Nerve Growth Factor and the Role of Inflammation in Tumor Development.

Nerve growth factor (NGF) plays a dual role both in inflammatory states and cancer, acting both as a pro-inflammatory and oncogenic factor and as an anti-inflammatory and pro-apoptotic mediator in a context-dependent way based on the signaling networks and its interaction with diverse cellular components within the microenvironment. This report aims to provide a summary and subsequent review of the literature on the role of NGF in regulating the inflammatory microenvironment and tumor cell growth, survival, and death. The role of NGF in inflammation and tumorigenesis as a component of the inflammatory system, its interaction with the various components of the respective microenvironments, its ability to cause epigenetic changes, and its role in the treatment of cancer have been highlighted in this paper.

Urine Dipstick Analysis on Automated Platforms: Is a Reliable Screening Tool for Proteinuria? An Experience from Umberto I Hospital in Rome.

Urinalysis is commonly used as a screening tool for kidney disease. In many cases, the dipstick urine assay includes the assessment of albumin/protein and creatinine; consequently, the value of their ratio is available on the urine section report. Identification of albuminuria/proteinuria at early stages is an important issue to prevent or at least delay the onset of chronic kidney disease (CKD), kidney failure, and the progression of cardiovascular damage linked to the kidney's loss of function. Sensitive and specific diagnostic methods are required for the assessment of such an important biomarker: urine albumin, creatinine, and their ratio (ACR) measured with quantitative assays are considered the gold standard. Routine dipstick methods (more rapid and at a lower cost) are intended for wide population screening. The aim of our study was to verify the reliability of an automated urinalysis dipstick method by comparing the results with the quantitative test of creatinine and albumin performed on a clinical chemistry platform. The first-morning voids of 249 patients who arrived from different departments were analyzed in the Central Laboratory of the University Hospital Policlinico Umberto I in Rome. We found a good correlation between the two assays, even though we observed that the dipstick assessment tends to overestimate the ACR's value, disclosing a higher number of false positives if compared to the reference method. As an important novelty in this study, we analyzed our data considering age (starting from pediatric to geriatric patients) and sex as variables for a sub-stratification of the participants. Our results show that positive values need to be confirmed with quantitative methods, especially in women and younger people, and that from samples that resulted as diluted at the dipstick assay, the ACR's values can be obtained if they are reanalyzed with quantitative assays. Moreover, patients with microalbuminuria (ACR 30-300 mg/g) or severe albumin urinary excretion (ACR > 300 mg/g) should be reanalyzed using quantitative methods to obtain a more reliable calculation of the ACR.

Alcohol and Head and Neck Cancer: Updates on the Role of Oxidative Stress, Genetic, Epigenetics, Oral Microbiota, Antioxidants, and Alkylating Agents.

Head and neck cancer (HNC) concerns more than 890,000 patients worldwide annually and is associated with the advanced stage at presentation and heavy outcomes. Alcohol drinking, together with tobacco smoking, and human papillomavirus infection are the main recognized risk factors. The tumorigenesis of HNC represents an intricate sequential process that implicates a gradual acquisition of genetic and epigenetics alterations targeting crucial pathways regulating cell growth, motility, and stromal interactions. Tumor microenvironment and growth factors also play a major role in HNC. Alcohol toxicity is caused both directly by ethanol and indirectly by its metabolic products, with the involvement of the oral microbiota and oxidative stress; alcohol might enhance the exposure of epithelial cells to carcinogens, causing epigenetic modifications, DNA damage, and inaccurate DNA repair with the formation of DNA adducts. Long-term markers of alcohol consumption, especially those detected in the hair, may provide crucial information on the real alcohol drinking of HNC patients. Strategies for prevention could include food supplements as polyphenols, and alkylating drugs as therapy that play a key role in HNC management. Indeed, polyphenols throughout their antioxidant and anti-inflammatory actions may counteract or limit the toxic effect of alcohol whereas alkylating agents inhibiting cancer cells' growth could reduce the carcinogenic damage induced by alcohol. Despite the established association between alcohol and HNC, a concerning pattern of alcohol consumption in survivors of HNC has been shown. It is of primary importance to increase the awareness of cancer risks associated with alcohol consumption, both in oncologic patients and the general population, to provide advice for reducing HNC prevalence and complications.

Anti-purkinje cell and natural autoantibodies in a group of psychiatric patients. Evidences for a correlation with the psychopathological status.

Phenomena of autoimmunity are frequent among psychiatric patients, but we don't know yet if they should be considered primary and linked to the pathophisiology of the disorder, or aspecific and associated to a general immune system activation. Paraneoplastic Cerebellar Degeneration (PCD) represents a well known model of specific autoimmunity. In order to better understand the abovementioned issues, we used this condition to compare a set of immune dysfunctions found in a group of psychiatric patients. For this reason we tested sera from 48 psychiatric patients (24 schizophrenics, 17 bipolars and 7 obsessive-compulsive), 22 PCD patients and 52 healthy controls for the presence of anti-Purkinje autoantibodies and of some natural autoantibodies (ANAs, AMAs, APCAs, ASMAs). Psychopatological status of the psychiatric patients was assessed with BPRS, SANS, SAPS, HAM-D, CGI-S. In the psychiatric group anti-Purkinje autoantibodies were identified in 11/48 (22,9%) patients, while they were present in 22/22 (100%) PCD patients and in 0/52 (0%) healthy controls. Among all anti-Purkinje autoantibody positive patients (in the PCD and psychiatric samples), only those belonging to the psychiatric sample, but not those with PCD, were frequently found positive also for natural autoantibodies, that are considered good markers of aspecific immune activation. In these patients, both anti-Purkinje and natural autoantibodies were found associated with acute/positive psychopathological symptoms. These results seem to point out that some phenomena of auto-immunity described in psychiatric patients could be aspecific, unrelated to the pathophysiology of the concomitant mental disorders and could be more frequent during phases of acute/positive symptoms.

PCNA immunoreactivity revealing normal proliferative activity in the brain of adult Lampetra planeri (Bloch, 1784).

It is now well known that the Teleosts among Osteichthyes, Urodele and Anuran Amphibians, Lacertilian Reptiles possess encephalic natural proliferative activities even into adulthood, as demonstrated by a great number of researches performed both under normal and various experimental conditions. Few years ago we have undertaken in adult heterothermic vertebrates a reappraisal on spontaneous cerebral proliferative events involving some organisms (Podarcis sicula, Triturus carnifex, Rana esculenta, Carassius carassius) representative of these vertebrates and belonging to the same or phylogenetically similar species used by previous researchers in studies having the same object. In our investigations, these performances were revealed by a proliferative immunocytochemical marker, the Proliferating Cell Nuclear Antigen (PCNA). At this point of our study in the scenario emerging from findings a missing piece is represented by Petromyzontidae. To fill up this gap in the present investigation, using our usual test, we have paid attention to adult specimens of Lampetra planeri. The obtained immunostaining panorama has revealed the presence of a considerable number of spontaneous proliferative activities. These events might differ in quantity, in various encephalic districts. PCNA-labelled cells appeared scattered in the cranial portion of olfactory bulbs, while the PCNA expression has been observed steadily localized with a distinctly continous distribution in cells interposed among the ependymal epithelium which lines the cavities of the proximal portion of the olfactory region and of the cerebral ventricles. DNA synthesis activity has been also found in cells scattered in the telencephalic, diencephalic, mesencephalic and medulla oblongata periventricular grey. This immunoreactivity was not revealable in the cerebellum. Our findings are discussed in the light of bibliographic news.

Antithrombin III metabolism in the pulmonary vessel endothelium.

In 85 patients undergoing aorto-coronary bypass for atherosclerotic coronary disease, we measured the antithrombin III activity levels and the thrombin-antithrombin III complex concentrations in blood from the pulmonary and the radial arteries, taken before the aorto-coronary bypass procedure, with the aim of investigating the role of the pulmonary endothelium in the metabolism of the inhibitor. Results showed significantly lower mean antithrombin III activity levels, expressed as a percentage of normal plasma, in blood from the radial artery with respect to levels from the pulmonary artery (0.78 +/- 0.12 versus 0.80 +/- 0.12, P<0.0001), while no significant difference was found in thrombin-antithrombin III complex concentrations. The results seem to show that the pulmonary endothelium contributes to the antithrombin III metabolism with a 0.023 breakdown rate, corresponding to about a 0.1 fraction of the reported 0.22-0.25 total body catabolic rate, as well as the pulmonary endothelial surface (50-70 m2) corresponding to about a 0.1 fraction of the peripheral vessels' endothelial surface (500-700 m2). The data support the hypothesis of a main endothelial catabolism of antithrombin III.

Expression of PCNA positivity in the brain of normal adult heterothermic vertebrates: further observations.

As part of our study of non-experimentally induced encephalic proliferation in unequivocally adult individuals of several heterothermic Vertebrates (Podarcis sicula, Triturus carnifex, Rana esculenta, Carassius carassius), we deal here with areas not considered in previous investigations, i.e. various encephalic regions (except the telencephalon) in Podarcis sicula, Triturus carnifex and Rana esculenta, the diencephalon and medulla oblongata in Carassius carassius, and the olfactory bulbs in the two Amphibians. In the previous and current research, we have used Proliferating Cell Nuclear Antigen (PCNA) as a marker. PCNA is a ubiquitous intracellular antigen of the cycline family (proteins that regulate the cell cycle), which acts as an auxiliary protein to DNA polymerase delta; it can be detected immunocytochemically with monoclonal antibodies to reveal cell cycle phases that coincide with DNA synthesis. Spontaneous proliferation events, revealed by PCNA positivity, were constantly present in this study, being substantial in the olfactory region and diencephalon, very modest in the mesencephalon and myelencephalon, and absent in the cerebellum. In particular, signs of proliferation were abundant in the epithelium lining the cavities of the olfactory bulbs, while they were of different magnitude in tracts (with multiple and comparatively different sites related to the dorsal and/or ventral thalami) of the ependyma that delimits portions of the III ventricle and also, in all the species examined, at the level of the preoptic and infundibular recesses. Such signs were rare in the ependymal epithelium of the mesencephalic ventricle in Podarcis sicula and the rhombencephalic ventricle in all four species examined. This immunoreactivity was also observed in extra-ependymal areas: in the internal granular layer of the olfactory bulbs in Triturus carnifex and Rana esculenta; in the diencephalic nuclei of the habenula in Podarcis sicula, in both Amphibians and in Carassius carassius; in the mesencephalic tectum in Podarcis sicula and in the two Amphibians. As in our previous studies, the current immunocytochemical picture revealed by PCNA positivity generally agrees with literature reports on the presence of normal proliferation in the areas investigated here. These literature sources consist primarily of the observations of Kirsche (1967), emerging from his preceding experimental investigations, and of confirmatory data from studies in subsequent decades by other researchers obtained with tests different from our marker. Nevertheless, the number of studies that deal with the species considered in the present research, or species closely related to them, is rather limited.

Signs of normal proliferation in the telencephalon of adult male songbirds (serinus serinus), as shown by PCNA-positivity.

The immunocytochemical expression of Proliferating Cell Nuclear Antigen (PCNA) (a cycline that coadjuvates DNA polymerase delta) becomes appreciable in the cell cycle when DNA synthesis occurs; hence cells in the S phase can be revealed by means of monoclonal antibodies. Therefore, PCNA can be considered a marker of proliferation, and numerous literature reports have demonstrated the reliability of the PCNA test. Since normal neurogenic events can still occur in the brain tissue of adult homeothermic vertebrates (especially songbirds), we evaluated if the persistence of spontaneous proliferation could be revealed in adult male songbirds (Serinus serinus) using the PCNA marker, the same test we used previously to study the persistence of natural proliferation in the encephalon of adult heterothermic vertebrates. The patterns of PCNA positivity showed normal proliferation in the telencephalon of the adult male Serinus serinus. This activity was shown by cells interposed among the epithelial cells lining the lateral side of each ventricular cavity, both in correspondence to the apical tracts and declivities of the ependyma and arranged, here and there, either in groups or slightly separated. As in our previous studies on PCNA expression and persistence of spontaneous encephalic proliferation in adult poikilothermal vertebrates (in the telencephalon of Podarcis, Triturus and Rana, and in the telencephalon, mesencephalon and cerebellum of Carassius), the results of the present research largely agree with the findings of previous Authors, usually obtained with different methods. This agreement confirms the reliability of the PCNA test used for this type of investigation.

PCNA-negativity in the telencephalon of adult female songbirds (Serinus serinus): absence of signs of normal proliferation.

In a recent study, we demonstrated the persistence of normal proliferation in the telencephalon of adult male songbirds (Serinus serinus), as shown by the expression of Proliferating Cell Nuclear Antigen (PCNA), a test that uses monoclonal antibodies to reveal cells in the S phase. Near the start of the breeding season, this proliferation was observed in circumscribed areas of the ventricular ependymal epithelium (zona germinativa dorsalis and zona germinativa ventralis) and in small masses of cells that were grouped or separated ("hot spots"). Therefore, we decided to extend this research to normal adult females of the species, using the same marker and the same time period as in the previous study. This time, however, we did not observe PCNA-positivity, and thus any sign of natural proliferation, in the telencephalic areas that showed it in the male songbirds. The immunocytochemical patterns recorded in females and males of Serinus serinus agree with the information reported in the literature, namely that in adult homeothermic Vertebrates the presence of spontaneous cerebral neurogenesis is limited to male songbirds.

Increased lymphocyte dopamine beta-hydroxylase immunoreactivity in Alzheimer's disease: compensatory response to cholinergic deficit?

There is growing interest in the characterization of peripheral blood lymphocytes (PBL) as a biological tool with which to investigate changes in the neurotransmitter-receptor system in neurodegenerative disorders. Here we show a slight decrease in acetylcholinesterase (AChE) and a significant increase in dopamine beta-hydroxylase (DBH) immunoreactivity in the PBL of patients with probable Alzheimer's disease (AD). Therapy with AChE inhibitors completely reversed the increase in DBH immunoreactivity. We hypothesize that the increase in DBH immunoreactivity may represent a compensatory response to cholinergic impairment. Our findings suggest that neurochemical interactions between the noradrenergic and cholinergic systems may be measured at a peripheral level in AD.

Expression of OP4 (ORL1, NOP1) receptors in vascular endothelium.

Endothelial cells from rat brain microvessels, human aortic artery and human umbilical vein were examined, together with ex vivo rat brain capillaries and rat aortic ring sections, for the expression of opioid receptor-like OP-4 mRNA and protein. High levels of mRNA expression and an immunopositive reaction for the receptor protein were detected in the endothelial cells from primary and from established in vitro cultures, as well as in the intima of ex vivo rat aortic rings, where the signal was limited to the endothelial layer. Interaction of the OP4 receptor with its physiological ligand nociceptin caused, in cultured endothelial cells, the activation of a mitogen-activated protein (MAP) kinase cascade. Taken together, these results show that the OP4 receptor is synthesised and functionally expressed in endothelial cells, presumably as a starting point for some vasoactive mechanism(s).

Myelin basic protein induces heme oxygenase-1 in human astroglial cells.

Heme oxygenase-1 (HO-1), also known as heat-shock protein 32 (HSP-32), is induced in many cells by a large variety of stimuli. Its induction in nervous system cells following toxic and oxidative stress was suggested to play a protective role. Its presence was recently detected by immunohistochemical studies at the level of inflammatory lesions of rat experimental autoimmune encephalomyelitis. In the present study, we demonstrate that myelin basic protein (MBP) induces HO-1 in human astroglial cells, as shown by Western blots and RT-PCR. Proteolytic fragments derived from the whole MBP show a different behavior in the HO-1 induction: MBP152-167 was able to produce a light but still significant increase in HO-1 mRNA and protein levels, whereas MBP68-84 was not. The increase in HO-1 production seems to be mediated by a Ca(2+)-dependent mechanism, since MBP addition to astrocytoma cultures induced a strong and immediate increment of [Ca(2+)](i) increase; MBP152-167 elicited a delayed and less pronounced [Ca(2+)](i) increase; no [Ca(2+)](i) changes were induced following cell treatment with MBP68-84. NO pathway involvement in the induction of HO-1 by MBP was ruled out since the expression of the inducible isoform of nitric oxide synthase was not upregulated in treated cells, neither nitrite levels were modified, as demonstrated by Greiss reaction. The possible significance of HO-1 induction following MBP stimulation is discussed.