bluuebunny/crossref_metadata_2025 huggingface dataset:
Pregabalin As Adjunctive Therapy for Partial Seizures
Authors: Brodie Martin J
Published: August 2004

Summary:  The efficacy and safety of pregabalin as adjunctive therapy for patients with partial epilepsy with or without secondary generalization has been studied in three randomized, double‐blind, placebo‐controlled trials involving 1,052 patients. Patients (≥12 years of age) participating in the trials were highly refractory to treatment, experiencing at least six seizures and no 4‐week seizure‐free period during the 8‐week baseline phase, even though 73% received at least two antiepileptic drugs and 23% received three. Each fixed‐dose study was 12 weeks in duration. In Study 1, patients received pregabalin 50, 150, 300, or 600 mg/day given two times daily with no titration period. Studies 2 and 3 had a 1‐week titration to dose levels of 150 and 600 mg/day given three times daily (Study 2), 600 mg/day given two times daily, and 600 mg/day given three times daily (Study 3). Pregabalin, at 150, 300, and 600 mg/day, was significantly superior to placebo in reducing seizure frequency with a clear dose–response relationship. Responder rates, defined as the percentage of patients with ≥50% reduction in seizure frequency from baseline, approached 50% at 600 mg/day. In the effective dose range (150–600 mg/day), seizure freedom in the last 28 days of treatment was 3–17%. There was no difference between two times daily and three times daily dosing regimens. Efficacy was apparent as early as week one. The most commonly reported adverse events were CNS related, and either mild or moderate in intensity and generally self limiting. Few patients (≤5% in any treatment group) discontinued due to lack of efficacy. These results indicate that pregabalin is highly effective as adjunctive therapy in the treatment of patients with partial seizures with or without secondary generalization.
Identity, narcissism and the emotional core
Authors: West Marcus
Published: August 2004

Abstract:  This paper describes the course of an analysis which demonstrates how borderline and narcissistic functioning can be understood in terms of a struggle with issues of identity. It shows how such functioning can come to exert a profound hold on the individual and why it seems, at times, a matter of life and death to the patient to avoid states of separation from the analyst. The paper suggests that these complex phenomena can be understood, perhaps surprisingly, in the simple terms of the nature of affect itself. The concept of the emotional core is introduced to embody and highlight that which lies beneath both Jungian and Freudian models (offering a potential bridge between the two models)—namely the psyche’s essentially affective nature. It is suggested that the emotional core can serve as an organ of perception giving the individual both their primary relation to reality and an emotional attachment to others. This emotional core is understood to function in a narcissistic manner to preference experiences of sameness and in aversion to experiences of difference—a view consonant with Stern’s understanding of infant development where the infant is able to distinguish self from other from the beginning of life (as Fordham also held); taking up Stern’s terminology, it gives the individual a ‘core’ sense of being. There is, however, no stable, on‐going sense of ‘I’ associated with this form of functioning as the individual is immersed in the latest affect to enter consciousness (as in the borderline state of mind) and consequently the individual comes to rely intensely on the other to determine their sense of being (the other becomes a self‐regulating other in Stern’s terms). The development of ego‐functioning gives a more stable and on‐going sense of ‘I’ to the individual, giving contact with the broader personality, allowing the individual to be less reliant on the other and orientating them to reality in a way more fitting to their overall needs.

The paper describes how consciousness, which is not seen as identical with the ego, moves between the mode of functioning of the ego and that of the emotional core, i.e., shifting in and out of states where projective identification predominates. It elaborates the range of self‐experience encompassing spiritual experience and states of disintegration (which are understood to have a similar structure) on one side, to ego‐based experience (which can itself be defensive and rigid at times) on the other. It explores the consequences of such a view for analytic technique and relates it to the Jungian view of the self and the Freudian unconscious.
The diversity and status of amphibians and reptiles in the Kyoga Lake Basin
Authors: Behangana M.
Published: August 2004

The research study was carried out between 1999 and March 2001 covering nine study sites, eight of which are satellite lakes of Lake Kyoga and the ninth is a bay on the main lake. Sampling took place during both the rainy and wet seasons. However, the resident time of each visit usually lasted a day or two, which definitely could affect the quality and quantity of the data collected. This was the first time the herpetofauna of the Kyoga Lake Basin was documented. A total of 18 amphibian species were recorded in the whole of the Kyoga Lake Basin. Lakes Nawampasa, Nakuwa and Iyingo on the main lake showed the highest species diversity (12, 13, and 12, respectively) followed by Kawi (10 spp.) and Agu (nine spp.). Lakes Bisina (six spp.) and Nyaguo (five spp.) were species poor.

Reptilian diversity was poor with L. Nakuwa being the most species rich site (six spp.). The Nile crocodile was reported in Iyingo, Nakuwa and Bisina only. However, the Nile monitor lizard was recorded in all the lakes sampled.
The distribution and diversity of amphibian fauna of Lake Nabugabo and surrounding areas
Authors: Behangana M. , Arusi J.
Published: August 2004

Six major vegetation types surrounding Lake Nabugabo were sampled for amphibians. These types were: shoreline zone (habitat type code A) representing vegetation on the shoreline about 2 m from the shoreline; Miscanthidium violaceum (B); Eragrostis‐Loudetia (C); swamp forest (D); Cyperus papyrus (E); and medium altitude evergreen forest (F). A total of 168 days were spent in the field with a total of 288 1 h counts. Forty‐eight counts were carried out in each vegetation type. Sampling was carried out between 17.00 and 24.00 h to coincide with the time amphibians are very active. Twenty‐four species were recorded during the study period. These account for about 27.9% of total number of amphibian species recorded in Uganda. Four families were identified, which included: Ranidae (12 spp.), Hyperolidae (8 spp.), Bufonidae (3 spp.) and Pipidae (1 sp.). There was substantial variation in observed species composition among vegetation types. The most species rich habitat was the Eragrostis‐Loudetia vegetation zone (17 spp.), followed by the swamp forest zone (14 spp.), then the M. violeceum swamp (11 spp.) and the shoreline (10 spp.). The most species poor sites were C. papyrus swamp (6 spp.) followed by the medium altitude forest (7 spp.). In terms of species abundance, the swamp forest and the M. violecium zones were the most important amphibian habitats for the common species while the Eragrostis‐Laudetia zone stood out as the most important for the rare species. From the data collected, it can be deduced that the three most important habitats for the conservation of amphibian fauna in the study area are the Eragrostis‐Laudetia, M. violeceum and the swamp forest zones.
Induction of vascular endothelial growth factor expression in human pulp fibroblasts stimulated with black‐pigmented Bacteroides
Authors: Yang L.‐C. , Tsai C.‐H. , Huang F.‐M. , Su Y.‐F. , Lai C.‐C. , Liu C.‐M. , Chang Y.‐C.
Published: August 2004

Aim  To investigate the effect of black‐pigmented Bacteroides on the expression of vascular endothelial growth factor (VEGF) gene in human pulp fibroblasts.

Methodology  The supernatants of Porphyromonas endodontalis, Porphyromonas gingivalis and Prevotella intermedia were used to evaluate VEGF gene expression in human pulp fibroblasts. The levels of mRNAs were measured by the quantitative reverse‐transcriptase polymerase chain reaction analysis.

Results  Black‐pigmented Bacteroides induced significantly high levels of VEGF mRNA gene expression in human pulp fibroblasts (P < 0.05). In addition, the expression of VEGF depended on the bacteria tested.

Conclusions  Black‐pigmented Bacteroides may be involved in developing pulpal disease through the stimulation of VEGF production that would lead to the expansion of the vascular network coincident to progression of the inflammation.
Survey of routines for sedation of patients on controlled ventilation in Nordic intensive care units
Authors: Guldbrand P. , Berggren L. , Brattebö G. , Mälstam J. , Rönholm E. , Winsö O. ,   Scandinavian Critical Care Trials Group
Published: August 2004

Background:  Sedation strategies and practice for patients on controlled ventilation is variable from place to place as well as over time. Less sedation results in shorter ventilation time and new ventilatory modes permit more awake patients. Previous works estimated sedative and analgesic use in Nordic ICUs some years ago, but current practice is not known. We therefore designed this study to describe pharmacological and practical routines for sedation of patients on controlled ventilation.

Material and methods:  We used an electronic questionnaire about characteristics of the participating ICUs and the routines for sedation of ventilator‐treated patients, and secondly, an Internet‐based 5‐day registration on the use of drugs for sedation and analgesia.

Results:  Eighty‐eight of 220 ICUs (36%) responded to the questionnaire and 47 out of these 88 units (53%) used a sedation scale. Written guidelines for sedation were used in 41% of the units. Both daily interruption of sedation infusions and guidelines for weaning from the ventilator were used in 15% of the units. Data on 202 patients (633 patient days) from 55 ICUs were reported. Among analgesics, fentanyl predominated (240/633 days), followed by ketobemidon (160/633 days) and morphine (115/633 days). Propofol and midazolam were the most commonly used agents for sedation (345 and 238/633 days, respectively).

Conclusion:  Most units used a sedation scale, although other strategies to reduce the sedation level had not yet been fully introduced. Differences in pharmacological strategies were found between the Nordic countries, and some favourite drugs could be identified.
Anti‐nociceptive and Sedative Effects of Sufentanil Long Acting during and after Sevoflurane Anaesthesia in Dogs
Authors: Polis I. , Moens Y. , Gasthuys F. , Hoeben D. , Tshamala M.
Published: August 2004

The purpose of the present study was to determine the most effective time interval between the administration of sufentanil long acting (LA) and the induction of sevoflurane anaesthesia in dogs. The occurrence of sedation, analgesia and other marked side‐effects were evaluated in addition to the possible dosage‐reducing effect of sufentanil on sevoflurane in dogs. Forty clinically normal beagles aged 1–2 years and weighing between 8.4 and 13.6 kg were included. Two control groups were used: one group of dogs (A) received sufentanil LA (50 μg/kg i.m.) and a second group (B) the sufentanil vehicle followed by standard inhalation anaesthesia of 90 min. After premedication with sufentanil LA immediately before (C0), 15 min (D15) or 30 min (E30) prior to induction with thiopental (i.v.) the dogs were anaesthetized for 90 min with sevoflurane in oxygen. Pain and sedation scores were evaluated every 10 min during sevoflurane anaesthesia and at 2 (T120), 4 (T240) and 24 h (T1440) after initiation of anaesthesia. The occurrence of adverse reactions such as hypothermia, lateral recumbency, ataxia, noise sensitivity, vomiting, defaecation, salivation, nystagmus and excitation was observed at the same time‐points. During the recovery period pain scores were lower and sedation scores higher in the sufentanil LA groups. In many dogs acceptable pain and sedation scores persisted during 24 h. Several dogs showed ataxia, lateral recumbency, arousal on auditory stimulation, defaecation, salivation and excitation at several time‐points after sufentanil LA administration. Sufentanil LA in addition to sevoflurane anaesthesia offered beneficial dosage‐reducing analgesic effects up to 69.8% for thiopental and 78.3% for sevoflurane; although several typical opioid side‐effects occurred. To achieve this advantageous dosage‐reducing effect 15 min should be respected between sufentanil LA administration and induction of sevoflurane anaesthesia.
Effects of Hyperosmolar Ionic and Low Osmolar Non‐ionic Contrast Media on Acid Base, Blood Gas and Electrolyte Status in Dogs
Authors: Izci C. , Ogurtan Z. , Ceylan C.
Published: August 2004

The dogs in groups I, II and III in equal numbers received diatrizoate, iohexol and ioxilan at a dose of 700 mgI/kg intravenously (i.v.) as a bolus, respectively. Blood samples were collected prior to contrast media (CM) administration and thereafter at 3, 15, 30, 60, 90 and 180 min to evaluate acid–base, venous blood gas status (pH, PCO2, PO2, HCO, BE, O2) and electrolytes (Na+, Ca++, K+). Values of pH, PCO2, BE, HCO, Na+ and K+ remained unchanged or within non‐significant fluctuations compared with the baseline values. PO2 was significantly different from the baseline values in group 1 up to 90 min after administration, significant alterations were found for O2 saturation in group 1 up to 90 min, and in group II at 3, 60 and 180 min; and for Ca++ in group 1 at all time points except at 90 min, and groups II and II at 3 and 15 min post administration. It was concluded that none of the CM are considered to cause long‐lasting and major effects on acid–base, blood gas and electrolyte status.
Mouse Bioassay to Assess Oestrogenic and Anti‐oestrogenic Compounds: Hydroxytamoxifen, Diethylstilbestrol and Genistein
Authors: Köhlerová E. , Škarda J.
Published: August 2004

Young intact and adult gonadectomized C3H male and female mice were utilized as bioassay models to detect endocrine disruption chemicals. Animals treated with oestradiol (E) or progesterone (Prog) or E plus Prog were used to assess steroid hormone agonist and antagonist activities of 4‐OH‐tamoxifen (TAM), diethylstilbestrol (DES) and genistein (GEN) by bioassay. The stimulation or inhibition of mammary growth by TAM depended on sex, the state of animal (intact or gonadectomized), hormonal treatment (Prog, E, E plus Prog) and dose of TAM. TAM stimulated mammary growth in untreated ovariectomized (OV‐X) females and in Prog‐treated intact males and OV‐X females. In intact males, mammary growth was increased by TAM at dose 0.1–1 μg day−1and decreased at dose 10 μg day−1. Mammary growth was inhibited by TAM in Prog‐treated intact females and in E or E plus Prog‐treated intact and gonadectomized males and females. Uterine weights were increased by TAM in both untreated and treated (E, Prog, E plus Prog) intact and OV‐X females; however, seminal vesicle weights were decreased by TAM in both untreated and treated (E, Prog, E plus Prog) intact males. DES alone affected mammary growth (an inverted‐U‐shaped dose–response curve) both in male and female mice. DES increased uterine weights; however, seminal vesicle weights were decreased. GEN increased mammary and uterine growth in OV‐X females, GEN plus Prog stimulated mammary growth in intact males. The results obtained in these studies show clearly that only a bioassay consisting of several endpoints reflective to the mechanism of oestrogen and anti‐oestrogen action has the ability to evaluate activities of a molecule.
Autopoietic Law and the ‘Epistemic Trap’: A Case Study of Adoption and Contact
Authors: Smith Carole
Published: August 2004

This paper examines autopoietic theory with reference to functionally differentiated social sub‐systems, particularly law, science, and politics. It sets out to ‘test’ the practical relevance of autopoietic theory in relation to ongoing debates about post‐adoption contact and personal identity issues. Law has resisted social scientific pressure to regulate post‐adoption contact in the context of a social policy approach, which emphasizes the relationship between identity development and genealogical continuity. I argue that law's response to this pressure relates to the particular nature of adoption as this is expressed through legislation and case law. Law's refusal to intervene in post‐adoption contact reflects its self‐referential operations and its attempts to avoid epistemic entrapment by a social scientific discourse. Applying autopoietic theory to law's practical operations in adoption clarifies its explanatory value, provides a conceptual framework for understanding the relationship between law, politics, and social science and indicates areas that require theoretical refinement.
Experts, Juries, and Witch‐hunts: From Fitzjames Stephen to Angela Cannings
Authors: Ward Tony
Published: August 2004

Angela Cannings's successful appeal against her convictions for murder has revived an old controversy about the competence of juries to evaluate expert evidence. In response to criticisms of the jury system in the wake of a series of controversial poisoning trials, the Victorian jurist J.F. Stephen argued that juries were well equipped to decide on behalf of the community which experts should be treated as authorities, whose opinions the lay public could accept for practical purposes as ‘beyond reasonable doubt’. Such practical decisions did not, Stephen argued, require that juries fully understand the experts’ reasons for their conclusions. This article draws on recent work in social epistemology to argue that Stephen's view of the jury remains tenable, and that his authoritarian arguments can be recast in more democratic terms. It also concurs in Stephen's blunt recognition that the courts’ need to make decisions despite the uncertainties of science renders some convictions of the innocent inevitable.
The Prior Authorisation Procedure Adopted for the Deliberate Release into the Environment of Genetically Modified Organisms: the Complexities of Balancing Community and National Competences
Authors: Brosset Estelle
Published: August 2004

Abstract: The cornerstone of the Community regulatory framework on this subject is EC Directive 90/220/EEC, as repealed by EC Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms (GMO), which is based on a fundamental premise: that in order to prevent the risks caused by the release of genetically modified organisms, GMOs can only be released subject to the grant of a prior authorisation, following a scientific assessment, which confirms that such organisms do not represent a risk to the environment and human health. Yet it is obvious that Member States are not willing to entirely transfer their sovereignty in relation to issues that are of such fundamental importance to environmental protection and human health; at the same time however, the objective of achieving free movement of GMOs implies the establishment of a genuine Community procedure for this sector. The need to strike a balance between these competing demands therefore requires an enormous collaborative effort from national and Community authorities, which, contrary to expectations, has not produced any concrete results as yet. The authorisation procedure that has been developed was intended to bridge the gap between the divergent interpretations of what actually constitutes a biotechnical risk; conversely, it has reached a stasis, and causing protracted delays in the procedures granting authorisations. Admittedly, some changes to the procedure were implemented in 2001, but they were not intended to restructure the general architecture of the procedure established in 1990. Yet are such remedies going to be effective? Or is it time to make a more radical revision of the existing procedures by redistributing the competences on this delay?
Community Regulation of Genetically Modified Organisms: a Difficult Relationship Between Law and Science
Authors: Forsman Zeynep Kivilcim
Published: August 2004

Abstract: The European Community legislator regulates the area of genetically modified organisms according to the precautionary principle, which implies keeping a distance from scientific results in decision‐making. However, a positivist approach continues to exist within the same legislation. Paradoxically, this approach is promoted by the means of implementation of the precautionary principle. But to a large extent, it takes root in the context of the interpretation of the principle by the Community legislator, who attempts to eliminate disparities between the national legislations and to conform to World Trade Organization norms.
Primary Cough Headache is Associated with Posterior Fossa Crowdedness: A Morphometric MRI Study
Authors: Chen Yen-Yu , Lirng Jiing-Feng , Fuh Jong-Ling , Chang Feng-Chi , Cheng Hui-Cheng , Wang Shuu-Jiun
Published: August 2004

The aetiology of primary cough headache (PCH) is obscure. The aim of this study was to investigate the magnetic resonance (MR) morphometric characteristics of the posterior cranial fossa (PCF) in patients with PCH. Eighteen consecutive patients with PCH (14M/4F, mean age 75.1 + 6.0 years) and 18 sex- and age-matched control subjects were recruited for study. Based on the midline sagittal MR images, parameters indicating posterior fossa crowdedness were measured. Compared with controls, patients with PCH had a similar size of hindbrain tissue area but a significantly smaller PCF area, resulting in a higher mean hindbrain/PCF ratio (0.78 + 0.04 vs. 0.73 + 0.06, P = 0.005). In addition, these patients also had a lower position of the cerebellar tonsillar tip, a shorter clivus length and shorter distances from the clivus to the mid-pons and from the basion to the medulla than the control group. Patients with PCH were associated with a more crowded PCF, which might be a contributing factor for the pathogenesis of this headache syndrome.
Effects of Sumatriptan on Capsaicin-Induced Carotid Haemodynamic Changes and CGRP Release in Anaesthetized Pigs
Authors: Arulmani U , Heiligers JPC , Garrelds IM , Sánchez-López A , Willems EW , Villalón CM , Saxena PR
Published: August 2004

It is suggested that during a migraine attack capsaicin-sensitive trigeminal sensory nerves release calcitonin gene related peptide (CGRP), resulting in cranial vasodilatation and central nociception. Hence, inhibition of trigeminal CGRP release may prevent the above vasodilatation and, accordingly, abort migraine headache. Therefore, this study investigated the effects of sumatriptan (100 and 300 μg/kg, i.v.) on capsaicin-induced carotid haemodynamic changes and on CGRP release. Intracarotid (i.c.) infusions of capsaicin (10 μg/kg/min, i.c.) increased total carotid, arteriovenous anastomotic and capillary conductances as well as carotid pulsations, but decreased the difference between arterial and jugular venous oxygen saturations. Except for some attenuation of arteriovenous anastomotic changes, the capsaicin-induced responses were not affected by sumatriptan. Moreover, i.c. infusions of capsaicin (0.3, 1, 3 and 10 μg/kg/min, i.c.) dose-dependently increased the jugular venous plasma concentrations of CGRP, which also remained unaffected by sumatriptan. The above results support the contention that the therapeutic action of sumatriptan is mainly due to cranial vasoconstriction rather than trigeminal (CGRP release) inhibition.
The Aura: A Tertiary Care Study of 952 Migraine Patients
Authors: Kelman L
Published: August 2004

The aim of this study was to document the frequency and types of aura symptoms, to define the relationship between aura symptoms and to define the aura frequency in different migraine types. In 952 migraine patients, aura frequency, duration, time to headache, characteristics and percentage of headaches with aura were analysed. Thirty-eight percent of IHS 1.1-1.5 patients reported aura, 38.1% of females and 33.0% of males. Average percentage of aura occurrence with headache was 19.7% of headaches, average aura duration 27.3 min and aura was followed by headache in 10.4 min on average. Visual disturbances occurred in 92.1% and aura without visual aura was rare. Aura frequency was headache-type dependent. The highest frequency of aura was seen in the more ‘full-blown’ migraine attack. Visual aura is the overwhelming aura symptom. Even in patients with aura the percentage of aura with migraine attacks is limited.
Purchasing Power Parity Across States and Goods Within Australia*
Authors: CHAUDHURI KAUSIK , SHEEN JEFFREY
Published: August 2004

Panel unit root tests show that intranational purchasing power parity cannot be rejected across major Australian cities from 1972:3 to 1999:1. The persistence of deviations in response to shocks is low, as measured by the estimated exact half‐life of between five and ten quarters. This is much lower than results for similar tests done on US cities, and for international purchasing power parity tests. The food CPI is largely responsible for the fast convergent results for city CPIs. Intranational purchasing power parity was rejected for the floating exchange rate period from 1984 to 1991 when inflation was high and not specifically targeted by the central bank.
Development of a two‐antibody model for the evaluation of copper‐64 radioimmunotherapy
Authors: Bryan J. N. , Lewis M. R. , Henry C. J. , Owen N. K. , Zhang J. , Mohsin H. , Jia F. , Sivaguru G. , Anderson C. J.
Published: August 2004

Copper‐64 emits β+ and β– particles suitable for positron emission tomography and radioimmunotherapy (RIT) of cancer. Copper‐64‐labelled antibodies have caused complete responses in laboratory animal RIT studies at far lower radiation doses than traditionally prescribed. The intracellular localization of copper radioisotopes may lead to cytotoxic effects by mechanisms beyond ionizing radiation damage. The purpose of this research was to develop a model using both internalizing and non‐internalizing antibodies for direct comparison in future RIT studies using the same animal model of cancer. The monoclonal antibodies, cBR96 and cT84.66, were conjugated with N‐hydroxysulfosuccinimidyl DOTA. All conjugates retained high immunoreactivity and labelled efficiently with 64Cu with high specific activity and radiochemical purity. Twenty‐four hour biodistributions determined in LS174T tumour‐bearing nude mice demonstrated low organ and high tumour uptakes for both monoclonal antibodies. This model constitutes a promising system for elucidating whether internalization of 64Cu is responsible for an enhanced tumour cytotoxicity in vivo.
Feline epitheliotrophic T‐cell lymphoma with paraneoplastic eosinophilia – immunochemotherapy with vinblastine and human recombinant interferon α2b
Authors: Cave T. A. , Gault E. A. , Argyle D. J.
Published: August 2004

A cat with epitheliotrophic T‐cell lymphoma with paraneoplastic eosinophilia is described. Initial attempts to control the disease with conventional therapies failed. The addition of recombinant human interferon α2b (rhINFα2b) resulted in a clinical, haematogenous and sonographic improvement for 49 days. The overall survival time from initial diagnosis was 100 days. Relapse was correlated with the development of serum antibodies directed against rhINFα2b. To our knowledge, this is the first report describing the clinical use of IFNα in the treatment of neoplasia in the cat.
Trends in veterinary cancer imaging
Authors: Wisner E. R. , Pollard R. E.
Published: August 2004

With the recent advances in diagnostic imaging technology, cancer imaging in veterinary medicine has become more specific for disease diagnosis, more accurate for determining tumour margins and more sensitive for detecting metastatic disease. Ultrasound provides highly detailed images of parenchymal masses and infiltrative lesions while providing a means for aspiration or biopsy using real‐time image guidance. Computed tomography and magnetic resonance‐imaging techniques provide exquisite anatomical resolution that improves diagnostic accuracy, provides an accurate means of radiation or surgical treatment planning and a quantitative means for monitoring response to therapy. In addition to traditional anatomic imaging, new techniques are being developed for estimating functional parameters such as tumour perfusion, cell metabolism and gene expression. While conventional planar scintigraphy has been available for some time, newer nuclear imaging modalities such as positron emission tomography promise to further improve the accuracy of initial tumour diagnosis and staging and determination of response to therapy. Although many of these functional techniques are not yet clinically available, it is highly likely that some will be integrated into routine clinical practice in the near future.
Correlation of histologic grading of canine mast cell tumors with Ki67/PCNA/AgNOR/c‐Kit scores: 38 cases (2002–2003)
Authors: Bergman P. J. , Craft D. M. , Newman S. J. , Baer K. , Camps‐Palau M. A. , McKnight J. A. , Leibman N. F. , Brenn S. , Finora K. , Hohenhaus A. E. , Bartick T. , Patnaik A. K. , Scase T.
Published: August 2004

Introduction:  The Patnaik grading system for canine cutaneous MCT is currently one of the best determinants of prognosis; however, clinical outcome does not always correlate with histologic grade. The development of molecular markers offers a potential advantage and may complement subjective grading. The primary purpose of this study was to correlate histologic grading to Ki67/PCNA/AgNOR/c‐Kit scores.

Methods:  Thirty‐eight dogs with cutaneous MCT underwent surgical resection. Tumors were graded, with expansion of grade II MCT to low, medium (or II only) and high. For statistical purposes, MCT grade I, II (low, medium, high) and III were assigned a score of 1, 2, 3, 4, or 5, respectively. Sections were processed for AgNOR staining and expression of PCNA, Ki67 and c‐Kit as previously described (modified biotin‐strepavidin with DAB substrate). Paraffin‐embedded canine tissue arrays were used as positive and negative controls (primary antibody replaced with pre‐immune sera). Parametric statistical testing was performed using Statview statistical software with P ≤ .05 as significant.

Results:  There were 12, 20, 5 and 1 grade II low, grade II medium, grade II high and grade III MCT, respectively. The mean Ki67 score was 9.114 (median 8.0, range 1–28), mean PCNA score was 26.25 (median 24.0, range 5–65), mean AgNOR score was 1.499 (median 1.35, range 1.02–2.76) and c‐Kit scores were +1 (9/37), +2 (19/37) and +3 (9/37). With parametric statistical testing, significantly positive correlations were found for Ki67/Grade, PCNA/Grade, AgNOR/Grade, Ki67/PCNA, Ki67/AgNOR and PCNA/AgNOR (all P < .0001). No significant correlation was found for c‐Kit and grade; however, +3 c‐Kit scores had statistically significantly higher grades than +2 c‐Kit scores (P = .0458).

Conclusions:  Ki67/PCNA/AgNOR scores are positively correlated to grade in dogs with MCT. Further studies to correlate Ki67/PCNA/AgNOR/c‐Kit scores with clinical variables are ongoing.
A two‐antibody model for copper‐64 radioimmunotherapy: biodistributions and tumor dosimetry in a mouse model of cancer
Authors: Bryan J. N. , Mohsin H. , Jia F. , Siegall C. B. , Anderson C. J. , Miller W. H. , Henry C. J. , Lewis M. R.
Published: August 2004

Objectives:  We previously presented preliminary data on pilot biodistributions of 64Cu‐labeled monoclonal antibodies (mAbs).1 The purpose of the present studies was to obtain comprehensive biodistributions using a two‐antibody model for direct comparison of an internalizing and a non‐internalizing mAb in the same animal model of cancer. These studies will allow evaluation of the efficacies of the two 64Cu‐labeled mAbs for radioimmunotherapy (RIT) and are part of a larger, ongoing project examining the unusual tumoricidal properties of internalized 64Cu.

Methods:  DOTA (1,4,7,10‐tetraazacyclododecane‐1,4,7,10‐tetraacetic acid) was conjugated to an internalizing antibody, cBR96, and a non‐internalizing antibody, cT84.66. Biodistributions of the 64Cu‐labeled conjugates in nude mice bearing LS174T human colorectal carcinoma xenografts were obtained at time points from 15 min to 48 h. Mouse tumor dosimetry was calculated using a Monte Carlo N‐Particle Transport Code.

Results:  The 64Cu‐DOTA‐cBR96 conjugate demonstrated rapid tumor uptake, reaching 20.2% injected dose per gram of tissue (% ID/g) at 3 h and peaking at 35.4% ID/g by 24 h. Tumor accumulation of 64Cu‐DOTA‐cT84.66 was more gradual, 8.19% ID/g at 3 h (p = 0.002), and reached 43.8% ID/g by 24 h, but maximum uptake was not statistically different from 64Cu‐DOTA‐cBR96 (p = 0.05). Liver, spleen, and kidney uptakes were statistically similar at 24 h for both conjugates (p = 0.05). Tumor absorbed radiation doses were estimated to be 1128 rad/mCi for 64Cu‐DOTA‐cBR96 and 1409 rad/mCi for 64Cu‐DOTA‐cT84.66.

Conclusion:  The biodistributions and tumor dosimetry of these two radiolabeled antibodies were sufficiently similar for direct comparison of the therapeutic efficacies of internalizing versus non‐internalizing 64Cu RIT agents in the same animal model of cancer. RIT studies using this two‐antibody model are currently in progress to test the hypothesis that internalization of 64Cu is necessary for enhanced cytotoxic properties at low tumor absorbed doses.
Investigating the molecular mechanism of pamidronate‐induced in vitro cytotoxicity in canine osteosarcoma cells
Authors: Fan T. M. , Charney S. C. , De Lorimier L. P.
Published: August 2004

Introduction:  Pamidronate is used to treat metastatic bone lesions in human cancer patients. By directly inhibiting the mevalonate pathway, pamidronate disrupts GTPase‐binding protein prenylation, resulting in osteoclast apoptosis. Pamidronate has been demonstrated to exert dose‐ and time‐dependent cytotoxic effects in several canine malignant osteoblastic cell lines. However, the exact cytotoxic mechanism remains speculative. The purpose of this study was to investigate and characterize the molecular mechanism of pamidronate‐induced cytotoxicity in canine malignant osteoblasts.

Methods:  The involvement of farnesyl or geranylgeranyl pyrophosphate synthase inhibition was evaluated by performing rescue experiments with cells incubated for 48 hours with cytotoxic concentrations of pamidronate (50 and 100 μM), with or without farnesol (FOH) and geranylgeraniol (GGOH). Cell lysates were fluorometrically‐assessed for caspase‐3‐like activity (R&D Systems) following incubation with varying concentrations of pamidronate (control, 50 and 100 μM) for 48 hours. The expression of a prenylated GTPase protein, Rap1 A, was qualitatively evaluated with immunoblotting using a polyclonal goat antibody (Santa Cruz Biotechnology) in cells incubated for 48 hours with varying concentrations of pamidronate (0, 1, 10, 50, and 100 μM).

Results:  In cells treated with lower cytotoxic concentrations of pamidronate (50 μM) for 48 hours, the addition of GGOH, but not FOH, was able to diminish the degree cytotoxicity, p ≤ 0.05. However, cells incubated with higher cytotoxic concentrations of pamidronate (100 μM), neither GGOH nor FOH were able to reduce the level of pamidronate‐induced cytotoxicity. Caspase‐3‐like activity directly correlated with the degree of pamidronate‐induced cytotoxicity. Cells treated with pamidronate at 50 μM and 100 μM increased caspase‐3‐like activity by 5.3 and 7.1‐fold, respectively over untreated controls. The detection of Rap1A by immunoblotting requires further optimization.

Conclusions:  Pamidronate‐induced cytotoxicity in canine osteosarcoma cells may share a similar mechanism as has been demonstrated for osteoclasts. Inhibition of the mevalonate pathway appears to contribute to the observed cytotoxicity at lower doses of pamidronate. In addition, caspase‐3‐like activity contributes to the apoptotic process induced by pamidronate in malignant canine osteoblasts.
Monitoring response to aminobisphosphonate therapy in canine osteosarcoma using dual energy x‐ray absorptiometry (dexa)
Authors: Lorimier L. P. , Fan T. M. , Charney S. C. , Lacoste H. I. , Heller D. A. , Endicott M. M. , Souza C. H.
Published: August 2004

Introduction:  Palliative therapy is essential to improve the quality of life of dogs with osteosarcoma (OSA), when definitive therapy is not considered a valid option. Bisphosphonates, a novel class of antiosteoclastic drugs, are widely used in humans for several painful osteolytic conditions. Dual energy x‐ray absorptiometry (DEXA) is recognized as a reliable tool to measure bone mineral density (BMD), and to monitor treatment response to bisphosphonates in humans. A prospective evaluation of pamidronate, an injectable aminobisphosphonate, is ongoing in dogs with appendicular OSA. The potential value of DEXA for objective evaluation of BMD variations with palliative therapies is concurrently being assessed.

Materials and Methods:  Dogs with naturally occurring appendicular OSA treated with pamidronate constitute the patient population. A DEXA scan (QDR‐4500 W, Hologic, Bedford, MA) is performed on day 0 (baseline) and on every treatment day with pamidronate thereafter (every 28 days). For each dog, a whole body scan is performed, followed by a scan of the tumor, and contralateral normal bone. Three regions of interest are subsequently analyzed for BMD changes in tumor and normal bone. Statistical analysis was performed using Student t‐test and paired t‐tests, with significance being set at p < 0.05.

Results:  Nineteen dogs have been enrolled to date. Seven responders and 6 non‐responders have suitable data for analysis. A significant difference is observed (p = 0.04) between tumor BMD variations of responders and non‐responders at day 28 (mean variations +18.0% and −4.6% respectively). The changes at day 28 are significant only in the responders (p = 0.038 vs p = 0.05 in non‐responders). When BMD of tumor and normal bone at day 84 is compared to day 0 in six responders, only tumor had a significant increase (p = 0.017 vs p = 0.279, respectively).

Conclusions:  Objective measurements of response to therapy are essential in pain palliation studies. Increased tumor bone BMD, as obtained by DEXA analysis, may correlate with subjective clinical improvement in pamidronate‐treated dogs with appendicular OSA.
RNA‐transfected CD40‐activated B cells as a vaccine strategy in canine malignancies
Authors: Overley B. , Coughlin C. M. , Leibowitz M. S. , Sorenmo K. U. , Vonderheide R. H.
Published: August 2004

Introduction:  Cell‐based vaccine strategies using dendritic cells as cellular adjuvant have entered phase III trials in humans and have been found to be safe, feasible, and potentially efficacious. Canine patients are generally smaller than adult human patients, which makes production of canine dendritic cell (DC) vaccines problematic, given patient size and the small number of available DC precursors. Here we describe feasibility studies of a novel cell‐based vaccine strategy which uses CD40‐activated B‐cells (CD40‐B) loaded with RNA. This strategy is based on our observations that RNA‐transfected human CD40‐B can drive anti‐tumor T cell responses. One advantage of using CD40‐B cells is the ability to expand this cell population ex vivo, allowing for the numbers of cells required for therapeutic vaccines.

Methods:  Twenty milliliters of blood were drawn from 6 normal dogs and 5 canine lymphoma patients. Peripheral blood mononuclear cells were separated by Ficoll centrifugation. Culture conditions for B cell activation were optimized using CD40‐ligand, canine IL‐4, and Toll‐like receptor stimulus with CpGoligodinucleotides (ODN). Cyclosporine was added to eliminate peripheral T lymphocytes. Proliferation and activation of CD40‐B cells were demonstrated by CFSE dilution of B cells quantified by flow cytometry. Gene transfer was achieved by mRNA electroporation.

Results:  Marked in vitro stimulation and proliferation of canine peripheral B cells were achieved with soluble trimeric CD40L, canine IL‐4, and ODN. CD40‐B cells showed dramatic upregulation of MHC class II molecules and CD21 (B‐cell activation marker). After two weeks in culture, cells were negative for CD3 and CD4. Canine CD40‐B cells were efficiently transfected with mRNA, with >60% of CD40‐B expressing green fluorescent protein after GFP mRNA electroporation.

Conclusion:  RNA‐transfected CD40‐B cells can be efficiently generated from normal and tumor‐bearing dogs. These results provide rationale to test tumor RNA‐transfected CD40‐B as a novel therapeutic approach to treating canine malignancies. Clinical trials in canine lymphoma have been proposed.
Differentiating effect of pamidronate on canine osteo‐sarcoma cell lines in vitro
Authors: Lacoste H. I. , Fan T. M. , De Lorimier L. P. , Charney S. C.
Published: August 2004

Introduction:  Pamidronate has been traditionally used to manage pathologic osteoclastic disorders. In addition to its effects on osteoclasts, pamidronate has also been demonstrated to promote phenotypic maturation and inhibition of proliferation in osteoblasts. Canine osteosarcoma (OSA) consists of malignant, undifferentiated osteoblasts. The objective of this study was to determine if micromolar concentrations of pamidronate could induce malignant osteoblastic differentiation as evaluated by an increase in alkaline phosphatase (ALP) activity and/or osteocalcin (OC) production, two specific markers of normal osteoblastic activity.

Methods:  Two canine OSA cell lines (HMPOS and COS 31) were used for all experiments. Cells were incubated for 48 or 72 hours with various pamidronate concentrations (0, 0.1, 1, 5, 10, and 20 μM). After incubation, the supernatants were sampled and the relative amounts of viable cells were determined with a cell proliferation assay (Cell Titer 96® AQuous, Promega). An ALP detection kit (Starbright®, Sigma®) was used to measure the ALP activity and an ELISA (Osteocalcin EIA kit, Biomedical Technologies) was used to determine the concentration of osteocalcin in the supernatants. The ALP and osteocalcin values were corrected for the amount of viable cells.

Results:  Pamidronate induced a dose‐dependent reduction in the number of viable COS 31 and HMPOS cells at both 48 and 72 hours. A dose‐dependent elevation in ALP activity from baseline was observed. At 20 μM, a 2.3‐fold increase was observed for HMPOS at 72 hours, while a 1.43‐fold increase was observed for COS 31 at 72 hours. Very low level (less than 2 ng/ml) of osteocalcin pre‐ and post‐pamidronate treatment was detected for both COS 31 and HMPOS.

Conclusion:  The data suggests that pamidronate increases alkaline phosphatase activity in canine OSA cells in a dose‐dependent manner. However, cytotoxic assays are needed in order to accurately characterize any concurrent decrease in the number of viable cells. The potential differentiating effect of pamidronate on malignant osteoblasts provides an additional argument for its use in the palliative treatment of OSA.
Noninvasive monitoring of doxorubicin cardiotoxicity: a pilot study of two dogs
Authors: Selting K. A. , Spier A. W. , Dodam J. R. , Skimming J. W. , Lattimer J. C. , Britt L. G. , Turk J. R. , Tyler J. W. , Durham H. E. , Garro M.
Published: August 2004

The cardiotoxicity of anthracycline anticancer agents, most notably doxorubicin, limits their use in treating a wide variety of cancers. Currently available methods for assessment of anthracycline‐induced cardiotoxicity (AIC) often lack specificity and sensitivity. The purpose of this pilot investigation was to determine noninvasive diagnostic methods that are predictive for AIC. Two mongrel dogs were anesthetized for injection of doxorubicin (22.5 mg) directly into the left coronary artery. Doxorubicin injection was repeated 2 weeks later. Dogs were followed for 12 weeks or until heart failure was documented. Dogs were monitored before and at serial time points after doxorubicin using echocardiography and signal‐averaged electrocardiography as well as ambulatory ECG monitoring. Blood was also collected for measurement of cardiac troponin I and T concentrations. Doxorubicin was readministered at 6 and 12 weeks after the second injection, as interim data analysis did not reveal adequate evidence of cardiomyopathy.

Dog‐1 appeared to be more sensitive than Dog‐2 to AIC based upon gradual changes in variables of cardiac function and earlier time to failure (113 days). In contrast, Dog‐2 had inconsistent changes in cardiac function and a longer time to failure (139 days). Dog‐1 also had greater peak plasma cardiac troponin I levels (17.96 ng/ml) as compared to peak levels in Dog‐2 (4.08 ng/ml). In addition, Dog‐1 had increased ventricular extrasystolic contractions (VE)(0/day at baseline, 443/day one week prior to death), whereas dog‐2 showed very few VE's throughout. Spectral analysis of ECG data also revealed a significant decrease over time in total spectral power in Dog‐1 from 37,148 at baseline to 603 msec2/Hz one week prior to death, indicating decreased parasympathetic tone. Endomyocardial biopsies revealed minor changes despite significant clinical abnormalities. These data indicate differences in susceptibility to AIC between Dog‐1 and Dog‐2, and suggest a role for these noninvasive measures in monitoring the heart during chemotherapy.
Novel concepts in the pathogenesis of HIV‐lymphoma
Authors: Zenger E.
Published: August 2004

Lymphoma remains a rapidly fatal consequence of HIV infection in developed countries despite the widespread use of highly active antiretroviral therapy. HIV‐associated lymphomas are predominantly of B‐cell origin, appearing to be outgrowths of antigen‐driven B‐cells with significant growth control influence provided by abnormal T‐cell and antigen‐presenting cells processes. Findings of clonal HIV integration within macrophages in a subset of these lymphomas provided evidence for abnormalities outside the B‐cell compartment playing roles in this disease. From this and other observations, the “sequential neoplasia” model, wherein a clonal macrophage would drive early stages of tumorigenesis, with sequential evolution of an independent tumor (1,2), emerged. To investigate this, we developed a pathogenesis‐based animal model of lymphoma wherein macrophages play an essential role in the lymphomagenesis process (3). Extensive phenotypic and genotypic analysis of human and murine tumors and associated macrophages revealed significant abnormalities providing compelling evidence that may explain the underlieing abnormalities and driving forces in AIDS‐associated lymphomas. Altered expression of specific cytokines, chemokines, and transcriptional activators have been detected that likely play a central role in the pathogenesis of this disease and possibly lymphoma in general.