My pigs are ok, why change? - animal welfare accounts of pig farmers.

Intensive pig production systems are a source of stress, which is linked to reduced animal welfare and increased antimicrobial use. As the gatekeepers of the welfare of the animals under their care, farmers are seen as the stakeholder responsible for improving animal welfare. The aim of this study was to explore the knowledge and attitudes of pig farmers towards pig welfare and the impact of such attitudes on farmers' selection of management strategies on the farm. We conducted in-depth semi-structured interviews with 44 pig farmers in one of the main pig producing regions of Brazil. Interviews covered knowledge and attitudes towards pig sentience and behaviour and welfare-related issues commonly observed in intensive pig farms (belly-nosing, fights, tail-biting, diarrhoea and castration without pain control) and farmers' conception and attitudes towards pig welfare. We identified many management and animal-based indicators of poor welfare, such as the use of painful and stressful management practices and use of environments that limit the expression of natural behaviours. However, most farmers were satisfied with animal welfare standards at their farms. Farmers' perceptions are aligned with their understanding of animal welfare. Although they identified all the dimensions that impact the welfare of a pig on a farm (affect, biological functioning and naturalness), their social reality, industry demands and available advice pushed them to perceive their range of action limited to biological and environmental aspects of the animals that do not necessarily benefit affective state. This precluded farmers from making associations between good health and the animal's ability to express a full behavioural repertoire, as well as from viewing abnormal behaviours as problems. The negative consequences for the welfare of the animals were commonly alleviated by routines that relied on constant use of medication, including high dependence on antibiotics. Expressions of estrangement from the production chain were common voices among the participants. This suggests that farmers may not be sufficiently informed or engaged in responding to consumers' expectations and commitments made by companies, which can pose a severe economic risk for farmers. The findings of this study indicate that economic, technical and social factors restrict farmers' autonomy and their ability to perform their role as stewards of animal welfare. (Re)connecting different human, animal and environmental interests may be a step to changing this scenario.

Nuevo coronavirus (COVID-19) en población general y pediátrica: una revisión epidemiológica: Chile 2020 / Novel coronavirus (COVID-19) in general and pediatric population: an epidemiological review: Chile 2020

Since December 2019, the outbreak of a novel coronavirus SARS Cov-2 has been reported in Wuhan, China. Currently, the new coronavirus disease has been declared a worldwide pandemic. Compared to adults, reporting of cases in pediatric patients has been significant smaller. The objective of this article is to provide epidemiological information of COVID-19, especially pediatric. Most of the confirmed cases of children are declared to be a cluster disease. The clinic is oligosymptomatic, less severe and with concentrated risk in children under 1 year of age and with comorbidity. In Chile, pediatric patients represent about 6% of the total number of infected and overall lethality is significantly lower than adults. The main control measures to reduce effective reproduction are mass testing, social distancing and school closure, without dismissing individual responsibility. The adequate supply of personal protection elements is key to avoid nosocomial infection and the compromise of healthcare providers.

A partir de diciembre del 2019, se ha reportado el brote de una nueva infección por SARS Cov-2 en Wuhan, China. Actualmente, la enfermedad por el nuevo coronavirus 2019 ha alcanzado el estatus de pandemia. El reporte de casos en pacientes pediátricos ha sido escaso. El objetivo de este artículo es entregar información epidemiológica del COVID-19, especialmente pediátrica. Los niños han presentado enfermedad en clusters, secundaria a contacto con parientes enfermos. La clínica es oligosintomática, menos severa y mayor riesgo concentrado en menores de 1 año y con comorbilidad. En Chile, los pacientes pediátricos representan cerca del 6% del total y la letalidad global es notablemente más baja que en adultos. Las principales medidas de control para la reducción de la reproducción efectiva son el testeo masivo, distanciamiento social y cierre escolar, sin desestimar la responsabilidad individual. El adecuado abastecimiento de elementos de protección personal es clave para evitar la infección nosocomial y del personal de salud.

Dairy calves' mortality survey and associated management practices in smallholding, pasture-based herds in southern Brazil.

Our retrospective, cross-sectional study aimed to identify the mortality rate of preweaned heifer calves and associated herd-management factors in family-run dairy holdings in the state of Rio Grande do Sul, Brazil. A survey was made available through an online platform to all the municipalities (n = 494) with a state official extension office (Emater), allowing for any milk producer in RS raising the calves on-farm to take part in the study. A total of 1451 farmers responded to the survey between October 2014 and December 2015. Yearly total mortality, stillbirths and post-natal mortality to weaning were calculated from responses. The association of herd characteristics and practices with herd mortality was evaluated using multilevel multivariable logistic regression models. Herd mortality was modelled as a two vector binded variable that included the number of failures (n = female calves born alive but dead before weaning) and successes (n = weaned female calves) on-farm in the past 12 months. Models included region as random effect. The herds had 13 lactating cows (median, range 1-130). Pure breed herds, especially Holstein, predominated (81 %). Milk production was 190 L/d (median, range 7-4000). Total mortality rate was 8.5 % (1065/12563, median 0.0 %, Q1 - Q3 0.0-14.3 %/farm), and stillbirths 1.7 % (207/12563; median 0.0 %, Q1 - Q3 0.0 - 0 %/farm). Post-natal mortality rate was 6.9 % (858/12356, median 0.0 %, Q1 - Q3 0.0-11.1 %/farm). In 89.2 % of the farms, no stillborn calf was reported, and in only 2.8 % there was more than one case. In 67 % of farms, no death calf born alive was reported; in 33 %, an average of 2 deaths/farm (range 1-12 deaths/farm) was reported. In 16 % of the farms, mortality was ≥ 20 %, and in 4.3 % it was ≥ 50 %. The main reported causes of death were diarrhoea and unknown causes. Higher mortality was associated with herds with mixed breeds (OR = 1.3, CI = 1.09-1.59), performing unattended calvings (OR = 1.2; IC = 1.04-1.40), leaving the calf for long periods with the cow (OR = 1.21, CI = 1.00-1.45), and housing various calves in a pen (OR = 1.4, CI = 1.20-1.60). Furthermore, reduced mortality was associated with medium size herds (i.e. 21-40 lactating cows; OR = 0.82, CI = 0.69-0.97). Although the mortality rate observed is conservatively lower or equal to other international reports, it is higher than could be desired. The factors identified as associated to mortality are understood as proxies for the poor quality of management of the practices adopted. Thus, reduction of mortality is at hand without representing major infrastructural changes.

Integrin-linked kinase mediates the hydrogen peroxide-dependent transforming growth factor-ß1 up-regulation.

Transforming growth factor type-ß1 (TGF-ß1) has been recognized as a central mediator in many pathological events related to extracellular matrix (ECM) proteins accumulation, where their locally increased expression has been implicated in the fibrosis process of numerous organs, including glomerular fibrosis in the kidney. We and others have reported the TGF-ß1 synthesis regulation by reactive oxygen species (ROS), and moreover we also described the implication of integrin-linked kinase (ILK) in the AP-1-dependent TGF-ß1 up-regulation. Thus, we propose here that hydrogen peroxide (H2O2)-dependent TGF-ß1 regulation may be mediated by ILK activation. First we confirmed the increase in TGF-ß1 expression in human mesangial cells (HMC) after treatment with H2O2 or with an alternative H2O2-generating system such as the glucose-oxidase enzyme (GOX). By using immunoblotting, immunofluorescence, and ELISA techniques, we demonstrate that extracellular H2O2 up-regulates TGF-ß1 transcription, as well as increases TGF-ß1 promoter activity. Furthermore, catalase-decreased intracellular H2O2 abolished TGF-ß1 up-regulation. The use of pharmacological inhibitors as well as knockdown of ILK with small interfering RNA (siRNA) demonstrated the implication of a PI3K/ILK/AKT/ERK MAPK signaling pathway axis in the H2O2-induced TGF-ß1 overexpression. Finally, we explored the physiological relevance of these findings by treating HMC with angiotensin II, a known stimuli of H2O2 synthesis. Our results confirm the relevance of previous findings after a more physiological stimulus. In summary, our results provide evidence that ILK activity changes may act as a mechanism in response to different stimuli such as H2O2 in the induced TGF-ß1 up-regulation in pathological or even physiological conditions.

Efficient gene expression from integration-deficient lentiviral vectors in the spinal cord.

Gene transfer to spinal cord cells may be crucial for therapy in spinal muscular atrophy, amyotrophic lateral sclerosis and spinal cord injury. Lentiviral vectors are efficient for transduction of a variety of cells, but like all integrating vectors they pose a risk of insertional mutagenesis. Integration-deficient lentiviral vectors (IDLVs) remain episomal but retain the transduction efficiency of standard integrating lentiviral vectors, particularly when the episomes are not diluted out through repeated cell division. We have now applied IDLVs for transduction of spinal cord in vitro, in explants and in vivo. Our results demonstrate similar efficiency of eGFP expression from integrating lentiviral vectors and IDLVs in most cell types analyzed, including motor neurons, interneurons, dorsal root ganglia (DRG) neurons and astroglia. IDLV-mediated expression of pro-glial-cell-derived neurotrophic factor (Gdnf) rescues motor neuron cultures from death caused by removal of exogenous trophic support. IDLVs also mediate efficient RNA interference in DRG neuron cultures. After intraparenchymal injection in the rat and mouse cervical and lumbar regions in vivo, transduction is mainly neuronal, with both motor neurons and interneurons being efficiently targeted. These results suggest that IDLVs could be efficient and safer tools for spinal cord transduction in future therapeutic strategies.

A reduction in milking frequency and feed allowance improves dairy cow immune status.

Twice-daily milking is the most common milking regimen used globally. A reduction in milking frequency to once daily, combined with a reduced feed allowance (FA), could reduce the physiological stress associated with the transition to peak milk production, and hence improve immune function. This study investigated how milking frequency and FA affect dairy cow immune status. Cows (n = 48) were milked once a day (OAD) or twice a day (TAD) on 1 of 2 FA: high (HFA) or low (LFA), in a 2 × 2 factorial arrangement of treatments. After the mean calving date of March 11, HFA cows were offered ad libitum grass silage and 7 kg of concentrates/cow per day until March 22, then 4 kg of concentrates/cow per day until April 17, and thereafter allocated 31.3 kg of dry matter (DM) grass/cow per day. The LFA cows were offered 4 kg of concentrates/cow per day, 1 kg of concentrates/cow per day, and allocated 19 kg of DM grass/cow per day for the same respective periods. Milk yield was recorded daily and body condition score weekly, and somatic cell count was performed at approximately 2-wk intervals. Blood samples were collected prepartum (d -7 to -1) and at d 1 to 7, d 14 to 21, and d 42 to 49 postpartum. Total and differential leukocyte percentage, IFN-γ production in response to concanavalin A and phytohemagglutinin, and cortisol, haptoglobin (Hp), and serum amyloid A (SAA) concentrations were evaluated. Cows milked OAD had reduced milk yield and body reserve mobilization, but higher somatic cell counts. Milking frequency and diet had no effect on total leukocyte counts. Cows milked OAD had a higher lymphocyte percentage and lower monocyte percentage, and tended to have a lower neutrophil percentage than cows milked TAD. In addition, the LFA cows had a higher eosinophil percentage than cows fed the HFA. Milking frequency and diet had no effect on IFN-γ, Hp, SAA, or cortisol production. Utilization of strategies to reduce milk yield at the beginning of the lactation could not only reduce body reserve mobilization, but also help to maintain a functioning immune system, and thus improve cow welfare.

Retrospective analysis of post-mortem findings in 1,444 aged donkeys.

The aim of this study was to describe and report the prevalence of conditions found at necropsy examination of UK donkeys. Records from 1,444 donkeys over a 7-year period were included in the analysis. Sixty-one categories of post-mortem finding were identified from 9,744 observations. The four most prevalent conditions noted were dental disorder (78.7%), vascular disease other than aneurysm (60.9%), arthritis (55.4%) and foot disorder (44.8%). Gastric ulceration was found in 42% of the donkeys and gastrointestinal impaction in 18.6%. The most frequent combination of two post-mortem findings in the same animal was arthritis and dental disorder. The most common disorders were associated with age, body weight and/or body condition post mortem and, for some disorders, gender. For many of the post-mortem findings, crude associations were found between the presence of one finding and the odds of also having certain other post-mortem findings. This study is the first to summarize all conditions noted at necropsy examination for a large group of donkeys. The findings increase knowledge of diseases and conditions of this species and may be useful when investigating the relevance of various pathological conditions in the live animal.

Tumor necrosis factor alpha and interferon gamma cooperatively induce oxidative stress and motoneuron death in rat spinal cord embryonic explants.

The accumulation of reactive microglia in the degenerating areas of amyotrophic lateral sclerosis (ALS) tissue is a key cellular event creating a chronic inflammatory environment that results in motoneuron death. We have developed a new culture system that consists in rat spinal cord embryonic explants in which motoneurons migrate outside the explant, growing as a monolayer in the presence of glial cells. The proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) have been proposed to be involved in ALS-linked microglial activation. In our explants, the combined exposure to these cytokines resulted in an increased expression of the pro-oxidative enzymes inducible nitric oxide synthase (iNOS), the catalytic subunit of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, gp91(phox) and cyclooxygenase-2 (COX-2), as compared to each cytokine alone. This effect was related to their cooperation in the activation of the transcription factor nuclear factor kappa B (NF-kappaB). TNF-alpha and IFN-gamma also cooperated to promote protein oxidation and nitration, thus increasing the percentage of motoneurons immunoreactive for nitrotyrosine. Apoptotic motoneuron death, measured through annexin V-Cy3 and active caspase-3 immunoreactivities, was also found cooperatively induced by TNF-alpha and IFN-gamma. Interestingly, these cytokines did not affect the viability of purified spinal cord motoneurons in the absence of glial cells. It is proposed that the proinflammatory cytokines TNF-alpha and IFN-gamma have cooperative/complementary roles in inflammation-induced motoneuron death.

Vascular endothelial growth factor protects motoneurons from serum deprivation-induced cell death through phosphatidylinositol 3-kinase-mediated p38 mitogen-activated protein kinase inhibition.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the selective degeneration and death of motoneurons in the spinal cord, brainstem and motor cortex which causes progressive muscle weakness and paralysis. Although the molecular mechanisms causing the disease remain unknown, excitotoxicity and loss of trophic support have been proposed as causes of degeneration. The present study was designed to elucidate the mechanisms of motoneuron death induced by serum deprivation and the potential neuroprotective effects of vascular endothelial growth factor (VEGF) in dissociated and organotypic rat spinal cord cultures. Serum withdrawal induced apoptotic cell death in dissociated spinal cord cultures, which was prevented in the presence of VEGF. In organotypic spinal cord cultures, low serum-induced motoneuron death was mediated by the stress-related kinase p38 mitogen-activated protein kinase (p38MAPK), as it was reversed by the p38MAPK inhibitor SB203580. In these cultures, exposure to VEGF blocked p38MAPK phosphorylation and prevented the demise of motoneurons. These effects of VEGF were mediated through the phosphatidylinositol 3-kinase/Akt (PI3-K/Akt) signal transduction pathway, as they were blocked in the presence of the PI3-K inhibitor LY294002. In addition, serum deprivation induced down-regulation of the anti-apoptotic protein Bcl-2 and this effect was prevented both by SB203580 and by VEGF via the PI3-K/Akt pathway. Therefore, Bcl-2 could also play an important role in the neuroprotection induced by VEGF in spinal cord cultures. Together, these findings indicate that VEGF prevents motoneuron death induced by serum deprivation blocking the activity of p38MAPK via the PI3-K/Akt signaling pathway.

Effect of genetic group and feed system on locomotion score, clinical lameness and hoof disorders of pasture-based Holstein-Friesian cows.

The aim of the present study was to determine the effect of the genetic group of the Holstein-Friesian (HF) and pasture-based feeding system (3 × 2 factorial arrangement) on locomotion score (six gait aspects scored from one to five), clinical lameness and hoof disorders within a seasonal calving milk production system. The three genetic groups compared had an average Economic Breeding Index (EBI) value of 40, 70 and 80: representing the Irish national average genetic merit (LOW-NA), high EBI genetic merit of North American ancestry (HIGH-NA) and high EBI genetic merit of New Zealand ancestry (HIGH-NZ), respectively. Two feed systems were compared: a high grass allowance, low-concentrate system typical of spring-calving herds in Ireland (control) and a high-concentrate system. Data from 126 cows collected across a complete lactation period were analysed using generalised estimating equations and survival analysis. Genetic group of HF had a significant effect on locomotion score, clinical lameness and hoof disorders. Higher EBI cows (HIGH-NA and HIGH-NZ) had lower hazard of poor locomotion score in some gait aspects (e.g. spine curvature) and lower odds of clinical lameness in the first 200 days post-calving (Odds ratios 0.08 and 0.24, respectively, relative to the LOW-NA) and some hoof disorders (e.g. traumatic lesions) compared with LOW-NA cows. The high-concentrate feed system showed a higher incidence and severity of digital dermatitis (P < 0.01). Thus, high EBI cows have better locomotion, fewer cases of clinical lameness and less-severe hoof disorders (i.e. digital dermatitis, white line disease and traumatic lesions) than low EBI cows. These findings have important implications for cow welfare and productivity.

Effect of Holstein-Friesian genetic group on peripartum and early lactation haematological and acute phase proteins profiles, health and fertility.

Pasture-based Holstein-Friesian cows from three genetic groups differing in the Irish 'Economic Breeding Index' (EBI) value and genetic background, namely North-American (NA) national average EBI genetic merit (LOW-NA, n = 42), North-American high EBI genetic merit (HIGH-NA, n = 42) and New Zealand (NZ) high EBI genetic merit (HIGH-NZ, n = 42), were studied. These genetic groups have been selected in different environments: pasture for NZ and confinement for NA. The objective was to determine the effect of genetic group on haematological and acute phase proteins profiles (white blood cell (WBC) counts, red blood cell (RBC) counts, acute phase proteins: serum amyloid A (SAA) and haptoglobin), health (rectal temperature (RT), clinical mastitis (CM) and somatic cell score), calving performance (stillbirth, calving assistance) and post-partum reproductive parameters (endometritis and ovarian cyclicity). Blood sampling and data recording took place 3 weeks pre-calving up to 7 weeks post-calving. Linear mixed models, logistic regression and generalised estimating equations were used for data analysis. HIGH-NZ animals had the highest (P < 0.05) RBC mean corpuscular volume (50.0 fl), exhibited a different WBC distribution pattern (P < 0.05) and had the lowest (P < 0.05) mean RT (38.4°C) for the first 10 days post-calving. These findings suggest enhanced reticulocyte turnover, peripartum response mechanisms and thermoregulation in the HIGH-NZ compared to the other two genetic groups. LOW-NA animals had the highest SAA peak throughout the peripartum period (55.12 mg/l, P < 0.05) and a tendency for higher somatic cell scores (P < 0.10) in early lactation. The HIGH-NA animals had the lowest incidence of udder quarter milk sample bacteria at calving, suggesting better udder health when commencing lactation. No differences were detected between genetic groups in calving performance, post-partum reproductive parameters or CM in the first 42 days post-calving. These results suggest that while inherited peripartum adaptation strategies have been developed by the different genetic groups selected in different environments (pasture = NZ v. confinement = NA), such differences have minimal impact on peripartum clinical health.

Transplacental exchange of moxidectin after maternal or fetal intravenous administration in sheep.

The transplacental exchange of moxidectin after maternal or fetal intravenous (i.v.) administration was studied using the chronically catheterized fetal sheep model. Nine pregnant Suffolk Down sheep of 65.7 +/- 5.9 kg body weight (bw) were surgically prepared to insert polyvinyl catheters in the fetal femoral artery and vein and amniotic sac. The ewes were randomly assigned to two experimental groups. In group 1 (maternal injection) five ewes were treated with an i.v. bolus of 0.2 mg of moxidectin/kg bw. In group 2, (fetal injection) an i.v. bolus of 1 mg of moxidectin was administered to the four fetuses by femoral vein catheters. Maternal and fetal blood and amniotic fluid samples were taken before and after moxidectin administration for a 144 h post-treatment period. Samples were analyzed by liquid chromatography. A noncompartmental pharmacokinetic analysis was performed and statistical differences were determined by mean of parametric and nonparametric statistical tests. Pharmacokinetic differences observed in maternal variables were shorter elimination half-life and mean residence time compared with values previously reported for ivermectin. Drug diffusion from maternal to fetal circulation (AUC(0-t) = 232.6 +/- 72.5 ng.h/mL) was statistically not different (P = 0.09) compared with fetal to maternal diffusion (AUC(0-t) = 158.0 +/- 21.6 ng.h/mL). Fetuses showed significantly (P = 0.008) lower drug body clearance values compared with those observed in the maternal side. Considering the observed transplacental passages between materno-fetal or feto-maternal circulations, we conclude that the placental barrier is not effective in preventing the moxidectin diffusion between mother and fetus.

[Clinical correlation of hypnagogic hypersynchrony during sleep in normal children and those with learning disability]. / Correlación clínica de la hipersincronía hipnagógica durante el sueño, en niños normales y con problemas de aprendizaje.

INTRODUCTION: One of the electroencephalographic (EEG) patterns that can be mistaken for paroxysmal clinical activity, when not taken into account and especially in children, is hypnagogic hypersynchrony (HH). This consists in generalised, paroxysmal, synchronic, symmetrical, slow, high voltage waves lasting 2 8 seconds, which appear in drowsiness and in stage I. It was observed that this pattern often appeared in children with learning disability (LD). AIMS. To correlate clinical data with the presence of HH during sleep in normal children and those with LD. PATIENTS AND METHODS: We assessed 180 children between the ages of 6 12 years with normal neurological development, 130 of which suffered LD and 50 who did not have LD. EEG was performed with sleep deprivation, following the International Federation of Clinical Neurophysiology guidelines. RESULTS: The presence or absence of HH, together with its characteristics, was assessed. Of the children with LD, 35.38% displayed HH and of the children without LD, only 4% displayed HH. Since the characteristics of HH in the children with LD were different to previous descriptions, we put forward criteria with which to evaluate those differences. CONCLUSIONS: HH appeared more often in children with LD than in normal children. Qualitative, quantitative (p< 0.05) and morphological changes were found in the paroxysmal activity of HH during the stages of sleep in children with LD.

Mu-opioid receptor activation prevents apoptosis following serum withdrawal in differentiated SH-SY5Y cells and cortical neurons via phosphatidylinositol 3-kinase.

Opioid peptides and alkaloids exert their effects via G protein-coupled receptors (GPCRs). It has been shown that, in addition to trophic factors, some GPCRs are able to activate the phosphatidylinositol 3-kinase/Akt (PI 3-K/Akt) signal transduction pathway, thus leading to cell survival. The aim of this study was to test whether activation of mu-opioid receptors has protective effects on serum withdrawal-induced cell death and to study the possible implication of PI 3-K in this process. In SH-SY5Y neuroblastoma cells fully differentiated by exposure to retinoic acid for five days, the enkephalin derivative selective mu-agonist DAMGO (0.1-2 microM) and the alkaloid morphine (0.1-10 microM) promoted cell survival after serum deprivation (MTT and trypan blue exclusion assays), without inducing cell proliferation. These effects were fully reversed by naloxone, by the selective mu-antagonist beta-funaltrexamine (beta-FNA) and also by the specific PI 3-K inhibitor LY294002. The two agonists stimulated Akt phosphorylation and the effect was also abolished by beta-FNA and by LY294002. In mouse primary cortical neurons, DAMGO reduced the percentage of apoptosis after 6, 12, 24 and 48 h of serum withdrawal; as determined by Hoechst staining. This effect was blocked by beta-FNA, by pre-treatment with pertussis toxin and by LY294002. DAMGO also stimulated Akt phosphorylation via PI 3-K in this primary neuronal culture. Together, these results indicate that stimulation of the mu-opioid receptor promotes neuronal survival in a G(i/o)-linked, PI 3-K-dependent signaling cascade and suggest that Akt may be a key downstream kinase involved in this anti-apoptotic effect.

[Intravenous treatment with immunoglobulins in epileptic syndromes which are difficult to control]. / Tratamiento intravenoso con inmunoglobulinas en síndromes epilépticos de difícil control.

INTRODUCTION: Several mechanisms have been found to be involved in the development of malignant epileptic syndromes in childhood and an immunological component has been shown in experimental studies of epilepsy. The effect of intravenous immunoglobulin (IgIV) has been reported in various cases of epilepsy which were difficult to control. OBJECTIVE: To show the usefulness of IgIV in West s syndrome (WS) and Lennox Gastaut syndrome (LGS) which were difficult to control. PATIENTS AND METHODS: We selected five patients who fulfilled the criteria for WS and LGS which were difficult to control. They were given IgIV for five days and subsequently six fortnightly doses of 0.5 g/kg/day. We recorded data on sex, age, onset of the disorder and laboratory investigations, including blood and cerebrospinal fluid (CSF) immunoglobulin levels. RESULTS: Four girls with symptomatic WS and one boy with LGS as a symptom of a neuro infection. Treatment was started at the age of 8 14 months. The spasms varied between 204 and 1,074 in 24 hours, and were markedly reduced after IgIV was given. The IgG in the CSF rose as the number of spasms fell (p< 0.05). CONCLUSIONS: There was a satisfactory response to treatment, as reported in previous publications. It is a coadjuvant treatment for cases which are difficult to control and may also be useful in cases of symptomatic epilepsy where goods results are also obtained.

Differential in vitro and in vivo behavior of mouse ascorbate/Fe3+ and diamide oxidized erythrocytes.

Chemical oxidation of mouse erythrocytes has been carried out using two different oxidizing systems namely: Diamide and Ascorbate/Fe3+ together with different concentrations of the oxidant. These oxidation treatments produced different extents of modification in membrane proteins as was observed by electrophoretic analyses that showed a possible formation of high molecular weight aggregates. Lipid peroxidation was also observed as the result of these chemical treatments. The action of these two oxidation treatments produced different extents of lipid peroxidation in which the effect Ascorbate/Fe3+ reached higher values than that shown by diamide treatments. To study the resulting in vitro behavior of such oxidized erythrocytes, we have evaluated the recognition of oxidized erythrocytes by peritoneal macrophages. In the conditions used, diamide oxidized erythrocytes were more highly recognized by macrophages than Ascorbate/Fe3+ treated erythrocytes. However, in both cases an influence of serum factors in the recognition process can be inferred. Additionally, we have correlated on one side the action of different oxidation systems on mouse erythrocytes with different in vivo behavior and organ uptake of the oxidized erythrocytes. On the other side, differential targeting of oxidized erythrocytes to a liver or spleen was observed on dependence of the oxidant used.

Influence of aerobic oxidation of mouse erythrocytes on their recognition by macrophages.

Membrane protein modification can change cell surface properties which can be correlated with altered macrophage-erythrocyte interactions. Mouse erythrocytes were incubated in phosphate buffer for different times to induce protein modification. Mouse erythrocyte membrane changes were analyzed by infrared analyses and gel electrophoresis. Proteolytic digestion of membrane proteins was observed. After 22 hours preliminary incubation, the number of erythrocytes adhering to a monolayer of macrophages reached a maximum, the majority of which had not been phagocytosed. Most of the erythrocytes incubated for 40 hours underwent phagocytosis after adhesion to the macrophages.

Induction of reactive astrocytosis and prevention of motoneuron cell death by the I(2)-imidazoline receptor ligand LSL 60101.

I(2)-imidazoline receptors are mainly expressed on glial cells in the rat brain. This study was designed to test the effect of treatment with the I(2)-imidazoline selective receptor ligand LSL 60101 [2-(2-benzofuranyl)imidazole] on the morphology of astrocytes in the neonate and adult rat brain, and to explore the putative neuroprotective effects of this glial response. Short-term (3 days) or chronic (7-10 days) treatment with LSL 60101 (1 mg kg(-1), i.p. every 12 h) enhanced the area covered by astroglial cells in sections of facial motor nucleus from neonate rats processed for glial fibrillary acidic protein (GFAP) immunostaining. Facial motoneurons surrounded by positive glial cell processes were frequently observed in sections of LSL 60101-treated rats. A similar glial response was observed in the parietal cortex of adult rats after chronic (10 days) treatment with LSL 60101 (10 mg kg(-1), i.p. every 12 h). Western-blot detection of the specific astroglial glutamate transporter GLT-1, indicated increased immunoreactivity after LSL 60101 treatment in the pons of neonate and in the parietoccipital cortex of adult rats. In the facial motor nucleus of neonate rats, the glial response after LSL 60101 treatment was associated to a redistribution of the immunofluorescence of the basic fibroblast growth factor (FGF-2) from the perinuclear area of motoneurons to cover most of their cytoplasm, suggesting a translocation of this mitogenic and neurotrophic factor towards secretion pathways. The neuroprotective potential of the above effects of LSL 60101 treatment was tested after neonatal axotomy of facial motor nucleus. Treatment with LSL 60101 (1 mg kg(-1), i.p. every 12 h from day 0 to day 10 after birth) significantly reduced (38%) motoneuron death rate 7 days after facial nerve axotomy performed on day 3 after birth. It is concluded that treatment with the I(2)-imidazoline selective receptor ligand LSL 60101 provokes morphological/biochemical changes in astroglia that are neuroprotective after neonatal axotomy.

Activation of I(2)-imidazoline receptors enhances supraspinal morphine analgesia in mice: a model to detect agonist and antagonist activities at these receptors.

This work investigates the receptor acted upon by imidazoline compounds in the modulation of morphine analgesia. The effects of highly selective imidazoline ligands on the supraspinal antinociception induced by morphine in mice were determined. 2. Intracerebroventricular (i.c.v.) or subcutaneous (s.c.) administration of ligands selective for the I(2)-imidazoline receptor, 2-BFI, LSL 60101, LSL 61122 and aganodine, and the non selective ligand agmatine, increased morphine antinociception in a dose-dependent manner. Neither moxonidine, a mixed I(1)-imidazoline and alpha(2)-adrenoceptor agonist, RX821002, a potent alpha(2)-adrenoceptor antagonist that displays low affinity at I(2)-imidazoline receptors, nor the selective non-imidazoline alpha(2)-adrenoceptor antagonist RS-15385-197, modified the analgesic responses to morphine. 3. Administration of pertussis toxin (0.25 microg per mouse, i.c.v.) 6 days before the analgesic test blocked the ability of the I(2)-imidazoline ligands to potentiate morphine antinociception. 4. The increased effect of morphine induced by I(2)-imidazoline ligands (agonists) was completely reversed by idazoxan and BU 224. Identical results were obtained with IBI, which alkylates I(2)-imidazoline binding sites. Thus, both agonist and antagonist properties of imidazoline ligands at the I(2)-imidazoline receptors were observed. 5. Pre-treatment (30 min) with deprenyl, an irreversible inhibitor of monoamine oxidase B (IMAO-B), produced an increase of morphine antinociception. Clorgyline, an irreversible IMAO-A, given 30 min before morphine did not alter the effect of the opioid. At longer intervals (24 h) a single dose of either clorgyline or deprenyl reduced the density of I(2)-imidazoline receptors and prevented the I(2)-mediated potentiation of morphine analgesia. 6. These results demonstrate functional interaction between I(2)-imidazoline and opioid receptors. The involvement of G(i)-G(o) transducer proteins in this modulatory effect is also suggested.