IGRP and insulin vaccination induce CD8+ T cell-mediated autoimmune diabetes in the RIP-CD80GP mouse.

Autoimmune diabetes is characterized by autoantigen-specific T cell-mediated destruction of pancreatic islet beta cells, and CD8(+) T cells are key players during this process. We assessed whether the bitransgenic RIP-CD80 x RIP-LCMV-GP (RIP-CD80GP) mice may be a versatile antigen-specific model of inducible CD8(+) T cell-mediated autoimmune diabetes. Antigen-encoding DNA, peptide-loaded dendritic cells and antigen plus incomplete Freund's adjuvant were used for vaccination. Of 14 pancreatic proteins tested by DNA vaccination, murine pre-proinsulin 2 (100% of mice; median time after vaccination, 60 days) and islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP) (77%, 58 days) could induce diabetes. Vaccination with DNA encoding for zinc transporter 8, Ia-2, Ia-2ß, glutamic acid decarboxylase 67 (Gad67), chromogranin A, insulinoma amyloid polypeptide and homeobox protein Nkx-2.2 induced diabetes development in 25-33% of mice. Vaccination with DNA encoding for Gad65, secretogranin 5, pancreas/duodenum homeobox protein 1 (Pdx1), carboxyl ester lipase, glucagon and control hepatitis B surface antigen (HBsAg) induced diabetes in <20% of mice. Diabetes induction efficiency could be increased by DNA vaccination with a vector encoding a ubiquitin-antigen fusion construct. Diabetic mice had florid T cell islet infiltration. CD8(+) T cell targets of IGRP were identified with a peptide library-based enzyme-linked immunospot assay, and diabetes could also be induced by vaccination with major histocompatibility complex (MHC) class I-restricted IGRP peptides loaded on mature dendritic cells. Vaccination with antigen plus incomplete Freund's adjuvant, which can prevent diabetes in other models, led to rapid diabetes development in the RIP-CD80GP mouse. We conclude that RIP-CD80GP mice are a versatile model of antigen specific autoimmune diabetes and may complement existing mouse models of autoimmune diabetes for evaluating CD8(+) T cell-targeted prevention strategies.

An analytical strategy for challenging members of the microplastic family: Particles from anti-corrosion coatings.

Potentially hazardous particles from paints and functional coatings are an overlooked fraction of microplastic (MP) pollution since their accurate identification and quantification in environmental samples remains difficult. We have applied the most relevant techniques from the field of microplastic analysis for their suitability to chemically characterize anti-corrosion coatings containing a variety of polymer binders (LDIR, Raman and FTIR spectroscopy, Py-GC/MS) and inorganic additives (ICP-MS/MS). We present the basis of a possible toolbox to study the release and fate of coating particles in the (marine) environment. Our results indicate that, due to material properties, spectroscopic methods alone appear to be unsuitable for quantification of coating/paint particles and underestimate their environmental abundance. ICP-MS/MS and an optimized Py-GC/MS approach in combination with multivariate statistics enables a straightforward comparison of the multi-elemental and organic additive fingerprints of paint particles. The approach can improve the identification of unknown particles in environmental samples by an assignment to different typically used coating types. In future, this approach may facilitate allocation of emission sources of different environmental paint/coating particles. Indeed, future work will be required to tackle various remaining analytical challenges, such as optimized particle extraction/separation of environmental coating particles.

Evaluation of continuous flow centrifugation as an alternative technique to sample microplastic from water bodies.

The scientific and public interest regarding environmental pollution with microplastic has considerably increased within the last 15 years. Nevertheless, up to now there is no widely applied standard operation procedure for microplastic sampling, resulting in a lack of inter-study comparability. In addition, many studies on microplastic occurrences do not indicate a sound methodological validation of the applied methods and procedures. This study presents an alternative volume-reduced sampling technique to sample the entire load of suspended particulate matter including microplastic particles in natural waters, based on continuous flow centrifugation. For the lab-scale validation of the proposed instrumental setup, six different microplastic types (PE, PET, PS, PVDC, EPS and PP) were used. The particles covered a size range from 1 µm to 1 mm and a density range from 0.94 g mL-1 to 1.63 g mL-1. Recoveries ranged from 95.0% ±â€¯2.3% - 99.1% ±â€¯0.3% for virgin powders and from 96.1% ±â€¯0.6% - 99.4% ±â€¯0.2% (1 SD, n = 2 - 3) for microplastic suspended in river water for 40 days. Gravimetric and microscopic analysis of the effluent indicates efficient removal of microplastic from the suspensions. Static light scattering analysis of the microplastic suspensions prior to and after centrifugation confirmed that no change of the particle size distribution has occurred - neither through aggregation nor through size-discrimination during centrifugation. Moreover, the system was tested in the field and used twice to sample suspended particulate matter from the Elbe estuary directly on site. Based on these first lab-scale experiments, continuous flow centrifugation proves a promising technique bearing potential to alleviate drawbacks such as contamination, filter clogging and particle size-discrimination of commonly used volume-reduced microplastic sampling approaches.

Glucocorticoid receptors and in vitro responses to glucocorticoid in acute nonlymphocytic leukemia.

Early clinical studies in which glucocorticoids were used alone in the treatment of acute nonlymphocytic leukemia (ANLL) reported a wide range of responses from remission in some patients to dramatic aggravation of the disease in others. In the hopes of identifying those patients likely to derive therapeutic benefit from glucocorticoids, we have studied glucocorticoid receptors and in vitro responses to glucocorticoids in 36 previously untreated adults with ANLL. The leukemic blasts of all patients contained glucocorticoid receptors (range, 4,300 to 28,400 total receptor sites per cell; median, 8,800). These receptors were similar in all respects studied to those from a variety of other normal and malignant tissues. There was little difference between receptor levels among the various French-American-British categories. In vitro responses to glucocorticoid were observed in leukemic blasts of 26 of 28 cases studied. These responses varied from near complete cell killing to stimulation of proliferation. Since the cells of patients with ANLL have about the same number of receptors as do cells from patients with acute lymphoblastic leukemia and these receptors are capable of mediating physiological responses in vitro, it is unlikely that qualitative or quantitative receptor defects underlie the relative resistance to glucocorticoid therapy of ALL compared to acute lymphoblastic leukemia. However, the broad range of in vitro responses and receptor levels suggests that these studies might be useful in identifying those patients with ANLL likely to derive benefit from steroid therapy.

Antioxidants vs carotenoids. Inhibitors or promoters of experimental colorectal cancers.

The organospecific, 1,2-dimethylhydrazine-induced murine tumor model was used to test the effects on tumor formation of the following dietary supplements: (1) ascorbic acid, 7% per weight; (2) alpha tocopherol, 1% per weight; (3) beta carotene, 1% per weight; and (4) canthazanthin, 1% per weight. Following a four-week dietary acclimation, a 16-week 1,2-dimethylhydrazine induction, and a four-week hiatus, the animals were killed, underwent autopsies, and tumor formation was recorded. The antioxidant supplements of ascorbic acid and alpha tocopherol resulted in a significant decrease in tumor formation when compared with control groups. In contrast, the beta carotene group showed no difference in tumor formation, and canthazanthin, a non-provitamin A carotenoid, resulted in an increase in tumor formation when compared with controls. In addition, the K-gel powder control diet (a carrier medium for alpha tocopherol acetate) had a significantly higher rate of tumor formation than the regular chow and placebo beadlet control diets. In sum, ascorbic acid and alpha tocopherol demonstrated a clear chemopreventive effect, whereas beta carotene had no effect, and canthazanthin appeared to function as a promoter in this organospecific tumor model.

Perioperative immunomodulation in cancer surgery.

Despite undergoing a curative resection, many patients with colorectal cancer will develop and die of metastatic disease. It has been shown clinically and experimentally that surgery causes a transient period of immunosuppression, and it is postulated that this may encourage both the implantation of surgically disseminated tumor cells and the growth of existing micrometastases. The present study used natural killer cell cytotoxicity (NKCC) and tumor burden to evaluate perioperative modulation of immunocompetence in a murine model. We measured NKCC and tumor burden responses to a standardized surgical stress (SSS) alone, and to either morphine sulfate (MS) (15 mg/kg subcutaneously x 4 doses), ketorolac (a prostaglandin synthetase--prostaglandin E2--inhibitor) (2.5 mg/kg subcutaneously x 4 doses), or interleukin 2 (2,000 units intraperitoneally x 3 doses) administration with the SSS. In this model, we found that both low-dose interleukin-2 (IL-2) and ketorolac reversed the NKCC suppression associated with surgery, whereas morphine resulted in further depression of NKCC. In addition, IL-2 significantly decreased tumor incidence, whereas continuous MS exposure markedly increased tumor burden after surgery. These data suggest that IL-2 and ketorolac may be effective agents for the restoration of perioperative immune competence, whereas the use of continuous morphine might have significant deleterious effects.

[The integration of psychiatry services in the general hospital]. / A inserção da psiquiatria no hospital geral.

This work consist of a bibliographic review about the inclusion of the psychiatry in general hospitals. Through it we were able to approach some historical aspects concerning the presence of mentally ill people in general hospitals, the creation and organization of psychiatric units in general hospitals, the hospitalization of psychiatric patients in institutions lacking specific units and the services of psychiatric interconsultation. The work consists of a part of a Master's dissertation in which we observed that there is a great amount of literature about the subject; however, all of them under the view of the nursing team. No references were found about the viewpoint of the mentally ill person about the hospitalization in a general hospital.

[Metastatic malignant melanoma with spinal cord compression and resulting recumbency ante partum in a mare. A case report]. / Metastasierendes malignes Melanom mit Festliegen ante partum infolge Rückenmarkskompression bei einer Stute. Ein Fallbericht.

The case report describes a narrowing of the spinal canal in the lumbar and sacroiliac regions with entailing acute recumbency in a nearterm 17-year-old Welsh B Pony mare. The reason for recumbency was metastases of a malignant melanoma in the spinal canal of the lumbo-sacral region, where the tumour had invaded the dura mater and applied severe pression on the spinal cord over a length of 10 cm. Clinical findings, laboratory diagnostics, pathological-anatomical and histopathological findings are presented.

Size matters more than chemistry for cloud-nucleating ability of aerosol particles.

Size-resolved cloud condensation nuclei (CCN) spectra measured for various aerosol types at a non-urban site in Germany showed that CCN concentrations are mainly determined by the aerosol number size distribution. Distinct variations of CCN activation with particle chemical composition were observed but played a secondary role. When the temporal variation of chemical effects on CCN activation is neglected, variation in the size distribution alone explains 84 to 96% of the variation in CCN concentrations. Understanding that particles' ability to act as CCN is largely controlled by aerosol size rather than composition greatly facilitates the treatment of aerosol effects on cloud physics in regional and global models.

Starch bioavailability in the upper gastrointestinal tract of colectomized rats.

Colectomized rats were used to compare the bioavailability of starch from canned peas, kidney beans, lima beans, corn, cooked rice and AIN-76A purified diet. Postoperative weight gains of colectomized and sham-operated rats were not different. Test meals of approximately 2 and approximately 3.3 g (dry weight) of each food containing chromic oxide (0.6 g/100 g) were administered to 4-6 rats by gastric intubation. Chromium recovery in ileal digesta 10 h after the meal ranged from 94 to 102%. Significant starch (11-15%) from peas, lima beans or kidney beans was recovered in the ileal digesta; 0.2-0.4% of starch from rice, corn or AIN-76A was in the ileal digesta. Pretreatment of kidney or lima beans with a heat-stable endo-alpha-amylase decreased the starch recovered in the ileal digesta to 2.7-4.3% of that fed; pretreatment with the amylase and pepsin had no further effect. Oligosaccharide extraction, the size of the test meal and the amount of starch did not affect starch bioavailability.

Geraniol, an inhibitor of mevalonate biosynthesis, suppresses the growth of hepatomas and melanomas transplanted to rats and mice.

Farnesyl-pyrophosphate is required for the posttranslational modification of G proteins including p21 ras, prelamin A and lamin B, each of which plays an essential role in cell proliferation. As a consequence, competitive inhibitors of mevalonate synthesis, the rate-limiting substrate for the synthesis of the prenyl-pyrophosphates, arrest cultured cells at the G1/S interface of the cell cycle and initiate apoptotic cell death. Geraniol, an acyclic monoterpenoid alcohol, suppresses 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity and concomitantly arrests the growth of cultured tumor cells. We evaluated the impact of dietary geraniol on the growth of two tumors. In the first study, geraniol (23 mmol/kg diet, 350 mumol/d) was fed to male buffalo rats for 14 d before and for 42 d after the transplant of Morris 7777 hepatomas. Tumor growth was suppressed (P < 0.001). In the second study, the dose-dependent impact of geraniol on the growth of B16 melanomas was assessed. Dietary geraniol (0.65, 6.5 and 65 mmol/kg diet) was fed to female C57BL mice for 14 d before and for 21 d after tumor transplant. Tumor growth was suppressed (P < 0.02) by 6.5 and 65 mmol geraniol/kg diet.

Utilization and preferred metabolic pathway of ketone bodies for lipid synthesis by isolated rat hepatoma cells.

Morris hepatoma 7777 cells freshly isolated from highly malignant tumors grown in the hindlimb of buffalo rats actively convert ketone bodies to cholesterol and fatty acids. On the basis of results obtained with (-)-hydroxycitrate, an inhibitor of the ATP citrate lyase enzyme, the metabolic pathway for acetoacetate conversion to lipids is exclusively cytoplasmic, whereas that for 3-hydroxybutyrate involves both extra- and intramitochondrial compartments. Subcellular distribution studies indicated accumulation and compartmentation of 3-hydroxybutyryl CoA primarily in the cytoplasm of hepatoma cells incubated with either ketone body. In contrast, the compartmentation of acetoacetyl CoA is dependent on whether the substrate is acetoacetate or 3-hydroxybutyrate. With acetoacetate, the acetoacetyl CoA is entirely cytoplasmic, whereas with 3-hydroxybutyrate, it is equally divided between the intra- and extramitochondrial compartments. The results are discussed in terms of the known and proposed metabolic pathways for lipid synthesis from ketone bodies, particularly that from 3-hydroxybutyrate.

[The significance of the columar structure of the stratum corneum for the effect of light on the skin]. / Uber die Bedeutung der Columnärstruktur des Stratum corneum für die Lichtwirkung auf die Haut

Autologous full thickness skin autografts were carried out on 60 male guineapigs in such a way that the hair roots were destroyed. The light sensitivity of the skin towards UV B was then tested on the hairless skin by means of the Xenon lamp and the corresponding filter. Exploratory excisions were taken from another area of the graft and used to demonstrate the structure of the stratum corneum according to the method of Christophers. It could be shown that light erythema developed significantly less than when the columnar structure of the stratum cornuem was well developed.

Acyl CoA and lipid synthesis from ketone bodies by the extramitochondrial fraction of hepatoma tissue.

The compartmentation of the pathway for the synthesis of lipids from ketone bodies was examined in a cell free, post-mitochondrial supernatant fraction of malignant Morris hepatoma 7777 tissue. A fortified supernatant system effectively incorporated radioactive D(-)-3-hydroxybutyrate and acetoacetate into cholesterol and fatty acids, and both ketone bodies were directly converted to their CoA thioesters. Furthermore, a microsome-free (100,000 x g) cytosolic fraction was also able to acylate 3-hydroxybutyrate to 3-hydroxybutyryl CoA. No previous identification of this enzyme activity has been described. These results which characterize a distinct extramitochondrial pathway for conversion of 3-hydroxybutyrate as well as acetoacetate into lipids also suggest the possibility of a previously undetected enzymatic activity for utilization of this ketone body.

Trauma and inflammation modulate lymphocyte localization in vivo: quantitation of tissue entry and retention using indium-111-labeled lymphocytes.

OBJECTIVE: Determine the in vivo localization pattern of indium-111-labeled lymphocytes after a standardized extremity injury or standardized laparotomy and after sterile inflammation of the central nervous system. DESIGN: Prospective animal study with concurrent controls. SETTING: Animal research laboratory. SUBJECTS: Male Lewis rats weighing 150-175 g. INTERVENTIONS: Indium-111-labeled splenic lymphocytes were injected into animals after a standardized hind limb trauma or laparotomy and after induction of sterile central nervous system inflammation. MEASUREMENTS AND MAIN RESULTS: Lymphoid and non-lymphoid organs were removed at fixed intervals after lymphocyte injection and the proportion of injected lymphocytes/gram of tissue was determined using a quantitative radionuclide calculation. Results from treated animals were compared with results from untreated control animals. Muscle injury caused early localization of lymphocytes to injured hind limbs, liver, and spleen compared with controls, whereas laparotomy decreased lymphocyte localization in the thymus and colon. Encephalitis increased localization to the central nervous system with no effect on other tissues. CONCLUSIONS: These results identify a sensitive method to track in vivo leukocyte localization and specifically demonstrate that lymphocyte localization is altered in both traumatic and nontraumatic models of inflammation.

Evaluation of the BOD POD for estimating percentage body fat in a heterogeneous group of adult humans.

The primary purpose of this investigation was to compare estimations of percentage body fat (%fat) using air displacement plethysmography (ADP) and hydrostatic weighing (HW) in a heterogeneous (age and %fat) sample of the population. Of secondary importance was to determine whether there were differences between the two methods among lean (n = 32), average (n = 34) and overweight (n = 29) subsets of this sample. A total of 95 adults (men 27, women 68) ranging in age from 18-52 years volunteered for this study. Test-retest reliability for %fat ADP (n = 16) was 0.99 with a technical error of 0.75%fat and a coefficient of variation of 3.4%fat. Mean body density using ADP [1.048 (SD 0.016) g.ml-1] was not significantly different when compared to HW [1.049 (SD 0.017) g.ml-1], which corresponded to a non-significant difference in %fat [22.5 (SD 7.3)% ADP compared to 22.0 (SD 7.6)% HW]. Regression analysis provided the equation: %fat HW = 0.9121%fat ADP + 1.5123; r = 0.88, SEE = 3.6, which did not differ significantly from the line of identity. Data for the subsets revealed a significant overestimation of %fat ADP [16.4 (SD 4.8)%] compared to HW [14.1 (SD 3.2)%] (P = 0.001) for lean individuals while no difference was found in the average [21.9 (SD 4.4)%fat ADP compared to 22.0 (SD 3.4)%fat HW] or overweight [29.9 (SD 5.5)%fat ADP compared to 30.8 (SD 4.1)%fat HW] subsets. Measuring %fat by ADP is a highly reliable method and valid when compared to HW for a heterogeneous sample of adults. The ADP method requires little expertise to operate, is quick to perform, and may be more accommodating for certain individuals compared to HW. However, in this study ADP was less valid for lean individuals. Further investigation is warranted to determine the bias of this method for subsets of the population which may be outside the average range of %fat (men 15.4%-22.0%, women 18.4%-28.5%).

The in vivo effects of general and epidural anesthesia on human immune function.

UNLABELLED: Impaired in vivo immunity is often observed after major surgery and is multifactorial. We conducted a randomized clinical study to determine the independent effects of general anesthesia (GA) and of lumbar epidural anesthesia (LEA) on human immune function in the absence of surgical trauma. Nineteen healthy volunteers were randomized to receive GA with thiopental and isoflurane, LEA with lidocaine, or no anesthesia (Control). Serial blood samples were tested for antibody responses to antigen inoculation, neutrophil and mononuclear cell antibody-dependent cell cytotoxicity (ADCC), natural killer cell cytotoxicity, and neutrophil phagocytic activity. Antibody responses were similar in the three groups. Mononuclear cell ADCC increased in the LEA group at the end of the anesthetic (P < 0.05 at effector/target [E/T] ratios of 10:1, 25:1, and 50:1). Natural killer cell cytotoxicity increased at the end of the anesthetic in both the LEA group (P < 0.05 at all E/T ratios) and the GA group (P < 0.05 at an E/T ratio of 5:1 and 10:1). No significant changes were observed for neutrophil ADCC or phagocytosis. General or epidural anesthesia alone, in the absence of surgery, seems to have only transient and minor effects on human immune function. IMPLICATIONS: General or epidural anesthesia alone, in the absence of surgery, seems to have only transient and minor effects on human immune function.

Morphine inhibits spontaneous and cytokine-enhanced natural killer cell cytotoxicity in volunteers.

BACKGROUND: Opioids are used by patients who have conditions ranging from the acute pain of surgery and chronic cancer pain to substance abuse. Despite their widespread use and considerable experimental data about them, little is known about how opioids may alter in vivo immunity in humans. This study was designed to evaluate the in vivo effect of morphine on human peripheral blood immune functions. METHODS: Healthy volunteers underwent continuous exposure to morphine for 36 h including a 24-h intravenous infusion in the hospital. Peripheral blood was drawn for immune function studies at five measurement times before, during, and after morphine exposure. Peripheral blood mononuclear cells were tested for acute and gamma-interferon-stimulated natural killer cell cytotoxicity (NKCC), antibody-dependent cell cytotoxicity, antibody Fc receptor expression, and human immunodeficiency virus infectivity. RESULTS: Significant suppression of NKCC was observed at 2 and 24 h after the onset of intravenous morphine exposure. Suppression of NKCC persisted for 24 h after termination of morphine infusion in a "high"-dose study group. gamma-Interferon-stimulated NKCC and antibody-dependent cell cytotoxicity were also decreased after 24 h of intravenous morphine exposure. No effect on Fc receptor expression was observed. Mean virus antigen production after lymphocyte infection with human immunodeficiency virus was not increased (p24 100 ng/ml after morphine vs. 43 ng/ml before morphine; P = 0.17). CONCLUSIONS: These results suggest that morphine administration, at doses within the range of analgesic use, can cause measurable suppression of some components of the human cellular immune system.

In-vitro glucocorticoid studies for predicting response to glucocorticoid therapy in adults with malignant lymphoma.

Neoplastic tissues from 28 adults with malignant lymphoma were examined for glucocorticoid receptors and in-vitro sensitivity to glucocorticoids. The patients were then treated with desamethasone for 5--14 days. 13 patients achieved at least a partial remission, and 15 had no significant tumour response. Lymphoma cells from patients who responded had more glucocorticoid-receptor sites per cell and greater in-vitro sensitivity as measured by glucocorticoid inhibition of incorporation of leucine and uridine than did tumour cells from non-responders. Study of tumour glucocorticoid receptors and glucocorticoid sensitivity in vitro may allow selection of those patients with lymphoma who should receive glucocorticoids as part of combination chemotherapy.