Luana Lipari

Cigarette smoke increases BLT2 receptor functions in bronchial epithelial cells: in vitro and ex vivo evidence

Leukotriene B4 (LTB4) is a neutrophil chemotactic molecule with important involvement in the inflammatory responses of chronic obstructive pulmonary disease (COPD). Airway epithelium is emerging as a regulator of innate immune responses to a variety of insults including cigarette smoke, the major risk factor for COPD. In this study we have explored whether cigarette smoke extracts (CSE) or soluble mediators present in distal lung fluid samples (mini‐bronchoalveolar lavages) from smokers alter the expression of the LTB4 receptor 2 (BLT2) and peroxisome proliferator‐activated receptor‐α (PPAR‐α) in bronchial epithelial cells. We also evaluated the effects of CSE on the expression of intercellular adhesion molecule 1 (ICAM‐1) and on the binding of signal transducer and activator of transcription 1 (STAT‐1) to ICAM‐1 promoter as well as the adhesiveness of neutrophils to bronchial epithelial cells. CSE and mini‐bronchoalveolar lavages from smokers increased BLT2 and ICAM‐1 expression as well as the adhesiveness of neutrophils to bronchial epithelial cells and decreased PPAR‐α expression. CSE induced the activation of STAT‐1 and its binding to ICAM‐1 promoter. These findings suggest that, in bronchial epithelial cells, CSE promote a prevalent induction of pro‐inflammatory BLT2 receptors and activate mechanisms leading to increased neutrophil adhesion, a mechanism that contributes to airway neutrophilia and to tissue damage.

Expression of cyclooxygenase-1 and cyclooxygenase-2 in normal and pathological human oral mucosa.

Cyclooxigenase (COX) is the rate-limiting enzyme for the conversion of arachidonic acid (AA) to prostaglandins (PGs). Two isoforms of COX have been identified: COX-1 is constitutively expressed in many cells and is involved in cell homeostasis, angiogenesis and cell-cell signalling; COX-2 is not expressed in normal condition however it is strongly expressed in inflammation. The oral cavity is constantly exposed to physical and chemical trauma that could lead to mucosal reactions such as hyperplasia, dysplasia and cancer. Early diagnosis is the most important issue to address for a positive outcome of oral cancer; therefore it would be useful to identify molecular markers whose expression is associated with the various stages of oral cancer progression. Since COX enzyme has been involved, with different mechanisms, in the development and progression of malignancies we decided to investigate the expression and localization of COX-1 and COX-2 in normal human oral mucosa and three different pathologies (hyperplasia, dysplasia and carcinoma) by immunohistochemistry and RT-PCR. COX-1 mRNA and protein have been detected already in normal oral mucosa and their expression progressively increases from normal samples towards hyperplasia, dysplasia and finally carcinoma. On the contrary, COX-2 is not expressed in the normal tissue, starts to be expressed in hyperplasia, reaches the maximum activation in dysplasia and then starts to be downregulated in carcinoma.

Expression of gelatinases (MMP-2, MMP-9) in human articular cartilage.

Osteoarthritis (OA) is a chronic degenerative joint disorder characterized by destruction of the articular cartilage, subchondral bone alterations and synovitis. Matrix metalloproteinases (MMPs) are expressed in joint tissues of patients with osteoarthritis (OA). The objective of this study was to define the steady state levels of two different MMPs to provide more insight into the role of MMPs in cartilage destruction in OA. We investigated the expression of gelatinases through immunohistochemistry Our results show that high levels of MMP-2 and MMP-9 are present in OA and suggest that once these MMPs are fully activated they may contribute to the cartilage destruction in OA.

Expression of MMP-2 and MMP-9 in odontogenic myxoma in a child: report of a clinical case

BackgroundOdontogenic myxoma (OM) is a benign, locally invasive, non-metastasizing neoplasm of the jaw bones. Despite the benign nature of these lesions, there is a high rate of recurrence and the current recommended therapy, depending on the size and behaviour of the lesion, can vary from curettage with peripheral ostectomy, segmental resection up to radical resections for more aggressive lesions. OM is a rare tumour which occurs predominantly in the third decade of life and it is rare in children. Matrix metalloproteinases (MMPs) are a family of extracellular endopeptidases responsible for the degradation and remodelling of extracellular matrix, they are known to be involved in the progression and invasiveness of many types of tumour. MMPs have been studied in OM because of their well-known role in extracellular matrix degradation, tumour invasion and recurrence.Clinical case reportWe report a case of OM in a 6-year-old boy. A conservative excision was accomplished. The mass was excised without affecting the mandibular bone and the inferior alveolar nerve. Curettage and removal of the first right inferior molar were performed. After 6-month follow-up, no evidence of recurrence was found.Experimental dataWe investigated the expression of MMP-2 and MMP-9 in this case of OM in a child. RT-PCR showed the expression of both MMP-2 and MMP-9 mRNAs. Immunohistochemistry showed a weak MMP-2 protein expression while MMP-9 protein was not detected.ConclusionIn this case of OM in a child, we report lack of recurrence after excision associated with low MMP-2 protein expression and absence of MMP-9. We believe it is worthy to deeply investigate the relationship between MMPs expression and OM behaviour with the aim to use MMPs as prognostic and/or therapeutic markers in OM.

Expression of Gelatinases (MMP-2, MMP-9) and Cyclooxygenases (COX-1, COX-2) in Some Benign Salivary Gland Tumors

Salivary gland tumors, most of which are rare benign tumors, represent a histologically heterogenous group with the greatest diversity of morphological and cellular features. The aim of this study is to analyse the expression and possible interactions between gelatinases (MMP-2, MMP-9) and cyclooxygenases (COX-1, COX-2) in some benign salivary gland tumors. We investigated the expression of gelatinases and cyclooxigenases in control salivary gland, Pleomorphic adenoma and Warthins tumor through immunohistochemistry and Reverse Transcription – Polymerase Chain Reaction (PCR). We identified the expression of both classes of enzyme in normal samples and in the two types of pathological samples without any quantitative differences. From the present data no significant differences emerge in the expression of these enzymes among the different pathologies examined. Nevertheless, due to the small number of samples included in this study, general statements regarding correlation between the degree of severity of the tumoral pathology and the quantitative expression of these potential tumoral markers can not be made.

Presence of Oxytocin, Vasopressin and Atrial Natriuretic Peptide and Their Modification in Rat Hypothalamic Paraventricular Nucleus During Resistance Training

Many studies have demonstrated the physiological effects of oxytocin (OT), atrial natriuretic peptide (ANP) and vasopressin (VP) in the homoeostasis of body fluids during physical exercise. However, a little information is available about the related immunohistochemical changes in hypothalamic magnocellular neurosecretory system during and after the training. The aim of the present work was to study the immunohistochemical changes in OT, ANP and VP levels in the hypothalamic paraventricular nucleus during and after resistance exercise protocol. Three groups of Wistar rats were trained by a rung ladder protocol for 15, 30 and 45 days, respectively; a fourth group was left to rest for 15 days after the training. Finally, four sedentary groups were used as controls. The results show that resistance training induces a significant reduction in the percentage of OT‐positive neurons, compared with sedentary controls. In contrast, this protocol did not induce any change in VP levels, and ANP levels did not change significantly. However, VP increased after the resting period of 15 days. Our work shows that neurons of the paraventricular nucleus are involved in body fluid homoeostasis during and after resistance exercise. The functional significance of these changes in OT and VP levels, during and after the protocol, needs to be further investigated.

Presence and interaction in tissues of atrial natriuretic peptide, oxytocin and vasopressin: new insights

Atrial natriuretic peptide, oxytocin and vasopressin are three well known and widely studied molecules since many years. They have been fully characterised from a genetic and biomolecular point of view and a number of receptor-dependent functions have been recognised for them. Nevertheless, in the last years our group has conducted morphologic studies, using an immunohistochemical approach complemented by molecular biology techniques, and could show non-canonical localization and co-localization of these peptides in normal and pathologic tissues, that permitted us to postulate that they may be involved in a wider range of functions than usually assumed and not yet fully understood. In this minireview we summarise some of the main results that open new scenarios in the comprehension of the biologic activities of these peptides and allow to postulate a role for them as diagnostic tools.

SOMATOTOPIC ORGANIZATION OF THE CRANIAL NERVE NUCLEI INVOLVED IN EYE MOVEMENTS: III, IV, VI.

The oculomotor nucleus is divided into: 1. somatic medial column innervating the superior rectus muscle and somatic lateral column, further subdivided into a lateral column innervating the inferior rectus, inferior oblique and medial rectus muscles; 2. central nucleus innervating the levator palpebrae superioris muscle; 3. visceral nucleus innervating the pupillary sphincter muscle, consisting of the following two groups of neurons: cholinergic, preganglionic neurons supplying the ciliary ganglion, termed the EdingerWestphal preganglionic (EWpg) neuron population, and the centrally projecting, peptidergic neurons, termed the Edinger-Westphal centrally projecting (EWcp) population. A detailed understanding of the position of both nuclei and their subgroups supplying the individual muscles is an essential prerequisite for eye movement assessment, offering a simple clinical method for diagnosing eventual dysfunctions of eye movement generation pathways.

Atrial natriuretic peptide (ANP) and oxytocin-expression in the adult rat and mouse cerebellum

BackgroundMany studies are in the literature on the ANP and oxytocin-presence in the brain, but very few studies with controversial results are reported on the presence of these peptides in the cerebellum. This immunohistochemical study investigates on the ANP and oxytocin-presence in the cerebellum of the adult rat and mouse rodents.ResultsThis study, firstly, evidences the ANP- immunopositivity in cerebellar cortex of both rat and mouse rodents. In rat the molecular layer presents some few immunopositive fibers, but no neuron resulted immunopositive; the granular and Purkinje cells are immunopositive. In mouse the cerebellar cortex ANP-immunopositivity is present in all layers. The oxytocin-presence in the rat the afferent fibers are immunopositive are in the granular layer; in mouse the OT-immunopositivity is in the molecular layer only.ConclusionsThis study, firstly, shows that ANP and OT are present in the cerebellar cortex both in rat and mouse rodents. In the mouse cerebellar cortex ANP-presence is more diffuse and OT- localization differences in the two species.

Atrial Natriuretic Peptide presence in parotid gland of human fetus at 13th week of development and in adult man

Our immunohistochemical study shows that atrial natriuretic peptide (ANP) is present in both human fetal and adult parotid gland. In the fetuses ANP is localized in cell clusters, while in adult parotid gland, the ANP is seen only in the wall of intra- and extralobular ducts. The results suggest that ANP might play a role in the differentiation of the parotid gland similarly as in brain and that during fetal growth ANP may play a role in the regulating the secretion of primitive salivary fluid, even if acini are still not developed.