Effects of cocaine and levamisole (as adulterant) on the isolated perfused Langendorff heart.

Cocaine-related deaths occur regularly in forensic routine work. In cases in which the detected concentration of cocaine is rather low and other causes of death apart from intoxication can be ruled out, the question arises if adulterants of cocaine might have played a crucial role. In the present study, cardiac effects of cocaine, of the adulterant levamisole and of mixtures of both were evaluated using the isolated perfused Langendorff heart. While exposed to the substances, functional parameters heart rate, left ventricular pressure and coronary flow were documented. Relevant alterations of these parameters were found for cocaine as well as for levamisole. Exposing the hearts to a mixture of both resulted in a combination of these effects; the emergence of new alterations or an obvious aggravation were not detected. Nevertheless, the results imply that the consumption of cocaine adulterated with levamisole bares an increased risk for cardiac complications, especially in the presence of preexisting cardiac pathologies.

Thymic stromal lymphopoietin deficiency attenuates experimental autoimmune encephalomyelitis.

In the present study we examined the role of thymic stromal lymphopoietin (TSLP) in experimental autoimmune encephalomyelitis (EAE). Here, we report that TSLP knock-out (KO) mice display a delayed onset of disease and an attenuated form of EAE. This delayed onset was accompanied by a reduced number of encephalitogenic T helper type 1 (Th1) cells in the central nervous system (CNS) of TSLP KO mice. In addition, CD4(+) and CD8(+) T cells from CNS of TSLP KO mice show a reduced activation status in comparison to wild-type mice. It is noteworthy that we could also show that lymph node cells from TSLP KO mice expanded less efficiently and that interleukin (IL)-6-, interferon (IFN)-γ and tumour necrosis factor (TNF)-α levels were reduced. Furthermore, CD3(+) T cells isolated in the preclinical phase from myelin oligodendrocyte glycoprotein peptide 35-55 (MOG(35-55))-immunized TSLP KO mice showed a reduced response after secondary exposure to MOG(35-55), indicating that differentiation of naive T cells into MOG(35-55)-specific effector and memory T cells was impaired in KO mice. The addition of recombinant TSLP enhanced T cell proliferation during MOG(35-55) restimulation, showing that T cells also respond directly to TSLP. In summary, these data demonstrate that expression of, and immune activation by, TSLP contributes significantly to the immunopathology of EAE.

Efficacy of a Brazilian calcium montmorillonite against toxic effects of dietary aflatoxins on broilers reared to market weight.

1. The protective effect of a natural Brazilian calcium montmorillonite (CaMont) against aflatoxins was studied in broiler chickens. 2. A total of 1056-d-old Cobb male broilers were housed in experimental pens (22 chickens per pen) for 42 d. Three levels of CaMont (0, 2.5 and 5 g/kg) and two levels of aflatoxins (0 and 3 mg/kg) were assayed. Each treatment had 8 replicate pens of 22 broiler chickens each. 3. Of all the chickens tested in the experiment, the ones treated with aflatoxins were the most adversely affected. CaMont treatment at concentrations of 2.5 and 5 g/kg improved body weight of chickens at 42 d of age by 13.3% and 22.7%, increased daily feed intake by 9.7% and 24.7%, and improved the productive efficiency index of chickens by 53% and 66.5%, respectively. 4. Dietary CaMont positively affected parameters such as weight of liver, heart and gizzard; however, serum potassium concentration decreased by 15.3% compared with that of chickens given only the aflatoxin-contaminated diet. 5. CaMont did not cause adverse effects in chickens that did not receive aflatoxins. 6. CaMont at pH 8.5 partially reduced the toxic effects of aflatoxins in broilers when included at levels of 2.5 and 5 g/kg in the diet.

Soluble CD83 ameliorates experimental colitis in mice.

The physiological balance between pro- and anti-inflammatory processes is dysregulated in inflammatory bowel diseases (IBD) as in Crohn's disease and ulcerative colitis. Conventional therapy uses anti-inflammatory and immunosuppressive corticosteroids to treat acute-phase symptoms. However, low remission rate and strong side effects of these therapies are not satisfying. Thus, there is a high medical need for new therapeutic strategies. Soluble CD83, the extracellular domain of the transmembrane CD83 molecule, has been reported to have interesting therapeutic and immunosuppressive properties by suppressing dendritic cell (DC)-mediated T-cell activation and inducing tolerogenic DCs. However, the expression and function of CD83 in IBD is still unknown. Here, we show that CD83 expression is upregulated by different leukocyte populations in a chemical-induced murine colitis model. Furthermore, in this study the potential of sCD83 to modulate colitis using an experimental murine colitis model was investigated. Strikingly, sCD83 ameliorated the clinical disease symptoms, drastically reduced mortality, and strongly decreased inflammatory cytokine expression in mesenteric lymph nodes and colon. The infiltration of macrophages and granulocytes into colonic tissues was vigorously inhibited. Mechanistically, we could show that sCD83-induced expression of indolamine 2,3-dioxygenase is essential for its protective effects.

c-Myc and EBV-LMP1: two opposing regulators of the HLA class I antigen presentation machinery in epithelial cells.

BACKGROUND: Epstein-Barr virus (EBV)-encoded latent membrane protein 1 (LMP1) up-regulates the human leukocyte antigen (HLA) class I antigen presentation machinery (APM). This appears counterintuitive with immune evasion in EBV-associated tumours like nasopharyngeal carcinoma (NPC). METHODS: Latent membrane protein 1-transfected epithelial cell lines were used as a model system to study the impact of LMP1 and c-Myc on HLA class I components. The expression of components of the HLA class I APM, c-Myc and Ki-67 was analysed in LMP1+ and LMP1- NPC by immunohistochemistry. RESULTS: In epithelial cells, LMP1 up-regulated HLA class I APM. This effect could be counteracted by c-Myc, which itself was up-regulated by LMP1 apparently through IL6 induction and Jak3/STAT3 activation. Studies of NPC biopsies revealed down-regulation of HLA class I APM expression. No difference was observed between LMP1+ and LMP1- NPC. However, expression of Ki-67 and c-Myc were up-regulated in LMP1+ tumours. CONCLUSION: These findings raise the possibility that c-Myc activation in NPC might antagonise the effect of LMP1 on HLA class I expression thus contributing to immune escape of tumour cells.

Twelve novel mutations in the tissue-nonspecific alkaline phosphatase gene (ALPL) in patients with various forms of hypophosphatasia.

Hypophosphatasia is a rare inherited disorder characterized by defective bone mineralization and deficiency of serum and tissue liver/bone/kidney tissue alkaline phosphatase (L/B/K ALP) activity. We report here the characterization of tissue-nonspecific alkaline phosphatase (TNSALP) gene mutations in a series of 11 families affected by various forms of hypophosphatasia. Nineteen distinct mutations were found, 7 of which were previously reported. Eleven of the 12 new mutations were missense mutations (Y11C, A34V, R54H, R135H, N194D, G203V, E218G, D277Y, F310G, A382S, V406A), the last one (998-1G>T) was a mutation affecting acceptor splice site.

Satellite videoconferencing for healthcare workers: audience characteristics and the importance of continuing education credits.

To assess the opinions of healthcare workers (HCWs) about a satellite videoconference as a means of earning continuing education credit, a telephone survey was conducted in September 1998, 1 month after a live interactive satellite videoconference on antimicrobial use and resistance. There were 180 registered sites in 45 states surveyed, representing 1,589 viewers: 764 nurses (48.1%), 201 physicians (12.6%), and 624 other HCWs (39.3%). Continuing education credit was requested by 51% of nurses, 31% of physicians, and 27% of all other HCWs. Although preferred learning formats varied, 70% of respondents said it was important to offer continuing education credit. Furthermore, 31% of the respondents stated that the videoconference influenced institutional strategies. We concluded that satellite videoconferences are a method to reach audiences around the world efficiently and effectively, provide the latest information, facilitate interaction, and meet some of the demand for continuing education credit for HCWs.

Treatment adherence and patient retention in the first year of a Phase-III clinical trial for the treatment of multiple sclerosis.

The purpose of this study was to examine the relationship between patient management strategies employed by study personnel, and patient retention and adherence to treatment in the first year of a Phase III clinical trial of interferon beta-1b for treatment of secondary progressive multiple sclerosis (MS). Study staff from each of 35 sites were interviewed regarding patient management practices. Sites which were rated as more empathetic, as instilling a sense of purpose in the patient, and promoting less formal relationships with patients had high rates of adherence to treatment. In addressing specific patient concerns, attention to patients' emotional status and patients' expectations of trial participation were related to better adherence.

Concurrent phase I trials of intravenous interleukin 6 in solid tumor patients: reversible dose-limiting neurological toxicity.

Interleukin 6 (IL-6) has antitumor activity comparable to IL-2 in murine models with less toxicity. Because the biological effects of intermittent and continuous infusions may differ, we conducted two concurrent Phase I trials of daily x5, 1-h, and continuous 120-h i.v. infusions to determine the toxicity, biological effects, and maximum tolerated dose of i.v. IL-6. Cohorts of six patients with advanced cancer received escalating doses (1, 3, 10, 30, 100, and 150 microgram/kg/day) of recombinant human IL-6 on days 1-5 and 8-12 of each 28-day course (1-h trial) or on days 1-5 of each 21-day course (120-h trial). Treatment was administered in regular inpatient wards and in outpatient clinics and was withheld in the event of grade 3 toxicity. Sixty-nine patients (1-h trial, n = 40; 120-h trial, n = 29) were enrolled, including 27 with renal cancer and 16 with melanoma. All were ambulatory, and 40 were asymptomatic. Fever (97%), anemia (78%), fatigue (56%), nausea or vomiting (49%), and elevated serum transaminase levels (42%) were the most frequent toxicities. Transient hypotension developed in 23 patients (33%). There were three deaths during the study due to progressive disease and/or infection. There were no objective responses. Dose-related increases in platelet counts and C-reactive protein levels were detected in most patients. Principal dose-limiting toxicities included atrial fibrillation (1 episode in the 1-h trial and 4 episodes in the 120-h trial) and neurological toxicities (3 episodes in the 1-h trial and 4 episodes in the 120-h trial). The neurological toxicities included confusion, slurred speech, blurred vision, proximal leg weakness, paraparesis, and ataxia. These effects were transient and reversed when IL-6 was discontinued. IL-6 can be given by i.v. infusion at biologically active doses with acceptable toxicity. Dose-limiting toxicities consisted mainly of a spectrum of severe but transient neurological toxicities and occasional episodes of atrial fibrillation. The maximum tolerated doses recommended for use with these i.v. schedules in Phase II trials are 100 microgram/kg/day by daily x5 1-h infusion and 30 microgram/kg/day by 120-h infusion. Phase II trials will be performed to determine the antitumor activity of IL-6 and better define its toxicity. Patients in these and other IL-6 studies should be monitored closely for neurological and cardiac effects.

Short-term effects of alcohol consumption on appetite and energy intake.

The relationship between alcohol intake and obesity remains uncertain. Evidence suggesting that alcohol-derived energy may be unregulated points to an inability to maintain appetite, energy balance and, hence, body weight when alcohol is introduced to the diet. This study investigated the short-term effects of alcohol on hunger and energy intake in 20 lean women. On 4 occasions, subjects were given a randomised preload drink ('alcohol', 'no alcohol', 'carbohydrate', 'water') followed by visual analogue scales (VAS) rating hunger and an ad lib test meal. There was no difference in hunger ratings (p > 0.05) nor in the amount of energy consumed during the test meal (F = 1.66, p > 0.05) following any of the 4 preloads. Consumption of the 2 high energy preload drinks ('alcohol', 0.91 MJ; 'CHO', 0.72 MJ) did not result in a compensatory decrease in the amount of energy subsequently eaten (ad lib intake: 'alcohol' = 2.62 MJ, 0.32 SEM; 'no alcohol' = 2.98 MJ, 0.28 SEM; 'CHO' = 2.93 MJ, 0.21 SEM; 'water' = 2.82 MJ, 0.25 SEM), suggesting either no physiological recognition or no regulation of energy consumed within a drink in quantities of less than 1 MJ. The addition of either alcoholic or CHO-containing carbonated beverages into the diet will result, in the short-term, to an overall increase in energy intake.

12-O-tetradecanoylphorbol-13-acetate promotion of transgenic mice expressing epidermal-targeted v-fos induces rasHA-activated papillomas and carcinomas without p53 mutation: association of v-fos expression with promotion and tumor autonomy.

Transgenic mice that expressed v-fos exclusively in the epidermis by means of a human keratin K1-based targeting vector (HK1.fos) developed preneoplastic epidermal hyperplasia and hyperkeratosis after long latency and an associated wound promotion stimulus. To assess the requirements for papilloma formation and malignant conversion and determine the sensitivity to a chemical promotion stimulus, HK1.fos mice were promoted with 12-O-tetradecanoylphorbol-13-acetate (TPA). HK1.fos mice were sensitive to TPA promotion but developed papillomas only after long latency (20-30 weeks of promotion) and in relatively few numbers per animal, suggesting the necessity of an additional genetic event prior to overt lesion formation. Consistent with this idea, at 60 weeks, on cessation of TPA promotion, these HK1.fos TPA-papillomas were found to be autonomous, TPA-independent tumors which persisted, grew larger, and converted to malignancy. Analysis of HK1.fos tumor RNA and DNA identified endogenous c-rasHa mutations at codons 12 and 61 in papillomas and carcinomas; however, no p53 tumor suppressor gene mutations were detected. These data indicate that epidermal expression of v-fos induces sensitivity to TPA promotion, but since additional genetic events, such as endogenous c-rasHa activation, appear to be required in tumorigenesis, v-fos may predominantly play a role in the mechanism of promotion to achieve papilloma autonomy and TPA independence. Furthermore, spontaneous malignant conversion in this model does not appear to involve mutations in the p53 tumor suppressor gene.

Inhibition of melanoma growth by adenoviral-mediated HSV thymidine kinase gene transfer in vivo.

To assess the potential of an in vivo, adenovirus-mediated gene therapy approach for the treatment of malignant melanoma, the efficacy of adenovirus-mediated herpes simplex virus thymidine kinase gene (HSV-Ek) transfer and administration of ganciclovir (GCV) was investigated using a nude mouse model. Initially, B16 murine melanoma cells were efficiently transduced in vitro by a recombinant replication-defective adenovirus containing the HSV-tk gene (ADV/RSVtk), and rendered sensitive to cell killing by 10 micrograms/ml GCV. A significant "bystander effect" was observed at low multiplicity of infection in comparison of cell killing to control B16 transduction by adenovirus containing the beta-galactosidase gene (ADV/RSV-beta-gal). In vivo, melanomas established from subcutaneous injection of 4 x 10(5) B16 cells were injected after 14 d with 1 x 10(10) ADV/RSV-tk viral particles. Subsequent treatment for 6 d with GCV resulted in an inhibition of melanoma growth, with an approximately 40-50% reduction in melanoma volume in comparison to controls in repeated experiments. These data demonstrate that adenovirus-mediated gene transfer can function as an efficient delivery system to reduce established tumor burden in vivo. This result may hold significant promise for the development of effective in situ gene therapy for melanoma in humans.

Phase I clinical trial of ormaplatin (tetraplatin, NSC 363812).

Ormaplatin is a platinum analog that was developed because of an altered toxicity profile and non-cross resistance to cisplatin in both in vitro and in vivo models. To determine the toxicities and maximum tolerated dose of ormaplatin on a daily times five schedule, patients with refractory solid tumors received ormaplatin on five consecutive days at nine dose levels ranging from 1.0 to 15.0 mg/m2/day. A total of 35 patients received 70 cycles of therapy. Nausea and vomiting and myelosuppression were moderate and not dose-limiting. Dose-limiting neurotoxicity, consisting of a sensory peripheral neuropathy, was seen in all five patients who received cumulative doses greater than or equal to 165 mg/m2. This neurotoxicity was symptomatic in all patients and caused significant functional impairment in four patients with inability to walk in two patients. A sensitive atomic absorption spectroscopy analysis performed for one patient at the 13.0 mg/m2/day dose level showed a Cpmax of 163 ng/ml and a t1/2 of 10.9 min for free platinum. A phase II dose could not be determined due to the onset of peripheral neuropathy at low cumulative doses and not at absolute dose levels.

Transgenic mice expressing targeted HPV-18 E6 and E7 oncogenes in the epidermis develop verrucous lesions and spontaneous, rasHa-activated papillomas.

In order to create a transgenic model for human papilloma virus (HPV)-associated carcinogenesis, we have used the regulatory elements of a human keratin K1 (HK1) gene to target the expression of the E6 and E7 oncogenes of HPV-18 exclusively to the epidermis. All murine expressors were viable and lived normal lifetimes; older mice (> 1 year) possessed numerous small lesions with a verrucous (wart-like) histotype. Analysis of newborn epidermis and lesions revealed that the HPV-18 E6/E7 genes were being expressed with a predominance of the E6*/E7 transcript over the full length E6/E7 message. The long latency in lesion appearance may reflect the low level of intact E6 transcripts and the requirement for additional genetic or epigenetic events before production of an overt lesion. In agreement with this proposal, spontaneous papillomas developed that expressed an activated rasHa oncogene (codon 61, A-->T; codon 13, G-->T). All lesions expressed keratin genes K1, K6, and K13 in a fashion characteristic of hyperproliferative or benign tumors with no evidence of malignant conversion. Our results demonstrate that the mouse epidermis represents a relevant in vivo model system to analyze the interaction between HPV and cellular genes in neoplasia.

Epidermal expression of transforming growth factor-alpha in transgenic mice: induction of spontaneous and 12-O-tetradecanoylphorbol-13-acetate-induced papillomas via a mechanism independent of Ha-ras activation or overexpression.

To assess the requirements for papilloma formation in transgenic mice that overexpress transforming growth factor-alpha (TGF-alpha) in the epidermis (HK1.TGF alpha), we tested the sensitivity of HK1.TGF alpha mice to tumor promotion with 12-O-tetradecanoylphorbol-13-acetate (TPA) and analyzed the resultant papillomas for synergic c-Ha-ras activation and overexpression. We observed that HK1.TGF alpha mice were highly sensitive to TPA promotion, exhibiting multiple papillomas as early as the third week of treatment. After 60 wk of promotion, malignant conversion was not observed and tumors regressed upon removal of the TPA promotion stimulus. Most of the TPA-induced papillomas did not have detectable c-Ha-ras mutations at codons 12, 13, or 61, but three papillomas arising after long-term TPA promotion (5-7 mo) exhibited c-Ha-ras activation at codon 61 (A-->T and A-->G). Conversely, spontaneous papillomas arising without TPA promotion, including persisting autonomous papillomas, were all negative for activating c-Ha-ras mutations. Both spontaneous and TPA-induced HK1.TGF alpha papillomas expressed c-Ha-ras message levels similar to those in normal, nontransgenic epidermis or HK1.TGF alpha hyperplastic epidermis. These data demonstrate that TGF-alpha overexpression can be an initiating event for TPA promotion, that papillomatogenesis in HK1.TGF alpha mice proceeds frequently via a pathway independent of Ha-ras activation or overexpression, and, thus, that other events are required for autonomous growth and malignant conversion.

Cooperation between v-fos and v-rasHA induces autonomous papillomas in transgenic epidermis but not malignant conversion.

Transgenic mice have been previously established that express v-rasHa or v-fos exclusively in the epidermis by means of a targeting vector based on the human keratin 1 gene (HK1). Epidermal expression of v-rasHa (HK1.ras) or v-fos (HK1.fos) resulted in hyperplasia, hyperkeratosis, and later, in benign tumors. To assess the potential for oncogene cooperation in vivo mating experiments were performed. Resultant HK1.fos/ras mice exhibited an obvious increase in the severity of neonatal and juvenile preneoplastic phenotypes, together with the immediate onset of tumorigenesis as compared to single oncogene sibling controls. The HK1.fos/ras tumors grew aggressively and often compromised the animals by 10-12 weeks. However, tumors remained benign as determined by histotype and specific keratin markers. These data indicate that v-fos can cooperate with an initiating v-rasHa phenotype to generate autonomous papillomas, but additional events are required for malignant conversion.

[Subtotal parathyroidectomy versus total parathyroidectomy with autotransplantation in secondary hyperparathyroidism. A randomized study]. / Subtotale Parathyreoidektomie versus totale Parathyreoidektomie mit Autotransplantation beim sekundären Hyperparathyreoidismus. Eine randomisierte Studie.

In this randomised study subtotal parathyroidectomy (PTX) is being compared with total parathyroidectomy including autotransplantation (PTX + AT) in 40 patients with secondary hyperparathyroidism. Both groups were followed up twice, first 19 +/- 6 months (PTX + AT) respectively 19 +/- 7 months (subtotal PTX) and secondly 43 +/- 9 respectively 40 +/- 7 months after surgery. In each group there were 17 patients alive at the time of the second postoperative follow-up. In the meantime 2 patients with subtotal PTX required reoperation because of recurrences originating from the remaining parathyroid tissue, another 2 patients of this group were hypercalcaemic. After PTX + AT serum-calcium normalized significantly more often (p less than 0.03), a corresponding result was seen in alkaline phosphatase. Reoperation was not required in this group. Also radiological alterations reformed significantly more often after PTX + AT, so did clinical signs like pruritus (p less than 0.005) and muscular weakness (p less than 0.04). Considering these results and the fact that in case of recidive a reoperation of the autograft in the forearm is much easier to perform than a reoperation in the neck, PTX + AT is recommended as method of choice in surgical treatment of sHPT.

The N-terminal 513 amino acids of the envelope glycoprotein gB of human cytomegalovirus stimulates both B- and T-cell immune responses in humans.

Host defense against human cytomegalovirus (HCMV) involves both humoral and cell-mediated immunity. In this report, human immune responses to glycoproteins encoded by the HCMV gB homolog gene have been examined by using glycoproteins purified by immunoaffinity from HCMV virions and recombinant proteins expressed by vaccinia viruses containing either the entire gB open reading frame or a C-terminal deletion mutant, gBm165, coding for the N-terminal 513 amino acids of gB. Neutralizing antibodies, helper T cells, and cytotoxic T cells reactive with epitopes on the N-terminal portion of gB were detected in some seropositive individuals, suggesting that this region of gB may be important in eliciting protective immunity during natural infection for some individuals.