Inhibition of HMGI-C protein synthesis suppresses retrovirally induced neoplastic transformation of rat thyroid cells.

Elevated expression of the three high-mobility group I (HMGI) proteins (HMGI, HMGY, and HMGI-C) has previously been correlated with the presence of a highly malignant phenotype in epithelial and fibroblastic rat thyroid cells and in experimental thyroid, lung, mammary, and skin carcinomas. Northern (RNA) blot and run-on analyses demonstrated that the induction of HMGI genes in transformed thyroid cells occurs at the transcriptional level. An antisense methodology to block HMGI-C protein synthesis was then used to analyze the role of this protein in the process of thyroid cell transformation. Transfection of an antisense construct for the HMGI-C cDNA into normal thyroid cells, followed by infection with transforming myeloproliferative sarcoma virus or Kirsten murine sarcoma virus, generated cell lines that expressed significant levels of the retroviral transforming oncogenes v-mos or v-ras-Ki and removed the dependency on thyroid-stimulating hormones. However, in contrast with untransfected cells or cells transfected with the sense construct, those containing the antisense construct did not demonstrate the appearance of any malignant phenotypic markers (growth in soft agar and tumorigenicity in athymic mice). A great reduction of the HMGI-C protein levels and the absence of the HMGI(Y) proteins was observed in the HMGI-C antisense-transfected, virally infected cells. Therefore, the HMGI-C protein seems to play a key role in the transformation of these thyroid cells.

A case of endobronchial paraganglioma.

Paragangliomas are rare lung tumours; endobronchial localisation is even more rare. This report describes the case of a 59-year-old patient with a symptomatic endobronchial paraganglioma successfully resected by means of pulmonary lobectomy. Recognition of this uncommon tumour can lead to a correct diagnosis and therapeutic strategy.

Preoperative assessment in patients with postintubation tracheal stenosis : Rigid and flexible bronchoscopy versus spiral CT scan with multiplanar reconstructions.

BACKGROUND: Postintubation stenosis remains the most frequent indication for tracheal surgery. Rigid bronchoscopy has traditionally been considered the technique of choice for the preoperative diagnostic assessment. However, this technique is not routinely available, and new techniques such as flexible videobronchoscopy and spiral computed tomography (CT) scan with multiplanar reconstructions have been proposed as alternatives to rigid bronchoscopy. The aim of this study was to compare these techniques in the diagnostic assessment of patients with tracheal stenosis submitted to surgical treatment. METHODS: Twelve patients who underwent airway resection and reconstruction for postintubation tracheal and laryngotracheal stenosis were preoperatively evaluated with rigid and flexible bronchoscopy and with spiral CT scan with multiplanar reconstructions. The following parameters were examined: involvement of subglottic larynx, length of the stenosis, and associated lesions. The results were compared with the intraoperative findings. RESULTS: The accuracy of rigid bronchoscopy, flexible bronchoscopy, and CT scan in the evaluation of the involvement of subglottic larynx was, respectively, 92%, 83%, and 83%. The evaluation of the length of the stenosis was correct in 83%, 92%, and 25% of the patients, respectively, with rigid bronchoscopy, flexible bronchoscopy, and CT scan. A significant correlation was observed between the length of the stenosis measured intraoperatively and preoperatively with rigid (p < 0.001) and flexible bronchoscopy (p < 0.05) but not with CT scan (p = 0.08). The three techniques correctly showed the presence of an associated tracheoesophageal fistula in two patients, but CT scan did not correctly show the exact location of the fistula in relation to the airway. Flexible bronchoscopy was the only effective technique in the assessment of laryngeal function. CONCLUSIONS: Rigid bronchoscopy remains the procedure of choice in the evaluation of candidates for tracheal resection and reconstruction for postintubation stenosis, and it should be available in centers that perform surgery of the airway. Flexible bronchoscopy and CT scan have to be considered complementary techniques in the evaluation of laryngeal function and during follow-up.

Expression of HMGI(Y) proteins in squamous intraepithelial and invasive lesions of the uterine cervix.

The expression of nuclear proteins high mobility group (HMG) I and HMGY was investigated in intraepithelial and invasive lesions of the uterine cervix. Human carcinoma cell lines C-41, ME-180, and CaSki were used for testing protein expression in neoplastic cells from the cervix. Morphological grading of the dysplasias (CIN 1, CIN 2, and CIN 3) and invasive carcinomas from formalin-fixed paraffin-embedded samples parallels the degree of nuclear immunostaining obtained using a polyclonal antibody raised against the amino-terminal region of HMGI(Y) proteins. The immunostaining obtained with HMGI(Y) antibody was compared with that observed using the antibody Ki-67, and the results were similar. We suggest the use of HMGI(Y) antibody in clinical oncology as a useful marker of intraepithelial lesions and invasive carcinomas.

Human colorectal carcinomas express high levels of high mobility group HMGI(Y) proteins.

A correlation has previously been demonstrated between the presence of the three HMGI proteins (HMGI, HMGY, and HMGI-C) and the expression of a highly malignant phenotype in epithelial and fibroblastic rat thyroid cells; this being subsequently extended to experimental thyroid, lung, prostate, mammary, and skin carcinomas. Recently, we have demonstrated that expression of HMGI and HMGY proteins, coded for by the HMGI(Y) gene, is associated with the malignant phenotype of human thyroid neoplasias. Here, we show that HMGI(Y) gene expression is present both at the RNA and protein level in human colorectal carcinoma cell lines and tissues examined in this study. Conversely, no HMGI(Y) proteins were detected in normal intestinal mucosa. Therefore, these results suggest an involvement of HMGI and HMGY proteins overexpression in colorectal tumorigenesis.

The expression of the high mobility group HMGI (Y) proteins correlates with the malignant phenotype of human thyroid neoplasias.

High Mobility Group I (HMGI) proteins are nuclear proteins involved in the regulation of chromatin structure and function. Elevated expression of the HMGI proteins (HMGI, HMGY and HMGI-C) has been correlated with the presence of a highly malignant phenotype in epithelial and fibroblastic rat thyroid cells, and in several experimental carcinomas. Here, we demonstrate that HMGI and HMGY proteins are expressed in human thyroid carcinomas and thyroid carcinoma cell lines, but not in adenomas, goiters, normal thyroid tissues and cells. These results indicate a correlation between HMGI and HMGY expression and the malignant phenotype of thyroid neoplasias, suggesting that these proteins may be used as markers in thyroid cancer.

Clinical application of exercise stress echocardiography: supine bicycle or treadmill?

Although exercise stress echocardiography is currently used to evaluate coronary artery disease (CAD) patients, the best exercise methodology is still undefined. The objectives of the study were: (1) to compare supine bicycle stress echocardiography (SBSE) and treadmill in the evaluation of CAD; and (2) to define, in normal subjects, the different behavior of factors determining MVO2 with treadmill and SBSE. We selected 10 male patients with CAD (group A), and 10 male control subjects (group B). Each patient underwent SBSE and treadmill testing in random order. We studied heart rate, systolic blood pressure, heart rate x systolic blood pressure, and end-diastolic and end-systolic volume indexes. In group A, we also studied wall motion score index (according to the American Society of Echocardiography) and in group B, systolic blood pressure/end-systolic volume index. The results were as follows: Group A: SBSE resulted in significantly lower work load, heart rate, and significantly higher systolic blood pressure, heart rate x systolic blood pressure, end-diastolic volume index, end-systolic volume index, and wall motion score index. SBSE showed wall motion abnormalities in each patient, whereas treadmill did not detect wall motion abnormalities in 4 patients (3 single-vessel; 1 multivessel); of the other 6 patients, 2 showed a lower wall motion score index and 4 did not show any difference in left ventricle kinetics with the 2 methodologies of exercise. Mean acquisition time for postexercise images was 72 +/- 6 seconds. Group B: SBSE resulted in lower work load, heart rate, heart rate x systolic blood pressure, systolic blood pressure/end-systolic volume index, and higher end-diastolic volume index and end-systolic volume index. Systolic blood pressure was similar with SBSE and treadmill testing. In conclusion, our experience suggests SBSE is a highly accurate diagnostic tool for evaluating CAD compared with treadmill testing; the maximum cardiovascular performance can be achieved with lower values of heart rate, suggesting the echo test is more feasible. Treadmill testing could lose important information about the existence, extension, and location of CAD; in contrast, SBSE detects even small, quickly reversible wall motion abnormalities.

QT dispersion variability and myocardial viability in acute myocardial infarction.

The aim of this study was to evaluate QT dispersion in acute and sub-acute stages of myocardial infarction and clarify the relationship between QT dispersion and myocardial viability. We studied 95 patients with acute myocardial infarction. The QT dispersion values were compared to those of a control group of 50 healthy subjects. In the patients with acute myocardial infarction dispersion of ventricular repolarization was evaluated on the standard electrocardiograms obtained at the time of admission and ten days later. Two-dimensional echocardiography examination was performed to assess and compare left ventricular wall motion at different stages. QT dispersion values were increased in patients with acute myocardial infarction and levels were higher in the early than in late phases. A better recovery of QT dispersion was found in those patients who demonstrated improvement of left ventricular contractility. The modifications of QT dispersion can reflect the alterations of myocardial contractility.

Mitochondrial localization of APE/Ref-1 in thyroid cells.

Mutations of mitochondrial DNA (mtDNA) are associated with different human diseases, including cancer and aging. Reactive oxygen species produced during oxidative phosphorylation are a major source of mtDNA damage. It is not clear, however, whether DNA repair mechanisms, able to abolish effects due to oxidative damage, are present in mitochondria. APE/Ref-1 is a nuclear protein possessing both redox activity (by which activates, "in vitro", the DNA-binding functions of several transcription factors) and DNA repair activity over apurinic/apyrimidinic sites. Immunohistochemical evidences indicate that in follicular thyroid cells, APE/Ref-1 is located in both nucleus and cytoplasm. Electronmicroscopy immunocytochemistry performed in the rat thyroid FRTL-5 cell line, indicates that part of the cytoplasmatic APE/Ref-1 is located in mitochondria. The presence of APE/Ref-1 inside mitochondria is further demonstrated by western blot analysis after cell fractionation. In the Kimol cell line (which is derived from FRTL-5, transformed by the Ki-ras oncogene) the amount of mitochondrial APE/Ref-1 is reduced by three to fourfold with respect to the normal FRTL-5 cells. These results suggest that: (i) a machinery capable of repairing DNA damaged by oxidative stress is present in mitochondria and (ii) mtDNA repair mechanisms may be impaired during cell transformation.

Ondansetron-induced headache. Our experience in gynecological cancer.

Chemotherapy-induced emesis is one of the major problems in the treatment of oncologic patients. Recently, a novel class of compounds, the selective 5HT3 receptor antagonists, has been introduced, achieving a dramatic improvement in the control of emesis. The absence of extrapyramidal side effects adds to their safety and good tolerability. The Authors herein analyse their experience on 269 cycles of chemotherapy in 47 patients treated for gynaecological and breast malignancies, with particular regard to adverse events such as headache. Their most frequent side-effects are headache and constipation, that are usually mild and self-limiting. Nevertheless, in some cases, severe, rebel headache has been reported, leading in our experience in 6.4% of cases to discontinuation of the antiemetic regimen. A previous history of recurrent or severe headache or migraine is not correlated with the occurrence of ondansetron-induced headache, as severe headache occurred after ondansetron only in 28.4% of the patients with positive anamnesis, and 70% of the patients that experienced had never suffered from severe headache before. In those patients complaining of severe headache, the Authors suggest an antiemetic association, with a loading dose of ondansetron i.v., followed by metoclopramide i.m. orally for the following days.

Infiltrating lobular carcinoma of the breast (ILC). Diagnostic and therapeutic aspects.

Invasive lobular carcinoma of the breast (ILC) is the second most common form of mammary neoplasm after ductal carcinoma. Due to its histological features, characterised by a diffuse infiltration of the tissue by malignant cells with scarce fibrotic reaction, lobular carcinoma often presents clinical and instrumental diagnostic difficulties. The Authors present a retrospective series of 28 patients with lobular carcinoma, with special regard to the diagnostic work-up. Clinical examination misdiagnosed 10 cases, in which only a mild thickening of the breast parenchyma was present. Mammographic false negatives were 21.4% (6 cases). Ultrasonography allowed correct diagnosis in 5 out of these 6 cases. Fine needle aspiration (FNA) had low sensitivity (33% false negatives). ILC constitutes a diagnostic challenge, often requiring the combination of multiple diagnostic tools.

Ondansetron in chemotherapy-induced emesis. Our experience.

UNLABELLED: Chemotherapy-induced nausea and vomiting is one of the major side effects of antiblastic treatment in cancer patients, seriously affecting both the compliance of the patient to therapy and his or her quality of life. OBJECTIVE: The Authors present their experience in the use of ondansetron in 47 patients receiving 186 cycles of chemotherapy for breast or genital neoplasms. RESULTS: Successful control of vomiting was achieved in the first 24 hours, in 74% of the cycles containing cisplatin and 82% of the cycles without cisplatin, if ondansetron was used. On delayed vomiting the difference in results was not as striking as on acute emesis. CONCLUSIONS: The major side effects with ondansetron were headaches (42.4%) and constipation (48.9%). As expected, no extra-pyramidal symptoms were observed in this group, versus 13.3% of the patients treated with metoclopramide.

[Gn-RH analogs in the treatment of benign gynecologic diseases: current trends]. / Gli analoghi del Gn-RH nel trattamento della patologia ginecologica benigna: orientamenti attuali.

Gn-RH analogues have been recently employed for the treatment of oestrogen-dependent benign gynaecological disorders, such as uterine myomata, endometriosis or metrorrhagia. They induce a "pharmacological castration", inducing a marked reduction of serum oestrogen levels. They proved more effective than other drugs used up to now in the medical treatment of these benign gynaecological diseases. Thus they were initially employed in every case. Later it became clear that Gn-RH analogues need a selective indication. The authors herein report their series of 70 patients with benign gynaecological disorders (45 uterine fibroids, 10 endometriosis, 15 metrorrhagia), treated with a Gn-RH analogue depot for 2-3 months preoperatively. They evaluated the efficacy of the treatment in the group with uterine fibroids in terms of disappearance of metrorrhagia, better haemoglobin level in anaemic patients, reduction of fibroids size allowing for a simpler and less extensive surgery (vaginal surgery, myomectomy, hysteroscopic resection). The authors discuss those cases when preoperative treatment with Gn-RH analogues is not indicated, or should be employed only under careful surveillance (in the preparation of multiple myomectomies, big submucosal myomas). In the group of 10 patients with endometriosis we observed the disappearance of pelvic pain and dyspareunia, whereas the size of endometriomas was only minimally reduced. The authors discuss the usefulness of this treatment in case of patients with endometriosis grade I or II (minimal or mild), with desire of children. In the group of 15 perimenopausal patients with metrorrhagia, 10 became amenorrhoic after termination of treatment, thus avoiding surgery. The major benefit for the other 5 patients was a better haemoglobin level at the time of surgery.

[Neoadjuvant chemotherapy in the treatment of breast cancer. Results and prospectives]. / La chemioterapia neoadiuvante nel trattamento del carcinoma della mammella. Risultati e prospettive.

Preoperative chemotherapy for breast cancer has been originally proposed in the treatment of locally advanced tumors (T3b-T4) in order to allow radiotherapy or radical mastectomy. Later, it has been employed also for less advanced stages of the disease (T2-T3), to allow conservative surgery. Personal series of 45 patients that underwent preoperative chemotherapy (FAC) for breast cancer stages T2-T3 is reported. A partial response in terms of reduction of tumor volume was obtained in 80% of these patients, a complete response in 6.6% of the cases. In 48.8% a quadrantectomy has been performed, as the lesion diameter was < 2.5 cm after chemotherapy. The survival rate was 70% at 10 years, and 80% for initially T2 tumors, compared with 50% 10 year survival rate in a group of patients with T2 tumors treated before the introduction of neoadjuvant chemotherapy at our Department. Neoadjuvant chemotherapy allows reduction of the initial volume of breast cancer and performance of conservative instead or radical surgery, with better cosmetic results. Moreover these data suggest that they may improve the plateau of the survival curve of patients with locally advanced breast cancer.

New positron emission tomography derived parameters as predictive factors for recurrence in resected stage I non-small cell lung cancer.

BACKGROUND: The recurrence rate for stage I non-small cell lung cancer is high, with 20-40% of patients that relapse after surgery. The aim of this study was to evaluate new F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) derived parameters, such as standardized uptake value index (SUVindex), metabolic tumor volume (MTV) and total lesion glycolysis (TLG), as predictive factors for recurrence in resected stage I non-small cell lung cancer. METHODS: We retrospectively reviewed 99 resected stage I non-small cell lung cancer patients that were grouped by SUVindex, TLG and MTV above or below their median value. Disease free survival was evaluated as primary end point. RESULTS: The 5-year overall survival and the 5-year disease free survival rates were 62% and 73%, respectively. The median SUVindex, MTL and TLG were 2.73, 2.95 and 9.61, respectively. Patients with low SUVindex, MTV and TLG were more likely to have smaller tumors (p ≤ 0.001). Univariate analysis demonstrated that SUVindex (p = 0.027), MTV (p = 0.014) and TLG (p = 0.006) were significantly related to recurrence showing a better predictive performance than SUVmax (p = 0.031). The 5-year disease free survival rates in patients with low and high SUVindex, MTV and TLG were 84% and 59%, 86% and 62% and 88% and 60%, respectively. The multivariate analysis showed that only TLG was an independent prognostic factor (p = 0.014) with a hazard ratio of 4.782. CONCLUSION: Of the three PET-derived parameters evaluated, TLG seems to be the most accurate in stratifying surgically treated stage I non-small cell lung cancer patients according to their risk of recurrence.

The surgical treatment of lung cancer in patients with previous lymphoproliferative disorders: a historical cohort study.

OBJECTIVES: We undertook a historical cohort study to compare, in terms of morbidity, mortality and long-term survival associated with lung cancer resection, a group of patients with previous lymphoproliferative disorders and a group without a hematological history. METHODS: We identified 29 patients with a previous lymphoproliferative disorder who underwent lung cancer resection. These subjects (Group-A) were matched with 87 patients without a hematological history who underwent pulmonary resection during the same period (Group-B). RESULTS: We found no significant difference between the two groups in length of hospitalization, comorbidities, spirometric parameters, type of surgery, histology, neoadjuvant chemotherapy, morbidity, mortality, median survival (Group-A = 37 months; Group-B = 52 months) and 5-year survival (Group-A = 37%; Group-B = 42%). The mean age of Group-A patients was significantly lower than that of Group-B patients (62 vs 66 years; p = 0.024). Group-A patients had a well differentiated lung cancer more frequently than Group-B patients (p = 0.001). Group-A patients had transitory bacteraemies more frequently than Group-B patients (p = 0.005). Multivariate Cox regression analysis showed that age (p = 0.01) and lung cancer stage (p = 0.04) were significantly associated with survival. CONCLUSIONS: Patients with lymphoproliferative disorders had a lower age and more differentiated lung cancers than those without lymphoproliferative disorders. Patients with lymphoproliferative disorders and those without a hematological history had similar morbidity, mortality and long-term survival after pulmonary resection. Distinguishing patients with and without a lymphoproliferative disorder seems to be of limited value in the decision-making process of evaluating the indications for surgical treatment of lung cancer.

Combined treatment of non-small cell lung cancer with synchronous brain metastases: a single center experience.

AIM: The aim of this study was to analyze our experience with combined treatment of non-small cell lung cancer with synchronous brain metastases. METHODS: Between 1992 and 2008, 31 patients were treated by performing neurosurgery (or stereotactic radiosurgery) and lung surgery. Patients were divided into two groups according to their preoperative mediastinal work-up: group A (CT scan) and group B (FDG-PET scan). RESULTS: Twenty-six patients had one brain metastasis and five had two. Neurosurgery was performed in 10 patients, stereotactic radiosurgery in 20 and both approaches in 1. Seven patients underwent chemotherapy after cerebral procedure. Pulmonary resection was complete in 27 cases and incomplete in 4. Histological findings showed: adenocarcinoma in 19 cases, squamous cell carcinoma in 8 and large cell carcinoma in 4. All patients underwent adjuvant chemotherapy. Overall 1, 2 and 5-year survival rates were 83%, 47% and 21%, respectively. The median survival was 22 months. Univariate analysis showed a better prognosis for complete resection (P=0.008), adenocarcinomas (P=0.015), N0 disease (P=0.038), and Group B (P=0.045). Multivariate analysis showed that only the radicality of the resection (P=0.027) and Group B (P=0.047) were independent prognostic factors. CONCLUSION: Our experience confirms that selected patients with non-small cell lung cancer and synchronous brain metastases may be effectively treated by combined therapy. Complete resection, adenocarcinoma histology and N0 disease were prognostic factors. The incorporation of FDG-PET scan into the preoperative work-up may translate into a survival benefit.

Surgical treatment of multiple primary adenocarcinomas of the lung.

INTRODUCTION: The incidence of lung adenocarcinomas has steadily increased over the last decades. The aim of this study was to assess the results of surgical treatment of multiple primary adenocarcinomas of the lung (MPAL) analyzing the radiological and histological features. METHODS: From 1988 to 2005, 26 patients underwent surgical treatment for MPAL at our department, for a total of 52 tumors. Three patients had synchronous and 23 had metachronous tumors. RESULTS: Thirty-seven tumors were classified as solid, two as ground-glass opacities (GGO) and 13 as mixed solid/GGO tumors on the basis of CT scan evaluation. Histology revealed 26 adenocarcinomas, five adenocarcinomas with a bronchioloalveolar (BAC) pattern and 21 BAC. There was no postoperative mortality. Five-year survival of patients with synchronous tumors was 66 %. Survival of patients with metachronous tumors was 95 % and 70 % from the first and second operation. Patients with stage II and III a tumors had significantly reduced survival rates ( P < 0.05). Survival was 60 % after lobectomy and 78 % after wedge resection. CONCLUSIONS: Surgical treatment of MPAL is associated with favorable results. Sublobar resections, when technically feasible, provide adequate oncological management.

Neoadjuvant chemotherapy and pneumonectomy in a lung cancer patient on hemodialysis.

Major pulmonary resections are rarely performed in non-small cell lung cancer patients on hemodialysis. To date only two cases of pneumonectomy performed in such patients are reported in the literature. Moreover, chemotherapy, as a treatment for advanced non-small cell lung cancer, is not routinely administered to patients with end-stage renal disease requiring hemodialysis. We present the case of a stage IIIB non-small cell lung cancer patient on hemodialysis who successfully underwent neoadjuvant chemotherapy followed by pneumonectomy. To our knowledge, this is the first case of non-small cell lung cancer patient on hemodialysis reported in the literature who successfully underwent this type of combined therapy.

[Supraventricular hyperkinetic arrhythmias in acute myocardial infarct: their prognostic assessment and correlation with the echocardiographic evolution]. / Aritmie ipercinetiche sopraventricolari nell'infarto miocardico acuto: valutazione prognostica e correlazione con l'evoluzione ecocardiografica.

To assess the prognostic significance of supraventricular tachyarrhythmias (SVTA) during acute myocardial infarction (AMI), we studied 388 patients with first AMI, without ventricular preexcitation or chronic atrial fibrillation. The prevalence of SVTA was 14% (56/388), including atrial fibrillation (57%), atrial flutter (22%), polyfocal atrial tachycardia (14%), monofocal atrial tachycardia (7%). The arrhythmia appeared within 72 hours from the onset of chest pain in 61% of patients (early SVTA < 72 hours), while in 39% appeared later (late SVTA > 72 hours). Patients with SVTA (Group I n = 56) and without SVTA (Group II n = 232) were similar regarding prevalence of hypertension, dyslipidemia, diabetes, site of infarction and fibrinolysis, but SVTA was associated with a significant increase in death (Group I 18% versus Group II 9%; p < 0.05) and complications as pulmonary oedema and cardiogenic shock (Group I 25% versus Group II 14%; p < 0.05). Left atrial dimensions (LAD), end-diastolic left ventricular volume (EDLVV), end-systolic left ventricular volume (ESLVV) and echo-score, evaluated at admission, were not different between Group I and II (LAD 41.3 +/- 6 mm versus 40.1 +/- 5 mm, NS; EDLVV 181 +/- 34 ml versus 173 +/- 30 ml, NS; ESLVV 80 +/- 21 ml versus 75 +/- 18 ml, NS; echo-score 6.7 +/- 3.1 versus 6 +/- 2.7, NS) while pre-discharge echo-grams in Group I showed a trend towards the increase in volumes and echo-score (EDLVV from 181 +/- 34 ml to 194 +/- 36 ml, p = 0.052; ESLVV from 80 +/- 23 ml to 88 +/- 23 ml, p = 0.051; echo-score from 6.7 +/- 3.1 to 7.8 +/- 3.3, p = 0.070).(ABSTRACT TRUNCATED AT 250 WORDS)