The epigenetic landscape in the cardiovascular complications of diabetes.

A growing body of evidence suggests that epigenetic modifications-changes to the genome that do not involve changes in DNA sequence-may significantly derail transcriptional programs implicated in angiogenesis, oxidative stress and inflammation, thus fostering cardiovascular damage in patients with diabetes. Notably, adverse epigenetic signals acquired over the life course can be transmitted to the offspring, and may contribute to early cardiovascular phenotypes in the young generations. Hyperglycaemia and insulin resistance-key hallmarks of diabetes-induce an array of epigenetic modifications (i.e., DNA methylation, histone marks, and non-coding RNAs) which are responsible for a long-lasting impairment of vascular and cardiac function, even after intensive glycemic control. Hence, unveiling the "epigenetic landscape" in patients with diabetes may provide a post-genomic snapshot of global cardiovascular risk, and may furnish the tools to design personalized, epigenetic-based therapies to alleviate the burden of cardiovascular disease in diabetic patients. The present review aims to acquaint the scientific community with the rapidly advancing field of epigenetics and its implications in the cardiovascular complications of diabetes.

Intrinsic resistance to selumetinib, a selective inhibitor of MEK1/2, by cAMP-dependent protein kinase A activation in human lung and colorectal cancer cells.

BACKGROUND: MEK is activated in ∼40% colorectal cancer (CRC) and 20-30% non-small cell lung cancer (NSCLC). Selumetinib is a selective inhibitor of MEK1/2, which is currently in clinical development. METHODS: We evaluated the effects of selumetinib in vitro and in vivo in CRC and NSCLC cell lines to identify cancer cell characteristics correlating with sensitivity to MEK inhibition. RESULTS: Five NSCLC and six CRC cell lines were treated with selumetinib and classified according to the median inhibitory concentration (IC(50)) values as sensitive (≤1 µM) or resistant (>1 µM). In selumetinib-sensitive cancer cell lines, selumetinib treatment induced G1 cell-cycle arrest and apoptosis and suppression of tumour growth as xenografts in immunodeficient mice. Evaluation of intracellular effector proteins and analysis of gene mutations showed no correlation with selumetinib sensitivity. Microarray gene expression profiles revealed that the activation of cAMP-dependent protein kinase A (PKA) was associated with MEK inhibitor resistance. Combined targeting of both MEK and PKA resulted in cancer cell growth inhibition of MEK inhibitor-resistant cancer cell lines in vitro and in vivo. CONCLUSION: This study provides molecular insights to explain resistance to an MEK inhibitor in human cancer cell lines.

Analyzing speckle contrast for HiLo microscopy optimization.

HiLo microscopy is a recently developed technique that provides both optical sectioning and fast imaging with a simple implementation and at a very low cost. The methodology combines widefield and speckled illumination images to obtain one optically sectioned image. Hence, the characteristics of such speckle illumination ultimately determine the quality of HiLo images and the overall performance of the method. In this work, we study how speckle contrast influence local variations of fluorescence intensity and brightness profiles of thick samples. We present this article as a guide to adjust the parameters of the system for optimizing the capabilities of this novel technology.

Development of a novel instrument to measure the pulsatile movement of ocular tissues.

We demonstrate an optical instrument that can measure the axial displacement of different eye tissues, including the cornea and the fundus. The instrument is based on spectral-domain low-coherence interferometry, which extracts displacement information from sequential axial scans of the eye with 100 Hz sampling frequency and with a precision of 400 nm. Longitudinal retinal and corneal movements were successfully measured in vivo in live rats, and Fourier analysis of the signal revealed the signature of the respiratory and cardiac cycles at 1.0 and 3.5 Hz, respectively. The effective amplitudes of retinal and corneal displacements at the cardiac frequency were found to be about 1.10 and 1.37 mum, respectively. The synchrony and direction of these two movements relative to the systole and diastole were found to be nearly the same. This novel instrument can be applied to assess biomechanical properties of the eye, which could be important for early diagnosis and for understanding the pathophysiology of glaucoma and other ocular diseases.

Can CAP guideline adherence improve patient outcome in internal medicine departments?

The impact of compliance with Italian guidelines on the outcome of hospitalised community-acquired pneumonia (CAP) in internal medicine departments was evaluated. All Fine class IV or V CAP patients were included in this multicentre, interventional, before-and-after study, composed of three phases: 1) a retrospective phase (RP; 1,443 patients); 2) a guideline implementation phase; and 3) a prospective phase (PP; 1,404 patients). Antibiotic prescription according to the guidelines increased significantly in the PP. The risk of failure at the end of the firstline therapy was significantly lower in the PP versus the RP (odds ratio (OR) 0.83, 95% confidence interval (CI) 0.69-1.00), particularly in Fine class V patients (OR 0.71, 95% CI 0.51-0.98). Analysis of outcome in the overall population (2,847 patients) showed a statistically significant advantage for compliant versus noncompliant therapies in terms of failure rate (OR 0.74, 95% CI 0.60-0.90) and an advantage in terms of mortality (OR 0.77, 95% CI 0.58-1.04). Antipneumococcal cephalosporin monotherapy was associated with a low success rate (68.6%) and the highest mortality (16.2%); levofloxacin alone and the combination of cephalosporin and macrolide resulted in higher success rates (79.1 and 76.7%, respectively) and significantly lower mortalities (9.1 and 5.7%, respectively). Overall, a low compliance with guidelines in the prospective phase (44%) was obtained, indicating the need for future more aggressive and proactive approaches.

Fluorescent two-photon nanolithography.

Improving the excitation conditions in two-photon fluorescent lithography reduces the size of the fabricated structures to nanometre scales. We demonstrate that a precise control of the illumination profile and the scanning speed of the laser beam is enough to decrease the photo-polymerization volume of resins by orders of magnitude. This work also shows experimental evidence of surface effects that yield a different polymerization intensity threshold compared with bulk. Such phenomenon enables to perform a non-linear optical nanolithography in a simple way, allowing fast-prototyping procedures. We present a detailed study of the polymer growth process using fluorescence and atomic force microscopy.

Non-invasive measurement of choroidal volume change and ocular rigidity through automated segmentation of high-speed OCT imaging.

We have developed a novel optical approach to determine pulsatile ocular volume changes using automated segmentation of the choroid, which, together with Dynamic Contour Tonometry (DCT) measurements of intraocular pressure (IOP), allows estimation of the ocular rigidity (OR) coefficient. Spectral Domain Optical Coherence Tomography (OCT) videos were acquired with Enhanced Depth Imaging (EDI) at 7Hz during ~50 seconds at the fundus. A novel segmentation algorithm based on graph search with an edge-probability weighting scheme was developed to measure choroidal thickness (CT) at each frame. Global ocular volume fluctuations were derived from frame-to-frame CT variations using an approximate eye model. Immediately after imaging, IOP and ocular pulse amplitude (OPA) were measured using DCT. OR was calculated from these peak pressure and volume changes. Our automated segmentation algorithm provides the first non-invasive method for determining ocular volume change due to pulsatile choroidal filling, and the estimation of the OR constant. Future applications of this method offer an important avenue to understanding the biomechanical basis of ocular pathophysiology.

Validation of a method for measuring (226)Ra in drinking waters by LSC.

A simple method for measuring (226)Ra in drinking waters has been validated and validation parameters are provided. It is based on the measurement by LSC of (222)Rn, at equilibrium with (226)Ra, which is absorbed into a water immiscible scintillation cocktail (Ultima Gold F) inside the counting vial. The validated application field ranges between the detection limit (2·10(-3) Bq/kg) to 150 Bq/kg. The method has proven to be reliable, effective and suitable for wide-range measuring campaigns. A summary of results obtained in recent years is also given.

Diagnosis of porcine trichinellosis: parasitological and immunoserological tests in pigs from endemic areas of Argentina.

In order to compare the reliability of serological and parasitological techniques for the diagnosis of porcine trichinellosis from endemic areas in Argentina, 116 pigs were studied: 61 animals from two separate outbreaks and 55 from a small abattoir. Direct diagnostic techniques included trichinoscopy and the artificial digestion method. Indirect diagnostic tests used in this study were the enzyme-linked immunosorbent assay (ELISA), employing the excretory-secretory products of muscle larvae (ML) as antigen, and the indirect immunofluorescence assay using as antigen ML in suspension (IIF-susp), cryostat sections of infected rat muscle or of free ML (IIF-slide). The percentage of parasitologically positive pigs was invariably lower than that of serologically positive animals (IIF-slide), even when digestion studies were carried out individually with a greater amount of muscle sample than required by current regulations. Close correlation was found between IIF using as antigen tissue sections and IIF using free ML sections, while IIF-susp proved unsuitable for diagnosis since this assay presented a high percentage of false negative results (20%). The IIF-slide technique proved positive in all parasitologically positive animals. ELISA rendered a lower percentage of positive reactions than IIF-slide, especially when worm burden was low. Since most parasitologically positive animals rendered at least two positive serological tests (two variations of IIF or IIF plus ELISA), those negative by digestion and positive by two serological methods were strongly suspected of having trichinellosis. Upon studying swine from a abattoir it was found that 9% of the pigs were positive when assayed by two serological techniques, but Trichinella spiralis infection could not be parasitologically confirmed. To sum up, serological methods may be used for screening all pigs and positive findings should be tested by the digestion method by analysing a greater quantity of pork than that required by current regulations, above all in areas with reported clinical trichinellosis in humans, to ensure that the pork is safe for human consumption.

Testicular function in men treated in childhood for undescended testes.

PURPOSE: The aim of the study was to evaluate testicular hormones and sperm counts of young men treated in childhood for cryptorchidism METHODS: Testicular volume, serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone as well as semen specimens were evaluated in 57 men (mean age, 19 years; range, 18 to 27 years) treated in childhood for unilateral (n = 47) and bilateral (n = 10) cryptorchidism. In 3 unilateral cases monorchidism was found. Thirty-seven patients underwent orchiopexy after hormonal treatment (luteinizing hormone releasing factor, 1.2 mg/d for 28 days followed by human chorionic gonadotropin, 500 IU intramuscularly 3 times a week for 3 weeks). The remainder underwent surgery. Mean age at surgical treatment was 5.4 years (range, 2 to 12 years). These patients were examinated again after a mean period of 13.3 years (range, 10 to 19 years). RESULTS: Reduced testicular volume (<12 mL) was found in 6 of 64 testes (9.3%). LH, FSH, and testosterone levels were found within the normal range in all patients. With linear regression, inverse relations were found between FSH and, respectively, testicular volume (P =.002), sperm concentration (P =.013), sperm motility (P =.023), and normally shaped sperms (P =.019). There were direct relations between testicular volume and sperm concentration (P =.02), sperm motility (P =.000), and normally shaped sperms (P =.001). We did not find any statistical correlation between age at surgery and semen quality. Significantly better results in terms of sperm counts were found in patients directly operated on in comparison to those treated with hormones before orchiopexy. CONCLUSIONS: Presented data indicate tubular impairment in young men operated on in childhood for cryptorchidism; FSH values increase and testicular volume decrease are related to sperm deterioration. Studies on children treated in the first 2 years of life are required to clarify the usefulness of early treatment of cryptorchidism.

Immunoelectrotransfer blot assay in acute and chronic human trichinellosis.

An immunoelectrotransfer blot assay (IETB) using excretory secretory products of muscle larvae of Trichinella spiralis (ML-ESP) and the avidin biotin system was developed in order to characterize reactivity against ML-ESP in sera from patients with acute and chronic trichinellosis. A complete pattern of up to 13 bands was developed by sera from individuals with trichinellosis where doublets, triplets, or single bands were shown to have molecular weights of roughly 66, 55, 45, 36, 29, 24, and 14 kDa. The bands at approximately 55, 36, 29, and 14 kDa proved specific for T. spiralis. The band at approximately 55 kDa was present in all trichinellosis sera, whereas the approximately 14-kDa band was present in only a small percentage of sera. The development of approximately 36- and 29-kDa bands suggests a modulation of the reactivity against ML-ESP over time. IETB proved more sensitive for the population of chronic trichinellosis under study than a conventional diagnostic enzyme-linked immunosorbent assay, allowing negative or borderline serum samples to be determined. Thus, this technique, when applied for human trichinellosis surveillance, should provide a useful tool in endemic areas.

Myiasis of a perineal fistula.

We report a case of myiasis of a rectocutaneous fistula in a subject suffering from bone metastases after surgical resection of a neoplastic bladder. The fistula was infested by maggots identified as third-instar larvae of flies belonging to the family Sarcophagidae, genus Sarcophaga. The infestation was neither intestinal nor pseudointestinal; it was probably caused by a fly ovipositing on the fistula of the patient while having wound care at home, even though the possibility that the infestation might have occurred during hospitalization cannot be ruled out.

Non-palpable testes. Orchiopexy in single stage.

From 1986 to 1990, 53 non-palpable testes were treated in the department of Pediatric Surgery and Endocrinology at the Regina Margherita Hospital, Turin, Italy. The reported patients were 45, 8 of them were bilateral with both testes non-palpable, 13 of the remaining patients were right cryptorchid and 24 were left. All patients received medical treatment, by LH-RH nasal spray and HCG, before surgery. None of the non-palpable testes descended by hormone therapy only, so that surgical exploration was always required. In the 53 reported cases, 10 gonads were not found, 13 were atrophic and 30 were abdominal. All abdominal gonads were positioned into the scrotum by orchiopexy in a single stage. Half of the patients underwent operation before the age of 2. Therefore, it is to be pointed out that the younger a patient the shorter his inguinal canal, this facilitates orchiopexy for high, undescended testes. In 10 patients a testicular biopsy was made during surgical procedure. Normal morphology and normal spermatogonia content were observed in the abdominal testes of the patients who had reached the age of 1 year. During the follow-up of the reported patients no upward displacement was observed. Eleven patients underwent postoperative testicular ultrasonography which showed that parenchymal structure of the operated testis was normal and the volume of the testis was slightly smaller than normal. Based on their experience, the authors suggest to correct cryptorchidism before the age of one year, surgical procedure follows immediately after ineffective hormone therapy.

Bulky-pedunculated hemolymphangioma of the esophagus: rare case in a two-years old girl.

Benign esophageal masses are rare. The authors present a rare case of bulky pedunculated hemolymphangioma of the esophagus in a two-year-old female. The symptomatology was characterized by acute episodes of dyspnea associated with the protrusion of the mass from the mouth. The mass was removed endoscopically.

Diagnosis of human trichinellosis: pitfalls in the use of a unique immunoserological technique.

Serum samples belonging to three outbreaks in Argentina (47 patients) taken at different times post-ingestion were analysed employing IIF and ELISA simultaneously. Results show that: a) the number of patients diagnosed by a unique technique, especially by ELISA (31 patients), was lower than the one obtained by the simultaneous use of both assays (38 patients); b) four patients out of the seven diagnosed by a unique technique were negative by the other assay over the period of time evaluated. Therefore, it can be concluded that the use of a sole immunoserological technique can not only lead to the delay in the detection but also to the misdiagnosis of this parasitic infection.

[The role of imaging diagnosis in the study of thyroid neoplasms]. / Ruolo della diagnostica per immagine nello studio delle neoplasie tiroidee.

The aim of this paper was to evaluate the role of imaging techniques and methods in the diagnosis of thyroid tumours. The main aim was to analyse the various imaging methods and to search for an integrated diagnosis based on clinical and laboratory evaluations and techniques such as fine needle aspiration biopsy for which imaging is used as a guide. The authors stress the importance of analysing the sensitivity of the various methods in preoperative staging. The methods examined here include: scintigraphy, ultrasound, fine needle aspiration biopsy, Doppler ultrasound, color-Doppler scan, TC and magnetic resonance.

[Preoperative instrumental diagnosis in secondary hyperparathyroidism]. / Diagnostica strumentale preoperatoria nell'iperparatiroidismo secondario.

There is a very high incidence of hyperparathyroidism secondary to chronic renal insufficiency in dialysed patients (67%). When medical therapy can no longer control the evolution of disease, an appropriate surgical strategy must be defined for the clinical case. Instrumental tests play an important role during the preoperative phase in the possible "localisation" of ectopic parathyroids (10% of cases) and the identification of those glands with a more pronounced hyperplastic appearance. The authors examine and describe the most useful tests for a preoperative study, namely ultrasonography, echo-colour Doppler, scintigraphy using a dual tracer and image subtraction, computerised xial tomography (CT), magnetic resonance (MR), digital angiography and intraoperative ultrasonography. In conclusion, it can be said that all the methods examined present similar sensitivity levels for the study of parathyroid, but the combination of more than one technique considerably improves diagnostic accuracy.

[Pseudoaneurysm in pancreatic pseudocysts. Report of a clinical case]. / Pseudoaneurisma in pseudocisti del pancreas. Descrizione di un caso clinico.

Pancreatitis in a percentage of 1.5% in a pseudocyst, 10% of which are represented as formations of the pseudoaneurysmatic type one of the visceral artery. The authors of the following study present the case of an 86 year old woman with positive anamnesis because of a chronic pancreatitis, who is affected by a symptomatology in the epigastric regione together with hematemesis and melena. An ecography shows at the head level a mass of cm 6.5 hypo ecogena, delimitated by a hyper ecogena band. A TAC, with and without contrast means and integrated by some RM and angio-RM sequences, shows that it is a vascular injury, with the characteristic area on enhancement after the infusion of the contrast means. The patients goes under surgical intervention, showing a trough supplied by a branch of the hepatic artery. The intervention is performed with the tying of the artery, with the drainage of the residual cavity plus the creation of a pancreatic, cystic and jejunum derivation.

[Serological, clinical, and epidemiological aspects of an outbreak of trichinosis in Azul, province of Buenos Aires]. / Aspectos serológicos, clínicos y epidemiológicos de un brote de triquinosis en Azul, Provincia de Buenos Aires.

An outbreak of human trichinellosis that took place in Southern Buenos Aires Province, an endemic area for this zoonosis, was studied. Eighteen individuals were infected by eating swine sausages lacking veterinary control. Clinical symptoms, as well as serological studies related to antibody production and cytotoxic capacity of the sera are described. Eight interdicted pigs and their products were analysed. In order to determine the transmission network of this zoonosis, wild rats were hunted in the farm where the swine were bred. The clinical symptoms of the infected individuals were: myalgia, edema, fever and cephalea. Sixty-one percent of the patients showed antibodies against muscle larvae 15-20 days after ingestion of the infected meat. Five out of eighteen patients showed anti-newborn larvae antibodies 40-60 days postinfection. The sausage burden was 5.3 muscle larvae/g. Two out of eight swine were diagnosed employing serological techniques, while parasitic techniques detected only one. The rat is an important reservoir in the domestic cycle of this zoonosis.

Liver focal lesions and hepatocellular carcinoma in cirrhotic patients: from screening to diagnosis.

This short review aims at pointing out the role of ultrasonography (US) as a valuable means for an early detection of liver focal lesions in cirrhotic patients. Among these lesions which may be benign and malignant, hepatocellular carcinoma (HCC) finds its place with an incidence in cirrhotic patients becoming higher as the time passes from the onset of cirrhosis. US is of fundamental importance in the regular screening of subjects with viral cirrhosis because of its sensitivity in evaluating focal lesions (79-82%); however, it is not yet the most valuable method for the diagnosis of HCC for which other diagnostic techniques are needed such as CT (Computerized Tomography), MR (Magnetic Resonance) and the US-guided fine-needle biopsy. In fact, the echo-structure of HCC varies with its dimensions and may mimic that of one of many other liver focal lesions. Lately, some new sonographic techniques have become available that can allow a more accurate investigation of HCC. Among them, the "Tissue Harmonic" and the "Pulse Inversion Imaging" techniques provide a better definition of the grey-scale image thus improving the conspicuity of focal lesions. Color and Power Doppler techniques, while allowing an accurate observation of the lesion vascularization, integrate the sonographic appearance of HCC. The enhancement of the US color Doppler signal obtainable by using echographic contrast agents, represents a further progress for a detailed observation of the focal lesions' micro-vascularization and, thus, for the HCC identification. Further advancements in terms of improvement of the image quality and characterization of the focal lesion are to be expected from the use of "Tissue Harmonic" and "Pulse Inversion Imaging" techniques associated with echographic contrast agents. However, the diagnosis of HCC still rests on the incontrovertible histological evidence obtained by echo-guided fine-needle biopsy.