John Wiley & Sons, Ltd.
p53 mediates interstitial cystitis antiproliferative factor (APF)‐induced growth inhibition of human urothelial cells
Authors: Kim Jayoung , Keay Susan K. , Dimitrakov Jordan D. , Freeman Michael R.
Published: July 2007

Antiproliferative factor (APF) is a sialoglycopeptide elevated in the urine of patients with interstitial cystitis, a urinary bladder disorder of unknown etiology that is characterized by chronic pelvic pain. The present study was directed toward uncovering a pathway through which APF signals. Treatment of human urothelial cells with native APF resulted in growth inhibition accompanied by blockade of cell cycle transit and increased p53. Reduced expression of p53 by RNA interference diminished, while ectopic expression of p53 mimicked, the effects of APF. These are the first findings implicating the network of p53 target genes in urothelial defects associated with interstitial cystitis.
Vaginal birth after cesarean delivery in the West African setting
Authors: Adanu R.M.K. , McCarthy M.Y.
Published: July 2007

Objective: To determine the fetal weight beyond which women with one previous cesarean delivery (CD) are most likely to have a repeat CD. Methods: A retrospective cohort study of 586 women who had one previous CD and were undergoing trial of labor was conducted in Accra, Ghana. Following delivery, the women were allocated to one of three groups according to whether they had a successful vaginal delivery, underwent a CD for cephalopelvic disproportion, or underwent a CD for another indication. The groups were then compared using analysis of variance or Kruskal‐Wallis tests. Multiple logistic regression was used to assess the effect of fetal weight on the odds of having a repeat CD. Results: A fetal weight greater than 3.45 kg tripled the odds of having a repeat CD, and the probability of having a repeat CD were 50% for a fetal weight of 3.70 kg. Conclusion: In settings similar to those in Ghana, women who have undergone a previous CD whose fetuses weigh more than 3.70 kg are likely to have less than a 50% chance of having a successful vaginal delivery.
Intraumbilical injection of uterotonics for retained placenta
Authors: Habek D. , Franičević D.
Published: July 2007

Objective

To assess the effect of injecting an uterotonic agent in the umbilical vein during the third stage of labor in women with retained placentas.

Methods

In this prospective clinical study, 75 women with retained placentas received 20 mL of a 0.9% saline solution with either 20 IU of oxytocin (n = 54), 0.5 mg of carboprost tromethamine (n = 7), or 0.2 mg of methylergometrine (n = 14) injected in the umbilical vein after clamping. The treatment success was determined by the clinical signs of placental ablation.

Results

There were no statistically significant differences among the 3 therapeutic groups regarding age, parity, risk factors, pregnancy duration, type of delivery (spontaneous, induced, or augmented), or possible early postpartum complications caused by the intraumbilical injection. The rates of therapeutic success were 76.9% in the oxytocin group, 85.7% in the synthetic prostaglandin group, and 64.2% in the methylergometrine group.

Conclusion

The intraumbilical injection of uterotonics is a noninvasive, effective, and clinically safe method of shortening the third stage of labor in women with retained placentas.
Facial nerve preservation in relation to tumour size in acoustic nerve tumour surgery
Authors: Morrison A. W. , King T. T.
Published: July 2007

A SERIES of twenty-five cases of acoustic nerve tumours operated upon by a translabyrinthine-transtentorial technique, employing micro-surgical techniques is analysed from the point of view of preservation of the facial nerve after total removal. The size of the tumour, as expected, has been the most important factor in determining preservation of the structure. If there is to be a high rate of facial nerve preservation after acoustic nerve tumour surgery, the diagnosis must be made by the otologist (rather than the neurologist) who has the opportunity of diagnosing the lesion when it is still small.
Invasive pleomorphic adenoma of hard palate
Authors: Joachims H. Z. , Altman M. M.
Published: July 2007

A case of pleomorphic adenoma of the hard palate is presented, which, in spite of its benign histological features, showed an unusual destructive capacity. The tumour invaded the nose and both maxillary sinuses. No similar cases have been reported in the literature. The tumour could be enucleated through a palatal approach.
Clinical value of tone burst vestibular evoked myogenic potentials at threshold in acute and stable Ménière's disease
Authors: Osei-Lah V , Ceranic B , Luxon L M
Published: July 2007

Introduction:

The objectives of this preliminary, prospective, cohort study were to ascertain the characteristics of vestibular evoked myogenic potentials at threshold levels in two groups of Ménière's disease patients – acute and stable – and to identify whether vestibular evoked myogenic potentials can provide any specific, objective information to distinguish acute from stable Ménière's disease.

Subjects and methods:

The study was based at a tertiary neuro-otology centre. Twenty adult patients who fulfilled the American Academy of Otolaryngology–Head and Neck Surgery1 criteria for Ménière's disease were divided into two groups: 11 patients with acute Ménière's disease and nine patients with stable Ménière's disease. Eighteen healthy adult volunteers served as controls. All subjects underwent vestibular evoked myogenic potential testing with ipsilateral, short tone burst stimuli at 500 Hz, as well as pure tone audiometry. The patients also underwent caloric testing.

Results:

Vestibular evoked myogenic potentials were present in all controls, and were present in 65 per cent of patients but absent in 35 per cent. The mean absolute threshold (Tvestibular evoked myogenic potential) ± standard deviation in normal controls was 116 ± 7.7 dBSPL; this did not differ statistically from that in patients, nor did it differ between acute and stable Ménière's disease. The p13/n23 latencies at the threshold levels in the normal, acute and stable groups (mean ± standard deviation) were respectively: 15 ± 2.2 ms/23.0 ± 2.5 ms; 15.7 ± 0.9 ms/23.7 ± 0.9 ms; and 15.3 ± 2.0 ms/24.2 ± 1.9 ms. The mean interaural amplitude difference ratio (IAD) ± standard deviation was significantly higher in the stable group compared with the acute group (0.54 ± 0.33 vs −0.15 ± 0.22; p = 0.007) and with the controls (0.54 ± 0.33 vs 0.1 ± 0.22; p = 0.05).

Conclusions:

The parameter that best differentiated acute from stable Ménière's disease at threshold was the interaural amplitude difference ratio. Therefore, this parameter may be used to monitor the clinical course of Ménière's disease.
Identification of levamisole resistance markers in the parasitic nematode Haemonchus contortus using a cDNA-AFLP approach
Authors: NEVEU C. , CHARVET C. , FAUVIN A. , CORTET J. , CASTAGNONE-SERENO P. , CABARET J.
Published: July 2007

A cDNA-AFLP (cDNA-Amplified Fragment Length Polymorphism)-based strategy has been used to identify levamisole (LEV) resistance markers in the nematode Haemonchus contortus. Transcript profiles of adult nematodes from two LEV-resistant and two susceptible isolates were compared. Among the 17 280 transcript-derived fragments (TDFs) amplified, 26 presented a polymorphic pattern between resistant and susceptible nematodes: 11 TDFs were present in both resistant isolates and absent from both susceptible isolates whereas 15 TDFs were present in both susceptible isolates and absent from both resistant isolates. 8 TDFs specifically present in resistant isolates were cloned and sequenced. Some of these TDFs could represent novel genes, as their sequences presented no homologies in databases. Interestingly, specific expression of one candidate (HA17) in resistant nematodes from different isolates was confirmed by RT-PCR experiments. The finding that HA17 expression correlates with LEV resistance in three H. contortus isolates vs five susceptible isolates strongly suggest that we identified a new potential marker of LEV resistance. This differential approach at the transcriptome level could be of great interest for the identification of the molecular mechanism involved in this phenotype.
Nematode ligand-gated chloride channels: an appraisal of their involvement in macrocyclic lactone resistance and prospects for developing molecular markers
Authors: McCAVERA S. , WALSH T. K. , WOLSTENHOLME A. J.
Published: July 2007

Ligand-gated chloride channels, including the glutamate-(GluCl) and GABA-gated channels, are the targets of the macrocyclic lactone (ML) family of anthelmintics. Changes in the sequence and expression of these channels can cause resistance to the ML in laboratory models, such as Caenorhabditis elegans and Drosophila melanogaster. Mutations in multiple GluCl subunit genes are required for high-level ML resistance in C. elegans, and this can be influenced by additional mutations in gap junction and amphid genes. Parasitic nematodes have a different complement of channel subunit genes from C. elegans, but a few genes, including avr-14, are widely present. A polymorphism in an avr-14 orthologue, which makes the subunit less sensitive to ivermectin and glutamate, has been identified in Cooperia oncophora, and polymorphisms in several subunits have been reported from resistant isolates of Haemonchus contortus. This has led to suggestions that ML resistance may be polygenic. Possible reasons for this, and its consequences for the development of molecular tests for resistance, are explored.
Associations between salivary bacteria and reported sugar intake and their relationship with body mass index in women and their adolescent children
Authors: Vågstrand Karin , Karin Lindroos Anna , Birkhed Dowen , Linné Yvonne
Published: July 2007

Objective

To study associations between reported sugar intake and salivary bacteria (mutans streptococci, MS and lactobacilli, LB), and sugar intake in relation to body mass index (BMI), in women and adolescents.

Design

Cross-sectional study. Habitual dietary intake was reported in a questionnaire and whole saliva samples were collected and cultivated.

Setting

Stockholm, Sweden.

Subjects

Four hundred and fourteen mothers and their 16-year-old children (57% girls).

Results

Reported total sugar intake was significantly associated with LB in mothers (P= 0.001), but not in daughters and sons (P= 0.03 for mother–son interaction). When excluding lactose, the associations with LB and MS were significant in daughters (P= 0.032 and 0.005, respectively). High BMI and sugar intake from milk were independently associated with MS counts in mothers in a multiple regression model (R2= 0.053). In daughters, sugar from beverages, candies/chocolates and fruits were all independently associated with MS counts (R2= 0.074), whereas in sons, only mothers’ MS counts were associated (R2= 0.084). High BMI, low intake from cooked meals and high intake of candies/chocolates were all independently associated with LB in mothers (R2= 0.076), while, except for candies/chocolates in daughters, no significant associations were found in children. BMI was significantly associated with bacteria counts in both mothers and children, whereas reported sugar intake was not.

Conclusion

The associations between reported sugar intake and bacteria were generally weak. In mothers and daughters, some significant associations were found, but not in sons. Bacteria counts, but not reported sugar intake, were positively associated with BMI.
Specific targeting of in vitro‐activated human antitumour effector cells using anti‐CD3 × anti‐c‐erbB‐2 bispecific antibody
Authors: TSUKAMOTO HIDEO , NAKAMURA YOSHIHIKO , MASUKO TAKASHI , HASHIMOTO YOSHIYUKI , HABU SONOKO , NISHIMURA TAKASHI
Published: July 2007

Bispecific antibody (BSAb) consisting of anti‐CD3 plus anti‐c‐cerbB‐2 Fab fragments for the application to adoptive tumour immunotherapy was prepared. This bifunctional hetero‐F(ab′)2 antibody reacted with both human CD3+ T cells and c‐erbB‐2 positive human tumour cells. Human CD8+ T cells activated with immobilized anti‐CD3 plus interleukin 2 showed marginal cytotoxicity against tumour cells. However, addition of the prepared BSAb into the culture resulted in a marked augmentation of the cytotoxicity by the activated CD8+ T cells in a dose‐dependent manner. The enhanced cytotoxicity of CD8+ T cells in the presence of BSAb was specific for c‐erbB‐2 positive tumour cells. Moreover, it was demonstrated that anti‐CD3 × anti‐c‐erbB‐2 BSAb was also effective for the specific targeting of various kinds of in vitro‐activated antitumour effector cells such as lymphokine‐activated killer cells, CD4+ helper/killer cells, γδ T cells and activated tumour‐infiltrating CD8+ T cells. These results indicated that BSAb consisted of anti‐CD3 and anti‐c‐erbB‐2 will become a useful tool for the adoptive tumour immunotherapy of human cancer expressing c‐erbB‐2 oncogene products.
Characterization of thymic involution induced by murine cytomegalovirus infection
Authors: PRICE PATRICIA , OLVER STUART D. , GIBBONS ANNE E. , TEO HUI KUN , SHELLAM GEOFFREY R.
Published: July 2007

Infection of BALB/c mice with murine cytomegalovirus (MCMV) decreased the numbers of cells recovered from the thymus by 80–90% after 4–7 days, although less than 10 thymocytes per million were productively infected with the virus. A loss of cortical thymocytes was evident in histologic sections and correlated with depletion of CD4+CD8+ cells.

Thymic involution was minimal in C57BL/6 mice. This resistance was not H‐2b‐associated, as BALB.B (H‐2b) mice were severely affected. In CXB recombinant inbred mice, thymic involution and MCMV replication co‐segregated with atrophy and infection of the spleen and bone marrow. This suggests common regulation by natural killer (NK)1.1+ cells, consistent with the enhanced thymic involution demonstrated in NK‐deficient bg/bg mice. However, CD4− CD8− cells were not depleted, so bone marrow hypoplasia may not be the proximal cause of thymic involution.

MCMV infection activated CD4+, CD8+ and CD4+ CD8+ thymocytes, as expression of MEL14, major histocompatibility complex class I (H‐2) and Sca‐1 antigens increased on these cells. In vitro lymphoproliferation and interleukin (IL)‐3 release were enhanced in unseparated and CD4+ ‐enriched thymus preparations. Maturation of the thymus population was also evident from the high frequencies of single positive CD4+ and CD8+ cells and the decline in Sca‐2 expression. However, unlike peripheral T cells, thymocytes from infected mice did not release IL‐2. The results suggest that thymic involution accelerates the transit of cells through the thymus. The possibility that this impairs the elimination of autoreactive T cells within the thymus and promotes the autoimmune manifestations of MCMV disease is discussed.
Differences in antigen‐binding affinity caused by single amino acid substitution in the variable region of the heavy chain
Authors: XIANG JIM , CHEN ZHINAN , DELBAERE LOUIS T. J. , LIU ENWU
Published: July 2007

Site‐directed mutagenesis has been used to examine the importance of histidine‐99 in the VH CDR3 region of a mouse/human chimeric anti‐TAG72 antibody, cB72.3‐1‐3. The expression vectors mpSV2neo‐EPl‐Vm4–10C.γ1, containing seven different mutant VH region fragments (Vm4–10) in association with the immunoglobulin enhancer (E), promoter (P1) and human genomic Cγ1 region fragments, were transfected into a heavy chain loss mutant cell line B72.3Mut(K), respectively. Mutant chimeric antibodies cB72.3m4–10 were purified from the transfectant supernates, and their binding affinities for the TAG72 antigen relative to that of the original cB72.3‐1‐3 antibody were compared. Substitution of histidine‐99 by glutamine resulted in a higher affinity antibody (cB72.3m4) whereas substitution by isoleucine resulted in a lower affinity antibody (cB72.3m9). The binding affinity of these mutant antibodies varied nearly eight‐fold. It was concluded that the residue at position 99 in the VH CDR3 region is in a ‘contact’ position in the B72.3/TAG72 antibody‐combining site. The polar side‐chains of glutamine and asparagine or the ionized side‐chains of histidine, arginine or glutamic acid contribute to higher binding affinity, whereas the hydrophobic side‐chains of isoleucine, leucine or phenylalanine resulted in a lower binding affinity for the TAG72 antigen.
Vitamin D‐binding protein (group‐specific component) is the sole serum protein required for macrophage activation after treatment of peritoneal cells with lysophosphatidylcholine
Authors: HOMMA SADAMU , YAMAMOTO MASASHI , YAMAMOTO NOBUTO
Published: July 2007

In vitro treatment of mouse peritoneal cells with 1 μg lysophosphatidylcholine (lyso–Pc)/mL in serum free‐0.1% egg albumin‐supplemented RPMI 1640 medium for 30 min, followed by 3 h cultivation in a medium supplemented with human serum, resulted in a greatly enhanced Fc‐receptor‐mediated phagocytic activity of macrophages. Vitamin D‐binding protein (group‐specific component [Gc]) of α2‐globulin fraction was shown to he the sole serum glycoprotein required for the generation of a potent macrophage‐activating factor. When a mixture of lysophosphatidylcholine (lyso‐Pc)‐treated nonadherent and adherent cells were cultured in a medium supplemented with a small amount of purified Gc protein (1 ng/mL), a greatly enhanced activation of macrophages was demonstrated. The generation of macrophage‐activating factor from purified Gc protein was far more efficient than that from whole serum, indicating that a serum component is inhibitory to the activation process of macrophages. While three other major serum glycoproteins (α2‐macrcglobulin, α2‐HS‐glycoprotein and haptoglobin) were neither stimulatory nor inhibitory to lyso‐Pc‐primed macrophage activation, serum albumin (competitively with Gc protein) appeared to be inhibitory to the process of macrophage activation.
Cytokine production in response to Epstein‐Barr virus infection of peripheral blood mononuclear cells in vitro
Authors: WHITTINGHAM SENGA , NASELLI GAETANO , HARRISON LEONARD C. , BOYD ANDREW W. , CEBON JONATHAN , JACK IAN
Published: July 2007

To obtain a better understanding of the immune response to Epstein‐Barr virus (EBV), we measured the cytokines tumour necrosis factor (TNF)‐α/β, interleukin‐2 (IL‐2), interferon‐gamma (IFN‐γ), IL‐6 and granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) in the conditioned medium of peripheral blood mononuclear cells from 10 healthy adults before and at 48 h and at 1, 2, 3 and 4 weeks following infection in vitro with EBV. Cultures were examined for regression of outgrowths of nascent virus‐transformed B cells, and populations of cells in the cultures were analysed by flow cytometry.

TNF‐α/β was not detected in infected or non‐infected cultures. In infected cultures assayed at the nominated times, the highest levels of IL‐2 were detected at 48 hours, IFN‐γ at 1 week, IL‐6 at 2 weeks and GM‐CSF between 2 and 4 weeks. IL‐6 and GM‐CSF, but not IL‐2 or IFN‐γ, were detected in non‐infected cultures but at lower levels than in infected cultures. Nine of the 10 healthy adults showed regression of outgrowths of virus‐transformed B cells and, of these, seven had antibodies to the EBV capsid antigen (VCA). Strong regression was associated with sequential increases in IL‐2, IFN‐γ, and low levels of IL‐6 and GM‐CSF. Absent or weak regression was associated with an undetectable level of IL‐2, a low level of IFN‐γ, high levels of IL‐6 and GM‐CSF and an increased frequency of cells bearing the phenotype CD20 and HLA‐DR in the final weeks of culture. The percentage of cells with a phenotypic marker for natural killer cells remained constant in all cultures over the 4 week period, and there was a modest decrease in the percentage of T cells in both infected and non‐infected cultures. The demonstration of different temporal profiles of cytokines is in accord with a programmed cellular immune response to EBV infection which could serve as a model to study the events that modulate the inflammatory response to EBV.
Multiple levels of post‐transcriptional regulation of collagenase (matrix metalloproteinase 1) in an epithelial cell line
Authors: WHITELOCK J. M. , PAINE M. L. , GIBBINS J. R. , KEFFORD R. F. , O'GRADY R. L.
Published: July 2007

Multiple levels of regulation of collagenase (matrix metalloproteinase 1; MMP‐1), have been demonstrated in a clonal rat epithelial cell line (A5P/B10). Secreted enzyme could not be demonstrated in culture medium from A5P/B10 cells but, using antibodies specific for collagenase, the enzyme was detected within the cytoplasm and on the surface of the cells. A probe for rat collagenase could not detect a signal for mRNA in the cytoplasm while nuclear run‐on data demonstrated that the gene for collagenase was being transcribed. Incubating the cells with 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA) significantly increased cytoplasmic mRNA levels and slightly increased the intensity of staining in permeabilized cells, but collagenase activity was still not detected in the conditioned medium. This indicated that the protein was being synthesized by the TPA‐treated cells but was not being secreted into the medium. These data suggest that the secretion of collagenase may be regulated both following transcription and after the completion of translation and it is suggested that multiple levels of control may be operating to determine the rate of collagenase release and hence, the rate of collagen turnover.
Immunosuppressive antibody treatment prolongs graft survival in two murine models of segmental pancreas transplantation
Authors: PURCELL LISA J. , MOTTRAM PATRICIA L. , MANDEL THOMAS E.
Published: July 2007

Successful pancreas transplantation requires the suppression of both allograft rejection and recurrence of autoimmune disease. In order to study treatments to suppress these two responses, separate models were developed for pancreas allograft rejection and autoimmune disease. In the first model, the diabetic state was induced with streptozotocin in CBA mice prior to the transplantation of pancreas grafts from BALB/c donors. In the absence of autoimmune disease, control mice rejected their grafts in 26 days (median). Antibody treatments (anti‐lymphocyte serum and anti‐CD4) significantly prolonged allograft survival beyond this time, but not to the extent we have previously reported in the heart graft model. NOD/Lt mice spontaneously developed autoimmune diabetes, and recurrence of disease was seen in isografts at 9.5 days (median). Antibody treatments significantly delayed disease recurrence, with anti‐CD4 being the most effective. Heart allografts (CBA donors) in NOD/Lt recipients were rejected within 17 days (median), and the anti‐CD4 treatment had a moderate effect in delaying graft survival (median 28 days). Anti‐lymphocyte serum did not prolong graft survival. Thus antibody treatment was effective in delaying both rejection and the recurrence of autoimmune disease in segmental pancreas grafts. However, the same doses were not effective in delaying heart rejection in the NOD/Lt model, so it would appear that treatments which inhibit autoimmune disease may not prevent allograft rejection.
Recombinant fowlpox virus vaccines for poultry
Authors: BOYLE D. B. , HEINE H. G.
Published: July 2007

The intensive poultry industries rely heavily upon the use of vaccines for disease control. Viral vector based vaccines offer new avenues for the development of vaccines for effective disease control in poultry. Techniques developed for the construction of recombinant vaccinia viruses have been readily adapted to the construction of recombinant viruses based on fowlpox virus (rFPV). The ability to insert several genes into the large genome of fowlpox may enable the development of multivalent vaccines and vaccines incorporating immune response modifiers such as lymphokines. Newcastle disease, avian influenza, infectious bursal disease and Marek's disease antigens expressed by rFPV have been shown to be effective vaccines in poultry. None appear, however, to provide a substantial improvement in vaccine efficacy. Recombinant FPV will be a valuable adjunct to conventional vaccines currently in widespread use. Whether rFPV or other vector based vaccines can circumvent the problems of vaccination in the presence of high maternally derived antibodies is yet to be resolved. The observation that avipoxvirus recombinants may be suitable for the vaccination of non‐avian species provides an added dimension to vaccines based on FPV or other avipoxviruses. Recombinant FPV will find a useful role in poultry disease control when used in conjunction with conventional vaccines.
Bovine herpesvirus‐1 vaccines
Authors: HURK SYLVIA VAN DRUNEN LITTEL‐van den , TIKOO SURESH K. , LIANG XIAOPING , BABIUK LORNE A.
Published: July 2007

Vaccination has been important in controlling a wide variety of viral and bacterial infections of man and animals. Vaccines to herpesvirus infection of cattle are no exception. The present review describes the different types of conventional vaccines that have been used to date and furthermore describes the novel approaches which are presently being implemented to develop more effective vaccines. These include subunit vaccines as well as genetically engineered modified live deletion mutants. Both these novel vaccine approaches appear to be more efficacious than conventional vaccines. Furthermore, these vaccines provide an additional dimension for control and eradication of infection by providing an opportunity to develop companion diagnostic tests to differentiate infected animals from vaccinated animals. This review summarizes these developments as well as present knowledge regarding the important host defence mechanisms required for preventing infection and aiding recovery from infection.
Vaccination against Lucilia cuprina: The causative agent of sheep blowfly strike
Authors: EAST IAIN J. , EISEMANN CRAIG H.
Published: July 2007

The sheep blowfly, Lucilia cuprina, is responsible for over 80% of cases of blowfly strike in Australia and the tosses in production and sheep deaths due to flystrike exceed S200 million per annum. Traditional methods of control are becoming less effective because of the blowfly's resistance to insecticides and thus other methods of control are necessary. In general, sheep develop very little immunity to flystrike even after repeated infestation, however vaccination against L. cuprina has shown considerable potential for controlling flystrike. The most successful sources of antigens have been the larvae's secreted proteases and several extracts from gut or peritrophic membrane. Immunization with these antigens results in retardation of larval growth and in some cases larval mortality. On sheep immunized with peritrophic membrane extracts, the growth retardation appears to be caused by a blocking of the peritrophic membrane which results in the larvae being starved of nutrients. The prospects for vaccine development and the remaining barriers to be overcome are discussed.
Immunological approaches to fertility regulation in domestic livestock
Authors: SCARAMUZZI R. J. , CAMPBELL B. K. , MARTIN G. B.
Published: July 2007

Antibodies to hormones have been used extensively to study endocrine regulation of reproduction and in recent times they have been commercially exploited as therapeutic agents to manipulate reproduction. Despite the release of two commercial products, little is known about the mechanism of action of immunoneutralization. The presence of hormone‐binding antibodies in the circulation and the detection of hormone antibody complexes in blood has led investigators to suggest a vascular site of action for immunoneutralization. However, a vascular site of action is difficult to reconcile with some of the known effects of immunoneutralization to the boar taint steroid (5α‐androst‐16‐en‐3‐one) to androstenedione and to gonadotrophin‐releasing hormone. In this paper we review some of our recent research into the mechanism of action of androstenedione immunoneutralization. We suggest that hormonal immunoneutralization of androstenedione may take place in the extracellular compartment or within the plasma membrane where antibodies to androstenedione may bind to a specific steroid‐binding protein in the plasma membrane of granulosa cells.
Effect of cationization on anti‐hapten antibody responses in sheep and mice
Authors: ALTMANN K. G.
Published: July 2007

The effect on anti‐hapten immune responses of altering the isoelectric point of hapten‐carrier conjugates by cationization was examined in sheep and mice. No adjuvants were employed in these studies. In sheep the cationization of ovalbumin‐dinitropheny1 (OVA‐DNP) resulted in an enhanced primary anti‐dinitropheny1 (DNP) response. The secondary anti‐DNP responses were not consistently greater for animals primed with cationized (CAT) OVA‐DNP than in animals primed with OVA‐DNP. Similar results were obtained with mice immunized with bovine serum albumin (BSA)‐DNP or CAT BSA‐DNP. Mice and sheep primed with OVA‐DNP failed to mount an anamnestic response following secondary immunization with CAT OVA‐DNP. Failure of a cationized hapten‐carrier to stimulate a secondary anti‐DNP response following priming with uncationized hapten‐carrier was also observed in mice using BSA‐DNP as the model antigen.
Interferon‐gamma and interleukin‐4 in human Leishmania donovani infections
Authors: KEMP MICHAEL , KURTZHALS JØRGEN A. L. , KHARAZMI ARSALAN , THEANDER THOR G.
Published: July 2007

Clinical and immunological similarities between Leishmania donovani infections in humans and L. major infections in mice suggest that some of the pathophysiological mechanisms are the same in the two conditions. Both infections can result either in a fatal systemic disease or in a self‐limiting infection with few and mild symptoms. In the murine model the outcome of the infection is critically related to the cytokines produced by T lymphocytes activated by leishmanial antigens. Activation of the IFN‐γ producing Th1 subset of CD4 positive T cells results in cure and survival, whereas activation of the IL‐4 secreting Th2 subset results in a progressive disease with fatal outcome. A similar Th1/Th2 dichotomy in the cytokine response to L. donovani may exist in humans, and may have influence on the outcome of infection. In murine leishmaniasis the levels of IL‐4 and IFN‐γ at the time of infection can direct the T cell response into Th1 or Th2 type. If similar mechanisms operate in humans, the outcome of L. donovani infections may depend on the local cytokine environment in which early activation of Leishmania specific T cells takes place. Cytokines secreted by cross‐reactive memory T cells, activated by antigens from the invading micro‐organism, may contribute to determine this environment.
Inhibitory mechanisms of antibody production by nitrogen oxides released from activated macrophages during the immune response: Relationship to energy consumption
Authors: TAKAGI KUNIAKI , NUKAYA IKUEI , YASUKAWA KIYOSHI , SUKETA YASUNOBU
Published: July 2007

We investigated the relationship between the sensitivity of mouse splenocytes in immune response to nitrogen oxides and energy consumption rate of the cells. Macrophage‐like cells (Mm 1) pretreated with lL‐6 served as the source of the nitrogen oxides. The antibody production of both 2,4,6‐trinitrophenyl‐keyhole limpet haemocyanin‐primed splenocytes and B cell hybridomas was markedly reduced; about 20–40% of splenocytes and B cell hybridomas were killed by co‐culture with IL‐6‐treated Mm 1. Cell viability and antibody production were completely restored by the addition of NG‐monomethyl L‐arginine to the culture medium. The cytotoxicity of the nitrogen oxides was correlated with the distance between effector and target cells. Under conditions of low cytotoxicity, antibody production by B cell hybridomas was suppressed by the nitrogen oxides, this suppression not being correlated with the reduction in cell growth. The sensitivity of the target cells differed in co‐cultures of antigen‐primed splenocytes and B cell hybridomas with IL‐6‐treated Mm 1. The nitric oxide‐sensitivity of the cells corresponded to their 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide reducing activity and ATP consumption rate. These findings suggest that nitrogen oxides act as regulatory molecules in immune response in three ways: cytostasis, reduction of cell growth and suppression of antibody synthesis.
Phenotype of T cells, their soluble receptor levels, and cytokine profile of human breast milk
Authors: EGLINTON BRETT A. , ROBERTON DONAL M. , CUMMINS ADRIAN G.
Published: July 2007

Human breast milk has important immunoprotective and immunosuppressive functions for an infant. The purpose of this study was to extend the phenotype of milk cells and to measure soluble T cell receptor levels and cytokines in milk, and to compare these with neonatal and adult blood. Milk T cells had a more equivalent CD4: CD8 ratio than blood; milk CD4 T cells mainly expressed the CD45RO (antigen primed/memory) phenotype; milk CD8 cells had an equivalent CDllb:CD28 suppressor; cytotoxic phenotype; and milk T cells had 2‐3‐fold higher percentages of activated CD4 IL‐2R and CD8 HML‐1 or CD8 VLA‐1 cells than blood. Soluble IL‐2R, CD4 and CD8 concentrations were lower in milk than adult blood, although relatively increased when compared to the lower T cell concentration in milk. Breast milk contained high levels of IFN‐γ but low levels of other measured cytokines compared to blood. These distinct differences of T cells and their soluble products are likely to influence an infant's immune system.
Measurement of cellular microbicidal activity against Pneumocystis carinii in vitro
Authors: CHEERS CHRISTINA , PAOLINI RITA , HOUSSAMI SOUHEIR
Published: July 2007

As Pneumocystis carinii cysts cannot be cultivated for enumeration of colony forming cells, two alternative approaches to measuring killing of P. carinii by mouse peritoneal cells were investigated. The cells tested were either normal resident peritoneal cells, or cells which were elicited by intraperitoneal injection of the bacterium Listeria monocytogenes. The latter population showed enhanced antibacterial activity against Listeria organisms and enhanced production of H2O2 in the presence of P. carinii, cysts from immunosuppressed rats. To assess killing of P. carinii, cysts were mixed with peritoneal cells at a ratio of 10:1, and after intervals of incuabation the peritoneal cells were lysed by saponin treatment. The viability of the cysts was assessed by staining with vital dyes or by uptake of tritiated uridine over 7 h incubation. Viability of cysts was unaffected by saponin treatment, and there was agreement between the two techniques that the elicited peritoneal cells killed approximately twice the number of cysts over a 20 h incubation period compared to normal resident peritoneal cells.
Cloning, sequencing, expression and inflammatory activity in skin of ovine interleukin‐8
Authors: SEOW H‐F. , YOSHIMURA T. , WOOD P. R. , COLDITZ I. G.
Published: July 2007

Ovine IL‐8 (oIL‐8) cDNA was obtained by probing a spleen cell cDNA library with human IL‐8 (hIL‐8) cDNA. The oIL‐8 cDNA was 1434 base pairs long with a single open reading frame encoding a 101 amino acid precursor protein of relative molecular mass 11 268. The inferred amino acid sequence has 78, 82, 84 and 67% similarity with human, rabbit, porcine and guinea‐pig IL‐8, respectively. By analogy with the most prevalent form of hIL‐8, a 72 amino acid form of oIL‐8 was expressed as a fusion protein containing glutathione‐S‐transferase and purified by affinity chromatography on a glutathione‐Sepharose column yielding 8 mg IL‐8/L broth culture. The fusion protein lacked chemotactic activity for ovine neutrophils, whereas the 72 amino acid form of oIL‐8 was equipotent with rhIL‐8. At 6 and 24 h after intradermal injection of 10−1 mol oIL‐8, there was intense accumulation of neutrophils, and very mild accumulation of eosinophils, CD5, CD4 and T19 (a σδ5 TCR subset) cells but not CD8 cells. The availability of roIL‐8 and its cDNA probes will permit the role of this important member of the IL‐8 family of chemotactic cytokines to be determined in inflammatory diseases of sheep.
A flow cytometric study of cell death: Failure of some models to correlate with morphological assessment
Authors: BRYSON GREGORY J , HARMON BRIAN V. , COLLINS RUSSELL J.
Published: July 2007

The balance between cell death and cell proliferation is a significant factor in the growth kinetics of normal and neoplastic tissues. Distinction between the two major forms of cell death, necrosis and apoptosis, is now recognized as important in understanding mechanisms regulating cell survival. A recent approach in the study of apoptosis has been the use of flow cytometry. with some reports indicating that, when stained with propidium iodide (PI), the DNA of apoptotic cells has decreased fluorescence compared with that of viable cells. In this study, we investigated a flow cytometric procedure which used the simultaneous analysis of DNA content and 90° light scatter (90LS). Significant differences in the PI staining pattern and a shift in 90LS were observed when apoptotic death, occurred at different stages of the cell cycle. Importantly, such differences only allowed accurate/quantification of apoptosis when it occurred in G1. While necrosis could be distinguished from apoptosis when examined during its early stages, a similar staining pattern to that found with apoptosis was observed when necrosis was examined during its latter stages. The results indicate that the measurement of DNA staining cannot be exclusively relied upon to detect apoptosis occurring in all models. However it is useful in the investigation of this process when the death occurs in G1, in that the method offers a rapid means for quantification.
Putative functional domain within ORF2 on the Mycobacterium insertion sequences IS900 and IS902
Authors: DORAN T. J. , DAVIES J. K. , RADFORD A. J. , HODGSON A. L. M.
Published: July 2007

Repeated DNA sequences have been identified in a range of mycobacterial species and have been implicated in the increased virulence of some of these species, namely, Mycobacierium paratuberculosis and M. avium subsp. silvaticum. Here we present a case to suggest that the insertion sequences IS900 and IS902 encode a protein from a putative gene positioned on the complementary strand to their transposase genes. Based on amino acid homology analyses, this open reading frame (ORF2) could encode a transport protein. The ORF2 protein thus IS900 and IS902, may have a role in the increased pathogenicity of M. paratuberculosis and M. avium subsp. silvaticum from an M. avium background.
β2 Integrin independent neutrophil recruitment and injury in anti‐GBM glomerulonephritis in rabbits
Authors: TIPPING P. G. , CORNTHWAITE L. J. , HOLDSWORTH S. R.
Published: July 2007

The role of the β2 integrin cell adhesion molecules (CAM) in directing neutrophil accumulation and injury in acute anti‐glomerular basement membrane antibody induced glomerulonephritis (anti‐GBM GN) was assessed in rabbits by in vivo inhibition of CD 18 dependent neutrophil/endothelial cell interactions using a monoclonal anti‐CD 18 antibody. Rabbits given horse anti‐rabbit GBM antibody developed significant glomerular neutrophil influx (2.9 ± 0.1 neutrophils per glomerular cross‐section [c/gcs], normal 0.1 ± 0.1 c/gcs, P= 0.002) and proteinuria (1389 ± 257 mg/16 h, normal 15± 4mg/16 h, P= 0.0015) after 16 h. Rabbits rendered neutropenic (< 500 neutrophils/μL) by mustine hydrochloride, or complement depleted by cobra venom factor, did not develop glomerular neutrophil accumulation and had markedly reduced proteinuria after anti‐GBM antibody, demonstrating complement‐induced neutrophil recruitment is a major mechanism of glomerular injury in this model. Anti‐CD 18 antibody treatment of rabbits developing anti‐GBM GN abrogated dermal neutrophil influx in response to intradermal injection of fMLP and zymosan activated serum. Treated rabbits achieved levels of anti‐CD 18 antibody in their serum which saturated the binding sites on rabbit neutrophils in vitro, and their circulating neutrophils had saturated anti‐CD 18 antibody binding in vivo. However, glomerular neutrophil influx (3.5 ± 0.4 c/gcs) and proteinuria (1210 ± 428 mg/16 h) were both unaffected. Thus, in this model of antibody‐initiated complement and neutrophil‐dependent glomerular injury, in which neutrophil transmigration across the endothelium is not required, effective functional inhibition of β2 integrin CAM in vivo did not reduce glomerular injury or glomerular neutrophil influx. These studies demonstrate that β2 integrin CAM are not a requirement for neutrophil accumulation and glomerular injury in anti‐GBM GN in rabbits.