RESUMEN
Presentation A 24-year-old newly graduated junior doctor presented to the emergency department with acute onset chest pain, haemoptysis and exertional dyspnoea following a dry needling session. Diagnosis Chest x-ray showed bilateral pneumothoraces, worse on the right side. Treatment The bilateral pneumothoraces were treated conservatively with supplemental oxygen initially. On the second day of admission, repeat chest x-ray demonstrated a worsening right sided pneumothorax. While vitally stable, the patient however had become increasingly dyspnoeic, and a needle aspiration was performed on the right side with good effect. Conclusion The anatomical location targeted along with the patients low-normal BMI makes her high-risk when considering the skin-to-pleura distance. Although the incidence of pneumothorax is low, it is imperative that we improve awareness both for the treating physician and the diagnosing clinician. We must begin to fill the distinct lack in available literature surrounding the potential adverse effects of dry needling.
Asunto(s)
Terapia por Acupuntura , Punción Seca , Neumotórax , Terapia por Acupuntura/efectos adversos , Adulto , Dolor en el Pecho/etiología , Disnea/etiología , Disnea/terapia , Femenino , Humanos , Neumotórax/diagnóstico por imagen , Neumotórax/etiología , Neumotórax/terapia , Adulto JovenRESUMEN
Protein is an expensive component of the dairy cow diet, and overfeeding protein can have adverse economic and environmental impacts. Our objective was to maintain milk production and components while decreasing dietary crude protein (CP) through use of a heat-treated, rumen-resistant sugar amino acid complex (SAAC) as the Schiff base, as an addition to low-protein diets. Dietary treatments included a negative control [NC, 146 g of CP/kg of dry matter (DM)], a positive control (PC, 163 g of CP/kg of DM), and the NC supplemented with SAAC in lieu of some barley grain (SAAD, 151 g of CP/kg of DM). Diets were fed to 30 multiparous Holstein-Friesian dairy cows for the first 50 d postpartum. Dry matter intake (DMI) was determined daily. Milk yield and content of fat, protein, lactose, and casein were recorded weekly from wk 2 to 7 of lactation. The fixed effects of treatment, week, treatment × week, month of calving, and BCS at calving, and a random effect of cow, were analyzed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). The SAAD treatment had greater energy-corrected milk yield than did NC. The PC treatment had greater DMI than did NC, and SAAD tended to have greater DMI than did NC. We found significant treatment effects for fat percentage and yield. The NC and SAAD treatments had higher fat percentages than did PC, and SAAD had a higher fat yield than did the NC and PC treatments. Treatment effects were found for casein yield and percentage. We discovered a treatment effect for protein percentage and yield. The PC treatment had higher protein percentage than did NC and SAAD. The PC treatment had a higher protein yield than did NC, and analysis revealed no difference in protein yield between PC and SAAD. The SAAD treatment had higher total milk solids than did the NC treatment. Lactose yield tended to be higher in PC than in NC, and no differences were found between PC and NC and SAAD treatments. The PC treatment had a higher casein percentage than did NC and SAAD; however, the SAAD and PC treatments had higher casein yields than did NC. The PC treatment had a higher casein:fat ratio than did the NC and SAAD treatments. The NC and SAAD treatments had higher Cheddar cheese yields than did PC. We found no treatment × week interactions for any parameter. Supplementing low-protein dairy cow diets with a heat-treated, rumen-resistant SAAC caused beneficial effects by improving milk components and increasing cheese yield to levels similar to those found when feeding expensive and environmentally damaging high-protein diets.
Asunto(s)
Bovinos/fisiología , Queso/análisis , Carbohidratos de la Dieta/metabolismo , Proteínas en la Dieta/metabolismo , Ingestión de Alimentos , Leche/metabolismo , Aminoácidos/administración & dosificación , Aminoácidos/metabolismo , Alimentación Animal/análisis , Animales , Dieta/veterinaria , Carbohidratos de la Dieta/administración & dosificación , Proteínas en la Dieta/administración & dosificación , Suplementos Dietéticos/análisis , Femenino , Distribución Aleatoria , Sacarosa/administración & dosificación , Sacarosa/metabolismoRESUMEN
Body condition score (BCS) change is an indirect measure of energy balance. Energy balance before calving may affect production and health in the following lactation. It is likely that cows may experience BCS loss before calving due to negative energy balance. The objective of this study was to determine if loss of BCS 15d before calving affected milk production, BCS profile, and metabolic status during the transition period and early lactation. On d -15 to d 0 relative to calving, BCS was assessed (1=emaciated, 5=obese) for 98 Holstein-Friesian cows. The cows were divided into 2groups: those that did not lose BCS between d -15 and d 0 (maintained, BCS-M, n=55) and those that lost BCS from d -15 to d 0 (lost, BCS-L, n=43, average loss of 0.29±0.11 BCS). The fixed effects of BCS group, parity, week (day when analyzing milk production records), their interactions, and a random effect of cow were analyzed using PROC MIXED of SAS (SAS Institute Inc., Cary, NC). Before calving, BCS-L cows tended to have higher concentrations of nonesterified fatty acids than BCS-M cows (0.88 vs. 0.78mmol/L). After calving, BCS-L cows had higher nonesterified fatty acid concentrations in wk 1 (0.93 vs. 0.71mmol/L), wk 2 (0.84 vs. 0.69mmol/L), and wk 4 (0.81 vs. 0.63mmol/L) than BCS-M cows. The BCS-L cows had higher concentrations of ß-hydroxybutyrate (BHB) in wk 1 (0.72 vs. 0.57mmol/L), wk 2 (0.97 vs. 0.70mmol/L), and wk 4 (0.94 vs. 0.67mmol/L) compared with BCS-M cows. We detected significant reductions in insulin concentrations in BCS-L cows from wk -1 (2.23 vs. 1.37 µIU/mL) to wk 2 (1.68 vs. 0.89 µIU/mL) and wk 4 (2.21 vs 1.59 µIU/mL) compared with BCS-M cows. Prevalence of subclinical ketosis increased in BCS-L cows in wk 3 and 4 when BHB was ≥1.4mmol/L and in wk 1, 3, and 4 when BHB was ≥1.2mmol/L. In wk 1, BCS-L cows tended to have lower levels of calcium than BCS-M cows (2.33 vs. 2.27mmol/L). We found no differences between the groups of cows for milk yield and energy-corrected milk. The BCS-L cows had lower BCS up to 75d in lactation. Overall, BCS-L cows had higher somatic cell scores with an elevated somatic cell score on d 45, d 60, and d 75. There was an overall tendency for BCS-L cows to have higher fat yield and an overall significant increase in fat percentage. Overall, BCS-L cows had lower lactose percentage, with a reduction on d 60. This work shows that BCS loss before calving may have significant consequences for metabolic status, milk composition, somatic cell score, and BCS profile in dairy cows.
Asunto(s)
Lactancia , Leche/metabolismo , Ácido 3-Hidroxibutírico/sangre , Animales , Bovinos , Metabolismo Energético , Ácidos Grasos no Esterificados/sangre , Femenino , ParidadRESUMEN
There is little research on injecting drug use in Nigeria. We investigated the prevalence of HIV, hepatitis B and C, and sexually transmitted infections (STIs) among male injection drug users (IDUs) in Lagos. Male IDUs (N = 328) were recruited through respondent-driven sampling. Participants completed an interview about their sexual and injecting risk behaviours and were tested for hepatitis B surface antigen (HBV), hepatitis C antibody (HCV), HIV and syphilis, as well as genital chlamydia and gonorrhoea infections. Three-quarters of IDUs (74%) reported injecting drugs in the past one month although most did not share needles (92%) and the majority obtained sterile needles from pharmacists (87%). Estimated HBV, HCV, HIV, syphilis, gonorrhoea and chlamydia prevalences were 7.8%, 7.7%, 0.9%, 1.9%, 0.0%, and 3.7%, respectively. The burden of HIV is presently low among IDUs in Lagos. Changes in accessibility to sterile needles at pharmacists would likely have a deleterious effect on IDUs' health. HBV vaccination and HCV prevention programmes for IDUs are urgently needed.
Asunto(s)
Infecciones por Chlamydia/epidemiología , Consumidores de Drogas/estadística & datos numéricos , Gonorrea/epidemiología , Infecciones por VIH/epidemiología , Hepatitis B/epidemiología , Hepatitis C/epidemiología , Abuso de Sustancias por Vía Intravenosa/epidemiología , Sífilis/epidemiología , Adolescente , Adulto , Chlamydia , Estudios Transversales , Antígenos de Superficie de la Hepatitis B/análisis , Anticuerpos contra la Hepatitis C/análisis , Humanos , Masculino , Persona de Mediana Edad , Nigeria/epidemiología , Vigilancia de la Población , Prevalencia , Factores de Riesgo , Conducta Sexual , Factores Socioeconómicos , Adulto JovenRESUMEN
The aim was to investigate 1) the relationship between the physical activity index created for each cow by activity monitoring systems and the identification of the preovulatory follicular phase, and 2) the influence of various production, health, and cow factors on the relationship between physical activity and estrous behavior. Eighty-nine spring calving cows, on pasture, were monitored during the breeding season using the neck-mounted estrous activity monitor Heatime (SCR Engineers Ltd., Netanya, Israel). Milk samples were collected twice weekly for progesterone assay to characterize resumption of reproductive activity. Reproductive tract health was assessed weekly by ultrasonography and vaginal mucus scoring. Body condition score and milk yield were assessed every 2 wk. Heatime identified 72% of preovulatory follicular phases from which 145 inseminations resulted in 69 conceptions; 32% of activity clusters were associated with high-progesterone states (i.e., false positives). By inclusion of a 6 to 8-h duration threshold and maintaining the borderline peak activity threshold, this was improved to 87.5% with 21.3% false positives. Mean (± standard error of the mean) peak activity and cluster duration (19.3±0.53 and 10.8±0.38, respectively) were highest for the second or subsequent preovulatory follicular phases followed, in descending order, by those during first preovulatory follicular phases (14.8±2.13 and 8.4±1.4, respectively) and high progesterone states (8.0±0.47 and 3.0±0.42, respectively). The odds of an activity cluster being in a preovulatory follicular phase rather than a high-progesterone phase improved by 29% for every 1-unit increase in peak activity and by 91% for every 2-h increase in duration. The probability of an activity cluster detecting a preovulatory follicular phase was improved if it was a second or subsequent follicular phase, if body condition score was 0.25 units higher, if milk yield was 10 kg lower, and uterine infection was absent. Conception rate was influenced by insemination on the same day (52%) or day after a cluster (34.3%); inseminations were carried out using the a.m.-p.m. rule. Advances in the development of more accurate automatic monitoring of the preovulatory follicular phase will aid the timing of insemination and, thus, improve conception rates.
Asunto(s)
Detección del Estro/métodos , Animales , Bovinos , Industria Lechera/métodos , Femenino , Leche/química , Monitoreo Fisiológico/veterinaria , Actividad Motora/fisiología , Periodo Posparto/fisiología , Embarazo , Progesterona/análisis , Factores de TiempoRESUMEN
The prospect of removing cellular deposits of lipofuscin is of considerable interest because they may contribute to age related functional decline and disease. Here, we use a decapod crustacean model to circumvent a number of problems inherent in previous studies on lipofuscin loss. We employ (a) validated lipofuscin quantification methods, (b) an in vivo context, (c) essentially natural environmental conditions and (d) a situation without accelerated production of residual material or (e) application of pharmacological compounds. We use a novel CNS biopsy technique that produces both an anti-ageing effect and also permits longitudinal sampling of individuals, thus (f) avoiding conventional purely cross-sectional population data that may suffer from selective mortality biases. We quantitatively demonstrate that lipofuscin, accrued through normal ageing, can be lost from neural tissue. The mechanism of loss probably involves exocytosis and possibly blood transport. If non-disruptive ways to accelerate lipofuscin removal can be found, our results suggest that therapeutic reversal of this most universal manifestation of cellular ageing may be possible.
Asunto(s)
Envejecimiento/fisiología , Encéfalo/metabolismo , Lipofuscina/metabolismo , Animales , Astacoidea , Biomarcadores , Encéfalo/ultraestructura , Estudios Transversales , Lateralidad Funcional , Ganglios de Invertebrados/metabolismo , Ganglios de Invertebrados/ultraestructura , Modelos Lineales , Microscopía Electrónica de Transmisión/métodos , Tejido Nervioso/metabolismo , Tejido Nervioso/ultraestructura , Reproducibilidad de los ResultadosRESUMEN
We studied the efficacy of botulinum toxin (BTX-A) injections in 167 patients, from a large cohort of 259 patients, presenting with writer's cramp (WC) and followed up to 10 years. The selection of the muscle was based on a careful physical examination, using up to 6 manoeuvres whilst attempting to write in order to bring out the original dystonic posture. The injection technique had to be precise, under EMG guidance, with a hollow recording needle to detect muscle or finger fascicle. The results showed a good efficacy and tolerance of this treatment in the long term with recovery of normal writing in 46 per cent, partial benefit in 10 per cent, failure in 21 per cent, and loss to follow-up after the first injection in 23 per cent. Among the responders, 27 per cent carried on the treatment every 9 months on average, with a duration of benefit of 6 months with follow-up between 3 and 9 years. Mirror dystonia had no prognostic value. Secondary dystonia, tremulous WC, long duration WC and progressive WC were associated with poor outcome.
Asunto(s)
Toxinas Botulínicas Tipo A/uso terapéutico , Trastornos Distónicos/tratamiento farmacológico , Fármacos Neuromusculares/uso terapéutico , Adulto , Anciano , Toxinas Botulínicas Tipo A/efectos adversos , Trastornos Distónicos/diagnóstico , Trastornos Distónicos/fisiopatología , Electromiografía , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Músculo Esquelético/fisiopatología , Fármacos Neuromusculares/efectos adversos , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
Patients with chronic hypoxaemia develop secondary polycythaemia that improves oxygen-carrying capacity. Therefore, normal haemoglobin and haematocrit values in the presence of chronic arterial hypoxaemia in cystic fibrosis constitute 'relative anaemia'. We sought to determine the cause of this relative anaemia in patients with cystic fibrosis. We studied haematological indices and oxygen saturation in healthy volunteers (n=17) and in adult patients with cystic fibrosis (n=15). Patients with cystic fibrosis had lower resting arterial oxygen saturation when compared with normal volunteers (P<0.0001), and exercise led to a greater reduction in arterial oxygen saturation (P<0.0001). However, haemoglobin and haematocrit values in patients with cystic fibrosis did not significantly differ from normal volunteers. Serum iron (P=0.002), transferrin (P=0.02), and total iron-binding capacity (P=0.01) were lower in patients with cystic fibrosis. There were no significant differences in serum ferritin, percentage iron saturation, serum erythropoietin or red cell volume between the groups. The data presented demonstrate a characteristic picture of anaemia of chronic disease in adult patients with cystic fibrosis, except for normal haemoglobin and haematocrit values. Normal haemoglobin and haematocrit values in patients with cystic fibrosis appear to represent a combination of the effects of arterial hypoxaemia promoting polycythaemia, counterbalanced by chronic inflammation promoting anaemia of chronic disease.
Asunto(s)
Anemia/etiología , Fibrosis Quística/complicaciones , Oxígeno/sangre , Adulto , Anemia/sangre , Anemia Ferropénica/etiología , Enfermedad Crónica , Fibrosis Quística/sangre , Pruebas Hematológicas , Humanos , Hipoxia/etiología , Policitemia/etiologíaRESUMEN
The ability to measure lipofuscin accumulation accurately is essential for understanding its role in physiological ageing and human disease, and for its recent use as an ecological tool for age determination. Existing quantification methods are problematic. In situ histological measurement by microscopy can be very precise but is labour intensive. Spectrofluorimetric measurement of whole lipid extracts is rapid but not sufficiently specific. A recent HPLC assay for the retinal pigment epithelium lipofuscin fluorophore, A2-E, is potentially both precise and rapid but not applicable to lipofuscin in other tissues, or from fixed samples. In this study, I explore the use of flow cytometry or fluorescence activated cell sorting (FACS) for specific quantification of lipofuscin granules in formalin-fixed CNS homogenates from lobsters (Homarus gammarus). Free neurolipofuscin granules were discriminated in FACS samples by their size distribution (forward scatter), distinctive orange autofluorescence (FL3) and refractive internal structure (side scatter). A quantitative neurolipofuscin index was developed, which was highly correlated with the microscopically measured neurolipofuscin concentration in the same tissue. Sample-processing rate was at least an order of magnitude greater for FACS than for quantitative microscopy but the latter yielded a much more precise estimate of neurolipofuscin concentration. While the FACS approach may be ideal where rapid handling and only semiquantitative results are required, loss of precision will preclude use in many ecological studies where the highest available resolution is needed. Further refinements to the FACS approach are possible but advanced histological methods for neurolipofuscin quantification remain the most reliable at this time.
RESUMEN
The available evidence for thermal modulation of neurolipofuscin deposition in poikilotherms is reviewed here and additional data are contributed. Mainly decapod crustacean models are employed and neurolipofuscin is treated as an index of physiological aging. In all cases, neurolipofuscin accumulation rate is positively correlated with environmental temperature but there appears to be lowered sensitivity in the thermal mid-range, an 'optimum' temperature for neurolipofuscin accumulation and possibly age-associated variation. The geographical position of the population within the species' thermal range may determine sensitivity of the response. There is seasonal oscillation of neurolipofuscin accumulation rate, providing preliminary evidence for neurolipofuscin turnover with net loss in winter. Spatial and temporal thermal variations of similar magnitude appear to have comparable effects on neurolipofuscin accumulation rate. Such effects may be extreme, suggesting important implications for physiological aging even in homeotherms. Inter-specific comparisons indicate that species-specific neurolipofuscin accumulation rates are positively correlated with habitat temperature and inversely correlated with maximum lifespan and age at maturity. These findings help explain some well-known bioclimatic trends in maturation- and maximum body size, such as Bergmann's rule. They also highlight the fact that global warming is likely to cause significant changes in life history parameters, population dynamics and responses to exploitation for many species.
RESUMEN
The childhood respiratory consequences of very low birth weight (birth weight < or =1,500 g) are incompletely understood, especially since the introduction of recent changes in neonatal care. To assess prevalence, trends, and risk factors for respiratory symptoms, the authors followed to age 8 years a cohort of 384 very low birth weight children from six regional neonatal intensive care units in Wisconsin and Iowa who were born between August 1, 1988, and June 30, 1991. A control group of 154 Wisconsin schoolchildren was also assembled. Respiratory symptoms in the past 12 months and history of asthma ("asthma ever") were reported by parents on a questionnaire used in the International Study of Asthma and Allergies in Childhood (ISAAC). Control group prevalence resembled ISAAC prevalence worldwide and in Canada, but respiratory symptoms were twice as common among very low birth weight children. With advent of the availability of pulmonary surfactants, the prevalence of wheezing at age 8 decreased from 50% to 16% (p = 0.002) among children with bronchopulmonary dysplasia, but it increased from 14% to 38% among those with milder neonatal respiratory disease. Bronchopulmonary dysplasia, family history of asthma, smoking in the household, and patent ductus arteriosus were predictive of wheezing in the previous 12 months. Antenatal steroid therapy had a borderline-significant protective association with wheezing (odds ratio = 0.56, 95% confidence interval: 0.29, 1.1). There were interaction effects between several of the predictors.
Asunto(s)
Asma/etiología , Recién Nacido de muy Bajo Peso , Enfermedades Respiratorias/etiología , Asma/epidemiología , Niño , Estudios de Cohortes , Femenino , Humanos , Recién Nacido , Masculino , Oportunidad Relativa , Prevalencia , Surfactantes Pulmonares , Ruidos Respiratorios , Enfermedades Respiratorias/epidemiología , Factores de Riesgo , FumarRESUMEN
The 51Cr release assay has traditionally been used to investigate effector cell cytotoxic function against labeled targets, but this method has inherent problems that include hazards associated with radioactivity, cell labeling and high spontaneous release. Here we describe a novel flow cytometric assay which addresses and improves upon the problems currently encountered with the 51Cr release assay. The fluorometric assessment of T lymphocyte antigen specific lysis (FATAL) assay employs dual staining (PKH-26 and CFSE) to identify and evaluate the target population. We found that the PKH-26/CFSE combination efficiently labeled target cells. Evaluation of the spontaneous leakage from dye labeled target cells was forty fold lower than the spontaneous leakage seen with the 51Cr release assay. The FATAL assay permitted a more accurate assessment of the effector: target ratio, and detected low levels of cytotoxic T lymphocyte (CTL) mediated lysis. There was a strong correlation between the 51Cr release and FATAL assays, when performed in parallel with identical effector and target cells (r(2)=0.998, P=<0.0001). This novel method of detecting cytolysis represents a qualitative and quantitative improvement over standard 51Cr release analysis. The FATAL assay will be of value to further investigate mechanisms of cytolysis by effector cell populations.
Asunto(s)
Pruebas Inmunológicas de Citotoxicidad , Linfocitos T/inmunología , Antígenos , Citometría de Flujo/métodos , Humanos , Sensibilidad y EspecificidadRESUMEN
This cross-sectional study investigated the effect of early highly active antiretroviral therapy (HAART) on human immunodeficiency virus (HIV) type 1-specific CD8 T cell responses in children. HIV-1-specific CD8 T cell responses were quantified using an enzyme-linked immunospot assay to measure interferon-gamma-secreting cells. HIV-1-infected children were classified by time of HAART initiation prior to age 1 year or after age 2 years as early (n=24) or late (n=28) treated. The magnitude and breadth of the HIV-1-specific CD8 T cell response was significantly lower in children receiving early compared with late HAART treatment (P=.0007 and.0001, respectively). However, total CD8 T cell responses in the early HAART treatment group did not differ significantly from those of age-matched non-HAART-treated controls (n=30). Thus, the reduced magnitude and breadth of the HIV-1-specific CD8 T cell response in early HAART-treated children is due to their younger age.
Asunto(s)
Fármacos Anti-VIH/farmacología , Linfocitos T CD8-positivos/efectos de los fármacos , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/inmunología , Adolescente , Fármacos Anti-VIH/uso terapéutico , Linfocitos T CD8-positivos/inmunología , Niño , Preescolar , Infecciones por VIH/virología , VIH-1/efectos de los fármacos , VIH-1/inmunología , Humanos , LactanteRESUMEN
CD4(+) T cells are thought to be critical in the maintenance of virus-specific CD8(+) cytotoxic T-cell (CTL) responses. In human immunodeficiency virus type 1 (HIV-1) infection, a selective decline in HIV-1-specific CTL as the CD4(+) T-cell count decreases has been reported. Using HLA-peptide tetrameric complexes, we show the presence at high frequency of HIV-1- and cytomegalovirus-specific CD8(+) T cells when the peripheral CD4(+) T-cell count was low or zero in three HIV-1-infected patients. No direct virus-specific CD8(+)-mediated effector activity was seen in these subjects, suggesting antigen unresponsiveness, although tetramer-sorted cells could be expanded in vitro in the presence of interleukin-2 into responsive effector cells. Thus, virus-specific CD8(+) T cells can be maintained in the peripheral circulation at high frequency in the absence of circulating peripheral CD4(+) T cells, but these cells may lack direct effector activity. Strategies designed to overcome this antigen unresponsiveness may be of value in therapies for the treatment of AIDS.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Citomegalovirus/inmunología , VIH-1/inmunología , Linfocitos T Citotóxicos/inmunología , Humanos , Factores de TiempoRESUMEN
Using three murine tumor models, we compared the antitumor efficacy and certain physiological effects of an in vivo interleukin-12 (IL-12) gene therapy protocol and a systemic IL-12 protein therapy protocol. An IL-12 cDNA gene construct was administered in situ into skin tissue via gene gun delivery, and recombinant IL-12 protein was administered subcutaneously at a dose of 1 microgram/mouse/treatment. Both treatment regimes induced a comparable level of regression of established intradermal MethA sarcomas. In B16 melanoma and P815 mastocytoma models, antitumor efficacy of IL-12 protein therapy appeared to be slightly higher than that of IL-12 gene therapy; however, the protein therapy protocol in this comparative study resulted in a high level of mortality of mice. It was also demonstrated that IL-12 gene therapy, in contrast to the IL-12 protein therapy, was not associated with weight loss, splenomegaly, increased Ly6 antigen expression in the spleen, or visible signs of toxicity, such as fur ruffling and lethargy. Moreover, serum levels of interferon-gamma (IFN-gamma) induced in response to IL-12 gene therapy were 300-1000 times lower than those induced by the systemic IL-12 protein administration. Together, these results suggest that gene gunmediated in vivo delivery of IL-12 cDNA may be considered as a safer alternative to IL-12 protein therapy for certain human cancers.
Asunto(s)
Biolística , Terapia Genética , Interleucina-12/genética , Interleucina-12/uso terapéutico , Neoplasias Experimentales/terapia , Animales , Antígenos Ly/sangre , Femenino , Interferón gamma/sangre , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Esplenomegalia , Pérdida de PesoRESUMEN
The circumsporozoite protein (CSP) is a target for effector Ab and cell mediated immunity against malaria parasites; DNA vaccination can induce both types of effector response. The immunogenicity and efficacy of two DNA plasmids expressing different amounts of Plasmodium berghei CSP were evaluated by immunizing BALB/c mice i.m. or epidermally and by varying the number of immunizations (one to three doses) and the interval between immunizations. Expanding the interval gave the strongest effect, increasing efficacy and antibody boosting, and, in the case of epidermal vaccination, promoting a switch in CSP-specific IgG isotypes from IgG1 to a balance with IgG2a. The strongest humoral immune response and the greatest level of protection were induced by vaccinating epidermally with high expresser plasmid, using a gene gun to administer three doses at 6-wk intervals. For this group, the mean, repeat-specific, prechallenge antibody titer among mice not infected after challenge was significantly higher than that in infected mice, but the mean prechallenge titers for antibody reactive with whole sporozoites were not significantly different. The interval-dependent induction of IgG2a antibodies by epidermal vaccination contradicts the widely held belief that antibody responses induced by this method are restricted to those that are Th2 dependent.
Asunto(s)
Anticuerpos Antiprotozoarios/biosíntesis , Plasmodium berghei/inmunología , Proteínas Protozoarias/biosíntesis , Proteínas Protozoarias/inmunología , Vacunas de ADN/administración & dosificación , Vacunas de ADN/inmunología , Animales , Bovinos , Epidermis , Femenino , Humanos , Esquemas de Inmunización , Inmunización Secundaria , Inmunoglobulina G/biosíntesis , Inmunoglobulina G/sangre , Inyecciones Intradérmicas , Inyecciones Intramusculares , Malaria/inmunología , Malaria/prevención & control , Ratones , Ratones Endogámicos BALB C , Plásmidos/biosíntesis , Plásmidos/síntesis química , Plásmidos/genética , Plasmodium berghei/genética , Proteínas Protozoarias/genética , Proteínas Recombinantes/biosíntesis , Análisis de Secuencia de ADNRESUMEN
A wide variety of approaches, all using gene transfer, have been tested experimentally as alternative means to vaccinate against cancer, either prophylactically or therapeutically. These include both ex vivo and in vivo gene transfer to tumour and/or non-tumour cells, using both viral and non-viral vectors. The transferred DNA has varied widely as well, including genomic or cDNA encoding tumour-associated or oncofoetal antigens, cytokines, histocompatibility molecules, and costimulatory molecules. Several of these approaches have been applied in human clinical trials. This review summarizes those approaches, then compares and evaluates various methods using cytokine DNA in conjunction with autologous tumour cells, with particular emphasis on particle-mediated gene transfer via a gene gun. Finally, prospects and needs for further development are discussed.
Asunto(s)
Vacunas contra el Cáncer , Neoplasias/terapia , Vacunas de ADN , Animales , Vacunas contra el Cáncer/síntesis química , Vacunas contra el Cáncer/uso terapéutico , Citocinas/genética , Técnicas de Transferencia de Gen , Terapia Genética/métodos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Humanos , Neoplasias/genética , Transfección/métodos , Vacunas de ADN/síntesis química , Vacunas de ADN/uso terapéuticoRESUMEN
The aim of this study was to determine if human melanoma cells could be molecularly modified by particle-mediated gene transfer with a "gene gun", using genes for interferon-gamma (IFN-gamma), the B7-1 costimulatory molecule (CD80), and human leukocyte antigen (HLA)-A2, to augment expression of both HLA molecules and B7-1. Established and early passage melanoma cells transfected with human IFN-gamma complementary DNA (cDNA) produced IFN-gamma (50-5,000 pg/mL). The biological effect of this IFN-gamma transgene included an upregulation, or de novo appearance, of HLA expression. These melanoma cells had no detectable baseline surface expression of the B7-1 costimulatory molecule, but 8% to 31% of these cells became B7-1 positive with no selection procedure after gene transfer with human B7-1 cDNA. After combination gene transfer with cDNAs for both IFN-gamma and B7-1, 9% to 33% of these cells expressed both HLA-DR and B7-1. In combination gene transfer experiments with cDNAs for both HLA-A2 and B7-1, dual expression of HLA-A2 and B7-1 was achieved in 10% to 17% of the melanoma cells. Thus, the molecular modification of human melanoma cells to increase expression of both HLA and B7-1 can be achieved by particle-mediated gene delivery and presents a promising strategy to stimulate antimelanoma T-cell immunity. Key words: Melanoma; T cells; B7-1 costimulatory molecule (CD80); major histocompatibility complex.
Asunto(s)
Antígeno B7-1/biosíntesis , Antígenos HLA/biosíntesis , Interferón gamma/biosíntesis , Melanoma , Transfección/métodos , Antígeno B7-1/genética , Biolística , Línea Celular , ADN Complementario , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Expresión Génica , Genes Reporteros , Terapia Genética/métodos , Antígeno HLA-A2/biosíntesis , Antígenos HLA-DR/biosíntesis , Humanos , Interferón gamma/genética , Complejo Mayor de Histocompatibilidad , Melanoma/terapia , Plásmidos , Proteínas Recombinantes/biosíntesis , Células Tumorales Cultivadas , beta-Galactosidasa/biosíntesisRESUMEN
The intensity of soluble 'lipofuscin-like' autofluorescences extracted from the brain of the freshwater crayfish, Cherax quadricarinatus, was compared with the concentration of morphological lipofuscin present in sections of the same tissue. This study represents the first quantitative demonstration that these soluble autofluorescences, previously attributable to lipofuscin or fluorescent age pigment (FAP), actually bear no quantitative relationship to it at all. This result confirms previous suspicions in the literature. In some cases the intensities of these unidentified soluble autofluorescences are positive linear or curvilinear functions of organ or body weight and, therefore, may give the misleading impression that they are related to age and derived from age pigment. It is strongly recommended that researchers, particularly on aquatic species, avoid using the original biochemical assay procedure for lipofuscin and its more recent modifications.