Control of carbapenemase-producing Klebsiella pneumoniae: a region-wide intervention.

Starting in 2010, there was a sharp increase in infections caused by Klebsiella pneumoniae resistant to carbapenems in the Emilia-Romagna region in Italy. A region-wide intervention to control the spread of carbapenemase-producing K. pneumoniae (CPKP) in Emilia-Romagna was carried out, based on a regional guideline issued in July 2011. The infection control measures recommended to the Health Trusts (HTs) were: phenotypic confirmation of carbapenemase production, active surveillance of asymptomatic carriers and contact isolation precautions for carriers. A specific surveillance system was activated and the implementation of control measures in HTs was followed up. A significant linear increase of incident CPKP cases over time (p<0.001) was observed at regional level in Emilia-Romagna in the pre-intervention period, while the number of cases remained stable after the launch of the intervention (p=0.48). Considering the patients hospitalised in five HTs that provided detailed data on incident cases, a downward trend was observed in incidence after the release of the regional guidelines (from 32 to 15 cases per 100,000 hospital patient days). The spread of CPKP in Emilia-Romagna was contained by a centrally-coordinated intervention. A further reduction in CPKP rates might be achieved by increased compliance with guidelines and specific activities of antibiotic stewardship.

Association between respiratory distress time and invasive mechanical ventilation in COVID-19 patients: A multicentre regional cohort study.

AIM: To determine whether the duration of respiratory distress symptoms in severe COVID-19 pneumonia affects the need for invasive mechanical ventilation and clinical outcomes. MATERIALS AND METHODS: An observational multicentre cohort study of patients hospitalised in five COVID-19-designated ICUs of the University Hospitals of Emilia-Romagna Region. Patients included were adults with pneumonia due to SARS-CoV-2 with PaO2/FiO2 ratio <300 mmHg, respiratory distress symptoms, and need for mechanical ventilation (invasive or non-invasive). Exclusion criteria were an uncertain time of respiratory distress, end-of-life decision, and mechanical respiratory support before hospital admission. MEASUREMENTS AND MAIN RESULTS: We analysed 171 patients stratified into tertiles according to respiratory distress duration (distress time, DT) before application of mechanical ventilation support. The rate of patients requiring invasive mechanical ventilation was significantly different (p < 0.001) among the tertiles: 17/57 patients in the shortest duration, 29/57 in the intermediate duration, and 40/57 in the longest duration. The respiratory distress time significantly increased the risk of invasive ventilation in the univariate analysis (OR 5.5 [CI 2.48-12.35], p = 0.003). Multivariable regression analysis confirmed this association (OR 10.7 [CI 2.89-39.41], p < 0.001). Clinical outcomes (mortality and hospital stay) did not show significant differences between DT tertiles. DISCUSSION: Albeit preliminary and retrospective, our data raised the hypothesis that the duration of respiratory distress symptoms may play a role in COVID-19 patients' need for invasive mechanical ventilation. Furthermore, our observations suggested that specific strategies may be directed towards identifying and managing early symptoms of respiratory distress, regardless of the levels of hypoxemia and the severity of the dyspnoea itself.

Computational Fluid Dynamics (CFD) Analysis of Subject-specific Bronchial Tree Models in Lung Cancer Patients.

Lung resection is the only potentially curative treatment for lung cancer. The inevitable partial removal of functional lung tissue along with the tumoral mass requires a careful and structured pre-operative condition of patients. In particular, the postoperative residual functionality of the lung needs to be predicted. Clinically, this is assessed through algorithms based on pulmonary function tests (PFTs). However, these approaches neglect the local airway segment's functionality and provide a globally averaged evaluation. CFD was demonstrated to provide patient-specific, quantitative, and local information on flow dynamics and regional ventilation in the bronchial tree. This study aims to apply CFD to characterize the flow dynamics in 12 patients affected by lung cancer and evaluate the effects of the tumoral masses on flow parameters and lobar flow distribution. Patient-specific airway models were reconstructed from CT images, and the tumoral masses were manually segmented. Measurements of lungs and tumor volumes were collected. A peripherality index was defined to describe tumor distance from the parenchyma. CFD simulations were performed in Fluent®, and the results were analyzed in terms of flow parameters and lobar volume flow rate (VFR). The predicted postoperative forced expiratory volume in 1s (ppoFEV1) was estimated and compared to the current clinical algorithm. The patients under analysis showed relatively small tumoral masses located close to the lung parenchyma. CFD results did not highlight lobar alterations of flow parameters, whereas the flow to the lung affected by the tumor was found to be significantly lower (p=0.026) than the contralateral lung. The estimation ppoFEV1 obtained through the results of the simulations showed a high correlation (ρ=0.993, p<0.001) with the clinical formula.Clinical Relevance- The proposed study establishes the efficacy and applicability of CFD for the pre-operative characterization of patients undergoing lobectomy surgery. This technique can provide additional information on local functionality and flow dynamics to support patients' operability.

Human fibroblasts and 900 MHz radiofrequency radiation: evaluation of DNA damage after exposure and co-exposure to 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5h)-furanone (MX).

The aim of this study was to investigate DNA damage in human dermal fibroblasts from a healthy subject and from a subject affected by Turner's syndrome that were exposed for 24 h to radiofrequency (RF) radiation at 900 MHz. The RF-radiation exposure was carried out alone or in combination with 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), a well-known environmental mutagen and carcinogen produced during the chlorination of drinking water. Turner's syndrome fibroblasts were also exposed for a shorter time (1 h). A signal similar to that emitted by Global System for Mobile Communications (GSM) mobile phones was used at a specific absorption rate of 1 W/kg under strictly controlled conditions of temperature and dosimetry. To evaluate DNA damage after RF-radiation exposure alone, the alkaline comet assay and the cytokinesis-block micronucleus assay were used. In the combined-exposure experiments, MX was given at a concentration of 25 microM for 1 h immediately after the RF-radiation exposure, and the effects were evaluated by the alkaline comet assay. The results revealed no genotoxic and cytotoxic effects from RF radiation alone in either cell line. As expected, MX treatment induced an increase in DNA migration in the comet assay, but no enhancement of the MX-induced DNA damage was observed in the cells exposed to RF radiation.

Exposure to 900 MHz radiofrequency radiation induces caspase 3 activation in proliferating human lymphocytes.

In this study, the induction of apoptosis after exposure to 900 MHz radiofrequency radiation (GSM signal) was investigated by assessing caspase 3 activation in exponentially growing Jurkat cells and in quiescent and proliferating human peripheral blood lymphocytes (PBLs). The exposure was carried out at an average specific absorption rate of 1.35 W/kg in a dual wire patch cell exposure system where the temperature of cell cultures was accurately controlled. After 1 h exposure to the radiofrequency field, a slight but statistically significant increase in caspase 3 activity, measured 6 h after exposure, was observed in Jurkat cells (32.4%) and in proliferating human PBLs (22%). In contrast, no effect was detected in quiescent human PBLs. In the same experimental conditions, apoptosis was also evaluated in Jurkat cells by Western blot analysis and in both cell types by flow cytometry. To evaluate late effects due to caspase 3 activity, flow cytometry was also employed to assess apoptosis and viability 24 h after radiofrequency-radiation exposure in both cell types. Neither the former nor the latter was affected. Since in recent years it has been reported that caspases are also involved in processes other than apoptosis, additional cell cycle studies were carried out on proliferating T cells exposed to radiofrequency radiation; however, we found no differences between sham-exposed and exposed cultures. Further studies are warranted to investigate the biological significance of our findings of a dose-response increase in caspase 3 activity after exposure to radiofrequency radiation.

Influence of trunk curl-up speed on muscular recruitment.

Although exercise speed is an acute variable to prescribe abdominal strengthening programs, current literature lacks studies analyzing the influence of speed on muscular activation in abdominal exercises. The aim of this work was to determine the influence of trunk curl-up speed on the amplitude of muscular activation and the way in which the trunk muscles were coactivated. Twenty recreationally trained volunteers (16 women and 4 men; age, 23.7 +/- 4.3 years; height, 166.2 +/- 6.3 cm; mass, 61.0 +/- 8.2 kg) participated in this study. Surface electromyographic data were collected from the rectus abdominis, external oblique, internal oblique, and erector spinae during 4 different curl-up cadences [1 repetition per 4 seconds (C4), 1 repetition per 2 seconds (C2), 1 repetition per 1.5 seconds (C1.5), 1 repetition per 1 second (C1)], and during maximum speed curl-ups (Cmax). The electromyographic amplitude was averaged and normalized using maximum voluntary isometric contractions (MVICs). Statistical analyses were performed using repeated-analyses of variance. Normalized electromyographic mean amplitudes of trunk muscles increased with curl-up speed. Although the rectus abdominis (ranged from 23.3% of MVICs at C4 to 49.6% of MVICs at Cmax) and internal oblique (ranged from 19.2% of MVICs at C4 to 48.5% of MVICs at Cmax) were the most active analyzed muscles at each speed, contribution of the external oblique increased appreciably with velocity (ranged from 5.3% of MVICs at C4 to 33.3% of MVICs at Cmax). Increasing trunk curl-up speed supposed greater trunk muscular coactivation, probably required for a faster performance and to ensure dynamic spine stability. On the basis of our findings, curl-up speed had an important effect on trunk muscular recruitment and must be taken into account when prescribing exercise programs for abdominal conditioning.

Straatsma syndrome: Satisfactory amblyopia treatment. Report of two cases. / Síndrome de Straatsma: tratamiento satisfactorio de ambliopía.Reporte de 2 casos.

CLINICAL CASES: The cases are presented on 2 female patients with Straatsma syndrome, with satisfactory treatment of amblyopia. DISCUSSION: The level of anisometropia and myelination of retinal nerve fibres were different in these two patients. However, both achieved 0.20 (logMAR) visual acuity with correction in both eyes following amblyopia treatment with ocular patching. Visual prognosis of amblyopia associated with myelination of retinal nerve fibres and anisometropia is poorer than anisometropic amblyopia without myelination. It is well known that the former is refractory to occlusive therapy. Despite having a poor prognosis, visual rehabilitation should be attempted. The two cases presented were successfully treated with eye-patching.

Cytogenetic observations in human peripheral blood leukocytes following in vitro exposure to THz radiation: a pilot study.

Emerging technologies are considering the possible use of Terahertz radiation in different fields ranging from telecommunications to biology and biomedicine. The study of the potential effects of Terahertz radiation on biological systems is therefore an important issue in order to safely develop a variety of applications. This paper describes a pilot study devoted to determine if Terahertz radiation could induce genotoxic effects in human peripheral blood leukocytes. For this purpose, human whole blood samples from healthy donors were exposed for 20 min to Terahertz radiation. Since, to our knowledge, this is the first study devoted to the evaluation of possible genotoxic effects of such radiation, different electromagnetic conditions were considered. In particular, the frequencies of 120 and 130 GHz were chosen: the first one was tested at a specific absorption rate (SAR) of 0.4 mW g-1, while the second one was tested at SAR levels of 0.24, 1.4, and 2 mW g-1. Chromosomal damage was evaluated by means of the cytokinesis block micronucleus technique, which also gives information on cell cycle kinetics. Moreover, human whole blood samples exposed to 130 GHz at SAR levels of 1.4 and 2 mW g-1 were also tested for primary DNA damage by applying the alkaline comet assay immediately after exposure. The results obtained indicate that THz exposure, in the explored electromagnetic conditions, is not able to induce either genotoxicity or alteration of cell cycle kinetics in human blood cells from healthy subjects.

Colonization by multidrug-resistant organisms in long-term care facilities in Italy: a point-prevalence study.

OBJECTIVES: To determine prevalence and risk factors for colonization by multidrug-resistant organisms (MDROs) in long-term care facility (LTCF) residents in Italy. Genotypes of MDRO isolates were investigated. METHODS: A point-prevalence study was conducted at 12 LTCFs located in four Italian cities (2 February to 14 March 2015). Rectal swabs, faeces and nasal/auxiliary swabs were cultured for extended-spectrum ß-lactamase (ESBL)- and/or carbapenemase-producing Enterobacteriaceae, Clostridium difficile and methicillin-resistant Staphylococcus aureus (MRSA) respectively. Antimicrobial susceptibility testing, detection of ESBL and/or carbapenemase genes and molecular typing of MDROs were performed. Risk factors for colonization were determined by univariate and multivariate analysis. RESULTS: A total of 489 LTCF residents aged ≥65 years were enrolled. The prevalence of colonization by ESBL-producing Enterobacteriaceae, MRSA and C. difficile was 57.3% (279/487), 17.2% (84/487) and 5.1% (21/409) respectively. Carriage rate of carbapenemase-producing Enterobacteriaceae was 1% (5/487). Being bedridden was a common independent risk factor for colonization by all MDROs, although risk factors specific for each MDRO were identified. ESBL-producing Escherichia coli carriage was associated with the sequence type (ST) 131-H30 subclone, but other minor STs predominated in individual LTCF or in LTCFs located in the same city, suggesting a role for intrafacility or local transmission. Similarly, MRSA from LTCF residents belonged to the same spa types/ST clones (t008/ST8 and t032/ST22) commonly found in Italian acute-care hospitals, but infrequent spa types were recovered in individual LTCFs. The prevalent C. difficile PCR ribotypes were 356/607 and 018, both common in Italian acute-care hospitals. CONCLUSIONS: MDRO colonization is common among residents in Italian LTCFs.

Insulin-like growth factor I receptor-mediated circuit in Ewing's sarcoma/peripheral neuroectodermal tumor: a possible therapeutic target.

The disappointingly low survival rate observed in Ewing's sarcoma (ES)/peripheral neuroectodermal tumor (PNET) despite the adoption of aggressive multimodal treatments prompted us to study the existence of autocrine circuits to be used as innovative therapeutic targets. Of the several circuits analyzed, only the insulin-like growth factor receptor (IGF-IR)-mediated loop was found to be constantly present both in cell lines and clinical samples, suggesting a role for this autocrine circuit in the pathogenesis of ES/PNET. The in vitro inhibition of the IGF-IR-mediated circuit by the specific IGF-IR binding antibody alphaIR3 suppressed the growth of ES/PNET cells by decreasing the proliferative rate and increasing apoptosis. alphaIR3 also significantly inhibited the ability of ES/PNET cells to grow in soft agar and to migrate following a chemotactic stimulus. Inactivation of the IGF-IR signaling pathway may therefore be considered as an effective therapeutic modality for patients with ES/PNET.

Expression of the myeloperoxidase gene in acute and chronic myeloid leukemias: relationship to the expression of cell cycle-related genes.

The expression of the myeloperoxidase (MPO) gene was studied, by means of Northern blot analysis in 14 cases of acute myeloid leukemia (AML), 11 cases of chronic myeloid leukemia (CML), and 6 cases of CML blast crisis, and in HL60 cells before and after induction of terminal differentiation with retinoic acid (RA), phorbol esters (TPA), or vitamin D. The expression of a panel of cell cycle-related genes, namely C-MYC, histone H3, ornithine decarboxylase, P53, vimentin, and calcyclin, was also studied in the same cell populations. Our results indicate that: (a) MPO gene expression (steady state mRNA levels) is strictly confined to the first stages of myeloid differentiation, reaching its peak at the promyelocyte stage and becoming undetectable in mature granulocytes and monocytes; (b) cells devoid of any detectable MPO enzymatic activity such as leukemic basophils have a high content of MPO mRNA; and (c) MPO gene expression is not related to the growth activity of the cell population. Finally, our results show that the pattern of expression of growth-regulated genes in the neoplastic myeloid disorders AML, CML, and CML blast crisis is remarkably different.

Quantitative comparison between the commercial software STRATOS(®) by Philips and a homemade software for voxel-dosimetry in radiopeptide therapy.

BACKGROUND: Targeted radionuclide therapy is a rapidly growing modality. A few commercial treatment planning systems are entering the market. However, some in-house systems are currently developed for a more flexible and customized dosimetry calculation at voxel-level. For this purpose, we developed a novel software, VoxelMed, and performed a comparison with the software STRATOS. METHODS: The validation of both of them was undertaken using radioactive phantoms with different volume inserts. A cohort of 10 patients was also studied after a therapeutic administration of (177)Lu-labelled radiopeptides. The activity, number of disintegrations, absorbed dose and dose-volume histogram (DVH) were calculated for the phantoms and the kidneys in patients, which were the main critical organs at risk in this study. RESULTS: In phantoms the absorbed doses computed with VoxelMed and STRATOS agree within 5%. In patients at the voxel-level the absorbed dose to kidneys (VoxelMed: mean 0.66 Gy/GBq) showed a limited difference of 5%, but with a remarkable range (-40%, +60%) between the two software packages. Voxel-dosimetry allows to estimate the dose non-homogeneities in volumes, which may be evaluated through DVHs. CONCLUSION: This study demonstrates that a fully 3D voxel-dosimetry with multiple SPECT images is feasible by using home-made or commercial software package and absorbed dose results obtained are similar. The main difference between the studied tools was observed in the activity integration method (effective vs physical half-time to time activity curve tail). We believe that an effective half-time integration method produces a more accurate approximation of clinical uptake and resultant dosimetry.

Analysis of P-glycoprotein expression in osteosarcoma.

Current treatment of high-grade osteosarcoma combines surgical removal of the lesion with chemotherapy. In this study we evaluated whether the expression of P-glycoprotein, a protein closely associated with multidrug resistance, may be helpful in identifying the patients whose tumours will be further resistant to specific agents. By using multidrug-resistant osteosarcoma cell lines as standards, the expression of P-glycoprotein was evaluated in 105 cases of primary and metastatic osteosarcoma by semiquantitative immunofluorescence. Overexpression of the protein was shown in 23% of primary and in 50% of metastatic lesions. In 38 cases, homogeneously treated and followed-up for at least 24 months, overexpression of P-glycoprotein appeared to be associated with a higher relapse rate and with a trend toward a worse outcome. These data support the role of P-glycoprotein in the response to chemotherapy and its involvement in the determination of the outcome of osteosarcoma patients.

Kappa light chain gene rearrangement in a T-cell lymphoma.

Forty-three patients were studied to determine whether light chain gene rearrangements may occur in hematopoietic cells not pertaining to the B-lineage. In only one patient, affected by T-cell lymphoblastic lymphoma, one kappa light chain allele was rearranged. Neither at the protein level nor at the RNA level the rearranged gene was expressed. These data confirm that, although rarely, kappa light chain gene rearrangements may occur in neoplastic T-cells. Furthermore, as in our patient Ig heavy chain genes retained a germline configuration, the present data demonstrate that kappa light chain gene rearrangements may occur regardless of Ig heavy chain gene arrangement.

Absence of stimulatory effect of g-csf on the growth of human sarcoma-cells.

The Colony-Stimulating Factors (CSFs) are undergoing clinical trials for their ability to stimulate the regeneration of bone marrow in patients receiving anticancer chemotherapy. However, the reported effects on the growth of tumor cell lines of different derivations, including osteosarcoma, raise the possibility that the use of these cytokines may induce proliferative effects also in residual tumor cells. In this study, we have used a panel of 12 human osteosarcoma (2 cell lines and 10 primary cultures) and 7 Ewing's sarcoma cell lines (5 cell lines and 2 primary cultures) to evaluate the presence of the G-CSF receptor by RT-PCR and the effects of recombinant Human (rHu) G-CSF on their in vitro growth ability. RT-PCR did not reveal the presence of G-CSF receptor band in any of the osteosarcoma or Ewing's cell lines examined. Moreover, after exposure to rHuG-CSF, no significant stimulatory or inhibitory effects were observed in any of the cell lines. Therefore, G-CSF may be safely used to stimulate marrow regeneration after high-dose chemotherapy both in osteosarcoma and in Ewing's sarcoma.

Value of immunohistochemical detection of noncollagenous proteins of bone for the diagnosis of bone tumours.

The expression of osteonectin, osteopontin, bone sialoprotein, and osteocalcin was evaluated by immunohistochemistry in 57 cases of osteoid-forming and non-osteoid-forming bone tumours using specific polyclonal antibodies and the avidin-biotin peroxidase complex method. A positive immunostaining was found in all of the osteoid-forming rumours (osteoblastoma and osteosarcoma), both in the cells and in the extracellular matrix. Among non-osteoid-forming tumours, immunoreactivity to noncollagenous proteins was present in the cells but not in the matrix of chondrosarcoma, malignant fibrous histiocytoma, and fibrosarcoma, as well as in the mononuclear component of giant-cell rumours. Contrary to small-cell osteosarcoma, Ewing's sarcoma was always negative for all of the noncollagenous proteins considered. These results suggest that the immunohistochemical evaluation of noncollagenous proteins of bone may be a useful tool for the differential diagnosis of bone neoplasms, particularly among the heterogeneous group of small round cell tumours.

Immunostaining of the p30/32MIC2 antigen and molecular detection of EWS rearrangements for the diagnosis of Ewing's sarcoma and peripheral neuroectodermal tumor.

The identification of Ewing's sarcoma (ES) and peripheral neuroectodermal tumor (PNET) among other small round cell tumors (SRCTs) is a critical issue in musculoskeletal pathology because of the lack of clearly distinctive morphological features. In this study, the authors have compared advantages and limits of two procedures that were recently suggested as additional tools for the identification of ES/PNET, the analysis of p30/32MIC2 antigen by immunohistochemistry, and the evaluation of the fusion products of two specific chromosomal aberrations, the t(11;22)(q24;q12) and the t(21;22)(q22;q12), by reverse transcriptase-polymerase chain reaction (RT-PCR). The authors have analyzed the expression of p30/32MIC2 in 28 cell lines and in 90 tumor samples. p30/32MIC2 was highly expressed in ES/PNET but was also present in all the other cell types. The broad spectrum of positivity for p30/32MIC2 in SRCTs of bone was substantially confirmed by the analysis of tissue samples. In the same material, the authors have evaluated the presence of t(11;22) or t(21;22) transcripts (EWS/FLI-1 and EWS/ERG, respectively) by RT-PCR. These transcripts were found in all the cell lines and tissue samples of ES/PNET, but not in other tumors. The authors' results question the use of p30/32MIC2 immunostaining alone for the identification of ES/PNET and suggest the adoption of RT-PCR as an advantageous alternative. Molecular diagnosis of ES/PNET by RT-PCR is highly specific and can be applied to small amounts of tissue. Moreover, RNA extracted from paraffin-embedded specimens was shown to be suitable for RT-PCR analysis, thus enabling analysis of archival material.

Long-term follow-up for patients with Ewing's sarcoma of bone treated with adjuvant and neoadjuvant chemotherapy.

The long-term results obtained in 252 patients with non-metastatic Ewing's sarcoma of bone treated between March 1972 and June 1988 according to three sequential protocols of treatment were evaluated. Primary tumor was treated with radiotherapy in 125 cases, with surgery in 52 and with surgical resection followed by radiotherapy in 75. In the first protocol (REA 1; 1972-78) chemotherapy was performed with a 3-drug regimen (VCR, CPM, ADM), whereas in the REA 2 protocol (1979-82) and in the REN 1 protocol (1983-88) a 4-drug regimen was used (VCR, CPM, ADM, ACTD). Chemotherapy was delivered as adjuvant treatment in REA 1 and 2, and as neoadjuvant in the last study. At a mean follow-up of 14.8 years, with the 95% of patients with a minimum FU of 10 years, 101 pts (40%) remained continuously free of disease, 144 patients relapsed, two patients died of adriamycin cardiotoxicity and 5 patients developed a second neoplasm. 6% of the patients relapsed 5 or more years after the diagnosis with the latest recurrence registered at the tenth year. Type of local treatment, LDH serum level, chemotherapy protocol and sex were predictive factors of DFS after a multivariate analysis. The possibility of late relapse in Ewing's sarcoma has been confirmed by our retrospective study and for patients with Ewing's sarcoma, a 10-year follow up should be recommended.

Response of the flexion-relaxation phenomenon relative to the lumbar motion to load and speed.

STUDY DESIGN: Nonrandomized control trial. OBJECTIVE: To determine if the variations of speed and loading conditions during trunk flexion-extension could influence the times of occurrence and disappearance of the electrical silence of the erector spinae muscles, the degrees of lumbar flexion at those instants, and the relative lumbar motion time. SUMMARY OF BACKGROUND DATA: It has been suggested that varying either the speed of movement or the load on the trunk during trunk flexion-extension movements may influence the flexion-relaxation phenomenon or the kinesiologic data. However, no study dealt with the simultaneous effect of the speed of movement on the spine rhythm and on the occurrence of the electrical silence of the erector spinae. METHODS: A total of 22 pain-free volunteers performed a series of trunk flexion-extension movements varying the speed and load. The motion of the lumbar spine ( degrees ) and the integrated electromyography (microV) of erector spinae muscles were simultaneously recorded. Two measures were calculated: the percentage of the maximum lumbar spine flexion at the instants when changes of electrical activity represented the beginning and end of the electrical silence and the relative lumbar spine motion time during trunk flexion and extension movements. RESULTS: The increase in speed of movement significantly increased the relative lumbar flexion time and significantly reduced the relative lumbar extension time (t = 2.49 and t = 2.25, P < 0.05); furthermore, it significantly delayed the appearance of the electrical silence in the range of flexion (t = 3.52, P < 0.01). There was no significant effect from a change in load. CONCLUSIONS: The relative spine motion time differed depending on the direction of movement, being longer during trunk flexion and shorter during extension. The increase in speed of movement produced greater differences in the relative time between trunk flexion and extension; furthermore, it delayed the appearance of the electrical silence of the erector spinae muscles in the range of flexion.

Biological activity and binding of estradiol to SK-Mel 23 human melanoma cells.

Patients expressing estradiol receptors in melanoma cells have been reported to have a better prognosis. We therefore decided to investigate the in vitro effects of beta-estradiol and tamoxifen on the growth and tyrosinase activity of SK-Mel 23 human melanoma cells. Twenty-four-hour treatment with 0.4 nM beta-estradiol inhibited cell proliferation in 30% (0.70 +/- 0.03 x 10(5) cells) and increased tyrosinase activity in 50% (7130.5 +/- 376.5 cpm/10(5) cells), as compared to untreated cells (1.0 +/- 0.05 x 10(5) cells and 4769 +/- 25.5 cpm/10(5) cells, respectively). Both responses were completely (100%) blocked by 1 microM tamoxifen. Higher concentrations (up to 1.6 nM) or longer treatments (up to 72 h) did not result in a larger effect of the hormone on proliferation or tyrosinase activity. Competition binding assays demonstrated the presence of binding sites to [2,4,6,7-3H]-beta-estradiol, and that the tritiated analogue was displaced by the unlabeled hormone (1 nM to 100 microM, Kd = 0.14 microM, maximal displacement of 93%) or by 10 microM tamoxifen (displacement of 60%). Beta-estradiol also increased the phosphorylated state of two proteins of 16 and 46 kDa, after 4-h treatment, as determined by Western blot. The absorbance of each band was 1.9- and 4-fold the controls, respectively, as determined with Image-Pro Plus software. Shorter incubation periods with beta-estradiol did not enhance phosphorylation; after 6-h treatment with the hormone, the two proteins returned to the control phosphorylation levels. The growth inhibition promoted by estradiol may explain the better prognosis of melanoma-bearing women as compared to men, and open new perspectives for drug therapy.