Buccal bone dimensions and aesthetic outcome at maxillary incisors replaced by conventional dental implants. A case series with a 5-year follow-up.

In the aesthetic field, successful replacement of a tooth with a dental implant requires blend and harmony within the existing dentition. The influence of the dimension of buccal bone at implant sites on aesthetic outcomes and the relation between buccal bone horizontal and vertical dimensions are unclear. The aim of the present study is to investigate the correlation between buccal bone thickness, buccal bone level and aesthetic outcome in conventionally placed implants ­ placed five or more years previously ­ supporting single maxillary incisors. Eight subjects with 8 implants and with periapical and parallel profile X-rays were clinically examined to assess the "Pink Aesthetic Score" (PES). Buccal bone level and thickness, together with the interproximal bone level, were measured. Implant survival was 100%. The mean time of implants function was 89.3 months (standard deviation 43, range 61-145). The mean PES value was 9.4. The mean interproximal bone level was located 1.3 mm apically to the implant abutment junction, while the corresponding buccal value was 1.6 mm. Buccal bone was mostly absent at the implant abutment junction; 2 and 4 mm apically respect to the junction the thickness was on average 0.44 and 0.89 mm, respectively. The dimension of buccal bone level was correlated to the buccal bone thickness at 2mm-level, to the interproximal bone level and to the soft tissue contour score. Conventional implant placement in pristine bone might lead to satisfactory long-term aesthetic results. The level of the facial mucosa and appearance of the alveolar process might emerge as critical aspects.

A novel CAD/CAM-based surgical template for mandibular osteoplasty and guided implant insertion.

Bone shaping is often a necessary procedure prior to implant insertion in mandibular full-arch rehabilitations. Adopting guided surgery procedures is necessary to use two distinct templates: one for bone shaping, a second for guided implant insertion. The present report describes the case of a 60-year-old patient requiring a full-arch, immediately loaded implant-supported mandibular rehabilitation. A CAD/CAM-bone supported surgical template for osteoplasty was used to develop a template for guided implant during an all-on-six immediate-loaded computer-aided implant surgery. The report describes the feasibility, accuracy and usefulness of this double, CAD/CAM developed, surgical template.

Treatment of skin defects with growth factors and biodegradable collagen carrier: histological evaluation in animal model.

The aim of the present study is to evaluate the effects on skin regeneration of a new collagen matrix (CM-10826) when used in different combination with or without growth factors, using skin regeneration without membrane as control. An area of 10x15 cm on rabbit back was shaved and three circular wounds on test side were covered with a differently soaked membrane. The first wound was soaked with Epidermal Growth Factor (EGF, 26mg/130mL) (Test EGF), the second with Platelet-Derived Growth Factor (PDGF, 6mg/120mL) (Test PDGF) and the third with EGF (13mg/65mL) and PDGF (3mg/60mL) (Test EGF+PDGF). On the control side, there was a dry membrane. After 7 days, the experiment was concluded. Healing process was evaluated at day 2 and 6 postoperatively. Analysis was made clinically and with light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Analyses with LM of Test EGF showed evidence of neoangiogenesis and good epithelium growth. Test PDGF resulted in moderate angiogenesis, less evident epithelial growth and more evident mesenchymal growth than Test EGF. Test EGF+PDGF showed rich angiogenesis, massive growth of epithelium and mesenchymal tissue. Control side showed weak angiogenesis, regenerating wound margin with normal epithelium and less dense mesenchymal layer. Analysis at TEM and SEM confirmed what was noticed at LM. In vivo studies on rabbits have shown that membrane CM10826 is well tolerated, it gives neither inflammation nor foreign body reactions and does not disturb healing process. CM10826 is safe, modulates angiogenesis and induces migration and proliferation of keratinocytes.

Bi-layered collagen nano-structured membrane prototype collagen matrix 10826® for soft tissue regeneration in rabbits: an in vivo ultra-structural study of the early healing phase.

Collagen Matrix (CM) 10826 is a nanostructured bi-layered collagen membrane obtained from type I and III porcine collagen, which in vitro has shown to have the potential to be a substitute and/or stimulant for soft oral tissue regeneration. The objective of this study was to evaluate the in vivo potential and safety of this membrane for soft tissue regeneration in the early stage of wound healing. Two soft tissue wounds (test and control) were created on the back skin of 5 rabbits (female New Zealand White Rabbits specific pathogen free). All wounds were protected by a special poly-tetra-fluoro-ethylene (PTFE) healing camera. On each rabbit on the test side CM-10826 was used, while on the control side conventional treatment (an autologous pedicle graft) was performed. The healing process was observed clinically after 2 and 6 days, and Magnetic Resonance Imaging (MRI) was performed after this period. After 7 days, animals were sacrificed and specimens were analyzed with light optic microscopy (LM), Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM). These in vivo trials on rabbits confirmed that CM-10826 is well tolerated, without signs of histological inflammatory reaction and proved to be able to accelerate the spontaneous repair of the skin defect taken as the control. The light-optic and ultra-microscopy of serial biopsies showed that the new matrix is biocompatible and is able to function as a scaffold inducing soft tissue regeneration. In conclusion this study demonstrates that CM-10826 promote early soft tissue regeneration and suggests it is a potential constituent for human autologous keratinocytes seeded derma bioequivalent. It protects the wound from injuries and bacterial contamination accelerating healing process. As a clinical relevance, we consider that the quality of life of patients will be improved avoiding the use of major autologous grafts, reducing the hospitalization time and morbidity.

Shaping and preserving ß-cell identity with microRNAs.

The highly sophisticated identity of pancreatic ß-cells is geared to accomplish its unique feat of providing insulin for organismal glucose and lipid homeostasis. This requires a particular and streamlined fuel metabolism which defines mature ß-cells as glucose sensors linked to an insulin exocytosis machinery. The establishment of an appropriate ß-cell mass and function during development as well as the maintenance of their identity throughout life are necessary for energy homeostasis. The small non-coding RNAs, microRNAs (miRNAs), are now well-recognized regulators of gene transcripts, which in general are negatively affected by them. Convincing evidence exists to view miRNAs as major actors in ß-cell development and function, suggesting an important role for them in the distinctive ß-cell 'identity card'. Here, we summarize key features that associate miRNAs and the establishment of the appropriate ß-cell identity and its necessary maintenance during their 'long life'.

Bioremediation of direct dyes in simulated textile effluents by a paramorphogenic form of Aspergillus oryzae.

Azo dyes are extensively used for coloring textiles, paper, food, leather, drinks, pharmaceutical products, cosmetics and inks. The textile industry consumes the largest amount of azo dyes, and it is estimated that approximately 10-15% of dyes used for coloring textiles may be lost in waste streams. Almost all azo dyes are synthetic and resist biodegradation, however, they can readily be reduced by a number of chemical and biological reducing systems. Biological treatment has advantages over physical and chemical methods due to lower costs and minimal environmental effect. This research focuses on the utilization of Aspergillus oryzae to remove some types of azo dyes from aqueous solutions. The fungus, physically induced in its paramorphogenic form (called 'pellets'), was used in the dye biosorption studies with both non-autoclaved and autoclaved hyphae, at different pH values. The goals were the removal of dyes by biosorption and the decrease of their toxicity. The dyes used were Direct Red 23 and Direct Violet 51. Their spectral stability (325-700 nm) was analyzed at different pH values (2.50, 4.50 and 6.50). The best biosorptive pH value and the toxicity limit, (which is given by the lethal concentration (LC(100)), were then determined. Each dye showed the same spectrum at different pH values. The best biosorptive pH was 2.50, for both non- autoclaved and autoclaved hyphae of A. oryzae. The toxicity level of the dyes was determined using the Trimmed Spearman-Karber Method, with Daphnia similis in all bioassays. The Direct Violet 51 (LC(100) 400 mg · mL(-1)) was found to be the most toxic dye, followed by the Direct Red 23 (LC(100) 900 mg · mL(-1)). The toxicity bioassays for each dye have shown that it is possible to decrease the toxicity level to zero by adding a small quantity of biomass from A. oryzae in its paramorphogenic form. The autoclaved biomass had a higher biosorptive capacity for the dye than the non-autoclaved biomass. The results show that bioremediation occurs with A. oryzae in its paramorphogenic form, and it can be used as a biosorptive substrate for treatment of industrial waste water containing azo dyes.

Electronic, mechanical and piezoelectric properties of glass-like complex Na2Si1-x Ge x O3 (x = 0.0, 0.25, 0.50, 0.75, 1.0).

Motivated by our previous work on pristine Na2SiO3, we proceeded with calculations on the structural, electronic, mechanical and piezoelectric properties of complex glass-like Na2Si1-x Ge x O3 (x = 0.0, 0.25, 0.50, 0.75, 1.0) by using density functional theory (DFT). Interestingly, the optimized bond lengths and bond angles of Na2SiO3 and Na2GeO3 resemble each other with high similarity. On doping we report the negative formation energy and feasibility of transition of Na2SiO3 → Na2GeO3 while the structural symmetry is preserved. Analyzing the electronic profile, we have observed a reduced band gap on increasing x = Ge concentration at Si-sites. All the systems are indirect band gap (Z-Γ) semiconductors. The studied systems have shown mechanical stabilities by satisfying the Born criteria for mechanical stability. The calculated results have shown highly anisotropic behaviour and high melting temperature, which are a signature of glass materials. The piezoelectric tensor (both direct and converse) is computed. The results thus obtained predict that the systems under investigation are potential piezoelectric materials for energy harvesting.

Angiogenesis in psoriatic skin and its modifications after administration of etanercept: videocapillaroscopic, histological and immunohistochemical evaluation.

Several studies suggest that microangiopathy plays a crucial role in the pathogenesis of psoriasis. TNFalpha up-regulates the genetic transcription of VEGF, a pro-angiogenetic cytokine over-expressed in psoriatic skin, which promotes micrangiopathic modifications in psoriatic plaque. Etanercept is a chimeric protein used in the treatment of psoriasis and other immunomediated disorders, which blocks inflammatory response by interfering in the binding of TNF-alpha to its receptors. Starting from this data, we retain that etanercept can improve microangiopathy in psoriatic skin by reducing the synthesis of pro-angiogenetic chemokine VEGF. The aims of the study are: to verify the effect of etanercept on cutaneous en plaque capillaries in vivo using intra-vital videocapillaroscopy analysis, to evaluate the relation between the en plaque videocapillaroscopic pattern and the immunohistochemical cutaneous expression of VEGF in psoriasis, and finally to correlate all these in data with clinical disease activity. Eighteen patients (10 male and 8 female, mean age 51, range 21-60) suffering from stable, en plaque type psoriasis, involving at least 10 percent of body surface area (BSA), and not responsive to conventional therapy were included in the study. All the enrolled patients received etanercept 50mg/twice/week, subcutaneously, for 12 weeks, and were carefully followed up for clinical response with PASI score and DLQI index both before (T0) and after 12 weeks (T12) of treatment with etanercept. A well demarcated psoriatic plaque of the extensor surface of upper extremities was chosen to perform an intra-vital videocapillaroscopy analysis (IVCP), and a skin biopsy for immunohistochemical study both at T0 and T12 in all the included patients, in order to evaluate the presence of microangiopathy and its modification after therapy. All the patients experienced a clinical improvement of cutaneous disease with a significant decrease of PASI score (p<0.0001) and DLQI level (p<0.0001), throughout the twelve weeks of treatment. On IVCP analysis, microangiopathy dramatically decreased (p<0.0001), this modification being significantly related with PASI and DLQI decrease at T12. Immunohistochemical expression of VEGF decreased significantly from T0 to T12 (p<0.0001), and was related with a reduction of psoriatic microangiopathy at T12. The results of our videocapillaroscopic and immunohistochemical investigation confirm that the therapeutic potentiality of etanercept is based also on its capability to promote the regression of psoriatic microangiopathy. Moreover, according to these considerations, videocapillaroscopic evaluation of psoriatic plaque, both before and after treatment with etancercept, may be a useful tool to objectively demonstrate its effect on microcirculation.

Complications, functional outcome and quality of life after intensive preoperative chemoradiotherapy for rectal cancer.

AIMS: To investigate early and late complications in 44 patients with locally advanced mid-low rectal cancer enrolled in a phase I-II study, who had received an aggressive chemoradiation treatment (50.4Gy/28F; 5-FU continuous infusion and weekly Oxaliplatin) followed by total mesorectal excision and 5-FU based postoperative chemotherapy. The aim of the present study is also to evaluate functional outcome and quality of life (QoL) in a sub-group of 22 patients. METHODS: Standardized forms for early and late surgical complications were completed for all patients. Anorectal function and QoL were also investigated in 22 patients who underwent surgery in the same surgical unit, using the fecal incontinence scoring system (FIS) and EORTC-QLQ-CR38 questionnaires, compiled before and after radiotherapy and at least 8 months after surgery. The differences over time in scores were analyzed using repeated measure ANOVA. RESULTS: The median age of patients (25 males and 19 females) was 58 (range: 34-73) years. A low anterior resection was performed in 39 cases, radical resection in 41, and 12 patients had a pathological complete response. There were no operative deaths; 4 and 9 patients required re-operation for early and late complications, respectively. FIS score did not present a significant worsening over time. According to data in the EORTC-QLQ-CR38 questionnaire, a significant improvement over time was found only for "future perspective". CONCLUSION: Our findings seem to indicate that this aggressive 5-FU-Oxalipaltin-based treatment implies no impairment of QoL and anorectal function, even if a high rate of late major complications was observed. Studies on larger series are required to confirm these results.

Bone marrow histopathological and molecular changes of small B-cell lymphomas after rituximab therapy: comparison with clinical response and patients outcome.

This study correlates bone marrow changes after Rituximab (RTX) treatment with the clinical characteristics and outcome of 26 patients with small B-cell lymphomas. The percentage, phenotypic profile and clonality pattern of bone marrow lymphoid infiltrate were analysed before and after RTX treatment. Clinical, histological and molecular responses to RTX were correlated to the clinical outcome of the patients. Sixteen out of twenty-six patients obtained a complete clinical remission (CR). A favourable histology--follicular lymphoma (FL), hairy cell leukaemia (HCL) and marginal zone lymphoma (MZL)--was associated with a higher frequency of clinical CR and histological remission (HR), in comparison with mantle cell lymphoma (MCL), chronic lymphocytic leukaemia (CLL) and lymphoplasmacytic lymphoma (LPL). Two patterns of bone marrow HR were observed: 1) complete lymphoid cell disappearance (9 patients); or 2) nodular/interstitial T-cell infiltration (10 patients). Three histological persistence (HP) patterns were observed: 1) persistence of CD20+ small lymphoid cells in 1 patient with MCL; 2) loss of CD20 antigen expression in 4 patients with CLL; or 3) persistence only of clusters of monotypic plasma cells in 2 patients with LPL. CR and HR were strongly correlated. The percentage of lymphomatous infiltrate after RTX was higher in patients who subsequently died of the disease. Molecular response showed no correlations with the further clinical course in 12 patients achieving a complete clinical remission. In conclusion, bone marrow morphological and immunohistochemical analysis with a restricted panel of antibodies is useful to avoid 42% false positive and 85% false negative interpretations. Persistence of monoclonality after RTX might have a role in evaluating the molecular pattern of CD20-negative clones that can emerge after RTX as a tumoral escape to therapy.

Ependymomas, malignant tumors of pancreatic islets, and osteosarcomas induced in hamsters by BK virus, a human papovavirus.

BK virus (BKV), a human papovavirus, was inoculated iv into 3-week-old Syrian golden hamsters. Between 2 1/2 and 9 months after inoculation, 82% of the animals developed tumors. The induced neoplasms were ependymoma, carcinoma of the pancreatic islets, osteosarcoma, adenocarcinoma, angiosarcoma, angioma, lymphoma, and seminoma. Hypersecretion of insulin, glucagon, C-peptide, and calcitonin was detected in tumors of pancreatic islets. BKV etiology of tumors was supported by the following evidence: 1) No tumors with BKV-specific markers appeared in animals given injections of buffer, animals inoculated with BKV neutralized by anti-BKV-specific serum, or uninoculated controls; 2) BKV tumor (T) antigen was detected by immunofluorescence and complement fixation tests in tumors of animals inoculated with infectious BKV and in transplanted tumors; 3) antibodies to BKV T-antigen were detected in sera of animals bearing primary or transplanted tumors; 4) BKV could be activated by Sendai virus-mediated fusion of neoplastic cells with susceptible Vero cells; and 5) no endogenous hamster oncornaviruses were found in tumors.

Immunocytochemical demonstration of estrogen receptors by monoclonal antibodies in human breast cancer: correlation with estrogen receptor assay by dextran-coated charcoal method.

Immunocytochemical demonstration of estrogen receptors in 115 human breast cancer specimens was performed using mouse monoclonal antibodies against estrogen receptor and avidin-biotin as the displaying system. The antibody indicated a highly heterogeneous endowment of neoplastic cells with estrogen receptor at both nuclear and cytoplasmic levels. The percentage of labeled cells within each tumor specimen was recorded to compare this immunocytochemical assay with the biochemical assay of estrogen receptors by the dextran-coated charcoal method. A significant correlation was observed between these two assays. The present results show that estrogen receptors can be confidently demonstrated at the single cell level, thus providing additional information to quantitative biochemical assays. Their prognostic and therapeutic predictive powers may be usefully integrated, particularly in view of the heterogeneous distribution of receptors among cancer cells.

Perioperative anemia and blood transfusions as prognostic factors in patients undergoing resection for non-small cell lung cancers.

We hypothesised that anemia could represent an important prognostic factor and perioperative blood transfusions do not reduce the risk of relapse. In order to explore this topic, we assessed the correlation of preoperative anemia and blood transfusions with survival in patients with resected non-small cell lung cancer (NSCLC). Patients who underwent radical surgery for NSCLC at the Department of Thoracic Surgery of Università Politecnica delle Marche from January 1996 through December 2001, were included in our study. Four hundred and thirty-nine patients were eligible for our analysis. Survival appeared worse in patients with haemoglobin (Hb) < or =10 g/dl versus Hb >10 g/dl (p=0.012). Stratifying patients in three groups on their Hb level (group 1: Hb < or =10 g/dl; group 2: Hb=10-12 g/dl; group 3: Hb > or =12 g/dl), we observed a worse prognosis in patients with lower Hb levels, too (p=0.0325) and also in the transfused population (p=0.046). At multivariate analysis, only the age of patients, pathological stage and Hb levels resulted indicators of prognosis. Our results suggested that anemia could represent an important prognostic factor in resected NSCLC and correction of anemia in the perioperative setting does not reduce the risk of relapse.

Estrogen receptor cytochemistry by fluorescent estrogen.

A recently synthesized fluorescein-labeled estrogen (17FE, 1-(N)-fluoresceinyl-estrone-thiosemicarbazone) interacts with estrogen-target cells like the native hormone and visualizes the uptake, transport, and distribution of estrogen in intact target cells. Moreover, estrogen binding sites are traced by 17FE in cryostat sections of estrogen target tissues as well. Cell and tissue 17FE binding sites fulfill the accepted criteria for specific estrogen receptors (finite binding capacity, high affinity, steroid and tissue specificity). This fluorescent probe allows estrogen receptors to be studied in a wide variety of cell and tissue preparations under varying conditions of physiologic and pathophysiologic interest.

Treatment of abacterial prostato-vesiculitis with nimesulide.

The efficacy and tolerability of nimesulide were assessed in the treatment of patients with prostato-vesiculitis. In a noncomparative investigation, 30 patients received oral nimesulide 100mg twice daily for three 10-day cycles. Micturition-related symptoms were resolved in 20 patients and clear amelioration of inflammatory signs was observed with transrectal ultrasound in 16 patients. Abnormal sperm forms decreased from 57 to 49% (p < 0.001). In a comparative investigation, 40 patients received nimesulide 200mg twice daily or ketoprofen 100mg twice daily via the rectal route. Patients and physicians expressed an overall opinion on efficacy in favour of nimesulide. In a pharmacokinetic study of healthy volunteers who received oral nimesulide 100mg as a single dose, the mean maximum nimesulide concentration (0.58 +/- 0.13 mg/L) in seminal fluid was achieved after 2 hours while the maximum seminal fluid: blood plasma ratio 0.32 +/- 0.02 was observed after 4 hours. These data suggest that nimesulide is an effective NSAID in the treatment of abacterial prostato-vesiculitis and also demonstrate that this drug has a favourable disposition within the genital tract.

Treatment of benign prostatic hyperplasia with 5-alpha-reductase inhibitor: morphological changes in patients who fail to respond.

AIMS: To describe the prostatic adenectomy specimens of six patients with symptomatic benign prostatic hyperplasia (BPH) who failed to respond to long term treatment with a 5-alpha-reductase inhibitor, finasteride. METHODS: Histological sections from six cases of BPH who had been treated with finasteride were investigated. Five patients were prescribed 5 mg finasteride daily for six months and one patient 5 mg daily for 12 months. The patients underwent adenectomy as their urethral obstruction failed to resolve. Twenty cases of untreated BPH served as controls. RESULTS: In patients taking finasteride for six months the prostatic adenectomy specimens showed a reduction in the size of the prostate and an increase in the stroma:epithelial and stroma:lumen ratios compared with controls. The size of the ducts and acini was not as uniform as in the controls. In particular, some ducts and acini were still lined by a bistratified epithelium similar to that found in controls but lacked undulations at the epithelial border; other ducts/acini were atrophic. Some scattered clusters of small acini with a focally fragmented basal cell layer were observed in two of the five treated cases. One prostatic adenectomy specimen, from the patient treated for one year, showed extensive lobular atrophy and diffuse squamous and transitional cell metaplasia. At the periphery of the transition zone, there was a complex intra-acinar papillary-cribriform proliferation of clear cells without nuclear atypia, similar to clear cell papillary hyperplasia. The periurethral region showed stromal nodules in both patients and controls. CONCLUSIONS: Morphological evaluation of finasteride treated BPH showed changes in the lobules of the transition zone, but not in the periurethral stroma.

Effects of combination endocrine treatment on normal prostate, prostatic intraepithelial neoplasia, and prostatic adenocarcinoma.

AIMS: To investigate the effect of combination endocrine treatment (CET) or luteinising hormone releasing hormone agonist and flutamide on non-neoplastic prostate, prostatic intraepithelial neoplasia, and prostatic adenocarcinoma. METHODS: The morphology, including the mitotic activity, of 12 radical prostatectomies from patients with prostatic adenocarcinoma pretreated for three months with CET was evaluated in haematoxylin and eosin stained sections and compared with an untreated age and stage matched control group. RESULTS: A differential effect on the non-neoplastic prostate was observed. In fact, the transition zone of the treated prostate showed simplification of the glandular lobules: the ducts and acini were small without undulations of the epithelial border and with a prominent basal cell layer. Within the peripheral zone there was inconspicuous branching of the ducts and acini which looked dilatated and lined by flattened atrophic epithelium. Prostatic intraepithelial neoplasia occurred in scattered ducts and acini in the peripheral zone of 10 of the 12 patients. The epithelial cell lining showed a prominent basal cell layer. A certain degree of secretory cell type stratification was always present. However, crowding was less evident than in the untreated prostate because of cytoplasmic clearing and enlargement as a result of coalescence of vacuoles. The treated adenocarcinomas had neoplastic acini which looked small and shrunken, and areas of individual infiltrating tumour cells separated by abundant interglandular connective tissue. The secretory cells of the nonneoplastic, prostatic intraepithelial neoplasia, and prostatic adenocarcinoma lesions had inconspicuous nucleoli, nuclear shrinkage, chromatin condensation, and cytoplasmic clearing. Apoptotic bodies were easily identifiable in all the cell layers. The lumina were rich in macrophages, sloughed secretory cells with degenerative features, and apoptotic bodies. Mitoses were not observed in any of the treated non-neoplastic prostate, prostatic intraepithelial neoplasia, or prostatic adenocarcinomas, whereas the mitotic frequency increased from non-neoplastic prostate through prostatic intraepithelial neoplasia up to prostatic adenocarcinomas in the untreated specimens. CONCLUSIONS: CET before radical prostatectomy causes regressive epithelial changes together with enhanced apoptosis and blocked mitotic activity.

Density of neoplastic lymphoid infiltrate, CD8+ T cells, and CD1a+ dendritic cells in mycosis fungoides.

BACKGROUND/AIMS: CD8+ T cells and epidermal/dermal dendritic cells expressing CD1a are found among neoplastic CD4+ T cells in mycosis fungoides (MF) lesions. This study analysed the relation of CD8+ tumour infiltrating lymphocytes (TILs), CD1a+ epidermal Langerhan's cells (LCs), and dermal dendritic cells (DDCs) to clinicopathological parameters in 46 MF cases. METHODS: Pretreatment diagnostic biopsy specimens of 46 MF cases were submitted to histological analysis and immunohistochemistry. Four histological grades were defined based on the density of the neoplastic infiltrate: grade 1 (mild superficial perivascular infiltrate), grade 2 (moderate superficial perivascular infiltrate with some tendency to confluence), grade 3 (pronounced superficial band-like infiltrate), and grade 4 (deep nodular infiltrate). Epidermotropism was scored as low, moderate, or high. Numbers of CD8+ T cells and of dermal and epidermal CD1a+ cells were scored as 1 (low), 2 (moderate), and 3 (high). Correlations between these parameters and clinical data (age, sex, clinical type of lesions, stage, response to treatment, and recurrence) were analysed by the chi(2) test. RESULTS: Numbers of TILs and DDCs were associated with subepidermal infiltrates, being lower in less dense infiltrates, whereas there was no association between epidermal CD1a+ cells and the analysed parameters. Complete remission in treated patients was related to subepidermal infiltrates but not to TILs, LCs, or DDCs. CONCLUSIONS: These results support the notion that CD8+ cells and dermal CD1a+ cells are active against tumour cells. MF with low numbers of TILs could represent an early stage of the disease, before TILs are activated against tumour specific antigens.

Role of nitric oxide in 2-deoxy-D-glucose-induced hypothermia in rats.

The present study was designed to test the hypothesis that nitric oxide (NO) plays a role in 2-deoxy-D-glucose (2-DG)-induced hypothermia. The body temperature of awake, unrestrained rats was measured before and after the administration of 2-DG, or N(G)-nitro-L-arginine methyl ester (L-NAME; a non-selective NOS inhibitor) or both treatments together. We observed a significant reduction in body temperature after 2-DG injection. L-NAME alone caused no significant change in body temperature. When the two treatments were combined, a reduction in the magnitude of 2-DG-induced hypothermia was observed. The neuronal NOS inhibitor 7-nitroindazole also inhibited 2-DG-induced hypothermia. The data indicate that NO, probably produced by neuronal NOS, plays a role in 2-DG-induced hypothermia.

Role of nitric oxide in rat locus coeruleus in hypoxia-induced hyperventilation and hypothermia.

The locus coeruleus modulates the ventilatory and thermoregulatory response to hypoxia and contains nitric oxide synthase. Therefore, we examined the effects of L-NAME unilaterally microinjected into the locus coeruleus on hypoxic hyperventilation and hypothermia. Ventilation and body temperature were measured before and after microinjection of L-NAME (100 nmol/0.5 microl) into the locus coeruleus, followed by hypoxia. Control rats received microinjection of D-NAME (an inactive enantiomer of L-NAME). Under normoxia, L-NAME treatment did not affect ventilation or body temperature. D-NAME did not affect hypoxia-induced hyperventilation and hypothermia. L-NAME treatment reduced the ventilatory response to hypoxia but did not affect hypoxia-induced hypothermia. These data suggest that nitric oxide in the locus coeruleus is involved in the ventilatory response to hypoxia, exercising an inhibitory modulation on the locus coeruleus neurons, but plays no role in hypoxia-induced hypothermia.