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BACKGROUND: Patients with anorexia nervosa (AN) may experience life-threatening malnutrition-related complications requiring inpatient medical stabilisation. Several management guidelines have been developed but discrepancies exist because of limited high-level evidence. AIMS: To review the evidence base for recommendations contained in Victorian health services guidelines for the nutritional management of inpatients with AN. METHODS: MEDLINE and Embase databases were searched for published studies on the nutritional management of inpatients with AN, combined with a manual search through citations. Studies including patients with AN aged 16 years and older were included. Case reports, small case series of <10 patients, studies of nonmedical management and studies with an exclusive paediatric population were excluded. The search results were compared with AN inpatient medical management guidelines sourced from large tertiary health services across Victoria, Australia. RESULTS: The search yielded 584 studies, subsequently reduced to nine studies using the inclusion and exclusion criteria. The results suggest that commencing refeeding at a higher caloric value allows faster weight gain and shorter hospitalisation. Enteral tube feeding is preferential to parenteral nutrition because of infrequent and milder complications. Zinc supplementation showed a doubled rate of body mass index increase compared with placebo. Comparison with Victorian health services guidelines revealed inconsistent recommendations for high-calorie refeeding and micronutrient supplementation. CONCLUSION: The evidence supports high-calorie refeeding of 2000 kcal/day in AN inpatient medical management and zinc supplementation in improving the rate of weight restoration. This is inconsistently reflected in different Victorian health services guidelines. Updated national consensus guidelines could assist in improving consistency of evidence-based health care.
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Anorexia Nerviosa , Síndrome de Realimentación , Humanos , Anorexia Nerviosa/complicaciones , Anorexia Nerviosa/epidemiología , Anorexia Nerviosa/terapia , Pacientes Internos , Síndrome de Realimentación/epidemiología , Síndrome de Realimentación/prevención & control , Victoria/epidemiología , Zinc , Adolescente , AdultoRESUMEN
A domestic supply chain to produce 238Pu fuel for radioisotope thermoelectric generators is critical for enabling future space exploration. A multi-laboratory effort has worked to establish a common target design to efficiently produce 238Pu in two research reactors. This approach ensures that the annual production goals set forth by NASA are met, while also establishing redundant production capabilities. This paper describes the effort to develop the common target design as well as considerations for future applications for the irradiation platform.
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STUDY DESIGN: Controlled animal study. OBJECTIVE: To assess the cellular contribution of autograft to spinal fusion and determine the effects of intraoperative storage conditions on fusion. SUMMARY OF BACKGROUND DATA: Autograft is considered the gold standard graft material in spinal fusion, purportedly due to its osteogenic properties. Autograft consists of adherent and non-adherent cellular components within a cancellous bone scaffold. However, neither the contribution of each component to bone healing is well understood nor are the effects of intraoperative storage of autograft. MATERIALS AND METHODS: Posterolateral spinal fusion was performed in 48 rabbits. Autograft groups evaluated included: (1) Viable, (2) partially devitalized, (3) devitalized, (4) dried, and (5) hydrated iliac crest. Partially devitalized and devitalized grafts were rinsed with saline, removing nonadherent cells. Devitalized graft was, in addition, freeze/thawed, lysing adherent cells. For 90 minutes before implantation, air dried iliac crest was left on the back table whereas the hydrated iliac crest was immersed in saline. At 8 weeks, fusion was assessed through manual palpation, radiography, and microcomputed tomography. In addition, the cellular viability of cancellous bone was assayed over 4 hours. RESULTS: Spinal fusion rates by manual palpation were not statistically different between viable (58%) and partially devitalized (86%) autografts ( P = 0.19). Both rates were significantly higher than devitalized and dried autograft (both 0%, P < 0.001). In vitro bone cell viability was reduced by 37% after 1 hour and by 63% after 4 hours when the bone was left dry ( P < 0.001). Bone cell viability and fusion performance (88%, P < 0.001 vs . dried autograft) were maintained when the graft was stored in saline. CONCLUSIONS: The cellular component of autograft is important for spinal fusion. Adherent graft cells seem to be the more important cellular component in the rabbit model. Autograft left dry on the back table showed a rapid decline in cell viability and fusion but was maintained with storage in saline.
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Fusión Vertebral , Animales , Conejos , Fusión Vertebral/métodos , Autoinjertos , Microtomografía por Rayos X , Columna Vertebral , Trasplante Autólogo , Trasplante Óseo/métodos , Vértebras Lumbares/cirugía , Ilion/cirugíaRESUMEN
Background: In Australia, clinical trial drugs are conventionally dispensed through clinical trial pharmacies only, while community pharmacies dispense drugs approved by Australia's regulatory body. A large HIV pre-exposure prophylaxis study aimed to deliver clinical trial drug through community pharmacies to improve convenience and mimic real world prescribing. This paper describes the process of making community trials compliant with good clinical practice and reports outcomes of delivering clinical trial drug through community pharmacies. Methods: Eight community and four clinical trial pharmacies across three Australian states were approached to participate. A good clinical practice checklist was generated and pharmacies underwent a number of changes to meet clinical trial pharmacy requirements prior to study opening. Changes were made to community pharmacies to make them compliant with good clinical trial practice including; staff training, structural changes, and implementing monitoring of study drug and prescribing practices. Study drug was ordered through standard clinical trial processes and dispensed from study pharmacies by accredited pharmacists. Throughout the trial, record logs for training, prescriber signature and delegation, temperature, participant, and drug accountability were maintained at each pharmacy. The study team monitored each log and delivered on-site training to correct protocol variations. Results: Each pharmacy that was approached agreed to participate. All community pharmacies achieved good clinical practice compliance prior to dispensing study drug. Over the course of the study, 20,152 dispensations of study drug occurred, 83% of these occurred at community pharmacies. Only 2.0% of dispensations had an error, and errors were predominantly minor. On five occasions a pharmacist who was not accredited dispensed study drug. Conclusions: Community based pharmacies can undergo training and modifications to achieve good clinical practice compliance and dispense clinical trial study drug. Community based pharmacies recorded few variations from study protocol. Community based pharmacies offer a useful alternative to clinical trial pharmacies to increase convenience for study participants and expanded use of these pharmacies should be considered for large clinical trials, including HIV prevention trials.
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PURPOSE: There are a significant number of patients with asymptomatic hormone-resistant prostate cancer who have increasing prostate-specific antigen (PSA) levels but little or no evaluable disease. The immunogenicity and minimal toxicity associated with cell-based vaccine therapy makes this approach attractive for these patients. EXPERIMENTAL DESIGN: We have evaluated a vaccine comprising monthly intradermal injection of three irradiated allogeneic prostate cell lines (8 x 10(6) cells each) over 1 year. The first two doses were supplemented with bacille Calmette-Guérin as vaccine adjuvant. Twenty-eight hormone-resistant prostate cancer patients were enrolled. Patients were assessed clinically and PSA levels were measured monthly. Radiologic scans (X-ray, computed tomography, and bone scan) were taken at baseline and at intervals throughout the treatment period. Comprehensive monthly immunologic monitoring was undertaken including proliferation studies, activation markers, cytokine protein expression, and gene copy number. This longitudinal data was analyzed through predictive modeling using artificial neural network feed-forward/back-propagation algorithms with multilayer perceptron architecture. RESULTS: Eleven of the 26 patients showed statistically significant, prolonged decreases in their PSA velocity (PSAV). None experienced any significant toxicity. Median time to disease progression was 58 weeks, compared with recent studies of other agents and historical control values of around 28 weeks. PSAV-responding patients showed a titratable T(H)1 cytokine release profile in response to restimulation with a vaccine lysate, while nonresponders showed a mixed T(H)1 and T(H)2 response. Furthermore, immunologic profile correlated with PSAV response by artificial neural network analysis. We found predictive power not only in expression of cytokines after maximal stimulation with phorbol 12-myristate 13-acetate, but also the method of analysis (qPCR measurement of IFN-gamma > qPCR measurement tumor necrosis factor-alpha > protein expression of IFN-gamma > protein expression of interleukin 2). CONCLUSIONS: Whole cell allogeneic vaccination in hormone-resistant prostate cancer is nontoxic and improves the natural history of the disease. Longitudinal changes in immunologic function in vaccinated patients may be better interpreted through predictive modeling using tools such as the artificial neural network rather than periodic "snapshot" readouts.
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Vacunas contra el Cáncer/uso terapéutico , Neoplasias de la Próstata/terapia , Anciano , Antígenos CD/análisis , Antineoplásicos Hormonales/administración & dosificación , Antineoplásicos Hormonales/uso terapéutico , Vacunas contra el Cáncer/efectos adversos , Vacunas contra el Cáncer/inmunología , Línea Celular , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Citocinas/genética , Diarrea/etiología , Progresión de la Enfermedad , Resistencia a Antineoplásicos , Citometría de Flujo , Humanos , Inmunoterapia/métodos , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Masculino , Persona de Mediana Edad , Náusea/etiología , Redes Neurales de la Computación , Neoplasias de la Próstata/inmunología , Neoplasias de la Próstata/patología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Linfocitos T/citología , Linfocitos T/efectos de los fármacos , Resultado del Tratamiento , Vómitos/etiologíaRESUMEN
BACKGROUND: CD4+CD28null T cells are present in increased numbers in the peripheral blood of patients with acute coronary syndrome (ACS) compared with patients with chronic stable angina (CSA). The triggers of activation and expansion of these cells to date remain unclear. METHODS AND RESULTS: Twenty-one patients with ACS and 12 CSA patients with angiographically confirmed coronary artery disease and 9 healthy volunteers were investigated. Peripheral blood leukocytes were stimulated with human cytomegalovirus (HCMV), Chlamydia pneumoniae, human heat-shock protein 60 (hHSP60), or oxidized LDL (ox-LDL). CD4+CD28null cells were separated by flow cytometry and assessed for antigen recognition using upregulation of interferon-gamma and perforin mRNA transcription as criteria for activation. CD4+CD28null cells from 12 of 21 patients with ACS reacted with hHSP60. No response was detected to HCMV, C pneumoniae, or ox-LDL. Incubation of the cells with anti-MHC class II and anti-CD4 antibodies but not anti-class I antibodies blocked antigen presentation, confirming recognition of the hHSP60 to be via the MHC class II pathway. Patients with CSA had low numbers of CD4+CD28null cells. These cells were nonreactive to any of the antigens used. Circulating CD4+CD28null cells were present in 5 of the 9 healthy controls. None reacted with hHSP60. CONCLUSIONS: We have shown that hHSP60 is an antigen recognized by CD4+CD28null T cells of ACS patients. Endothelial cells express hHSP60 either constitutively or under stress conditions. Circulating hHSP60-specific CD4+CD28null cells may, along other inflammatory mechanisms, contribute to vascular damage in these patients.
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Angina de Pecho/inmunología , Linfocitos T CD4-Positivos/inmunología , Chaperonina 60/inmunología , Enfermedad Coronaria/inmunología , Subgrupos de Linfocitos T/inmunología , Enfermedad Aguda , Presentación de Antígeno , Antígenos CD28/análisis , Recuento de Linfocito CD4 , Linfocitos T CD4-Positivos/metabolismo , Separación Celular , Chaperonina 60/sangre , Chlamydophila pneumoniae/inmunología , Citomegalovirus/inmunología , Citometría de Flujo , Antígenos HLA-D/inmunología , Humanos , Inflamación/inmunología , Interferón gamma/biosíntesis , Interferón gamma/genética , Lipoproteínas LDL/inmunología , Activación de Linfocitos , Glicoproteínas de Membrana/biosíntesis , Glicoproteínas de Membrana/genética , Infarto del Miocardio/inmunología , Perforina , Proteínas Citotóxicas Formadoras de Poros , ARN Mensajero/biosíntesis , Subgrupos de Linfocitos T/metabolismoRESUMEN
There is clear evidence that certain forms of immunotherapy can be successful against certain cancers. However, it would appear that cancerous cells of various origin are exceptionally adept at subverting the immune response. Consequently, it is probable that the most efficacious therapy will be one in which multiple responses of the immune system are activated. There is currently an embarrassment of riches with regard to multiple vaccine strategies in the clinic, although no single method seems to hold the solution. Here, we draw together several of the humoral- and cellular-activating strategies currently under clinical investigation.
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Inmunoterapia , Neoplasias/terapia , Antígenos de Neoplasias/inmunología , Vacunas contra el Cáncer/inmunología , Humanos , Neoplasias/inmunología , Escape del TumorRESUMEN
Real-time PCR is an extremely powerful technique, however, often its results are open to interpretation since there is no convention for data presentation. This anomaly has arisen because many applications rely on non-standard calibration genes, which themselves often change in value during experimental manipulation. We present a novel method for absolute quantification of cDNA species using a combination of extremely accurate double-stranded DNA quantification and a plasmid reference curve. PicoGreen and reference standards are used to measure the amount of cDNA present in a sample using fluorescence. Real-time PCR products are cloned into plasmids and then used to calibrate unknown samples. This cloning is achieved using the same primers necessary for real-time PCR and thus does not involve a second design stage. Results are expressed as copy number per microgram of oligo-dT primed cDNA and consequently may be compared between both inter and intra-experimentally. We show results from a sample human system in which absolute levels of interferon-gamma, TNF-alpha, interleukin-2 and interleukin-10 are measured. We further compare the copy numbers of these genes with levels of released protein and find remarkable correlation. Although our interest has been cytokine quantification, we believe that this technique is widely applicable to the majority of real-time PCR applications.
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Citocinas/análisis , ADN Complementario/análisis , Perfilación de la Expresión Génica/métodos , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Citocinas/genética , Cartilla de ADN , Colorantes Fluorescentes , Humanos , Separación Inmunomagnética , Linfocitos/fisiología , Compuestos Orgánicos , Plásmidos/genética , Estándares de Referencia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Sensibilidad y EspecificidadRESUMEN
The optimal sterilization method for load bearing allografts remains a clinical concern. Recently, supercritical carbon dioxide (SCCO2) treatments have been shown to be capable of terminally sterilizing a range of bacteria and viruses, while preserving the static mechanical properties of cortical bone. This study evaluated the effect of SCCO2 treatment compared with two doses of gamma irradiation, on clinically relevant dynamic mechanical properties of cortical bone. Quasi-static testing was also performed to compare the impairment of treatment. Whole paired adult rabbit humeri were dissected and randomly assigned into either SCCO2 Control, SCCO2 Additive or gamma irradiation at 10 or 25kGy treatment groups. The bones were treated and mechanically tested in three-point bending, with the lefts acting as controls for the treated rights. Maximum load, energy to failure and stiffness were evaluated from static tests. The number of cycles to failure was determined for fatigue at 6-60% of the ultimate load. This study found that SCCO2 treatment with or without additive did not alter static or dynamic mechanical properties. Gamma irradiation had a deleterious dose dependent effect, with statistically significant (p<0.05) reductions in all static mechanical parameters at 25kGy. This effect was increased in fatigue with statistically significant decreases in both the 10 and 25kGy dose groups. This study highlights the expediency of SCCO2 treatment for load bearing bone allograft processing as terminal sterilization can be achieved while maintaining both the quasi-static and dynamic mechanical properties of the graft.
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Húmero/fisiología , Esterilización , Animales , Fenómenos Biomecánicos , Trasplante Óseo , Dióxido de Carbono/farmacología , Rayos gamma , Húmero/efectos de los fármacos , Húmero/efectos de la radiación , Masculino , Conejos , Soporte de Peso/fisiologíaRESUMEN
We are reporting a case of nonabsorbable suture-induced osteomyelitis in patient who had an open rotator cuff repair with nonabsorbable Ethibond anchor suture. Patient in this case presented with very subtle clinical features of osteomyelitis of the left proximal humerus 15 years after initial rotator cuff repair surgery. Literature had shown that deep infection following rotator cuff repairs, although rare, can be easily missed and can cause severe complications. Absorbable suture had been demonstrated to be more superior, in terms of rate of deep infection, as compared to nonabsorbable suture when used in rotator cuff repair surgery. Both absorbable and nonabsorbable suture had been demonstrated to have similar mechanical properties by several different studies. The case demonstrated that initial presentation of deep infection can be subtle and easily missed by clinicians and leads to further complications.
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Antarctic microbial biodiversity is the result of a balance between evolution, extinction and colonization, and so it is not possible to gain a full understanding of the microbial biodiversity of a location, its biogeography, stability or evolutionary relationships without some understanding of the input of new biodiversity from the aerial environment. In addition, it is important to know whether the microorganisms already present are transient or resident - this is particularly true for the Antarctic environment, as selective pressures for survival in the air are similar to those that make microorganisms suitable for Antarctic colonization. The source of potential airborne colonists is widespread, as they may originate from plant surfaces, animals, water surfaces or soils and even from bacteria replicating within the clouds. On a global scale, transport of air masses from the well-mixed boundary layer to high-altitude sites has frequently been observed, particularly in the warm season, and these air masses contain microorganisms. Indeed, it has become evident that much of the microbial life within remote environments is transported by air currents. In this review, we examine the behaviour of microorganisms in the Antarctic aerial environment and the extent to which these microorganisms might influence Antarctic microbial biodiversity.
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Microbiología del Aire , Biodiversidad , Regiones Antárticas , Bacterias , Frío , HongosRESUMEN
A Gram-negative, non-spore-forming, motile, moderately halophilic, aerobic, rod-shaped bacterium, designated strain FP2.5(T), was isolated from the inland hypersaline lake Fuente de Piedra, a saline-wetland wildfowl reserve located in the province of Málaga in southern Spain. Strain FP2.5(T) was subjected to a polyphasic taxonomic study. It produced colonies with a light-yellow pigment. Strain FP2.5(T) grew at salinities of 3-15 % (w/v) and at temperatures of 20-40 degrees C. The pH range for growth was 5-9. Strain FP2.5(T) was able to utilize various organic acids as sole carbon and energy source. Its major fatty acids were C(16 : 0), C(18 : 1)omega9c and C(16 : 1)omega9c. The DNA G+C content was 58.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain FP2.5(T) appeared to be a member of the genus Marinobacter and clustered closely with the type strains of Marinobacter segnicrescens, Marinobacter bryozoorum and Marinobacter gudaonensis (levels of 16S rRNA gene sequence similarity of 98.1, 97.4 and 97.2 %, respectively). However, DNA-DNA relatedness between the new isolate and the type strains of its closest related Marinobacter species was low; levels of DNA-DNA relatedness between strain FP2.5(T) and M. segnicrescens LMG 23928(T), M. bryozoorum DSM 15401(T) and M. gudaonensis DSM 18066(T) were 36.3, 32.1 and 24.9 %, respectively. On the basis of phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness data, strain FP2.5(T) is considered to represent a novel species of the genus Marinobacter, for which the name Marinobacter lacisalsi sp. nov. is proposed. The type strain is FP2.5(T) (=CECT 7297(T)=LMG 24237(T)).
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Conservación de los Recursos Naturales , Agua Dulce/microbiología , Marinobacter/clasificación , Cloruro de Sodio , Humedales , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/análisis , Ácidos Grasos/análisis , Genes de ARNr , Genotipo , Marinobacter/genética , Marinobacter/aislamiento & purificación , Marinobacter/fisiología , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Fenotipo , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , España , Especificidad de la EspecieRESUMEN
A moderately halophilic, Gram-negative bacterium (strain CG4.1(T)), which was isolated from a solar saltern at Cabo de Gata, a wildlife reserve located in the province of Almería, southern Spain, was subjected to a polyphasic taxonomic study. This organism was an aerobic, motile rod that produced colonies with a yellow pigment. Strain CG4.1(T) grew at salinities of 3-25 % (w/v), at 15-45 degrees C and at pH 5-9. The organism reduced nitrate, hydrolysed starch and had phenylalanine deaminase activity. The major fatty acids were C(18 : 1)omega7c, C(16 : 0) and C(19 : 0) cyclo omega8c. The DNA G+C content was 63.6 mol%. On the basis of phenotypic and phylogenetic data, strain CG4.1(T) appears to be a member of the genus Chromohalobacter and clustered closely with Chromohalobacter species, with 95-96 % similarity between their 16S rRNA gene sequences. However, DNA-DNA relatedness between the isolate and the type strains of Chromohalobacter species was low. Therefore, it is proposed that strain CG4.1(T) represents a novel species, Chromohalobacter salarius sp. nov. The type strain is strain CG4.1(T) (=CECT 5903(T)=LMG 23626(T)).
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Halomonadaceae/clasificación , Halomonadaceae/aislamiento & purificación , Agua de Mar/microbiología , Aerobiosis , Aminoácido Oxidorreductasas/análisis , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Ácidos Grasos/análisis , Genes de ARNr , Halomonadaceae/genética , Halomonadaceae/fisiología , Concentración de Iones de Hidrógeno , Locomoción , Datos de Secuencia Molecular , Nitratos/metabolismo , Hibridación de Ácido Nucleico , Filogenia , Pigmentos Biológicos/biosíntesis , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico , España , Almidón/metabolismo , TemperaturaRESUMEN
A moderately halophilic bacterium, strain CG2.1T, isolated from a solar saltern at Cabo de Gata, a wildlife reserve located in the province of Almería, southern Spain, was subjected to a polyphasic taxonomic study. This organism was an aerobic, motile, Gram-negative rod that produced orange-pigmented colonies. Strain CG2.1T was able to grow at salinities of 3-25 % (w/v) and at temperatures of 15-40 degrees C. The pH range for growth was 5-9. Strain CG2.1T was a heterotroph capable of utilizing various carbohydrates as carbon sources. The organism reduced nitrate and showed phenylalanine deaminase activity. The major fatty acids were C(18 : 1)omega7c, C(16 : 0) and C(19 : 0) cyclo omega8c. The DNA G+C content was 60.9 mol%. On the basis of the phenotypic and phylogenetic data, strain CG2.1T appeared to be a member of the genus Halomonas and clustered closely with Halomonas marisflavi (97.1 % 16S rRNA gene sequence similarity). However, the level of DNA-DNA relatedness between the novel isolate and the most closely related Halomonas species was low. On the basis of these data, strain CG2.1T represents a novel member of the genus Halomonas, for which the name Halomonas indalinina is proposed. The type strain is CG2.1T (=CECT 5902T=LMG 23625T).
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Halomonas/clasificación , Halomonas/aislamiento & purificación , Agua de Mar/microbiología , Aerobiosis , Aminoácido Oxidorreductasas/análisis , Antibacterianos/farmacología , Técnicas de Tipificación Bacteriana , Composición de Base , Metabolismo de los Hidratos de Carbono , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Ácidos Grasos/análisis , Ácidos Grasos/química , Genes de ARNr , Halomonas/citología , Halomonas/fisiología , Concentración de Iones de Hidrógeno , Datos de Secuencia Molecular , Movimiento , Nitratos/metabolismo , Oxidación-Reducción , Filogenia , Pigmentos Biológicos/biosíntesis , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Solución Salina Hipertónica/farmacología , España , Temperatura , Microbiología del AguaRESUMEN
Allogeneic whole tumour cell vaccines are inherently practical compared with autologous vaccines. Cell lines are derived from allogeneic tumour, grown in bulk and then administered as a vaccine to the patient, following irradiation, which not only prevents any replication but also enhances antigen presentation. Protection is believed to occur through the presentation of antigens shared between the syngeneic and allogeneic tumours. Although cytokine-transfected tumour whole cell vaccines have been used clinically, little data is available comparing the effects of immunomodulatory cytokine-transfection directly on the same cells when used as both an allogeneic and autologous vaccine. To address this, weakly immunogenic B16-F10 (H-2b) murine melanoma was transfected to secrete either GM-CSF, IL-4 or IL-7. Prophylactic vaccination of both syngeneic C57/BL6 (H-2b) (B6) and allogeneic C3H/Hej (H-2k) (C3H) mice showed the effects of transfected cytokine varied between models. Both GM-CSF and IL-7 significantly (P<0.05) increased the levels of protection within syngeneic B6 mice, but had a diminished effect (P>0.05) within C3H allogeneic mice. Allogeneic B16-F10 cells and syngeneic K1735 cells generated CTL against K1735 suggesting cross-reactive immunity. Using cells labeled with fluorescent dye we demonstrate that irradiated vaccines, of either syngeneic or allogeneic origin, appear to generate potent immune responses and fragments of either vaccine remain at the injection site for up to 9 days. This study shows that protection can be enhanced in vivo by using transfected cytokine, but suggests that irradiated whole cell vaccines, of either tissue-type, are rapidly processed. This leads to the conclusion that the cytokine effects are transient and thus transfection with cytokine may be of limited long-term use in situ.
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Vacunas contra el Cáncer/efectos de la radiación , Citocinas/metabolismo , Melanoma Experimental/prevención & control , Trasplante Homólogo/inmunología , Trasplante Isogénico/inmunología , Animales , Trasplante de Células , Citotoxicidad Inmunológica , Femenino , Citometría de Flujo , Ratones , Trasplante de Neoplasias , Transfección , Células Tumorales CultivadasRESUMEN
We have applied bioluminescent ATP detection methods to microbial enumeration in Antarctic Dry Valley mineral soils, and validated our ATP data by two independent methods. We have demonstrated that ATP measurement is a valid means of determining microbial biomass in such sites, and that the desiccated surface mineral soils of the Antarctic Dry Valleys contain cell numbers over four orders of magnitude higher than previously suggested.