Showing papers by "University of Modena and Reggio Emilia published in 1997"

Effect of testosterone and estradiol in a man with aromatase deficiency.

Abstract: Recent reports of disruptive mutations of the genes for the estrogen receptor or for cytochrome P-450 aromatase1–6 have shed new light on the role of estrogen. In females the lack of estrogen due to aromatase deficiency leads to pseudohermaphroditism and progressive virilization at puberty, whereas in males pubertal development is normal. In members of both sexes epiphyseal closure is delayed, resulting in a eunuchoid habitus, and osteopenia is present.6 These findings suggest a crucial role of estrogen in skeletal maturation.1–6 We describe the responses to androgen and estrogen in a man with a novel, homozygous inactivating mutation of . . .

Granulocyte macrophage colony-stimulating factor is overproduced by keratinocytes in atopic dermatitis. Implications for sustained dendritic cell activation in the skin.

Abstract: Lesional skin of atopic dermatitis (AD) harbors high numbers of dendritic cells with enhanced stimulatory capacity for T lymphocytes. In this study, lesional AD skin was shown to stain heavily in both epidermal and dermal compartments for GM-CSF, a cytokine crucial to dendritic cell functions. Keratinocyte cultures established from uninvolved skin of AD patients exhibited markedly increased spontaneous and PMA-stimulated release of GM-CSF compared with keratinocytes from nonatopic controls. Correspondingly, keratinocytes from AD patients showed higher constitutive as well as PMA-induced GM-CSF gene expression. Larger amounts of GM-CSF were produced by AD keratinocytes, also in response to IL-1alpha, but not after stimulation with LPS, lipoteichoic acid, or staphylococcal enterotoxin B. Hydrocortisone reduced GM-CSF gene expression and protein release in both atopic and control keratinocytes. Supernatants from atopic keratinocytes were able to strongly stimulate PBMC proliferation in a GM-CSF-dependent manner. Moreover, conditioned medium from PMA-treated AD keratinocytes, together with exogenous IL-4, could support phenotypical and functional maturation of peripheral blood precursors into dendritic cells. Enhanced production of GM-CSF by keratinocytes may contribute relevantly to the establishment and chronicity of AD lesions, in particular to the increased number, sustained activation, and enhanced antigen-presenting functions of dendritic cells.

Mitochondria alterations and dramatic tendency to undergo apoptosis in peripheral blood lymphocytes during acute HIV syndrome.

Abstract: Objective To study alterations of mitochondrial membrane potential (delta psi) and the propensity to undergo apoptosis in peripheral blood lymphocytes (PBL) from subjects with acute HIV syndrome; and to evaluate possible modulations of these phenomena by antioxidants that can be used in therapy, such as N-acetyl-cysteine (NAC), nicotinamide (NAM), or L-acetyl-carnitine (LAC). Methods Mitochondrial function and the tendency of PBL to undergo spontaneous apoptosis were studied on freshly collected PBL from patients with symptomatic, acute HIV-1 primary infection, which were cultured for different durations in the presence of absence of NAC. NAM or LAC. By a cytofluorimetric method allowing analysis of delta psi in intact cells, we studied the function of these organelles under the different conditions. PBL apoptosis was evaluated by the classic cytofluorimetric method of propidium iodide staining, capable of revealing the typical DNA hypodiploid peak. Results Significant delta psi alterations and tendency to undergo apoptosis were present in PBL from the subjects we studied. Indeed, when cultured even for a few hours in the absence of any stimulus, a consistent number of cells died. However, the presence of even different levels of NAC, NAM or LAC was able to rescue most of them from apoptosis. Both a fall in delta psi and apoptosis were evident in PBL collected in the earliest phases of the syndrome (before seroconversion), and changed significantly after a few days. A significant correlation was found between spontaneous apoptosis and tumour necrosis factor (TNF)-alpha or p24 plasma levels, as well as between apoptosis and the percentages of circulating CD4+ or CD8+ T cells. Conclusions PBL from patients with acute HIV syndrome are characterized by both significant mitochondrial alterations and a dramatic tendency to undergo apoptosis. The use of NAC, NAM or LAC seems to rescue cells through a protective effect on mitochondria, a well-known target for the action of TNF-alpha and for reactive oxygen species, the production of which is strongly induced by this cytokine. Thus, our data could provide the rationale for the use of such agents in addition to antiviral drugs in primary infection.

Resistance to apoptosis in CTLL-2 cells constitutively expressing c-Myb is associated with induction of BCL-2 expression and Myb-dependent regulation of bcl-2 promoter activity

Abstract: c-Myb, the cellular homologue of the transforming gene of the avian myeloblastosis virus, is preferentially expressed in all hematopoietic lineages, including T and B lymphocyte lineages. In T lymphocytes, c-Myb expression appears to be required for cell cycle progression and proliferation. To further investigate the role of c-Myb in T cell proliferation and survival, interleukin (IL) 2-dependent CTLL-2 cells were transfected with a constitutively active c-myb or with a c-myb antisense construct able to down-regulate endogenous Myb levels, and the transfectants were assessed for proliferation and survival in low concentrations of IL-2 and for susceptibility to dexamethasone-induced apoptosis. Compared with control cells, CTLL-2 cells constitutively expressing c-Myb proliferate in low concentrations of IL-2 and are less susceptible to apoptosis induced by IL-2 deprivation or treatment with dexamethasone. In contrast, cells transfected with an antisense c-myb construct do not proliferate in low concentrations of IL-2 and undergo apoptosis upon IL-2 deprivation or dexamethasone treatment more rapidly than parental cells. Overexpression of c-Myb was accompanied by up-regulation of BCL-2 expression. In transient transfection assays, the murine bcl-2 promoter was efficiently transactivated by c-Myb, but such effect was observed also in cells transfected with a DNA binding-deficient c-myb construct. Moreover, in gel retardation assays, a 38-bp oligomer in the shortest bcl-2 promoter segment regulated by c-Myb formed a specific complex with nuclear extracts from c-Myb-transfected CTLL-2 cells. Thus, these results strongly suggest that c-Myb, in addition to regulating T cell proliferation, protects T lymphocytes from apoptosis by induction of BCL-2 expression, which involves a c-Myb-dependent mechanism of promoter regulation.

Pro-Opiomelanocortin-Derived Peptides, Cytokines, and Nitric Oxide in Immune Responses and Stress: An Evolutionary Approach

Abstract: In vertebrates, including man, the study of stress has contributed substantially to unravelling the complex relationship between immune-neuroendocrine interactions and the systems involved. On the basis of data on the presence and distribution of the main actors (POMC products, cytokines, biogenic amines, and steroid hormones) in different species and taxa from invertebrates to vetebrates, we argue that these responses have been deeply connected and interrelated since the beginning of life. Moreover, the study of nitric oxide suggests that the inflammatory reaction is located precisely between the immune and stress responses, sharing the same fundamental evolutionary roots.The major argument in favor of this hypothesis is that the immune,stress, and inflammation responses appear to be mediated by a common pool of molecules that have been conserved throughout evolution and that form a network of adaptive mechanisms. One cell type, the macrophage, appears to emerge as that most capable of supporting this network critical for survival;it was probably a major target of selective pressure. All these data fit the Unitarian hypothesis we propose,by which evolution favors what has been conserved, rather than what has changed,as far as both molecules and functions are concerned.

Human herpesvirus 6 (HHV-6) ORF-1 transactivating gene exhibits malignant transforming activity and its protein binds to p53

Abstract: The 357 amino acid open reading frame 1 (ORF-1), also designated DR7, within the SalI-L fragment of human herpesvirus 6 (HHV-6) exhibited transactivation of the human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) promoter and increased HIV-1 replication (Kashanchi et al., Virology, 201, 95-106, 1994). In the current study, the SalI-L transforming region was localized to the SalI-L-SH subfragment. Several ORFs identified in SalI-L-SH by sequence analysis were cloned into a selectable mammalian expression vector, pBK-CMV. Only pBK/ORF1 transformed NIH3T3 cells. Furthermore, cells expressing ORF-1 protein produced fibrosarcomas when injected into nude mice, whereas control cells, expressing either no ORF-1 protein or C-terminal truncated (after residue 172) ORF-1 protein, were not tumorigenic. Western blot analysis of proteins extracted from the tumors revealed ORF-1 protein. Additional studies indicated that ORF-1 was expressed in HHV-6-infected human T-cells by 18 h. Co-immunoprecipitation experiments showed that ORF-1 protein bound to tumor suppressor protein p53, and the ORF-1 binding domain on p53 was located between residues 28 and 187 of p53, overlapping with the specific DNA binding domain. Functional studies showed that p53-activated transcription was inhibited in ORF-1, but not in truncated ORF-1, expressing cells. Importantly, the truncated ORF-1 mutant also failed to cause transformation. Analysis of several human tumors by PCR revealed ORF-1 DNA sequences in some angioimmunoblastic lymphadenopathies, Hodgkin's and non-Hodgkin's lymphomas and glioblastomas. The detection of ORF-1 sequences in human tumors, while not proof per se, is a prerequisite for establishing its role in tumor development. Taken together, the results demonstrate that ORF-1 is an HHV-6 oncogene that binds to and affects p53. The identification of both transforming and transactivating activities within ORF-1 is a characteristic of other viral oncogenes and is the first reported for HHV-6.

Persistent high MR signal of the posterior pituitary gland in central diabetes insipidus

Abstract: We describe three cases of central diabetes insipidus, each with a different pathogenesis, in which unexpected hyperintensity of the posterior pituitary gland was seen on T1-weighted MR images obtained at the time of presentation. In the first case (idiopathic), the posterior pituitary signal persisted more than 10 years; in the second case (Langerhans cell histiocytosis), the signal disappeared within 3 months, despite early specific chemotherapy with etoposide; and in the third case (transient), the posterior signal disappeared within 1 year, but it was documented at the time of spontaneous reversal of polyuria and polydipsia.

All-trans retinoic acid in hematological malignancies, an update. GER (Gruppo Ematologico Retinoidi).

Abstract: BACKGROUND AND OBJECTIVE: During the past ten years, the study of retinoids has undergone a total transformation. The Italian Society of Experimental Hematology decided to discuss these advances at a meeting in Florence on April 18, 1996. INFORMATION SOURCES: The material examined in the present review includes articles and abstracts published in journals covered by the Science Citation Index and Medline. In addition, all the authors of the present article have been actively working in the field of retinoids and have contributed several papers. Summaries of their oral presentations at the Florence meeting are reported in the Appendix to this review article. STATE OF ART AND PERSPECTIVES: One of the most important advances has been the elucidation of new molecular mechanisms of control of gene expression by retinoids. A number of new retinoids have been synthesized by chemists, some of which are being screened for potential clinical use, and a few have already had a tremendous impact on clinical practice. The most important achievements have been obtained in acute promyelocytic leukemia. In 1988 a Chinese group working in Shanghai showed that using all-trans retinoic acid (ATRA) alone 94% of acute promyelocytic leukemic patients obtained complete remission through differentiation of the leukemic clone. This result transformed a dream into reality and allowed researchers to move from laboratory experience to clinical applications of this differentiating therapy. Expanding the spectrum of hematological malignancies that may respond to ATRA remains a challenge; however, several results show some activity of retinoids alone or in combination with other drugs in juvenile chronic myeloid leukemia (CML), myelodysplastic syndrome, cutaneous T-cell lymphoma and CML. Particularly interesting are the studies that explored the potential clinical synergism of ATRA-based combination therapies with growth factors, other differentiating agents such as vitamin D3, immunomodulators like interferons, or chemotherapeutic agents, in particular Ara-C, all of which show promising in vitro effects when used in combination with retinoids.

Lesion load quantification in serial MR of early relapsing multiple sclerosis patients in azathioprine treatment : A retrospective study

Abstract: Azathioprine (AZA) has a slight but consistent effect on clinical outcome in multiple sclerosis (MS), but very few data are available on magnetic resonance imaging (MRI) changes. We performed a retrospective study aimed to quantify changes of lesion load in two serial proton density weighted MRI sequences (TR 2500, TE 30, 1.5 T) at a mean interval of 2.5 years in 36 relapsing-remitting (RR) MS patients: 19 had been treated with AZA, beside steroids after relapses (AZA group), and 17 had been treated with steroids only (control group). All but 3 patients were in the early phase of the disease. Total lesion area (TLA) was measured by manual outlining method and the arbitrary score proposed by Ormerod (total score) was also calculated from the number and diameter of lesions. Lesion load was the same at baseline, but median percentage difference of TLA between first and second scan was + 15.6% in control, -43.7% in the AZA group (p < 0.05, Mann-Whitney test). The distribution of patients according to TLA change, assuming that an increase or decrease was significant if larger than 50%, was found to be significantly different in favor of AZA-treated patients (chi(2) = 35.92, p < 0.001). These results suggest an effect of AZA treatment on MRI lesion load in early RR MS: a larger prospective study is worthwhile.

Hodgkin's disease presenting below the diaphragm. The experience of the Gruppo Italiano Studio Linfomi (GISL)

Abstract: BACKGROUND AND OBJECTIVE: Infradiaphragmatic Hodgkin's disease is rare, making up 5-12% of cases in clinical stages I and II; consequently, several questions concerning prognosis and treatment strategy remain to be answered. The aim of this study was to analyze the clinical and prognostic characteristics and outcome of this condition. METHODS: A series of 282 patients with CS I-II Hodgkin's disease (HD) was investigated. In 31 patients the disease was confined below the diaphragm (BDHD), and in the remaining above the diaphragm (ADHD). The presenting features and outcomes were compared in the two groups. RESULTS: The BDHD group was older (p < 0.0002), had a higher frequency of males (p < 0.08) and a different histological subtype group distribution (p < 0.0001). Stage II BDHD patients had a worse overall survival rate (OS) than stage II ADHD patients (68.8% vs 86.6% at 8 years, p < 0.01) if age is not considered; patients with more than 40 years of age, in fact, had the same survival rates as those with ADHD. BDHD patients with intra-abdominal disease alone had worse prognostic factors and OS (p = 0.12) than patients with inguinal-femoral nodes. INTERPRETATION AND CONCLUSIONS: Although BDHD patients present distinct features, they have the same OS and relapse-free survival rate as age-adjusted ADHD patients. According to our experience patients with stage I peripheral BDHD respond well to radiotherapy-based regimens. Those with stage II and or intra-abdominal disease are more challenging; chemotherapy or a combined therapy seem to be more suitable approaches for these patients.

Impairment in dating and retrieving remote events in patients with early Parkinson's disease.

Abstract: Remote memory has been studied in a group of 25 non-demented patients with Parkinson's disease and their performance has been compared with that of 22 healthy control subjects. Only patients who scored > or = 27 on the mini mental state examination and with no anticholinergic treatment were included in the sample. A remote memory questionnaire was given, to evaluate memory for public events that occurred from 1966 to 1990. Each event was probed with five questions concerning its content and one for the date. Compared with healthy subjects, patients with Parkinson's disease were significantly impaired both in recalling the content and in dating remote events. These results support the claim that remote memory in patients with Parkinson's disease is disrupted independently of dementia. This impairment might result from a dysfunction at the level of the circuit connecting the basal ganglia to the frontal lobes.

Effects of a yeast-based dietary supplementation on premenstrual syndrome : A double-blind placebo-controlled study

Abstract: A dietary approach has proven to be effective in alleviating symptoms of premenstrual syndrome. In our previous studies, magnesium improved premenstrual irritability and mood scoings. In this double-b

Treatment of allergic diseases during pregnancy.

Abstract: Pregnancy may variously modify the natural history of allergic disorders through occurring endocrinologic, functional and immunological changes. A pharmacologic treatment of allergic diseases (mainly asthma) is often necessary during pregnancy. On the other hand, a drug should be not potentially teratogenic and should not have serious side effects, both for the mother and the fetus. This paper reviews current knowledge about allergic diseases during pregnancy, considering the points of view of the different specialists involved in their management. Topical mucosal agents seem to be the safest, due to their minimal or absent absorption which should reflect reduced side effects. Preferred agents should be topical antihistamines (for rhinitis and conjunctivitis), and cromones and topical steroids (for asthma), as they are both safe and effective.

Self gamma2a(b) protein is presented in vivo by gamma2a(b) B cells but not by dendritic cells.

Abstract: We have previously shown that IgG2a(b) Ig does not induce tolerance in MHC class II restricted CD4 T cells in a TCR transgenic model and that anti-IgG2a(b) transgenic T cells specific for peptide 435-451 are indeed present in the periphery where they interact with gamma2a(b)-positive B cells. We also observed that because T cell tolerance depends on the presentation of self peptides, it was probable that IgG2a(b) was not easily processed and presented in vivo. In this study, we have investigated the presentation of naturally processed gamma2a(b) (435-451) determinants to specific T cells. Dendritic cells, macrophages, and B cells were purified from the spleens of Igh-1b mice. These cells were then functionally tested for the presence of specific peptide-MHC complexes. The results showed that, in vivo, gamma2a(b)-producing B cells, but not dendritic cells, are the only APCs able to present this self peptide. This indicates that recognition of the IgG2a(b)-self peptide is exclusively mediated by T-B cell interaction.

Environmental and biological monitoring of occupational exposure to perchloroethylene in dry cleaning shops

Abstract: Occupational exposure to perchloroethylene (PCE) was studied in a total of 106 workers in 78 dry cleaning shops in the province of Pavia, Northern, Italy. Environmental monitoring was performed by personal passive sampling. The median time weighted average (TWA) level of PCE was 57 mg/m3, i.e., about 30% of the current Threshold Limit Value (TLV) proposed by the American Conference of Governmental Industrial Hygienists (ACGIH). However, in 12 workers exposure exceeded this limit. Biological monitoring was performed via measurement of urinary trichloroacetic acid (TCA), i.e. the exposure index currently used in Italy, and urinary excretion of unmodified perchloroethylene (PCE-U) in samples collected at the end of the half-shift. Median levels of TCA and PCE were 1.03 mg/l and 17.7 micrograms/l respectively. The correlation coefficient between environmental TWA concentrations of perchloroethylene and PCE-U was 0.755 (0.809 after logarithmic transformation), compared to 0.660 for TCA values. The subjects were then classified as "low exposed" and "heavily exposed" according to whether personal exposure was lower or higher than 57 mg/m3, the median TWA value of the whole group. PCE-U levels were significantly correlated to exposure in both subgroups whereas TCA was correlated only in the "heavily exposed subjects", but not in those with lower exposure. The results of the study show that in the majority of dry cleaning shops exposure to PCE was well below the current occupational limits. Nevertheless surveillance of dry cleaners is recommended as nearly 10% of the workers exceeded the environmental and biological limits. Urinary excretion of unmodified PCE appears to be a very reliable indicator for biological monitoring of PCE exposure in dry cleaning and is also significantly correlated to exposure at low levels. The estimated biological equivalent exposure level (BEEL) for PCE-U, corresponding to the current TLV-TWA proposed by the ACGIH, is 55 micrograms/l. Urinary TCA seems to be less suitable for assessment of individual exposure to perchloroethylene in dry cleaners as it is poorly representative of exposure to low levels of the solvent, which is a very common occurrence in this occupational group nowadays.

Effect of heparin on cell proliferation : Lack of correlation with heparin binding sites on cell membrane

Abstract: In this study an attempt was made to correlate the in-vitro anti-proliferative effect of heparin with the heparin binding on the cell surface. Cells with different sensitivities to the anti-proliferative effect of heparin (BHK-21, FAO, SMC, BAEC, A-431, V-79, and skin fibroblasts) were incubated with [ 3 H]heparin either in the presence or in the absence of unlabelled heparin. A saturable binding was found only in BHK-21, FAO, SMC, BAEC and V-79. Scatchard analysis revealed the presence of a single class of binding sites. The binding of [ 3 H] heparin was efficiently displaced by unlabelled heparin, pentosan polysulfate and low-molecular-weight heparin, but not by dermatan sulfate. Although the sensitivity to the anti-proliferative effect of heparin varied considerably among the cell types (BHK-21 >SMC, FAO >BAEC > V-79), there was no correlation between the reduction of proliferation of these cells and either their heparin binding capacity or the number of binding sites per cell.