Non-lactose-fermenting uropathogenic Escherichia coli from dogs: virulence profile characterization.

Non-lactose-fermenting Escherichia coli (NLFEC) has a few descriptive studies restricted to human infections. In the present study, isolates of NLFEC obtained from urine samples of dogs with hyperadrenocorticism were characterized regarding their virulence ability, biofilm formation capacity and antimicrobial susceptibility profile. Escherichia coli lactose-fermenting strains from urinary infection in dogs with the same conditions were analysed to provide comparisons. The non-lactose-fermenting E. coli strains were classified as belonging to clade I E. coli, whereas the lactose-fermenting strains were classified in phylogroup B2. All strains presented virulence markers to adhesion, iron acquisition, toxins, colicin and cytotoxin production, and biofilm regulation. Components of the extracellular matrix in addition to the in vitro biofilm formation ability were observed in the strains. Multidrug resistance (MDR) profiles were observed by in vitro susceptibility tests to all NLFEC strains. In summary, non-lactose-fermenting uropathogenic E. coli from dogs behaves similar to lactose-fermenting E. coli, exhibiting MDR profile, and pathogenic potential of promote animal infections.

Molecular detection of Salmonella serovars Enteritidis, Heidelberg and Typhimurium directly from pre-enriched poultry samples.

1. Salmonella is one of the most important pathogens in public health and it is usually associated with food-borne diseases. Salmonella serovars Enteritidis and Typhimurium are widespread in the world with outbreaks frequently associated with consumption of poultry products; furthermore, there is an increasing public health concern with the wide dissemination of the serovar Heidelberg in poultry flocks. 2. The aim of the experiment was to develop and to validate rapid methods to detect Salmonella serovars Enteritidis, Typhimurium, and Heidelberg by real-time PCRs and test isolates from pre-enriched poultry samples. 3. Three real-time PCRs were developed and used in combination to detect the serovars Enteritidis, Typhimurium and Heidelberg. These assays were validated by the analysis of 126 Salmonella isolates, eight other enteric bacterial species and 34 naturally contaminated poultry samples after pre-enrichment with buffered peptone water (BPW). 4. Real-time PCRs detected the isolates of the most important poultry serovars (Enteritidis, Typhimurium and Heidelberg) with 100% inclusivity and exclusivity in each assay. The PCR identified monophasic variants of the serovars Typhimurium and Heidelberg. All PCRs were validated in detecting these specific serovars directly from pre-enriched poultry samples. The whole analytical procedure was performed in less than 24 h in a veterinary diagnostic laboratory.

Laparoscopic transperitoneal hernia repair (TAPP) in emergency: long-term follow-up in a high volume centre.

PURPOSE: Aim of this study was to analyse feasibility, safety and effectiveness of laparoscopic transperitoneal hernia repair (TAPP) approach to inguinal hernias in the emergency setting, with a longer follow-up than the studies present in literature. METHODS: We retrospectively analysed all patients who underwent emergency TAPP hernia repair in San Marco Hospital (Zingonia, Italy), from September 2010 to June 2020. A prospectively collected database of 685 consecutive TAPP hernia repair was reviewed. Feasibility and safety were evaluated through operative time, conversion rate, perioperative mortality, morbidity and prosthesis infection rate. Effectiveness was assessed by recurrence and complication rate, acute (during admission) and chronic (during follow-up) pain by Visual Analogic Scale (VAS), and recovery to normal activity in a long-term follow-up (mean period 1718 days). RESULTS: The final study group included 47 patients. The mean age was 59.6 years (range 22-89), 29 patients were male and 18 were female. The mean operation time was 64 min. Two cases were converted to open approach (4%). In four cases bowel resection became necessary (8.5%). Appendicectomy and omentectomy occurred once and twice, respectively (2% and 4%), and orchiectomy was required in two patients (4%). Seroma was the only postoperative complication that we registered in four patients. During a mean follow-up period of 1718 days, there was no recurrence of the hernia or other complications. Five patients referred mild occasional groin pain (VAS < 3), with quick relief without taking any pain killers. CONCLUSION: TAPP approach is a safe, feasible and effective therapeutic option for groin hernias in emergency setting. No recurrence or severe complications were reported in over 4 years of follow-up.

Insights on the genetic features of endometrial pathogenic Escherichia coli strains from pyometra in companion animals: Improving the knowledge about pathogenesis.

Endometrial pathogenic E. coli (EnPEC) isolates are involved in endometrial infections in animals and humans. Besides the high aggressiveness of the endometrial infections, the EnPEC virulence profile and pathogenesis are still little known. In this study, we have sequenced and analyzed an EnPEC strain from canine pyometra (E. coli_LBV005/17), following a molecular characterization of the virulence profile and phylogenetic evolution of an EnPEC collection from canines and felines (45 strains). Most of the strains belonged to phylo-group B2, and display a high virulence profile. In particular we highlight the classification of the E. coli_LBV005/17 as sequence type 131 (ST131), in addition to other five strains, as observed by gyrB phylogenetic analysis. Also, the phylogenetic position of EnPEC strains from pyometra in companion animals suggests that their origins are from both extraintestinal and commensal E. coli strains. Accordingly to Principal Coordinates Analysis (PCoA) and phylogenetic analysis we can propose that EnPEC strains have neither the same genetic profile, nor a unique common ancestral. In summary, the present work characterize an EnPEC genome from bitch pyometra and the genetic profile of 45 EnPEC strains from companion animals pyometra, being the commonest virulence pattern: fimA, papC, hlyA, hlyE, cnf1, entB, iroN, irp1, bssS, bssR, and hmsP. These data improving the background knowledge of this E. coli pathotype related to pyometra in companion animals and may support new methods to prevent the disease evolution.

Asthma induced by inhalation of flour in adults with food allergy to wheat.

BACKGROUND: Wheat is one of the major food allergens and it is also an inhalant allergen in workers exposed to flour dusts. Food allergy to wheat in adulthood seems to be rare and has never been reported to be associated with asthma induced by flour inhalation. OBJECTIVE: The study aimed at detecting adults with food allergy to wheat and screening them for the presence of specific bronchial reactivity to inhaled wheat proteins. METHODS: Adults with a history of adverse reactions to ingestion of wheat underwent skin prick test with commercial wheat extract and were assessed for the presence of specific wheat IgE in the sera. Food sensitivity to wheat was confirmed by double-blind, placebo-controlled food challenge (DBPCFC). Specific bronchial reactivity was investigated through a specific bronchial challenge with wheat proteins. RESULTS: In nine patients with evidence of specific IgE response to wheat, a diagnosis of food allergy was made by DBPCFC. Only two subjects had asthma as disease induced by ingestion of wheat. Seven subjects reported a history of respiratory symptoms when exposed to flour dusts. A significant reduction of forced expiratory volume in 1 s (FEV(1)) was detected in these seven patients when a specific bronchial challenge with flour proteins was performed. Only three out of seven subjects with asthma induced by flour could be considered occupationally exposed to flour dusts. CONCLUSION: For the first time, it has been shown that specific bronchial reactivity to wheat proteins can be detected in patients with different disorders associated with food allergy to wheat. The presence of asthma induced by inhaled flour is not strictly related to occupational exposure and it may also occur in subjects not displaying asthma among symptoms induced by wheat ingestion.

Stapled hemorrhoidopexy: day surgery or one day surgery?

Hemorrhoidal disease is one of the most common anorectal disorders, from 10% to 20% of all patients admitted at a clinical investigation need to undergo surgery, stapled haemorrhoidopexy is gaining wide acceptance as an interesting, safe and less painful technique, but hemorrhage is one of the most serious early complications and is a severe complication in day surgery. In our day surgery proctology, surgical procedures represent about 32%. Of these, 24% are for hemorrhoidal disease, we present our protocol and experience for early and safe discharge, 6h after stapled hemorrhoidopexy surgery.

Outcomes of laparoscopic Miles' operation in very low rectal adenocarcinoma. Analysis of 32 cases.

AIMS: Minivasive techniques for excision of low rectal tumours have spread worldwide with good results, but their employment is still under discussion. The purpose of this study is to assess short term results and survival of laparoscopic abdominoperineal resection (LAPR) in very low rectal cancers. METHODS: The charts of 32 patients undergoing LAPR for very low rectal adenocarcinoma (0-2cm from dentata line) were reviewed retrospectively. Outcomes were evaluated considering surgical procedure, short and long-term results and survival. RESULTS: A thorough LAPR was performed in 31 patients and conversion to laparotomy was required in 1 patient. Mean operating time was 244min. The length of hospital stay (LOS) was 13,3days. The mean number of nodes collected was 12 and the distal margin was 3,6cm on average. There was 1 post-operative death. In the follow up no pelvic recurrence was observed, while metachronous metastases were observed in 5 patients and peritoneal carcinosis in 2 patients. No port site metastasis was registered. Cumulative 5year survival probability was 0,50. CONCLUSIONS: The outcomes of this study suggest that LAPR in very low rectal cancer is a reliable procedure, operating time and LOS were acceptable. Oncologic principles were respected: length of specimen, distal margin and number of nodes retrieved were quite acceptable. Pelvic recurrence frequency was nil. Long term results were comparable with those of other series.

Hypertension and abnormalities of carbohydrate metabolism possible role of the sympathetic nervous system.

To investigate the relationships between the sympathetic nervous system (SNS) and parameters of glucose metabolism in arterial hypertension, daily urinary excretion of catecholamines and plasma glucose, insulin, and C-peptide response to an oral glucose load (OGL) have been evaluated in 77 untreated patients with mild-to-moderate essential hypertension and in 31 normotensive controls. Urinary excretion of norepinephrine (UNE) was positively correlated with body mass index and with plasma glucose levels both at fast and after OGL. No correlations were found between urinary excretion of catecholamines and plasma insulin and C-peptide levels both at fast and in response to OGL. Because the frequency distribution of UNE was bimodal, hypertensive subjects were separated into two subgroups using an arbitrary cutoff, and the parameters of glucose metabolism were compared. Subjects with UNE > 205 microg/day had greater levels of fasting glucose and greater glycemic response to OGL than subjects with UNE < 205 microg/day, whereas no significant differences between the groups were found in fasting and stimulated plasma insulin and C-peptide. Thus, activation of SNS is related to glucose tolerance but not hyperinsulinemia and insulin hypersecretion in essential hypertension. Plasma glucose levels, independent of insulin, may contribute to the relationship between SNS activity and blood pressure in essential hypertension.

Glucose metabolism and insulin receptor binding and mRNA levels in tissues of Dahl hypertensive rats.

Increased insulinemic response to an oral glucose load has been demonstrated in Dahl salt-sensitive hypertensive rats. To determine whether this abnormality is mediated at the level of the insulin receptor, we compared insulin receptor binding and mRNA levels in tissues of Dahl salt-sensitive rats (DS) and in their normotensive controls, Dahl salt-resistant rats (DR). To evaluate possible influences of dietary sodium intake, rats were fed either low (0.07% NaCl) or high salt (7.5% NaCl) chow until the DS became hypertensive, and then were killed by decapitation. Fasting plasma glucose and plasma insulin levels did not differ between DR and DS rats and were not affected by salt intake. In response to an oral glucose load, plasma glucose had a similar increase in DR and DS rats, but the increase in plasma insulin was significantly greater in DS rats. Scatchard analysis of binding was obtained from in situ autoradiographic studies performed in frozen skeletal muscle and kidney sections, and insulin receptor mRNA levels were measured by slot-blot hybridization. Number and affinity of insulin receptors were comparable in skeletal muscle and kidney of DR and DS rats and, in both groups, binding parameters were not affected by dietary sodium chloride. Hepatic and renal insulin receptor mRNA levels were also comparable in DR and DS rats fed either low or high salt chow. Thus, increased plasma insulin response to oral glucose load is associated with normal insulin receptor binding and gene expression in peripheral tissues in rats with Dahl hypertension. A postreceptor defect is likely responsible for the decreased sensitivity to insulin in this model of genetic hypertension.

Abnormalities of autonomic nervous system in hyperadrenergic orthostatic hypotension: description of a case.

Hyperadrenergic orthostatic hypotension was diagnosed in a 27-year-old man because of recurrent episodes of hypotension associated with high plasma noradrenaline levels. In this patient, laboratory tests were performed to evaluate autonomic nervous system function. Decreased response to Valsalva maneuver and carotid sinus massage indicated decreased baroreflex and vagal responsiveness, respectively. Cardiovascular response to the handgrip was reduced in comparison to controls. Passive leg raise showed normal reduction of plasma norepinephrine, indicating normal responsiveness of cardiopulmonary receptors. 'Non-dipper' profile in the ambulatory blood pressure monitoring provided further evidence for an impaired autonomic control of cardiovascular function in this patient. This report suggests the presence of autonomic dysfunction in hyperadrenergic orthostatic hypotension.

[Angiotensin II receptors in the heart conduction tissue]. / I recettori dell'angiotensina II nel tessuto di conduzione cardiaco.

Angiotensin II exerts a positive chronotropic effect on mammalian heart. This effect can be mediated either by activation of the sympathetic nervous system or binding to specific receptors located in cardiac conduction tissue. Recently, two angiotensin II receptor subtypes have been identified in rat tissues. In the present study, we have investigated tissue distribution of angiotensin II receptor subtypes in conduction tissue obtained from rat hearts, employing an in situ autoradiographic technique. Receptor subtypes were identified by use of the peptide antagonists losartan (AT1) and PD123,177 (AT2). Angiotensin II binding was homogeneously distributed in right and left ventricle, and interventricular septum, with each subtype accounting for approximately 50% of the specific binding. Receptor density in the conduction tissue, identified by acetylcholinesterase histochemistry, was quantitated by counting the silver grains on tissue sections which, following incubation, were dipped in photographic emulsion and developed. In both sinoatrial and atrioventricular node AT1 and AT2 receptors were homogeneously distributed. Receptor density in sinoatrial node was comparable to myocardium, whereas it was significantly higher (p < 0.02) in atrioventricular node. In conclusion, both AT1 and AT2 receptors are homogeneously distributed in cardiac conduction tissue. Binding density is higher in the atrioventricular node than in the other structures examined in our study, suggesting a possible role of angiotensin II in the atrioventricular conduction.

Palatal obturators in patients after maxillectomy.

Prosthodontic management of palatal defects is fundamental to improve patient's life undergoing to a maxillary surgical treatment. A lot of maxillary defects are a direct consequence of surgical treatment of malformations, neoplasms or trauma. The obturators are prosthesis used to close palatal defects after maxillectomy, to restore masticatory function and to improve speech. The primary goals of the obturator prosthesis are to preserve the remaining teeth and tissue and to provide comfort, function, and aesthetics to the patients. Different materials and retention methods are a characteristic of new types of obturators.

Laparoscopic resection for rectal adenocarcinoma.

AIMS: Laparoscopic surgery for rectal cancer is still under discussion, but there is evidence that minimal access surgery can be feasible and safe also in this field. The aim of this study was to confirm that laparoscopic resection for rectal cancer can afford good results in terms of recurrence rate and survival. PATIENTS AND METHODS: Since June 1998 through December 2007 as many as 252 patients underwent laparoscopic resection for rectal cancer. Laparoscopic anterior resection (LAR) was performed in 209 and laparoscopic abdominoperineal resection (LAPR) in 43. Neoadjuvant radiochemotherapy (nCRT) was administered in 48 patients with mid-low rectal cancer stage II and III with evidence of nodal involvement in preoperative work up. RESULTS: Patients who received nCRT showed a significant longer duration of surgery compared to patients who did not (p=0.004). Conversion to laparotomy was needed in 24 cases, (21 LAR and three LAPR) but no patient receiving nCRT needed conversion. Postoperative surgical complications occurred in 38 patients, 20 of which were represented by anastomotic leak after LAR. Six patients died postoperatively, in half the cases for surgery related causes. Downstaging after nCRT was seen in 40 patients, and complete histological response was observed in six cases. The mean number of lymph nodes harvested was 12, also in patients receiving nCRT. The mean follow-up was 48+/-33 months (range 0.1-120.4), and 10 patients experienced local recurrence. Cumulative 5 year survival was 73.7%. CONCLUSION: Laparoscopic resection for rectal cancer is feasible and safe, with morbidity and long-term results quite acceptable also in patients receiving neoadjuvant treatment.

In situ characterization of renal insulin receptors in the rat.

Insulin regulates carbohydrate metabolism, and water, sodium, potassium, and phosphate reabsorption in the kidney by binding to specific receptors. Insulin receptors have been identified in the kidney using membrane preparations obtained from both glomeruli and tubules. In this study, an autoradiographic technique was used to characterize insulin receptors in the rat kidney. Frozen tissue sections were preincubated to remove endogenously bound insulin, incubated in a buffer containing 200 microM 125I-Tyr-insulin, washed, and dried before exposure on Ultrofilm. Binding density was assessed by computerized microdensitometry. In the cortex, binding density was comparable in glomeruli and tubules. In the medulla, bound radioligand was found primarily in longitudinal structures traversing the outer portion, presumably corresponding to vascular bundles, and in the inner portion. Scatchard analysis of competition binding data resulted in curvilinear profiles, indicating either two classes of receptors with different affinity or the presence of a single class of receptors with a negative cooperative hormone-receptor interaction. Data analyzed for a two-site model showed one receptor site with Kd of 0.39 +/- 0.14 nmol/l and Bmax of 3.5 +/- 1.0 x 10(10) receptors/mm3 and another site with Kd of 0.30 +/- 1.1 pmol/l and a Bmax of 3.2 x 10(13) receptors/mm3. Thus, in situ autoradiography can be used to determine distribution and binding characteristics of insulin receptors in rat kidney and might be employed in receptor studies on rat models of human disease.

Right atrial myxoma with pulmonary embolism.

Right atrial myxomas are rare tumors that often go undiagnosed. Pulmonary embolism originating from the tumor mass is a potentially fatal complication. We report the case of a young man in whom a right atrial myxoma with recurrent episodes of pulmonary embolism was diagnosed 1 month after the beginning of symptoms by two-dimensional transthoracic and transesophageal echocardiography. Early diagnosis of cardiac myxomas is important since surgical treatment is usually resolutive with low recurrence rates and good long-term survival.

Benign familial macrocytosis.

We have identified a 52-year-old woman and her 27-year-old daughter with macrocytosis, normal haemoglobin and mean corpuscular haemoglobin concentration. Macrocytosis could be demonstrated from the age of 40 and 25 respectively. All blood tests were normal including vitamin B12 and folic acid. Bone marrow investigation showed rare macroblasts without other abnormalities. Endoscopy of the upper gastrointestinal tract and ultrasonography of the abdomen were normal. Thus, persistent macrocytosis was present without evidence of diseases that might account for it. In these subjects, macrocytosis is likely to be related to the presence of a genetic defect.

[Angiotensin receptors in the rat myocardium during pre- and postnatal development]. / Studio dei recettori dell'angiotensina nel miocardio di ratto durante lo sviluppo pre- e post-natale.

Angiotensin II exerts positive inotropic and chronotropic effects on mammalian heart by binding to specific membrane receptors. Recently, 2 subtypes of angiotensin II receptors (AT1 and AT2) have been distinguished using the nonpeptide antagonists losartan and PD123177. Because angiotensin II has been reported to have growth potentiating effects in several tissues, we examined angiotensin II receptors in fetal (embryonic day 16 and 19), neonatal (1, 2, 3 and 10 days), and adult (10 and 16 weeks) rats. We performed an 125I-[Sar1,Ile8]-angiotensin II in situ binding assay on tissue sections obtained from Sprague-Dawley rats. Binding specificity was verified by competition with unlabeled [Sar1]-angiotensin II. Distribution of AT1 and AT2 receptors was determined by competition with losartan and PD123177, respectively, and the density of receptors quantitated by emulsion autoradiography. Angiotensin II receptors were widely distributed throughout the heart, with each receptor subtype accounting for approximately 50% of the specific binding. Binding density was comparable in right and left ventricles, and interventricular septum. Throughout cardiac development a significant increase (p < 0.005) in the density of both receptor subtypes was found immediately after birth, reaching a maximum on day 2, and decreasing toward prenatal values thereafter. No variation in the proportion of the 2 receptor subtypes was observed during development. Thus, in rat heart, AT1 and AT2 receptors are equally distributed over the myocardium. The density of these angiotensin II receptors is developmentally regulated, suggesting a possible role of the cardiac renin-angiotensin system in heart growth and in the adaptation of the heart to postnatal circulatory conditions.

[The role of G-regulatory proteins in the intracellular mechanism of cardiac angiotensin-II receptors]. / Ruolo di proteine-G regolatrici nel meccanismo intracellulare dei recettori cardiaci dell'angiotensina II.

High-affinity angiotensin II receptors have been identified in cardiac tissue of many animal species. In the heart, angiotensin II exerts positive inotropic and chronotropic effects, constricts coronary vessels, and stimulates cell growth. In vascular smooth muscle and adrenal cortex angiotensin II interacts with guanidine nucleotide regulatory proteins because GTP-gamma-S causes dissociation of the radioligand from its receptor. To investigate whether angiotensin II interacts with guanidine nucleotide regulatory proteins also in cardiac tissue, we studied the effects of GTP-gamma-S on [Sar1, Ile8]-angiotensin II binding to angiotensin II receptor subtypes (AT1 and AT2) in hearts obtained from 16- to 20-week-old Sprague-Dawley rats. We employed an in situ technique performed on frozen tissue sections. Competition experiments performed with the nonpeptide inhibitors losartan and PD123177 allowed identification of both AT1 and AT2 angiotensin II receptors in rat heart. These receptors were present in comparable amounts. In a different set of experiments the effects of GTP-gamma-S (100 microM) on radioligand displacement from AT1 and AT2 receptors were studied. GTP-gamma-S caused a progressive dissociation of the radioligand from the AT1 receptor indicating that this receptor interacts with guanidine nucleotide regulatory proteins. In contrast, the AT2 receptor does not appear to directly interact with guanidine nucleotide regulatory proteins. In summary, the study shows that both angiotensin II receptor subtypes are present in rat heart and that guanidine nucleotide regulatory proteins are implicated in the signal transduction mechanism of the cardiac AT1 receptor.