Road traffic deaths: A retrospective analysis (2009-2019) in the north of the Italian region Emilia Romagna.

OBJECTIVE: International literature and several national studies demonstrate that alcohol and illicit drugs impair driving abilities, diminishing the level of attention, and cause traffic accidents. In Italy, driving under the influence of alcohol or drugs is regulated by Articles 186 and 187 of the National Street Code, which defines penalties and fines for the convicted. The aim of this study was the collection of all available data from 2009 to 2019 focusing on deaths related to road accidents in the Unit of Legal Medicine of Department of Medicine and Surgery at the University of Parma, in order to assess any consumption of alcohol, illicit drugs, and medicinal drugs among drivers. METHODS: Data were retrieved from autopsy reports found at the Unit of Legal Medicine of Parma University related to 327 subjects who died following road accidents in the Italian areas of Parma, Reggio-Emilia, and Piacenza. The population was divided into subgroups according to age, gender, crash time, and drug positivity. RESULTS: Those in the age group 46 to 65 years old were involved in the most accidents, whereas the category with fewest members included subjects under 26 years old. The majority of road accidents occurred during the daytime and on weekends. Among the toxicological investigations carried out (only for drivers), the highest prevalence was found for alcohol (43.1%), followed by illicit drugs (14.4%) and medicinal drugs (7.8%). The prevalence of alcohol and illicit drugs in combination was 11.8%. Regarding subjects positive for alcohol and illicit drugs in combination, 44.4% had a blood alcohol concentration (BAC) > 1.5 g/L and overall, in 61.1% of the total cases a BAC > 0.81 g/L was detected. CONCLUSIONS: Our results are in line with national and international studies highlighting the prevalence of high BAC levels in most of the cases. Confirmation analyses on blood collected from people who died following road accidents showed levels of BAC above 0.8 g/L (threshold for penal sanctions) in the majority of the subjects who tested positive for alcohol. They also revealed cocaine, cannabis, and benzodiazepines as the most common illicit drugs and medicinal drugs used, respectively, as demonstrated in several international studies.

Biofilm in voice prosthesis: A prospective cohort study and laboratory tests using sonication and SEM analysis.

OBJECTIVE: The objective of the study was to compare the biofilm growing pattern and its morphological extent on silicone and a teflon-like material using a sonication process and a Scanning Electron Microscope (SEM). DESIGN: A prospective cohort study and a laboratory study. SETTING: Otolaryngology -Head and Neck surgery Department and the Microbiology Institute. PARTICIPANTS: The participants included fifteen laryngectomised patients with phonatory prostheses, which were removed because of device failure, and two different kinds of phonatory prostheses from the laboratory (Provox 2 and ActiValve) that were artificially colonised by Candida albicans. MAIN OUTCOME MEASURES: Tracheo-oesophageal puncture (TEP) is currently considered the gold standard for post-laryngectomy voice rehabilitation. "Leakage" represents the most common cause of substitution and is generated by biofilm colonisation of the prosthesis by mixed mycotic and bacterial agents. New biomaterials have been developed that are deemed to be more resistant to the colonisation of micro-organisms and material deformation. RESULTS: The devices showed colonisation by mixed bacterial flora (Staphylococci 13%, Streptococci 9%, and Haemophilus influenzae 5%) and by yeasts (Candida albicans 12%). Moreover, we observed a different distribution of biofilm layers in Provox ActiValve (22.56%) compared to Provox 2 (56.82%) after experimental colonisation by the previously isolated Candida strain. CONCLUSION: Resident microbiological species from the upper airways unavoidably colonise the polymer surfaces, and no strategies have been effective except for the manipulation of the chemical-physical properties of the device's polymer. Our study confirms that Provox ActiValve, which is made with a fluoroplastic material (teflon-like), is less subject to in vitro colonisation by Candida, and thus showed a higher clinical resistance to biofilm and a longer lifespan. The sonication seems to significantly improve the knowledge of bacterial and mycotic flora in biofilm colonisation. The design of a device for the daily cleaning capable to reach and brush the oesophageal flange of the prosthesis preserving the valve mechanism could represent a practical and simple help in this still unsolved problem.

Role of biofilms in chronic inflammatory diseases of the upper airways.

UNLABELLED: The objective of our studies was to document the presence of bacterial biofilms in recurrent and chronic infectious diseases of the upper airways (UA) (adenoiditis, tonsillitis, chronic rhinosinusitis) and to assess the association between the presence of biofilm and the maintenance of a chronic inflammation. METHODS: 16 surgical samples of tonsils and adenoids from patients with UA infections and 24 samples of ethmoid mucosa from patients who underwent endoscopic sinus surgery for chronic rhinosinusitis (CRS) were cultured using conventional methods and subjected to scanning electron microscopy (SEM) to detect evidence of biofilm. RESULTS: Bacterial biofilms were observed in 57.5% of chronically infected UA mucosa; in 41.7% of ethmoid mucosa of CRS patients they were significantly (p<0.001) associated with a marked destruction of ciliated epithelium. DISCUSSION: Our studies confirm that biofilm formation plays a role in UA infections, it not only explains the resistance of these infections to antibiotic therapy, but also represents an important element that contributes to the maintenance of a chronic inflammatory reaction.

Potential applications of human saliva as diagnostic fluid.

The use of human saliva as a diagnostic and prognostic fluid has until recently been somewhat disregarded. Although sample collection is non-invasive, physiological and genetic variations were largely responsible for its infrequent application in the past. Recently, several proteomic studies contributed to partial elucidation of the salivary proteome (more than 2400 protein components have been characterized), both in terms of composition, contributions to whole saliva and genetic/physiological variability. On this basis, is not too optimistic to believe that in the near future human saliva could become a relevant diagnostic fluid. In this review, the characterization by proteomic approaches of new salivary markers in oncology, head and neck carcinoma (oral cavity, oropharynx, larynx, and salivary glands), breast and gastric cancers, salivary gland function and disease, Sjögren syndrome, systemic sclerosis, dental and gingival pathology, systemic, psychiatric and neurological diseases, is described.

Vitamin K, bone fractures, and vascular calcifications in chronic kidney disease: an important but poorly studied relationship.

Vitamin K denotes a group of lipophilic vitamins determining post-translational modification of proteins. There are 2 main forms of vitamin K: vitamin K1 (phylloquinone, found in vegetables); vitamin K2 (menaquinone, produced by bacteria in the intestine and in fermented foods). Vitamin K stores are limited in humans, but it can be recycled. Vitamin K1 is principally transported to the liver, regulating the production of coagulation factors. Vitamin K2, instead, is also transported to extra-hepatic tissues, such as bone and arteries, regulating the activity of matrix Gla-protein (MGP) and osteocalcin [bone Gla-protein (BGP)]. In patients with chronic kidney disease (CKD), cardiovascular mortality is the first cause of death. Some pathogenetic mechanisms of vascular calcification (such as hyperparathyroidism, hyperphosphatemia, hypercalcemia, role of vitamin D) have been widely investigated, but the potential role of vitamin K is still uncertain. Vitamin K could play a key role, as it transforms glutamic acid residues into γ-carboxyglutamic acid, through a carboxylation process, makings both MGP (cMGP) and BGP (cBGP) biologically active. cMGP inhibits vascular calcifications (VC), while cBGP has an important role for a proper mineralization process. Uncarboxylated MGP and BGP (ucMGP and ucBGP) concentrations are indirect markers of vitamin K2 deficiency. The purpose of this review is to analyze the current literature to understand the relationship between vitamin K2 status, fragility fractures and VC in CKD patients. This analysis could be of help in planning investigations of Vitamin K status and its possible supplementation in CKD patients to avert fragility fractures and VC.

Oxidative stress-related proteins in a Conn's adenoma tissue. Relevance for aldosterone's prooxidative and proinflammatory activity.

BACKGROUND AND AIM: Angiotensin II (Ang II) induces oxidative stress (OxSt), which is essential for cardiovascular remodeling. Aldosterone also induces fibrosis and remodeling through direct effect on non-classical mineralocorticoid (MR) target tissues. However, studies on the role of aldosterone on OxSt and related factors in humans are lacking. MATERIALS AND METHODS: We assessed gene and protein expression of p22phox (RT-PCR and Western blot), NAD(P)H oxidase subunit essential for superoxide production and gene expression of transforming growth fator (TGF) beta, plasminogen activator inhibitor (PAI)-1, and heme oxygenase (HO)-1, effectors of OxSt (RT-PCR), in a Conn's adenoma, removed from a patient with primary hyperaldosteronism. Ang II type 1 (AT1R) and MR receptors expression were also evaluated (RT-PCR). The normal adrenal tissue adjacent to the adenoma was used as control. RESULTS: p22phox gene and protein expression were higher (31% and 53%, respectively) in the adrenal adenoma. TGFbeta, PAI-1, and HO-1 gene expression were also higher (25%, 129%, and 25%, respectively) in the adrenal adenoma while AT1R gene expression was similar (8%). The expression of MR in the adenoma was documented. CONCLUSIONS: This report demonstrates in a human model that the increased aldosterone production has effects on enzyme systems related to OxSt, enhancing the systemic fibrogenic effects of aldosterone excess through TGFbeta and PAI-1 expression which was previously demonstrated only indirectly in vitro and in animal models. The presence of MR expression in the adenoma may link the hormone with the adenoma growth. Therefore, the results of this study derived from a single case might represent an important working hypothesis for further research in a larger number of cases to clarify the role of aldosterone overproduction on OxSt and its clinical relevance.

[Bacterial biofilm identification in the rhinopharingeal mucosa of children with recurrent infection of the upper respiratory tract and otitis media]. / Identificazione di biofilm batterici nella mucosa rinofaringea di bambini affetti da infezioni ricorrenti delle vie aeree superiori associate ad otite media.

OBJECTIVE: The objective of the present study was to identify bacterial biofilms in tissue samples obtained from paediatric patients undergoing surgical treatment, for recurrent upper airway infections, frequently associated to effusive otitis media, unresponsive to repeated cycles of selective medical antibiotic and anti-inflammatory therapy and assay the ability of Haemophilus influenzae strains, most frequently identified in our cultural examinations, to grow as biofilm in vitro. METHODS: We examined 18 surgical specimens (18 adenoids) from the upper respiratory tract, obtained from 18 paediatric patients. Tissues were cultured using conventional methods and subjected to scanning electron microscopy for detection of biofilm. Haemophilus influenzae strains, were cultured on 96-sterile well polystyrene microtiter plates (CELLSTAR-greiner bio-one) and stained with 1% crystal violet to quantify biofilm production. RESULTS: Bacterial cocci attached to the tissue surface and organized in colonies, with a morphology consistent with bacterial coccoid biofilms, were observed in all adenoid (18/18) samples. Haemophilus influenzae isolates from 11/18 (61.1%) of our tissue samples scored a percentual transmittance (%Tbloc) > 50, identifying a high capacity to form biofilms (level 4). CONCLUSIONS: Bacterial biofilms identified in adenoid tissue of paediatric patients with recurrent upper airway inflammatory processes, associated to chronic effusive otitis media, may represent a bacterial "reservoir" responsible of the maintenance of chronic inflammatory mucosal reactions, resistant to selective antibiotic therapy and requiring surgical treatment.

Wegener's granulomatosis: an update on diagnosis and therapy.

Wegener's granulomatosis (WG) is a unique clinicopathological disease characterized by necrotizing granulomatous vasculitis of the respiratory tract, pauci-immune necrotizing glomerulonephritis and small-vessel vasculitis. Owing to its wide range of clinical manifestations, WG has a broad spectrum of severity that includes the potential for alveolar hemorrhage or rapidly progressive glomerulonephritis, which are immediately life threatening. WG is associated with the presence of circulating antineutrophil cytoplasm antibodies (c-ANCAs). The most widely accepted pathogenetic model suggests that c-ANCA-activated cytokine-primed neutrophils induce microvascular damage and a rapid escalation of inflammation with recruitment of mononuclear cells. The diagnosis of WG is made on the basis of typical clinical and radiologic findings, by biopsy of involved organ, the presence of c-ANCA and exclusion of all other small-vessel vasculitis. Currently, a regimen consisting of daily cyclophosphamide and corticosteroids is considered standard therapy. A number of trials have evaluated the efficacy of less-toxic immunosuppressants and antibacterials for treating patients with WG, resulting in the identification of effective alternative regimens to induce or maintain remission in certain subpopulations of patients. Recent investigation has focused on other immunomodulatory agents (e.g., TNF-alpha inhibitors and anti-CD20 antibodies), intravenous immunoglobulins and antithymocyte globulins for treating patients with resistant WG.

Biofilm formation by Haemophilus influenzae isolated from adeno-tonsil tissue samples, and its role in recurrent adenotonsillitis.

Aim of the present study was to identify bacterial biofilms in tissue samples obtained from paediatric patients undergoing surgical treatment, for chronic and recurrent adeno-tonsillitis, not responding to repeated cycles of selective medical antibiotic and anti-inflammatory treatment and to assay the ability of Haemophilus influenzae strains, most frequently identified in the culture examinations, to grow as biofilm in vitro. Overall, 25 surgical specimens (15 adenoids, 10 tonsils) were examined from the upper respiratory tract, from 15 paediatric patients (mean age 6 years). All patients were affected by recurrent and/or chronic adenoiditis and adenotonsillitis unresponsive to selective antibiotic and anti-inflammatory therapy. Tissues were cultured using conventional methods and subjected to scanning electron microscopy for detection of biofilm. Haemophilus influenzae strains, were cultured on 96-sterile well polystyrene microtitre plates (CELLSTAR-greiner bio-one) and stained with 1% crystal violet to quantify biofilm production. Bacterial cocci attached to the tissue surface and organized in colonies, with a morphology consistent with bacterial coccoid biofilms, were observed in all adenoid (15/15) and in 6/10 tonsil samples. Haemophilus influenzae isolates from 12/25 (48%) of our tissue samples scored a percent transmittance (%T(bloc)) > 50, displaying a high capacity to form biofilms (level 4). In conclusion identification of bacterial biofilms in chronic and/or recurrent paediatric upper airway inflammatory processes and the capacity to produce biofilm in vitro, demonstrated by Haemophilus influenzae (the most frequently identified bacteria in our samples), could be related to the aetiopathogenic role of biofilms in chronic inflammatory mucosal reactions and to the resistance of these infections to selective antibiotic therapy.

Recurrent upper airway infections and bacterial biofilms.

BACKGROUND: Bacterial biofilms identified in various medical devices used in otorhinolaryngology, including tympanostomy tubes, voice prostheses, and cochlear implants, can directly colonise mucosal tissues. The upper airways seem to be at high risk for this type of colonisation. Chronic and/or recurrent upper airway infections may be related to the complex structural and biochemical (quorum sensing) organisation of the biofilm which interferes with the activity of antibiotics (including those with proven in vitro efficacy), thus promoting the establishment of a chronic infection eradicable only by surgical treatment. Biofilm formation plays a role in upper respiratory infections: it not only explains the resistance of these infections to antibiotic therapy but it also represents an important element that contributes to the maintenance of a chronic inflammatory reaction. OBJECTIVES: To document the presence of biofilms in surgical tissue specimens from patients with recurrent infection diseases, and identify their possible role in the chronicity of these infectious processes. METHOD: We examined 32 surgical specimens from the upper respiratory tract (tonsils, adenoids, mucosa from the ethmoid and maxillary sinuses) of 28 patients (20 adults, eight children) with upper airway infections that had persisted despite repeated treatment with anti-inflammatory agents and antibiotics with demonstrated in vitro efficacy. Tissues were cultured using conventional methods and subjected to scanning electron microscopy for detection of biofilm formation. RESULTS: Over 80 per cent (26/32; 81.3 per cent) of the tissue specimens were culture-positive. Bacterial biofilms (associated in most cases with coccoid bacteria) were observed in 65.6 per cent of the tissue samples.

Interactions between ephrin-B and metabotropic glutamate 1 receptors in brain tissue and cultured neurons.

We examined the interaction between ephrins and metabotropic glutamate (mGlu) receptors in the developing brain and cultured neurons. EphrinB2 coimmunoprecipitated with mGlu1a receptors, in all of the brain regions examined, and with mGlu5 receptors in the corpus striatum. In striatal slices, activation of ephrinB2 by a clustered form of its target receptor, EphB1, amplified the mGlu receptor-mediated stimulation of polyphosphoinositide (PI) hydrolysis. This effect was abolished in slices treated with mGlu1 or NMDA receptor antagonists but was not affected by pharmacological blockade of mGlu5 receptors. An interaction among ephrinB2, mGlu1 receptor, and NMDA was supported by the following observations: (1) the NR1 subunit of NMDA receptors coimmunoprecipitated with mGlu1a receptors and ephrinB2 in striatal lysates; (2) clustered EphB1 amplified excitatory amino acid-stimulated PI hydrolysis in cultured granule cells grown under conditions that favored the expression of mGlu1a receptors; and (3) clustered EphB1 amplified the enhancing effect of mGlu receptor agonists on NMDA toxicity in cortical cultures, and its action was sensitive to mGlu1 receptor antagonists. Finally, fluorescence resonance energy transfer and coclustering analysis in human embryonic kidney 293 cells excluded a physical interaction between ephrinB2 and mGlu1a (or mGlu5 receptors). A functional interaction between ephrinB and mGlu1 receptors, which likely involves adaptor or scaffolding proteins, might have an important role in the regulation of developmental plasticity.

Effect of epoetin on HO-1 mRNA level and plasma antioxidants in hemodialysis patients.

OBJECTIVE: Patients with renal failure and undergoing hemo- (HD) or peritoneal dialysis are under oxidative stress which is thought to contribute to the long-term complications noted in this patient population. One effect of HD-induced oxidative stress is via red blood cell (RBC) membrane lipid peroxidation leading to RBC destruction and anemia. Interaction of this oxidative stress with epoetin (EPO) treatment to increase RBC number and Hb concentration remains unexplored. PATIENTS AND METHODS: This preliminary study used RT-PCR as well as colorimetric based assay approaches to evaluate the effect of EPO-alpha treatment on markers of oxidative stress in hemodialysis patients. Eighteen patients (12 males, 6 females, age range 45 - 68), were treated with EPO-alpha (Eprex) 50 UI/kg thrice weekly over an 8-month study period. Monocytes were isolated at baseline, then monthly thereafter, monocyte heme-oxygenase-1 (HO-1) and plasma Hb and antioxidant power (AOP) were determined. RESULTS AND CONCLUSIONS: Treatment with EPO increased Hb (9.4 +/- 0.7 g/dl to 10.9 +/- 0.5, mean +/- SD p < 0.001). In addition, both monocyte HO-1 mRNA (0.34 +/- 0.08 vs. 0.59 +/- 0.02 d.u. p < 0.001) and plasma AOP (1,379.8 +/- 175 micromol/l to 1,624 +/- 170, p < 0.04) increased. While AOP changes showed no correlation with other indices, increases in HO-1 and Hb were positively correlated using 2 different measures: delta Hb (peak Hb - baseline Hb) vs. delta HO-1 (peak HO-1 mRNA - baseline HO-1 mRNA) as well as delta Hb(5 months-baseline) vs. delta HO-1 (5 months - baseline) mRNA (r = 0.81, p < 0.001 and r = 0.76, p < 0.001; respectively). In conclusion, the increases upon EPO treatment of both HO-1 gene expression and plasma AOP as well as the significant correlation between delta Hb and delta HO-1 mRNA suggest that EPO treatment reduces oxidative stress via a combination of effects. These could potentially include effects on oxidative stress directly as well as effects on the levels and types of antioxidants present in plasma.

Bile reflux as possible risk factor in laryngopharyngeal inflammatory and neoplastic lesions.

Laryngopharyngeal reflux is now of major interest as an aetiologic factor in chronic inflammatory and neoplastic lesions of upper digestive tract. However, reports in the literature refer only to the irritating action of the acid component of reflux, while possible damaging action of other reflux components remains unknown. Aim of this study was to verify the hypothesis that alkaline-bile reflux could also be involved in onset of inflammatory, precancerous and neoplastic laryngeal lesions. A total of 40 consecutive gastrectomized patients coming to our Clinic from Gastroenterology Outpatient Unit for an anamnestic and clinical evaluation with videolaryngoscopy of upper digestive airways, entered the study. All presented bile or alkaline reflux as a direct consequence of gastroduodenal anastomosis (Billroth I) and gastrojejunal anastomosis (Billroth II) performed over a time span > 20 years. Oesophagogastroduodenoscopy revealed the presence of bile in the residual gastric cavity in all operated patients objectively confirming duodenogastric reflux. Examination of data showed that 3 patients (7.5%) had undergone CO2 laser cordectomy in the 3 years prior to the study for squamous cell laryngeal carcinoma, 3 patients (7.5%) had leukoplakia, 8 (20%) vocal cord chronic oedema with signs of chronic diffuse laryngitis, 6 (15%) posterior laryngitis, 8 (20%) interarytenoid oedema while only 12 (30%) showed no ENT lesions. Statistical analysis revealed a significant correlation between incidence of inflammatory and neoplastic laryngeal lesions and type of surgery (Billroth II and total gastrectomy) with respect to other types of gastric resection. There was also a significant increase in presence and severity of laryngopharyngeal lesions in relation to time elapsed after surgery. These results, although preliminary, seem to confirm that some components of reflux (duodenal content), other than the acid component, play a damaging role involved in the onset of multiple clinical signs and symptoms of laryngopharyngeal reflux disease. It is concluded that systematic use of bile measurement, together with 24-hour pH monitoring, is advisable in subjects with clinical signs and symptoms of laryngopharyngeal reflux, but unresponsive to classic medical treatment, and in gastrectomized patients in order to confirm, on larger series, this fascinating aetiopathogenetic hypothesis.

The role of acid and alkaline reflux in laryngeal squamous cell carcinoma.

HYPOTHESIS: At present, main factors considered responsible for the onset of squamous cell carcinoma are tobacco smoking, alcohol abuse, and exposure to viral and toxic agents. In last years, great interest has been focused on gastroesophageal reflux as independent carcinogenic factor and co-carcinogen in association with smoking and alcohol assumption. STUDY DESIGN: Initially, the aim of this study was to objectively evaluate the presence of distal and proximal esophageal reflux with multielectrode pH measurement in patients with cancer of the larynx and/or hypopharynx (group A). However, in the course of the study, pharyngolaryngeal cancer was also observed in 4 patients with achlorhydria; therefore, the hypothesis that alkaline reflux might be involved in the onset of laryngeal cancer was tested (group B). METHODS: Twenty-one consecutive patients with laryngeal or hypopharyngolaryngeal squamous cell carcinoma (group A) entered the study. Twenty-one patients without laryngo-pharyngeal diseases were used as control subjects. A validated questionnaire of the clinical history was completed by all patients who underwent 24-hour pH monitoring. Group B included 40 consecutive gastrectomized patients (28 males and 12 females) in whom biliary or alkaline reflux was directly consequent to Billroth I or Billroth II operation. The control group was composed of 40 non-gastrectomized dyspeptic patients. The clinical history was controlled and obtained; EDGS and ENT examination with videolaryngoscopy was performed in all patients. RESULTS: In group A, pH measurement showed pathological reflux in 80.9% (17 of 21) of patients with no typical symptoms in 63.7% of them. The difference was significant with respect to the control group. In group B, 6 of 40 (15%) had preneoplastic lesions or a history of laryngeal tumor. The difference was significant with respect to the control group. A total of 7.5% of group B patients had previously undergone CO2 laser cordectomy for laryngeal squamous cell carcinoma and 7.5% had leukoplakia. We found a significantly higher incidence (<.05) of neoplastic and preneoplastic disease of the larynx in patients undergoing Billroth II and total gastrectomy than in those undergoing Billroth I and Roux-en-Y resection. We also found a significant increase (<.01) in ENT lesions in the group of patients who had undergone gastrectomy more than 20 years previously. CONCLUSIONS: In agreement with literature reports, results obtained in group A confirmed that gastroesophageal reflux is often present in patients with neoplastic lesions of the pharynx and larynx. Furthermore, gastric resection is indicated for the first time as an additional risk factor or cofactor of precancerosis and squamous cell carcinoma of the pharynx or larynx. Further studies are necessary to establish the cause and effect relationship between biliary reflux and pharyngo-laryngeal tumors.

Diabetic ketosis activates lymphomonocyte-inducible nitric oxide synthase.

AIMS: Inappropriate production of nitric oxide (NO) may be responsible for the haemodynamic disturbances of diabetic ketoacidosis. We investigated whether this metabolic condition is associated with increased plasma nitrate (the stable oxidation product of NO) levels and NO synthase gene expression in lymphomonocytes. RESEARCH DESIGN AND METHODS: Plasma nitrate concentrations, lymphomonocyte-inducible nitric oxide synthase (iNOS) gene expression, tumour necrosis factor-alpha (TNF-alpha) and soluble thrombomodulin were measured in 11 Type 1 diabetic patients at baseline, during mild ketosis and after euglycaemia was re-established. RESULTS: During diabetic ketosis plasma nitrate concentrations were higher (18 (16-21) vs. 9 (7-11) micro mol/l; (95% lower-upper confidence interval) P < 0.05) than at baseline. At baseline lymphomonocyte iNOS mRNA expression and iNOS protein levels were undetectable, but in ketosis both were increased (both at P < 0.0001). After recovery from ketosis, NO3 concentration, iNOS mRNA, and iNOS expression (270 +/- 36%, mean +/- sd) decreased but not significantly. No significant changes were observed in either TNF-alpha or soluble thrombomodulin levels between the three conditions. CONCLUSIONS: Diabetic ketosis is associated with increased nitrate levels and the activation of iNOS expression in circulating lymphomonocytes, but it does not affect either the proinflammatory cytokine TNF-alpha or a marker of endothelial dysfunction such as thrombomodulin. Our data support the hypothesis that, during diabetic ketosis, alterations in NO homeostasis are present in circulating lymphomonocytes.

Warm hepatic ischemia in pigs: effects of L-arginine and oligotide treatment.

Reperfusion injury represents a key event leading to graft nonfunction. Maintaining adequate nitric oxide levels and stimulating vasodilator synthesis can probably minimize endothelial damage. The aim of this study was to investigate the effect of L-arginine, a substrate of nitric oxide synthesis, and oligotide, a promoter of prostacyclin synthesis, on liver function and morphology after warm ischemia-reperfusion injury. After constructing a side-to-side portacaval shunt, ischemia was induced by clamping the hepatic hilum for 2 h above the shunt, in 19 female pigs divided into a control group (n = 7), an L-arginine treatment group (n = 6), and an oligotide treatment group (n = 6). Liver function tests and measurements of serum and red blood cell malondialdehyde and plasma nitric oxide levels were performed before reperfusion and at 1, 10, 60, and 120 min after reperfusion. Liver biopsies, taken before reperfusion and at 30 min and 7 days after reperfusion, were analyzed for tissue malondialdehyde, histological-ultrastructural features, and apoptosis evaluation. Thirty minutes after reperfusion, liver malondialdehyde, sinusoidal congestion, necrosis, and apoptosis were significantly lower in the L-arginine group than in the controls (p < .05). On postoperative day 7, tissue malondialdehyde decreased, while plasma nitric oxide and hepatocyte glycogen content were increased in the L-arginine group compared to controls (p < .05). This study demonstrates the protective effect of L-arginine on hepatic lipoperoxidation and liver morphology in a pig model of warm ischemia-reperfusion injury. The increased plasma levels of nitric oxide a week after ischemia-reperfusion injury support the hypothesis that it has a role in preventing liver damage. The same beneficial effect was not confirmed for oligotide.

[Role of gastroesophageal reflux in precancerous conditions and in squamous cell carcinoma of the larynx: our experience]. / Ruolo del reflusso gastroesofageo nelle precancerosi e nel carcinoma squamocellulare della laringe: nostra esperienza.

The major risk factors for the onset of precancerous lesions and squamous cell carcinoma of the larynx are, above all, tobacco smoke, alcohol abuse and exposure to viral and toxic agents. In recent years, however, gastro-esophageal reflux (GER) has also aroused significant interest not only as carcinogen but also as co-carcinogen in association with smoking and alcohol consumption. The purpose of the present work is to provide an objective evaluation of the presence of distal and proximal esophageal reflux using multi-electrode pH monitoring in patients with precancerous lesions of the larynx and laryngeal and pharyngolaryngeal neoplasms. A total of 24 patients consecutively hospitalized during 2000 were evaluated: 20 with squamous cell carcinoma of the larynx and/or pharynx-larynx and 4 with precancerous vocal cord lesions. All the patients provided a case history using a validated questionnaire and underwent electronic videolaryngoscopy and 24-hour pH monitoring. Data analysis showed that in 83.3% of the cases (20/24) pH monitoring was indicative of pathological GER and 63.7% of these patients had no complaints related to reflux. Moreover, 7/24 patients have had a previous gastrectomy (Billroth II) strictly related to the presence of bile reflux. The Authors conclude that, according with recent literature, pH monitoring data further indicate that patients with precancerous or neoplastic pharyngeal-laryngeal lesions present often a positive history of GER, even though no direct cause-effect relationship can yet be established because of the co-existence of other important risk factors such as smoking and alcohol consumption. Additionally, the achlorhydria found in 7/24 patients with an history of gastrectomy could suggest a possible, as yet unproven, role of other reflux components--in particular, of bile (alkaline) and chlorhydro-peptic (acid) components--as a risk factor or co-factor for tonset of precancerous and/or neoplastic lesions of the larynx.

[Gastro-esophageal reflux and laryngeal phlogistic disorders: clinical evaluation and multi-electrode pH monitoring]. / Malattia da reflusso gastroesofageo e patologia flogistica laringea: valutazione clinica e mediante pHmetria multielettrodo.

Gastro-esophageal reflux (GER) is a complex pathology characterized by many clinical symptoms: gastroenterological (typical symptoms) and odontostomatological, pneumological and, more frequently, otorhinolaryngological (atypical symptoms). For this last manifestations no cause-effect relationship has yet been proven. Using a validated questionnaire about the typical reflux symptoms and the use of substances of abuse (tobacco and alcohol) and with the application of instrumental means such as electronic videolaryngoscopy and multi-electrode 24 hours esophageal pH monitoring, the Authors have studied 34 selected patients (16 female, 18 male) suffering from chronic clinical laryngeal symptoms and presenting a peculiar pharyngeal-laryngeal objectivity. Both these manifestations could be ascribed to atypical GER symptoms. Analysis of the data showed that 67.6% of the subjects presented pathological reflux, with a significant prevalence of inflammatory lesions in the posterior portion of the larynx (in 47.8% laryngitis and retrocricoid edema). The validated questionnaire on typical reflux symptoms showed that 69.5% did not complain of the typical gastro-esophageal symptoms thus indicating that pharyngeal-esophageal reflux can be totally asymptomatic. While confirming the strong association between inflammatory pathologies and GER, not enough cases have been studied to identify a direct cause-effect relationship and this is complicated by the presence of other concomitant risk factors such as cigarette smoking or alcohol consumption. Therefore the Authors underline the importance of an in-depth, multi-discipline approach in the evaluation of pharyngeal-laryngeal phlogoses. This approach must make use of all available modern diagnostic techniques (Electronic videolaryngoscopy, multi-electrode pH monitoring) in order to obtain as much information as possible to throw light on the pathogenic mechanism responsible for the damage and find more effective forms of anti-reflux therapy.