Expression of TP53 mutation-associated microRNAs predicts clinical outcome in head and neck squamous cell carcinoma patients.

BACKGROUND: TP53 mutation is associated with decreased survival rate in head and neck squamous cell carcinoma (HNSCC) patients. We set out to identify microRNAs (miRNAs) whose expression associates with TP53 mutation and survival in HNSCC. PATIENTS AND METHODS: We analyzed TP53 status by direct sequencing of exons 2 through 11 of a prospective series of 121 HNSCC samples and assessed its association with outcome in 109 followed-up patients. We carried out miRNA expression profiling on 121 HNSCC samples and 66 normal counterparts. miRNA associations with TP53 mutations and outcome were evaluated. RESULTS: A TP53 mutation was present in 58% of the tumors and TP53 mutations were significantly associated with a shorter recurrence-free survival. This association was stronger in the clinical subgroup of patients subjected to adjuvant therapy after surgery. The expression of 49 miRNAs was significantly associated with TP53 status. Among these 49, we identified a group of 12 miRNAs whose expression correlates with recurrence-free survival and a group of 4 miRNAs that correlates with cancer-specific survival. The two groups share three miRNAs. Importantly, miRNAs that correlate with survival are independent prognostic factors either when considered individually or as signatures. CONCLUSIONS: miRNAs expression associates with TP53 status and with reduced survival after surgical treatment of squamous cell carcinoma of the head and neck.

The potential prognostic role of cardiovascular autonomic failure in α-synucleinopathies.

Cardiovascular autonomic failure is the second most common dysautonomic feature of α-synucleinopathies and has significant impact on daily activities and quality of life. Here we provide a systematic review of cardiovascular autonomic failure in α-synucleinopathies, emphasizing its impact on cognitive functions and disease outcomes. Articles spanning the period between January 1985 and April 2012 were identified from the PubMed database using a keyword-based search. Epidemiological studies highlight the negative prognostic effect of cardiovascular autonomic failure on cardiovascular and cerebrovascular outcomes and overall mortality in all α-synucleinopathies. Altered cerebral perfusion, vascular pressure stress, and related disruption of the blood-brain barrier may also contribute to the white matter hyperintensities and cognitive dysfunction frequently found in patients affected by neurocardiovascular instability. These findings support the hypothesis that cardiovascular autonomic failure may play a negative prognostic role in α-synucleinopathies and suggest that precocious screening and therapeutic management of cardiovascular autonomic failure may positively impact disease course.

Mapping pathophysiological features of breast tumors by MRI at high spatial resolution.

Magnetic resonance imaging (MRI) is a noninvasive method that reveals anatomical details in vivo and detects lesions for diagnosis. Although standard breast MRI cannot clearly delineate breast cancer, contrast-enhanced MRI enables the detection of breast masses with high sensitivity. Dynamic studies demonstrated that malignant lesions were characterized by a faster signal enhancement rate than benign ones. Dynamic MRI of human breast cancer in mice revealed high heterogeneity in the distribution of contrast-enhanced curves and derived pathophysiological features, indicating the importance of high spatial resolution. With clinical MRI, it is difficult to achieve simultaneously high spatial and temporal resolution. In previous dynamic studies, the emphasis was on high temporal resolution and mainly empiric analyses. We describe here a new model-based method that optimizes spatial resolution by using only three time points, and yet characterizes tumor heterogeneity in terms of microvascular permeability and extracellular fraction. Mapping these pathophysiological features may aid diagnosis and prognosis assessment, while the high spatial resolution may improve the capacity to detect smaller lesions. The method was tested in human breast tumors implanted in mice and in a limited number of benign and malignant breast lesions of patients.

Mutant p53: an oncogenic transcription factor.

Inactivation of tumor-suppressor genes is one of the key hallmarks of a tumor. Unlike other tumor-suppressor genes, p53 is inactivated by missense mutations in half of all human cancers. It has become increasingly clear that the resulting mutant p53 proteins do not represent only the mere loss of wild-type p53 tumor suppressor activity, but gain new oncogenic properties favoring the insurgence, the maintenance, the spreading and the chemoresistance of malignant tumors. The actual challenge is the fine deciphering of the molecular mechanisms underlying the gain of function of mutant p53 proteins. In this review, we will focus mainly on the transcriptional activity of mutant p53 proteins as one of the potential molecular mechanisms. To date, the related knowledge is still quite scarce and many of the raised questions of this review are yet unanswered.

Autonomic cardiovascular function and baroreflex sensitivity in patients with cervical dystonia receiving treatment with botulinum toxin type A.

OBJECTIVE: To investigate possible changes in autonomic cardiovascular regulation and cardiopulmonary baroreflex sensitivity in patients with primary cervical dystonia receiving chronic treatment with botulinum toxin type A. METHODS: Short-term power spectral analysis of heart rate and systolic blood pressure variability, high-frequency and low-frequency oscillations of heart rate variability, low frequency/high frequency ratio and baroreflex sensitivity (alpha index) were measured in 12 patients with cervical dystonia before and 2-4 weeks after botulinum toxin type A injection and compared with normative data. RESULTS: Before treatment, at rest, patients had significantly lower high frequency power than healthy subjects (p < 0.01), whereas no differences were found in low frequency power. Botulinum toxin injection in patients induced no changes in either power frequency. In patients before treatment and healthy subjects the low frequency oscillatory components increased similarly from rest to tilt (p < 0.01), but tilt induced lower low frequency values in patients than in healthy subjects (p < 0.01). In patients before treatment, the high frequency variations from rest to tilt remained unchanged, whereas in healthy subjects they decreased significantly (p < 0.01). Botulinum toxin type A injection in patients induced no changes in low frequency or high frequency powers. In patients before treatment the low frequency/high frequency ratio increased slightly from rest to tilt, but in healthy subjects increased significantly (p < 0.01). Botulinum toxin type A left the pretreatment low frequency/high frequency ratio unchanged. The alpha-index measured at rest in patients before treatment was lower than in healthy subjects (p<0.05), whereas during tilt was similar in both groups. The alpha-index measured after botulinum toxin injection in patients remained unchanged at rest and during tilt. CONCLUSIONS: Patients with cervical dystonia receiving treatment with botulinum toxin type A have mild, subclinical abnormalities in autonomic cardiovascular regulation and cardiopulmonary baroreflex sensitivity. These changes do not worsen after acute botulinum toxin type A injection.

Combined transcranial Doppler and EEG recording in vasovagal syncope.

BACKGROUND: In neurally mediated syncope a 'typical' EEG pattern during hyperventilation (HV) may be observed. This study aimed to investigate transcranial Doppler (TCD) and EEG variations in response to hyper- and hypocapnia using simultaneous recording. METHODS: Syncope patients with a typical EEG pattern during HV (SEEG+, n = 15) and those without abnormalities (SEEG-, n = 16) were compared with healthy controls (n = 20). Simultaneous TCD and EEG recordings were performed at rest (baseline), during 2 apnea tests and during HV. Cerebrovascular vasoreactivity, index for hypocapnia, total vasomotor reserve and time to flow velocity normalization after HV (t-norm) were recorded. RESULTS: With TCD, a reduction in Vasomotor reserve was observed in SEEG+ compared with the other 2 groups (control: 67 +/- 8%; SEEG-: 67 +/- 10%; SEEG+: 57 +/- 8%; p < 0.0001). t-norm was longer in all syncopal patients and in particular in SEEG+ (control: 20.2 +/- 3 s; SEEG-: 40 +/- 7 s; SEEG+: 123 +/- 45s; p < 0.0001). Quantitative EEG showed an increase in slow bands in all subjects during HV, small and nonsignificant in controls and SEEG-, higher and significant in SEEG+, related with flow reduction. CONCLUSIONS: Changes in the sympathetic modulation of cerebral vasoconstriction may explain both the pathophysiology of vasovagal syncope and the typical paroxysmal EEG findings.

Leopard syndrome.

The L.E.O.P.A.R.D. syndrome is an autosomal, dominant disorder with characteristic features that include: multiple lentigines, café au lait spots, electrocardiographic conduction abnormalities, ocular hypertelorism, obstructive cardiomyopathy, pulmonary stenosis, abnormal (male) genitalia, retardation of growth, and deafness. Patients do not usually present all the clinical features traditionally associated with the disorder. Indeed, several features are not present until late in life and do not become clinically manifest until puberty. It has been observed that this syndrome is caused by a "missense" mutation in PTPN11, a gene encoding the protein tyrosine phosphatase SHP-2 located on chromosome 12q22. A diagnosis of LEOPARD syndrome may be established exclusively on the basis of clinical criteria. In our case, the patient was diagnosed with the syndrome late in his life when he was already exhibiting all its distinctive clinical features. We have reported the case of a LEOPARD syndrome patient exhibiting extremely elongated vertebral and basilar arteries previously undescribed in the literature.

Mutant p53 gain of function: reduction of tumor malignancy of human cancer cell lines through abrogation of mutant p53 expression.

Mutations in the TP53 tumor suppressor gene are the most frequent genetic alteration in human cancers. These alterations are mostly missense point mutations that cluster in the DNA binding domain. There is growing evidence that many of these mutations generate mutant p53 proteins that have acquired new biochemical and biological properties. Through this gain of function activity, mutant p53 is believed to contribute to tumor malignancy. The purpose of our study was to explore mutant p53 as a target for novel anticancer treatments. To this aim, we inhibited mutant p53 expression by RNA interference in three different cancer cell lines endogenously expressing mutant p53 proteins, and evaluated the effects on the biological activities through which mutant p53 exerts gain of function. We found that depletion of mutant p53 reduces cell proliferation, in vitro and in vivo tumorigenicity, and resistance to anticancer drugs. Our results demonstrate that mutant p53 knocking down weakens the aggressiveness of human cancer cells, and provides further insight into the comprehension of mutant p53 gain of function activity in human tumor.

S100A2 gene is a direct transcriptional target of p53 homologues during keratinocyte differentiation.

The p53 paralogues p73, p63 and their respective truncated isoforms have been shown to be critical regulators of developmental and differentiation processes. Indeed, both p73- and p63-deficient mice exhibit severe developmental defects. Here, we show that S100A2 gene, whose transcript and protein are induced during keratinocyte differentiation of HaCaT cells, is a direct transcriptional target of p73beta and DeltaNp63alpha and is required for proper keratinocyte differentiation. Transactivation assays reveal that p73beta and DeltaNp63alpha exert opposite transcriptional effects on S100A2 gene. While DeltaNp63alpha is found in vivo onto S100A2 regulatory regions predominantly in proliferating cells, p73beta is recruited in differentiating cells. Silencing of p73 impairs the induction of S100A2 during the differentiation of HaCaT cells. Moreover, silencing of p73 or S100A2 impairs the proper expression of keratinocyte differentiation markers. Of note, p53 family members do not trigger S100A2 gene expression in response to apoptotic doses of cisplatin and doxorubicin.

The transcriptional repressor ZEB regulates p73 expression at the crossroad between proliferation and differentiation.

The newly discovered p73 gene encodes a nuclear protein that has high homology with p53. Furthermore, ectopic expression of p73 in p53(+/+) and p53(-/-) cancer cells recapitulates some of the biological activities of p53 such as growth arrest, apoptosis, and differentiation. p73(-/-)-deficient mice exhibit severe defects in proper development of the central nervous system and pheromone sensory pathway. They also suffer from inflammation and infections. Here we studied the transcriptional regulation of p73 at the crossroad between proliferation and differentiation. p73 mRNA is undetectable in proliferating C2C12 cells and is expressed at very low levels in undifferentiated P19 and HL60 cells. Conversely, it is upregulated during muscle and neuronal differentiation as well as in response to tetradecanoyl phorbol acetate-induced monocytic differentiation of HL60 cells. We identified a 1-kb regulatory fragment located within the first intron of p73, which is positioned immediately upstream to the ATG codon of the second exon. This fragment exerts silencer activity on p73 as well as on heterologous promoters. The p73 intronic fragment contains six consensus binding sites for transcriptional repressor ZEB, which binds these sites in vitro and in vivo. Ectopic expression of dominant-negative ZEB (ZEB-DB) restores p73 expression in proliferating C2C12 and P19 cells. Thus, transcriptional repression of p73 expression by ZEB binding may contribute to the modulation of p73 expression during differentiation.

The teriparatide in the treatment of severe senile osteoporosis.

The osteoporosis is a systemic disease of multicausal etiopathogenesis. A progressive bone loss and qualitative alterations in the macro- and micro-architecture of the remaining bones, resulting in a loss of strength of bones to such an extent that even very modest traumas will cause fractures characterize it. Three forms are defined (i) postmenopausal appearing after the menopause, (ii) senile appearing with advancing age, and (iii) the idiopathic forms. Severe osteoporosis is declared when the patients suffer vertebral or femoral fractures without any trauma during a treatment with anti-reabsorptive medicines of at least 1-year. The treatment of osteoporosis is based on various categories of pharmaca, such as bisphosphonates, selective estrogen receptor modulators (SERMs), diaminobutyric acid (DABA), parathyroid hormone (PTH), estrogens and non-hormonal drugs. The teriparatide, the recombinant human (rh)PTH(1-34), is identical in amino acid sequence until the 34th (N-terminal) amino acid of the endogenous, human PTH. It is produced in E. coli using the recombinant DNA technology. It is a pharmacon having a strong trophic-anabolic action on the bone tissue, assuring both the inhibition of the bone loss, and the formation of new bones of good quality. It acts as a stimulant of the osteoblast functions, and at the same time, increases the absorption of calcium from the intestine, and also the renal reabsorption of calcium, and decreases the excretion of phosphates in the kidney. This study summarizes our own experience with the use of rhPTH(1-34) in the treatment of senile patients with severe osteoporosis. Our sample consisted of 40 elderly women of the mean age of 78+/-5 years, having severe osteoporosis. They displayed a columnar T-score>-3.5 and femoral T-score>-2.5, had been under antireabsorptive treatment since at least 12 months. In particular, 15 patients were treated with Alendronate (70 mg/week), 10 of them with Risedronate (35 mg/week), and 15 of them with Raloxifene (60 mg/day). These patients in our study were treated for 1 year with 20 microg/day of rhPTH (1-34), injected subcutaneously, and supplemented also with a daily dose of 1g of calcium and 800 IU of Vitamin D, per os. At start of this treatment (time t(0)), after 6 months (time t(6)) and after 12 months (time t(12)) patients underwent a bone mineral density (BMD) analysis (Dexa-Lunar-DPX-P) on the lumbar vertebral column, (L1-L4 zone), as well as a femoral BMD. We applied also quality of life (QoL) questionnaire of the European Foundation for Osteoporosis (QUALEFFO), and evaluated also the use of non-steroidal anti-inflammatory drugs (NSAIDs). Our final considerations are that the teriparatide therapy increases significantly the bone mass density, expressed in terms of T-Score, reduces the occurrence of new fractures, improves the QoL, and decreases also the consumption of NSAIDs.

From p63 to p53 across p73.

Most genes are members of a family. It is generally believed that a gene family derives from an ancestral gene by duplication and divergence. The tumor suppressor p53 was a striking exception to this established rule. However, two new p53 homologs, p63 and p73, have recently been described [1-6]. At the sequence level, p63 and p73 are more similar to each other than each is to p53, suggesting the possibility that the ancestral gene is a gene resembling p63/p73, while p53 is phylogenetically younger [1,2].The complexity of the family has also been enriched by the alternatively spliced forms of p63 and p73, which give rise to a complex network of proteins involved in the control of cell proliferation, apoptosis and development [1,2,4,7-9]. In this review we will mainly focus on similarities and differences as well as relationships among p63, p73 and p53.

Respiratory sinus arrhythmia and cardiovascular neural regulation in athletes.

Studies using spectral analysis of cardiovascular variability as a noninvasive means for assessing autonomic nervous system activity have provided controversial results in athletes. One reason is that a slow breathing rate--a common feature in athletes--affects spectral estimation because it causes the low-frequency (LF) and high-frequency (HF) components to overlap. Low-frequency power increases during sympathetic activation; high-frequency corresponds to respiratory sinus arrhythmia. In this study, to assess how controlled respiration influences autonomic nervous system activity, we determined the effect of controlled and uncontrolled breathing conditions on cardiovascular variability. Our aim was to identify a standard respiratory rate for spectral estimation of cardiovascular neural control in athletes. During electrocardiographic recordings, subjects lay supine and breathed at their spontaneous frequency and at rates of 15, 12, and 10 to 14 (random) breaths x min(-1). Uncontrolled and random breathing rates significantly altered spectral sympathetic indices; conversely, 15 and 12 breaths x min(-1) redistributed respiratory related power through the HF, thus yielding correct LF power estimation. None of the breathing conditions significantly changed mean heart rate, arterial blood pressure, or spectral total power of cardiovascular variability. In conclusion, when power spectral analysis is used for assessing autonomic activity in athletes, respiration should be standardized at 15 breaths x min(-1). Controlled respiration at this rate leaves autonomic nervous system activity unchanged.

Nocturnal headache-hypertension syndrome: a chronobiologic disorder.

The study of blood pressure (BP) monitoring in essential hypertensive patients recurrently suffering from nocturnal headache revealed a rhythmic elevation of sphygmomanometric values during the night. Such a finding was not detected in essential hypertensive patients suffering from occasional headache. The nocturnal elevation of BP was seen to be paralleled by the circadian peak of heart rate, suggesting that the disorder is a systemic phenomenon. Importantly, the headache episodes were seen to disappear after antihypertensive therapy that was adjusted to lower the nocturnal increase of BP. The therapeutic results suggested that the nocturnal headache was dependent on the phasic elevation of BP. The beneficial effects further suggested that the nocturnal headache and the nocturnal elevation of BP may represent a particular syndrome with a cause-effect relationship. The term "nocturnal headache-hypertension syndrome" is proposed.

BCL-2: the pendulum of the cell fate.

The homeostasis of normal tissues is a balance between cell proliferation and cell death. Alterations of both pathways contribute to a clonal expansion of cancer cells. Bcl-2 and its family play an important role in the regulation of the apoptotic pathway. Apoptosis or programmed cell death is an active form of cell suicide that is characterized by specific morphological and biochemical events. These include cleavege of genomic DNA into oligonucleosome-length DNA fragments by endonucleases, chromatin condensation and marginalization, nuclear fragmentation, plasma membrane blebbing, and cell shrinkage. Though the role of apoptosis is clearly defined in the maintaining of physiological tissue homeostasis, several pathological conditions and external factors cause apoptosis. Anticancer drugs and radiation, two of the most important tools in the cancer treatment, cause apoptotic cell death. The understanding of the mechanisms underlying the regulation of the apoptosis in response to different types of DNA damage might provide relevant information to improve cancer treatment. In this review we mainly discuss bcl-2 and its partners in human cancers and how their disregulation might contribute to the development and the difficult treatment of cancer.

Pressor effects from daily events and laboratory complex stimuli relating personality factors.

Our preliminary research has attempted to establish a series of methods to study the complex interactions occurring between pressor reactivity and personality profile. Ten untreated mild hypertensives (age 42.9 +/- 8) without damaged target organs were recruited from an outpatient hypertension center along with an equal number of normotensive volunteers (age 38.2 +/- 8.1). We performed a sequence of stressor types under laboratory conditions (sensory perceptual activities, psychomotor responses, and cognitive behavior) following an order ranging from inferior levels to superior levels of systemic integration. The subjects also underwent a 24-h automatic noninvasive blood pressure recording which took into account the situational reactivity. They filled in MMPI and STAI questionnaires before and after the stressor batteries. Only the sensory-perceptual test (Stroop color test modified), the arithmetic test, and the psychomotor test provoked a significant increase in blood pressure and, in the latter test, also a significant increase of the heart rate. The test batteries' mean differences were not significant between the two groups. Similarly, the answers to the trait-anxiety questionnaires did not allow us to make a substantial division between normotensive and hypertensive subjects. On the contrary, the situational anxiety questionnaires showed a significant difference in the score reading preceding and following a task performed by the hypertensive subjects. We observed significant differences for both systolic and diastolic 24-h blood pressure data in transition from a working situation to the sleeping period. However, there was not a significant difference in hypertensive blood pressure readings recorded during work and at home.(ABSTRACT TRUNCATED AT 250 WORDS)

A multivariate time-variant AR method for the analysis of heart rate and arterial blood pressure.

This paper approaches the problem of short-term mechanisms that regulate heart rate and blood pressure variability signals, by focusing the evident changes of their frequency content during transients (dynamic situations in which the behaviour of these control mechanisms may vary on a beat-to-beat basis). In this study, we suggest an autoregressive time-variant spectral estimation method, which is able to follow such dynamic changes in the signals. This method has also been extended to a multivariate approach in order to take into account more than one process at a time, and to assess the mutual influences between the different controlling systems. The algorithms successfully tested on simulated series have also been used to analyse series recorded during a vaso-vagal syncope episode in a tilt manoeuvre and a physical exercise stress test protocol. The results show how this method is able to follow the changing dynamics of the signals on the basis of a closed-loop model of their interaction on a beat-to-beat basis. After a proper identification procedure of the blocks forming the model, it is possible, therefore, to obtain the classical spectral parameters and the gain of the transfer function between the signals. Such parameters constitute new time series that describe the physiopathology of the cardiovascular control systems, even during non-stationary epochs.

The management of clinically occult breast lesions. A Johannesburg learning experience.

There is an increase in demand for pre-operative radiologically guided percutaneous localisation of occult breast lesions. Specific problems in the progression from mammographic identification to histological diagnosis are cited. Accurate pre-operative localisation should always be followed by specimen radiography.

[Retrospective study of 41 cases of pulmonary carcinoma diagnosed from 1983 to 1986]. / Studio retrospettivo su 41 casi di carcinoma polmonare diagnosticati dal 1983 al 1986.

In their retrospective study of 41 cases of lung carcinoma diagnosed in their department in the years 1983-86, the authors aimed at assessing the dynamics of their clinical, instrumental and therapeutic approach to this widespread and socially relevant pathology. Among the aspects considered are: the patients' mean age and sex, relationship to smoking, clinical and cytologic diagnosis, instrumental exploration, presenting symptoms, therapeutic approach, mean survival time. On the basis of their findings, the authors come to the conclusion that their experience coincides with that reported in national and international studies with the same dramatic picture (usually late onset of symptoms, short survival, highly invasive growth). They therefore hope that in the near future the scientific community will engage in a more decisive manner and with better support from the health authorities and more adequate financial resources in ways aimed at reducing the incidence of this pathology, at acquiring better knowledge of the relationship with the environment, and at an improvement of diagnostic and therapeutic strategies.

[Phyllodes tumor of the breast: diagnostic and therapeutic problems]. / Tumore fillode della mammella: problematiche diagnostiche e terapeutiche.

Phyllodes tumour (PT) is a rare breast tumor with mixed connective and epithelial components, which usually relapses topically after excision. Five patients with PT took part in the study (4 were positive for benign PT and one was positive for malignant PT at extemporary histological examination). The patients (3) with 3 cm or minus benign lesion had mass excision with a large resection of the surrounding mammary tissue. In the remaining patient with benign neoplasm (8 cm lesion) simple mastectomy was carried out. Finally the patient with malignant PT (5 cm lesion) underwent a simple mastectomy without following chemotherapy or radiotherapy. Follow-up (at 6 and 12 months) consisted of mammography and clinical examination. One out of the 3 patients treated with large resection had a local relapse which was then treated surgically. There was no evidence of relapse at 12 months in the remaining patients with benign tumor. The patient with malignant PT returned to our attention with multiple bone metastases in the thoracic lumbar tract two years later. She received multiple courses of combined radiotherapy and chemotherapy. Local relapses do not appear to affect survival: as a consequence, wide excision should be the primary treatment of benign phyllodes tumors. Total mastectomy has been indicated for malignant phyllodes tumors and for very large tumors.