Developing PI3K Inhibitors for Respiratory Diseases.

A number of different experimental models using both non-selective and selective PI3K inhibitors have shown that many pathogenic steps of respiratory disorders, such as bronchial asthma, Chronic Obstructive Pulmonary Disease (COPD), Idiopathic Pulmonary Fibrosis (IPF), Acute Respiratory Distress Syndrome (ARDS) and Lung Cancer (LC) are, at least in part, regulated by the PI3K signaling pathway, suggesting that the inhibition of PI3K could represent an ideal therapeutic target for the treatment of respiratory diseases. This chapter summarizes the current state of the therapeutic strategies aimed to exploit the inhibition of PI3K in this context. In animal models of asthma, selective δ and γ inhibitors have shown to be effective, and when administered by inhalation, reasonably safe. Nevertheless, very few clinical trials have been performed so far. The efficacy of current traditional therapies for allergic bronchial asthma has likely diminished the need for new alternative treatments. Surprisingly, in COPD, where instead there is an urgent need for new and more effective therapeutic approaches, the number of clinical studies is still low and not capable yet, with the exception for an acceptable safety profile, to show a significant improvement of clinical outcomes. In IPF, a disease with a disappointing prognosis, PI3K inhibitors have been bound to a FAP ligand with the aim to selectively target myofibroblasts, showing to significantly reduce collagen production and the development of lung fibrosis in an animal model of lung fibrosis. Due to its role in cell activation and cell replication, the PI3K pathway is obviously largely involved in lung cancer. Several studies, currently ongoing, are testing the effect of PI3K inhibitors mainly in NSCLC. Some evidence in the treatment of cancer patients suggests the possibility that PI3K inhibitors may enhance the response to conventional treatment. The involvement of PI3Kδ in the modulation of airway neutrophil recruitment and bronchial epithelial functional alterations also suggest a potential role in the treatment of ARDS, but at the current state the ongoing trials are aimed to the treatment of ARDS in COVID-19 patients. In general, few clinical trials investigating PI3K inhibitors in respiratory disorders have been performed so far. This relatively new approach of treatment is just at its beginning and certainly needs further efforts and additional studies.

Subclinical carotid atherosclerosis in elderly patients with primary Sjögren syndrome: a duplex Doppler sonographic study.

A growing body of evidence presents a link between chronic inflammatory rheumatic diseases and atherosclerosis. To evaluate subclinical carotid atherosclerosis in an elderly group of patients with primary Sjögren syndrome compared with a control group matched for age, sex, ethnicity and cardiovascular risk factors, we enrolled 18 patients with Primary Sjögren Syndrome (mean age 65 ± 5.93 SD) and 18 mild Ostheoarthritic patients (mean age 66 ± 5.94 SD) from the outpatient department of Rheumatology, University “Campus Bio-Medico, Rome, Italy, matched for age, sex, ethnicity and cardiovascular risk factors. A duplex Doppler sonographic study of carotids was performed in order to evaluate intima-media thickness (IMT), stiffness and haemodynamic parameters [resistivity and pulsatility indices (RI and PI, respectively)]. No significant difference was found between primary Sjögren syndrome and control patients in IMT, stiffness and haemodynamic parameters. The lack of significant difference in subclinical atherosclerosis between elderly primary Sjögren syndrome and control matched patients, indicates that traditional cardiovascular risk factors, immunologic alterations and chronic inflammation do not influence the progression of vascular damage in the carotid circulation of patients with median disease duration of 6.5 years.

Hepatic PPARs: their role in liver physiology, fibrosis and treatment.

Complex molecular and cellular mechanisms are involved in the pathway of liver fibrosis. Activation and transformation of hepatic stellate cells (HSCs) are considered the two main reasons for the cause and development of liver fibrosis. The peroxisome proliferator-activated receptors (PPARs) belonging to the family of ligand-activated transcription factors play a key role in liver homeostasis, regulating adipogenesis and inhibiting fibrogenesis in HSCs. Normal transcriptional function of PPARs contributes to maintain HSCs in quiescent phase. A reduced expression of PPARs in HSCs greatly induces a progression of liver fibrosis and an increased production of collagen. Here, we discuss role and function of PPARs and we take into consideration molecular factors able to reduce PPARs activity in HSCs. Finally, although further validations are needed, we illustrate novel strategies available from in vitro and animal studies on how some PPARs-agonists have been proved effective as antifibrotic substances in liver disease.

Systemic lupus erythematosus patients with and without neuropsychiatric manifestations: a neck and transcranial duplex sonography study.

Neuropsychiatric manifestations are not rarely associated with systemic lupus erythematosus (SLE). Magnetic resonance angiography and positron emission tomography can provide excellent images of cerebral perfusion and metabolism whereas information is still lacking on a possible diagnostic role of ultrasound. In this study we aim to assess whether duplex sonography of neck and intracranial vessels may be useful in distinguishing patients with and without neuropsychiatric SLE (NPSLE). Neck and transcranial duplex sonography was performed by a single operator on 33 women affected by SLE (mean age +/- SD: 47.69+/-8.17 years) and on 15 healthy control subjects. Nineteen patients presented NPSLE. Pulsatility and resistivity indices (PI and RI) were automatically calculated by the ultrasound instrument in internal carotid (ICA) and middle cerebral artery (MCA), on both sides, according to standard methods. No significant haemodynamic differences were found in mean and median PI and RI values of ICA and MCA comparing SLE with NPSLE patients and with healthy control subjects. No correlation was found between MCA and ICA parameters in the same group of patients. Duplex sonography of cerebral vessels is unable to distinguish SLE and NPSLE patients. Heterogeneity of causes in the pathogenesis of NPSLE and the different vascular adaptation of cerebral macrocirculation as opposed to cerebral microcirculation may represent possible reasons that explain the inability of ultrasound to differentiate SLE patients from NPSLE patients.

ENT assessment in the integrated management of candidate for (maxillary) sinus lift.

As stated at the 1996 Consensus Conference at Babson College, a (maxillary) sinus lift is a "safe and predictable" procedure for increasing alveolar bone height in the postero-superior alveolar regions in order to allow oral rehabilitation and restore masticatory function by means of the insertion of a dental implant even in the case of an atrophic maxilla. However, the procedure has a well-known impact on the delicate homeostasis of the maxillary sinus: the concomitant presence of systemic, naso-sinusal or maxillary sinus disease may favour the development of post-operative complications (particularly maxillary rhino-sinusitis), which can compromise a good surgical outcome. On the basis of these considerations, the management of sinus lift candidates should include the careful identification of any situations contraindicating the procedure and, if naso-sinusal disease is suspected, a clinical assessment by an ear, nose and throat specialist, which should include nasal endoscopy and, if necessary, a computed tomography scan of the maxillofacial district, particularly the ostio-meatal complex. This first preventive-diagnostic step should be dedicated to detect presumably irreversible and potentially reversible contraindications to a sinus lift, whereas the second (preventive-therapeutic) step is aimed at correcting (mainly with the aid of endoscopic surgery) such potentially reversible ear, nose and throat contraindications as middle-meatal anatomical structural impairments, phlogistic-infective diseases and benign naso-sinusal neoplasms the removal of which achieves naso-sinusal homeostasis recovery, in order to restore the physiological drainage and ventilation of the maxillary sinus. The third (diagnostic-therapeutic) step is only required if mainly infective and sinusal complications arise after sinus lift surgery, and is aimed at ensuring early diagnosis and prompt treatment of maxillary rhino-sinusitis in order to avoid, if possible, implant loss and, in particular, the related major complications. The purpose of this report is to describe these three steps in detail within the context of a multidisciplinary management of sinus lift in which otorhinolaryngological factors may be the key to a successful outcome.

Protein biochip array of adhesion molecule expression in peripheral blood of patients with nasal polyposis.

Nasal polyposis is a chronic non-infectious inflammatory disease of the nasal and paranasal cavity mucosa of unknown multifactorial origin in which inflammatory cells, and in particular eosinophils, seem to play a pivotal role. Eosinophil migration from the bloodstream to nasal polyps is considered to be specific and is a complex process involving several different molecules such as ICAM-1, VCAM-1, and L-, P- and E-selectins. The aim of this study was to investigate, using a protein biochip array technology, the concentrations of these molecules in the peripheral blood of a group of patients affected by nasal polyposis. Patients exhibited a significantly higher expression of VCAM-1, E-selectin, and L-selectin compared to healthy controls, and Spearman's rank correlation test limited to the molecules with significant betweengroup differences demonstrated a significant correlation between VCAM-1 and E-selectin, VCAM-1 and L-selectin, and Eselectin and L-selectin. The results of this investigation are in line with those coming from various imunohistochemical analyses, and seem to confirm the role of inflammation in the pathogenesis of nasal polyposis. These molecules may also represent novel therapeutic targets in the treatment of nasal polyps, and may allow the selection of pharmacological prophylactics that would allow effective inhibition of the inflammation induced by a given allergen.

Do mesenchymal stem cells play a role in vocal fold fat graft survival?

OBJECTIVES: Adipose tissue in vocal fold lipoinjection is currently used to treat patients affected by laryngeal hemiplegia or anatomical defects. The aim of this study has been to evaluate the efficacy of this clinical strategy, by long-term follow-up of the patients and to investigate whether the fat samples used to treat them contain a stem cell population with a wide differentiation potential. MATERIALS AND METHODS: Fat samples harvested from 12 patients affected by severe breathy dysphonia who had undergone vocal fold lipoinjection were analysed by immunocytochemistry, by flow cytometry and reverse transcription-polymerase chain reaction, and the isolated adipose derived mesenchymal stem cells (ADMSCs) were evaluated in order to define their ability to produce soluble factors possibly involved in tissue regeneration, and to differentiate towards different lineages. RESULTS: ADMSCs were efficiently and successfully isolated from all of the samples. They were positive for SSEA-4, an embryonic marker recently identified on bone marrow MSCs and which could explain their high differentiation plasticity. Molecular analysis showed that these cells also expressed Oct-4, Runx-1 and ABCG-2, which characterize the stem cell state, and a number of other specific lineage markers. Flow cytometry revealed mesenchymal markers expressed on ADMSCs and identified a subpopulation characterized by CD146(+)/34(-)/45(-) cells consistent with perivascular/pericyte-like cells. Osteogenic, adipogenic and endothelial tissue differentiation were obtained. CONCLUSIONS: Our results confirmed the therapeutic efficacy of this clinical approach and showed that adipose tissue, administered to patients in order to restore glottic competence, contains mesenchymal stem cells.

Modern management of obstructive salivary diseases.

Over the last fifteen years, increasing public demand for minimally-invasive surgery and recent technological advances have led to the development of a number of conservative options for the therapeutic management of obstructive salivary disorders such as calculi and duct stenosis. These include extracorporeal shock-wave lithotripsy, sialoendoscopy, laser intra-corporeal lithotripsy, interventional radiology, the video-assisted conservative surgical removal of parotid and sub-mandibular calculi and botulinum toxin therapy. Each of these techniques may be used as a single therapeutic modality or in combination with one or more of the above-mentioned options, usually in day case or one-day case under local or general anaesthesia. The multi-modal approach is completely successful in about 80% of patients and reduces the need for gland removal in 3%, thus justifying the combination of, albeit, time-consuming and relatively expensive techniques as part of the modern and functional management of salivary calculi. With regard to the management of salivary duct anomalies, such as strictures and kinkings, interventional radiology with fluoroscopically controlled balloon ductoplasty seems to be the most suitable technique despite the use of radiation. Operative sialoendoscopy alone is the best therapeutic option for all mobile intra-luminal causes of obstruction, such as microliths, mucous plugs or foreign bodies, or for the local treatment of inflammatory conditions such as recurrent chronic parotitis or autoimmune salivary disorders. Finally, in the case of failure of one of the above techniques and regardless of the cause of obstruction, botulinum toxin injection into the parenchyma of the salivary glands using colour Doppler ultrasonographic monitoring should be considered before deciding on surgical gland removal.

Clinico-prognostic value of D-type cyclins and p27 in laryngeal cancer patients: a review.

Despite recent improvements in surgical and radiation therapy, failures still occur in patients with laryngeal squamous cell carcinomas, which may have a very different clinical outcome even when their clinical and histopathological characteristics are similar. The apparent inadequacy of "traditional" prognostic factors in predicting the clinical evolution of laryngeal squamous cell carcinomas has led to attempts to develop additional markers capable of distinguishing patients with a good prognosis from those who are more likely to relapse. A number of studies have demonstrated a relationship between tumourigenesis and alterations in the expression of cyclins, cyclin-dependent kinases and cyclin-dependent kinase inhibitors, but the data regarding laryngeal squamous cell carcinomas are somewhat conflicting. Herein a review is made of the published literature concerning the clinico-prognostic role of cyclin D1, D3 and p27, and personal data are described concerning laryngeal squamous cell carcinoma patients who underwent surgical resection at the ENT Department of the University of Milan. The results of our multivariate analyses demonstrated that cyclin D1, p27 and cyclin D3 overexpression are statistically significant predictors of disease-free survival (p = 0.0238, p = 0.0001 and p = 0.0217, respectively); the statistical correlation with overall survival was significant in the case of p27 (p = 0.0009) and cyclin D3 (p = 0.0189), and borderline in the case of cyclin D1 (p = 0.0622). In relation to cyclin D1/p27 coexpression, the patients with a cyclin D1-/p27+ phenotype showed the best prognosis, those with a cyclin D1/p27+ or cyclin D1-/p27- phenotype, an intermediate prognosis, and those with a cyclin D1+/p27- phenotype, the poorest prognosis (p = 0.0001 and p = 0.0001 for trend for disease-free survival; p = 0.0015 and p = 0.0008 for trend for overall survival). In the case of cyclin D1/cyclin D3 coexpression, the patients with cyclin D1+/cyclin D3+ tumours had the poorest overall survival, those with cyclin D1/cyclin D3+or cyclin D1+/cyclin D3- tumours showed intermediate course, and those with cyclin D1 /cyclin D3- tumours had the most favourable outcome (p = 0.0002). The findings of this review indicate that both types of cyclin D and p27 are involved in the genesis of laryngeal squamous cell carcinomas, and that immunohistochemical evaluations of biopsy samples may provide useful additional markers capable of identifying subgroups of patients with a poor prognosis who can be treated by means of more aggressive surgery, adjuvant radiotherapy and chemotherapy, as well as those requiring a closer and more prolonged follow-up. Finally, preliminary results suggest that the administration of new molecular therapies that exert their antitumoural activities by functionally subverting the pathways regulated by D-type cyclins and their cyclin-dependent kinase counterparts may represent a further therapeutic modality for patients with refractory head and neck squamous cell carcinomas [corrected]

Microsatellite analyses of recurrence or second primary tumor in head and neck cancer.

BACKGROUND: In head and neck squamous cell carcinoma, distinguishing second primary tumours and recurrences may help to orient clinical decisions concerning therapy. PATIENTS AND METHODS: A panel of eight microsatellite markers was used to analyse the loss of heterozygosity and genomic instability in a selected group of 32 patients experiencing a recurrence after having undergone surgery for oral or oropharyngeal carcinoma, in order to establish the clonality and origin of the recurrence. RESULTS: Twenty-three patients showed genetic changes in primary and/or relapsing tumour DNA: clonally-related patterns were detected in six cases, whereas the different patterns between paired tumours indicated the presence of a second primary tumour in 17 cases. None of the markers was informative in nine cases. CONCLUSION: Our observations suggest that only a small proportion of patients have primary and secondary tumours developing from a single contiguous altered field (thus indicating a common clonal origin), whereas the metachronous tumour arises in unrelated fields in the majority of cases.

Prognostic significance of P27 and cyclin D1 co-expression in laryngeal squamous cell carcinoma: possible target for novel therapeutic strategies.

Tumour cells are characterised by uncontrolled growth due to alterations in the genes that play a key role in cell repair systems and apoptosis: pro-mitotic oncogenes such as cyclin D1, and tumour suppressor genes such as p27. Recent studies have demonstrated that these genes are involved in different epithelial neoplasms and that their expression is generally associated with prognosis. The aim of this immunohistochemical study was to analyse the clinical relevance of cyclin D1/p27 co-expression in a homogeneous series of 132 laryngeal squamous cell carcinomas. Multivariate analysis showed that cyclin D1 and p27 were the only statistically significant predictors of disease-free and overall survival. In relation to the simultaneous expression of p27 protein and cyclin D1, the patients with a cyclin D1+/p27-phenotype had the poorest disease-free and overall survival rates. On the basis of these immunohistochemical results, it was possible to select a subgroup of patients with a high risk of recurrence and poor prognosis to undergo more extended surgical treatment and/or combination antitumoral therapeutic procedures.

[Arytenoid subluxation. Experience with a new voice therapy]. / La sub-lussazione aritenoidea. Esperienze con una nuova terapia vocale.

The systematic use of video-fibro-endoscopy techniques has made it possible to precisely define an uncommon laryngeal pathology of traumatic origin: arytenoid subluxation. This pathology general arises after anesthesiological intubation and is characterized by a pathological anteromedial shift of one arytenoid resulting in cord immobility. The symptoms of this disorder are dysphonia similar to recurrent nerve palsy. Differential diagnosis is not always easy. The treatment of choice indicated in the literature is based on invasive approaches (reduction under direct laryngoscopy or, less frequently, even more aggressive surgery) while voice treatment plays a marginal role. The authors offer an overall picture of arytenoid subluxation in view of the anatomophysiological aspects of the crico-arytenoid articulation and describe a new approach to voice rehabilitation. While watching himself in the video-fibroscopy monitor (visual feedback), the patient performs a particular exercise enabling him to raise the larynx, lifting the arytenoids and causing hyperadduction and vibration of the arytenoid hood. After having learned the exercise the patient continues with regular speech therapy sessions and voice therapy at home. Periodically the patient undergoes video-endoscopy and phoniatric-speech therapy check-ups until vocal function and cord motility have been restored. This type of approach was applied in 4 patients, all of whom responded relatively quickly with reactivation of the articulation and recovery of the voice. The authors conclude that voice therapy can achieve full healing in patients with arytenoid subluxation and should be given priority over other forms of treatment. The excellent results are explained asserting that particular movement of the larynx can achieve a reduction in subluxation by rearranging the articulatory surface.

Falls in the elderly: the development of a risk questionnaire and posturographic findings.

In elderly people, owing to a perturbation at several levels, including the motor, sensory and cognitive levels, a condition of dizziness and unsteadiness complicated by frequent falls often appears. In this article, we review the most recent information about clinical and instrumental tools available for preventing mobility-related accidents and report the results of a comparative study of postural control, carried out through tetraataxiometry (by Tetrax, Tel Aviv, Israel), in two samples of elderly women belonging to two different populations: 24 Italian women (11 reporting falls and 13 without falls) having a mean age of 73.1 years, and 37 Israeli women (12 with falls and 25 without falls) having a mean age of 72.5 years. The posturographic findings show that the falling subjects, to maintain postural control, are highly dependent on somatosensory inputs and have a weaker "systeme postural fin" (fine postural system), according to Gagey. They also show that an elderly subpopulation exists that, for unknown reasons, is immune to destabilization and falls.

Plasmatic cytokine network in patients with laryngeal carcinoma after surgical treatment.

Alterations in host immunity, inflammation, angiogenesis and metabolism are all prominent clinical features in patients with laryngeal squamous cell carcinoma (LSCC). Although the origin of the signals and mechanisms underlying these responses are not well understood, their local and systematic nature suggest that squamous cell carcinoma-produced cytokines with proinflammatory and immunoregulatory activity may contribute to the pathogenesis of LSCC. In order to gain a better insight into the roles and relationships of the cytokines, we investigated serum IL-6, IL-10 and IL-12 concentrations in LSCC patients under baseline conditions and after surgery. In comparison with controls, all the patients had higher plasma IL-10 concentrations before surgical treatment (T0), while plasma IL-6 and IL-12 concentrations were higher in 22 (84.6%) and 24 patients (92.3%). The differences in plasma IL-6, IL-10 and IL-12 concentrations at T0 and T1 were statistically significant (p<0.001, p<0.0046 and p<0.011). Our finding suggest that plasma cytokines are overexpressed in LSCC patients. There was an independent increase in plasma IL-6 levels before and after surgical treatment. Furthermore, the up- and down-regulation of plasma IL-10 and IL-12 suggest a regulatory relationship between them.

99mTc-sulfur colloid gastroesophageal scintigraphy with late lung imaging to evaluate patients with posterior laryngitis.

UNLABELLED: The aim of this study was to use gastroesophageal and pulmonary scintigraphy to evaluate the prevalence of gastroesophageal reflux and airway involvement among patients with posterior laryngitis. METHODS: The study included a total of 201 patients (131 females, 70 males; age range, 15-77 y; mean age +/- SD, 49 +/- 16 y). All patients had posterior laryngitis documented by laryngoscopy and symptoms such as a dry cough, painful swallowing, and hoarseness. A control population of 20 healthy volunteers (13 females, 7 males; age range, 19-74 y; mean age, 53 +/- 13 y) was also evaluated. After a 12-h fast, all subjects underwent gastroesophageal scintigraphy through administration of 300 mL orange juice labeled with 185 MBq 99mTc-sulfur colloid. After 18 h, planar anteroposterior thoracic images were acquired with the subjects supine. RESULTS: Sixty-seven percent of patients (134/201) had scans positive for gastroesophageal reflux; of these, 30 (22%) had distal reflux and 104 (78%) had proximal reflux. In addition, the scans of 31 patients were positive for proximal reflux-associated pulmonary uptake. The frequency, duration, and degree of reflux episodes were significantly greater in patients with proximal reflux than in patients with distal reflux (P < 0.001). The 67 patients in whom reflux was not detected had diseases or reflux-associated cofactors that could account for laryngeal symptoms. No statistically significant difference in symptoms or esophageal motility parameters could be identified among the patient groups, but patients with proximal reflux had significantly prolonged gastric emptying times compared with healthy volunteers. CONCLUSION: Most patients with posterior laryngitis had detectable proximal gastroesophageal reflux. Exposure of the proximal part of the esophagus to acid, by setting the stage for microaspiration of gastric material into the larynx, remains a major cause of damage to the laryngeal mucosa. Slowed gastric emptying may be a predisposing factor. Moreover, symptoms such as a dry cough, painful swallowing, or hoarseness may not be reliable predictors of the presence of gastroesophageal reflux or of associated airway involvement.

Comparative study of middle-latency responses and auditory brainstem responses in elderly subjects.

Auditory senescence is studied through an electrophysiological investigation of the brainstem, midbrain and thalamocortical tracts of auditory pathways. For this purpose, comparative electrophysiologic studies were carried out by recording middle-latency responses (MLR) and auditory brainstem responses (ABR) in elderly subjects in the 7th-8th and 9th decades of life and in a control group consisting of young subjects in the 3rd decade of life. All these subjects were free of otological, neurological, vascular and metabolic pathologic states. In elderly subjects, the statistical analysis of the results, especially of MLRs, showed a deterioration in the progression of information from the periphery to the center due to changes in morphology, increased latency, reduced amplitude and poorer reproducibility. Furthermore, a different behavior was noticed in females versus males. Indeed, the shorter latency of evoked potentials in the female control group was remarkably reduced in the elderly subjects. MLRs scan a wide tract of auditory pathways and give evident results even by adopting a juxtaliminal stimulation; therefore, they are a more complete and sensitive test than ABRs for the study of involutional processes.

Xibornol: multiple dose pharmacokinetics and diffusion in lung, tonsillar tissue and laryngeal mucosa.

In ten patients with severe chronic bronchitis and in a further 23 with planned resection of lung, tonsils or larynx, 500 mg doses, single or multiple, of xibornol (6-isobronyl-3, 4-xylenol) were administered for an antibacterial effect. The pharmacokinetics and diffusion of the drug in the tissues were studied. A high diffusion and distribution value of xibornol was observed, with levels in the tissues constantly higher than that in the serum. The concentrations reached within the respiratory tract were adequate for their antibacterial effect.