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Aim: To evaluate serum ghrelin and motilin concentration in infants with infantile colic., Methods: A case-control study was conducted on fasting blood venous samples obtained from 18 infants with infantile colic and 20 healthy infants to measure ghrelin (RIA test) and motilin (RIA test)., Results: Colicky infants showed higher ghrelin serum levels (2534.2 +/- 600.0 pg/ml; ln 7.8 +/- 0.2) than controls (2126.1 +/- 281.3 pg/ml; ln 7.6 +/- 0.1) (p = 0.011). Serum motilin concentration was significantly higher in colicky infants (94.6 +/- 23.2 pmol/l) than in controls (64.1 +/- 30.1 pmol/l) (p = 0.001). Motilin concentrations were higher in formula-fed colicky infants (104.5 +/- 20.4 pmol/l) than in breastfed ones (82.2 +/- 21.3 pmol/l) (p = 0.038)., Conclusion: Our finding shows that ghrelin and motilin concentrations are higher in infants with colic than in controls, supporting an organicistic aetiopathogenesis of this disorder. Furthermore, the role of ghrelin on gastrointestinal motility may open new doors to better understand the aetiology of infantile colic.

Advanced liver disease is characterized by decreased arterial blood pressure and peripheral vascular resistances, increased cardiac output and heart rate in the setting of a hyperdynamic circulatory pattern favoured by total blood volume expansion, circulatory overload and overactivity of the endogenous vasoactive systems. Reduced heart responses to stressful conditions such as changes in loading conditions of the heart in presence of further deterioration of liver function such as refractory ascites, hepatorenal syndrome, spontaneous bacterial peritonitis and bleeding esophageal varices have been recently identified and the knowledge of the cirrhotic cardiomyopathy syndrome has gained the dignity of a new clinical entity. Facing the availability of therapeutic interventions (paracentesis, transjugular intrahepatic portosystemic shunt, peritoneovenous shunt, orthotopic liver transplantation) currently employed to manage the life-threatening complications of the most advanced phases of cirrhotic disease, the knowledge of their impact on cardiovascular function is of paramount relevance.

Methemoglobinemia is not a rare condition arising from the exposure to hemoglobin-oxidizing agents such as nitrates-nitrites present in well water or vegetables. Infants < 3 months of age are more susceptible than adults because of lower amounts of a key enzyme, NADH-cytochrome b5 reductase, which converts methemoglobin back to hemoglobin. We report 2 infants, aged respectively 2 and 1 months, suffering from methemoglobinemia, fed with a formula that was reconstituted with a high concentration of courgette soup to resolve constipation. They developed a severe cyanosis with methemoglobinemia (respectively 30.4 and 27%) and were hospitalized and treated with methylene blue at 1%. After 12 h the syndrome was completely resolved. Home-prepared infant foods containing vegetables are potential causes of methemoglobinemia. It is important not to feed infants with vegetables having a high nitrate content (e.g., courgette, spinach, beets and green beans) to resolve constipation since, particularly in the first months of life, they may cause severe methemoglobinemia., (Copyright 2006 S. Karger AG, Basel.)

The presence of a hyperdynamic circulation in cirrhotic liver is currently a well established concept. The first studies of cardiac function in patients with cirrhosis suggested the existence of an alcoholic cardiomyopathy. More recently, altered left ventricular response to physiological and/or pharmacological stimuli in patients with post-viral liver cirrhosis has been established, and clinically insignificant diastolic cardiac function has also been observed. Neurohumoral hyperactivity and hyperdynamic circulation, which are associated with chronic exposure to the cardiodepressant substances present in advanced liver disease, play a decisive role in the genesis of this process. The lack of relaxation of the left ventricle and alteration in the pattern of transmitral flow, both of which are characteristics of this clinical entity, are easily detected by echocardiography. The growing evidence of diastolic dysfunction in liver cirrhosis, particularly in decompensated cirrhosis, suggests the clinical importance of the problem, thus introducing the concept of "cirrhotic cardiomyopathy". Greater insight into this phenomenon could help to decrease cardiovascular risk, especially during maneuvers commonly used in the treatment of the complications of liver cirrhosis, such as paracentesis, transjugular intrahepatic portosystemic shunt stent implantation, and liver transplantation.

Aim: A hyperdynamic circulatory pattern in advanced liver disease is known since a long time. The first studies evaluating cardiac function in cirrhosis were performed in patients with alcoholic liver disease and thus this condition was attributed to the toxic effects of ethanol. A reduced performance of the left ventricle after physical and pharmacological strains along with an altered diastolic function has been demonstrated also in postviral cirrhosis. Many factors are involved in advanced cirrhosis whereas little is known in the earlier stages of disease., Methods: To this aim we have investigated patients with different stages of hepatitis C virus (HCV)-related liver disease to detect the time-course of diastolic dysfunction. An impaired relaxation and increased thickness of left ventricular walls along with an altered pattern of transmitral flow can be easily detected by means of echocardiography., Results: In chronic hepatitis diastolic function is preserved but increased thickness of left ventricle parietal walls can be detected in patients with fibrosis on liver biopsy. The typical pattern of diastolic dysfunction is observed in Child A cirrhotic patients and in Child C ascitic patients but thickness of parietal walls is more relevant in the former group. Chronic aldosterone blockade could exert favourable effects in heart remodeling suggesting a potential role of these drugs in cirrhotic cardiomyopathy., Conclusions: The presence of increased thickness of left ventricle parietal walls in chronic hepatitis C in the precirrhotic stage point to a putative role of HCV in this heart structural abnormality that can become a co-factor in the more advanced stages of cirrhosis when portal hypertension and its deleterious effects on systemic hemodynamics, cardiac function and structure become manifest.

The microbial community associated with ascocarps of the ectomycorrhizal fungus Tuber borchii Vittad. was studied by both cultivation and direct extraction of bacterial 16S rRNA gene (rDNA) sequence approaches. The inner part of six T. borchii ascoma collected in North-Central Italy was used to establish a bacterial culture collection and to extract the total genomic DNA to obtain a library of 16S rDNAs representative of the truffle bacterial community. Most of the isolates were affiliated to the gamma-Proteobacteria, mainly Fluorescent pseudomonads; some isolates were members of the Bacteroidetes group and Gram-positive bacteria, mostly Bacillaceae. The majority of the clones from the library were alpha-Proteobacteria showing significant similarity values, of greater than 97%, with members of the Sinorhizobium/Ensifer Group, Rhizobium and Bradyrhizobium spp. not previously identified as Tuber-associated bacteria. Only a few bacterial strains belonging to this bacterial subclass were found in the culture collection and isolated on a medium specific for Rhizobium-like organisms. A few clones were members of the beta- and gamma-Proteobacteria; as well as low and high G+C Gram-positive bacteria. Our findings clearly indicate that a dual approach increases the information obtained on the structural composition of a truffle bacterial community as compared to that derived via cultivation or direct recovery of 16S rDNA sequences alone.

We present here a high-performance liquid chromatographic method for the evaluation of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity. The automated method was applied to fungal and mouse liver extracts and validated by the addition of mevastatin to the reaction mixture and by several intra- and inter-day assays. This method offers important advantages over those previously reported because no radiolabeled substrates or expensive techniques such as mass spectrometry are required, and the time of analysis is relatively short. Moreover, the method can be successfully applied to different biological samples; hence, it should be very useful in evaluating potential inhibitors of the HMG-CoA enzyme and investigating cholesterol metabolism, cell growth and differentiation processes.

The formation of the fruit body represents the final phase of the ectomycorrhizal fungus T. borchii life cycle. Very little is known concerning the molecular and biochemical processes involved in the fructification phase. 2-DE maps of unripe and ripe ascocarps revealed different protein expression levels and the comparison of the electropherograms led to the identification of specific proteins for each developmental phase. Associating micropreparative 2-DE to microchemical approaches, such as N-terminal sequencing and 2-D gel-electrophoresis mass-spectrometry, proteins playing pivotal roles in truffle physiology were identified.

Fruit body development is a particular phase of the Tuber life cycle, characterised by the aggregation of different types of hyphae, i.e., vegetative hyphal cells and highly specialised reproductive hyphae (asci). In order to identify the volatile organic compounds (VOCs) produced in different stages of the Tuber borchii ripening fruit body, solid-phase microextraction with gas chromatography and mass spectrometry was used. The volatile organic compounds were extracted using a DVB/CAR/PDMS 50/30 microm fiber placed for 10 min at room temperature in the truffle headspace. The results obtained reveal 49 compounds each of which was present only in a particular stage of maturation. 1-octen-3-ol, aromadendrene, alpha-farnesene and other terpenoid compounds were of particular interest, and their possible biological roles are discussed. The production of aromadendrene in the completely unripe fruit body suggests the existence of communication events in the early stage of ascomata formation between the fungus and the host plant. alpha-Farnesene could represent a chemotactic attractant to saprophytic organisms in order to disperse the fungal spores in the environment. The identification of the VOCs produced by truffles during their maturation could give information about the processes underlying this phase of Tuber life cycle., (Copyright 2003 John Wiley & Sons, Ltd.)

From 1992 through 1997, there were 41 deaths by homicidal blunt impact head trauma in Hillsborough County, Florida. Twenty-one cases were excluded from the study because of putrefaction or survival beyond the emergency department doors, leaving 20 cases for the study. One of the 15 nonputrefied victims found dead at the scene and 1 of the 5 victims pronounced dead in the emergency department had definite venous air embolism. Victim 1 was found dead, bludgeoned with a concrete block, and had open vault and comminuted basilar skull fractures. The dura forming the right sigmoid sinus at the jugular foramen was lacerated. A preautopsy chest radiograph and examination under water documented gas in the pulmonary artery and right ventricle. Victim 2 was bludgeoned with a steel stake and was pronounced dead on arrival in the emergency department. He had open comminuted vault fractures, a transverse basilar skull fracture, and lacerations of the brain. Direct examination and preautopsy chest radiography revealed air in the right side of the heart. A third victim, with basilar fractures, had a small gas bubble in the pulmonary artery not detected by the case pathologist. A fourth victim, with a basilar skull fracture, had an unusual radiographic finding that was thought to be air in the posteromedial aspect of the lower lobe of the left lung but could not be excluded as an air embolus. Optimal postmortem documentation of venous air embolism includes the demonstration of the embolus and the site of air ingress. This study demonstrates that venous air embolism occurs in some victims of homicidal bludgeoning and suggests that when significant, it is easily demonstrated in the absence of putrefactive gas formation. The presence of venous air embolism can serve as evidence that a victim was alive and breathing at the time of the infliction of head wounds. In the belief that venous air embolism might be underdiagnosed in many medical examiner offices, the authors have sought to bring attention to the entity by publishing their experience with it in cases of bludgeoning.

SNF5/INI1 is a component of the ATP-dependent chromatin remodeling enzyme family SWI/SNF. Germ line mutations of INI1 have been identified in children with brain and renal rhabdoid tumors, indicating that INI1 is a tumor suppressor. Here we report that disruption of Ini1 expression in mice results in early embryonic lethality. Ini1-null embryos die between 3.5 and 5.5 days postcoitum, and Ini1-null blastocysts fail to hatch, form the trophectoderm, or expand the inner cell mass when cultured in vitro. Furthermore, we report that approximately 15% of Ini1-heterozygous mice present with tumors, mostly undifferentiated or poorly differentiated sarcomas. Tumor formation is associated with a loss of heterozygocity at the Ini1 locus, characterizing Ini1 as a tumor suppressor in mice. Thus, Ini1 is essential for embryo viability and for repression of oncogenesis in the adult organism.

The ascomycete Pulvinula constellatio was found to form competing ectomycorrhizal relationships with plants that had been inoculated with Tuber spp. and other edible ectomycorrhizal fungi. In order to be able to distinguish P. constellatio mycorrhizae from those of the Tuber spp., we determined the morphological and bio-molecular characteristics of P. constellatio. The complete sequence of the ITS regions was determined, in order to select specific primers. The ITS region was also studied using restriction fragment length polymorphism analyses with several restriction enzymes that allowed an unambiguous identification of the species.

Species-specific primers selected from the internal transcribed spacer region sequence were used to set up a multiplex polymerase chain reaction (PCR) able to simultaneously identify the white truffle species Tuber magnatum, Tuber borchii, Tuber maculatum and Tuber puberulum. Furthermore, a primer specific for the competitive fungus Sphaerosporella brunnea was designed and added to the multiplex PCR set, allowing the detection of the Tuber species and the contaminant fungus in a one-step reaction.

ATP-dependent chromatin-remodeling complexes are conserved among all eukaryotes and function by altering nucleosome structure to allow cellular regulatory factors access to the DNA. Mammalian SWI-SNF complexes contain either of two highly conserved ATPase subunits: BRG1 or BRM. To identify cellular genes that require mammalian SWI-SNF complexes for the activation of gene expression, we have generated cell lines that inducibly express mutant forms of the BRG1 or BRM ATPases that are unable to bind and hydrolyze ATP. The mutant subunits physically associate with at least two endogenous members of mammalian SWI-SNF complexes, suggesting that nonfunctional, dominant negative complexes may be formed. We determined that expression of the mutant BRG1 or BRM proteins impaired the ability of cells to activate the endogenous stress response gene hsp70 in response to arsenite, a metabolic inhibitor, or cadmium, a heavy metal. Activation of hsp70 by heat stress, however, was unaffected. Activation of the heme oxygenase 1 promoter by arsenite or cadmium and activation of the cadmium-inducible metallothionein promoter also were unaffected by the expression of mutant SWI-SNF components. Analysis of a subset of constitutively expressed genes revealed no or minimal effects on transcript levels. We propose that the requirement for mammalian SWI-SNF complexes in gene activation events will be specific to individual genes and signaling pathways.

The alignment of the 28S gene of several species of Pezizales allowed to select two pairs of primers able to amplify the internal transcribed spacer region of ribosomal DNA in mycorrhizal fungi, such as truffles. The higher yield of the amplification product demonstrates a better annealing of the new primers to the rDNA, as compared to the universal primers internal transcribed spacer 1 and internal transcribed spacer 4. Therefore, the new primers can be used as an easier and more sensitive tool for the identification of truffle species in any stage of their life cycle, including the mycorrhizal phase.

Purpose: We assess and clarify diagnostic features for making the prenatal diagnosis of cloacal exstrophy., Materials and Methods: We evaluated 9 patients born with cloacal exstrophy at our institutions (2 prospectively and 7 retrospectively) for diagnostic features on prenatal ultrasound studies. We also thoroughly reviewed the literature on 13 previous prenatally diagnosed cloacal exstrophy cases. Diagnostic criteria were assessed by combining the findings in our patients and those in previous reports., Results: Of the 22 patients with prenatal ultrasound studies and cloacal exstrophy whom we analyzed 1 of our 9 and 2 in the literature had a cloacal membrane that persisted at 22 weeks of gestation. Major ultrasound criteria for diagnosing cloacal exstrophy prenatally are nonvisualization of the bladder, a large midline infraumbilical anterior wall defect or cystic anterior wall structure (persistent cloacal membrane), omphalocele and lumbosacral anomalies. Seven less frequent or minor criteria include lower extremity defects, renal anomalies, ascites, widened pubic arches, a narrow thorax, hydrocephalus and 1 umbilical artery., Conclusions: We propose major and minor criteria to assist in the prenatal diagnosis of cloacal exstrophy. Despite these major and minor criteria the certainty of establishing a prenatal diagnosis remains challenging. Persistence of the cloacal membrane beyond the first trimester in 1 patient was an exception to the classic concept of cloacal exstrophy embryogenesis. An accurate prenatal diagnosis requires validation of these criteria by further correlation of prenatal and postnatal observations.

The role of radiotherapy in breast cancer is examined, based on a review of literature. Its impact on survival is discussed in the framework of a multidisciplinary treatment of the disease.

Aims and Background: The management of patients with T1 carcinoma of the glottic larynx is controversial, because surgery and radiation therapy are reported to be effective treatments. Several studies have shown radiotherapy to be safe and effective, with a high percentage of voice preservation and minimal complications, but most Italian physicians prefer to surgically treat such patients., Methods: From 1980 to 1990, 36 patients with stage I squamous cell glottic carcinoma were treated with radiotherapy alone at the institute of Radiology of University of Rome "La Sapienza". In all patients the irradiation fields were limited to the larynx, with field size ranging from 4 x 4 cm to 7 x 7 cm. Total tumor doses ranged between 51 and 70 Gy (median 60 Gy) with a mean number of 30 fractions of 2 to 3 Gy per fraction (3 or 5 fractions per week)., Results: After a median follow-up of 98 months, we observed an overall survival rate at 5 years of 91.4% and actuarial 10-year survival of 85.7%. Local control was achieved in 97.1% of cases, with an event-free survival of 94.2% at 5 and 10 years. No major complications like necrosis or persistent edema of the larynx were observed. Minor complications like dysphonia (8%) and dysphagia (5.5%) were temporary; laryngeal function was completely preserved at the end of therapy. Final voice quality ranged from good to excellent., Conclusions: Our series confirms that radiation therapy has a major role in the management of early glottic cancer, with results comparable to surgical approaches and with better voice preservation.

The authors analyse the results of treatment with radiotherapy with therapeutic doses and traditional fractionation scheme 60 patients affected by bladder cancer, from 1980 through 1991, at the Institute of Radiology of the University "La Sapienza" in Rome. We evaluated the association of radiotherapy (RT) with trans-urethral resection and biopsy (TURB) and the association of RT with cystectomy (total or partial), both in T1-T2 and T3-T4 bladder stages. The mean follow-up was 85 months (from 24 months to 11 years). The survival at 5 years after completing the treatment was 33.3%; the survival of stages T1-T2 was of 40% at 5 years while that of stages T3-T4 was of 28.5%. Our study shows that the association of radiotherapy and TURB for T1 or T2 bladder cancer is an alternative to mutilating surgery like total or partial cystectomy; in this group, indeed, we showed a survival of 54.5% at 5 years as compared to the 28.5% of the group treated with cystectomy + RT. For T3 and T4 bladder cancer we had better results with surgery (total or partial cystectomy) and RT: in these patients the survival at 5 years was of 31%, while the group treated with TURB + RT showed a survival of 19%.

Percutaneous endoscopic gastrostomy (PEG) is a worldwide accepted non surgical technique for enteral feeding in patients with prolonged swallowing difficulties or inabilities. PEG was recently introduced in Italy, and experience confirmed that such a technique is easy to perform, cheap, safe, well accepted by patients and carries a very low incidence of complications. In the present work technical procedure and our personal experience are reported; thirteen PEGs were performed our centre and no short nor long-term complications were observed.

The prevalence and characteristics of atrial septal aneurysm were identified by transesophageal echocardiography (TEE) in a multicenter prospective study. One hundred and seventy-seven consecutive patients were evaluated in 2 years and 2 groups were compared: Group 1, 51 patients with documented cerebral ischemia event; Group 2, 126 patients affected by cardiac disease referred for other reasons. Group 1 included patients selected among 352 patients admitted to the Neurological and Geriatric Division of our Hospital in the period of this study. Patients with stroke-related carotid lesions and patients with a negative TC scan were excluded from this study. Atrial septal aneurysm was identified in 15 patients: 8 in Group 1 (16%), and 7 in Group 2 (5%), with a significant statistical difference between the groups (p = 0.02). All patients with atrial septal aneurysm underwent before TEE transthoracic echocardiography, leading to a correct diagnosis in 10 of 15 cases (66%); all patients underwent 24-hours ECG monitoring. A right to left shunt was detected by contrast echocardiography in 9 patients, 6 in Group 1 and 3 in Group 2, (NS). A more pronounced shunt was evident in Group 1. There was no difference between the 2 groups with regard to associated cardiac disease, arrhythmias and type of atrial septal aneurysm. The thickness of the septum was greater in Group 1, with significant statistical difference (p = 0.002). It is concluded that atrial septal aneurysm, diagnosed by TEE, is a potential source of embolic events.

Untreated patients suffering from tuberculoid, lepromatous and indeterminate leprosy, their domiciliary contacts, and healthy controls, all living in Guadeloupe, West Indies, were tested by an ELISA for detecting IgM antibodies to the terminal disaccharide of the phenolic glycolipid-I antigen of Mycobacterium leprae. On most subjects, a Mitsuda test was also performed. A large majority of the tuberculoid patients and healthy subjects were Mitsuda positive. The seropositivity rate reached 44% among tuberculoid patients, and 6% among healthy subjects, with low antibody levels. Lepromatous patients were all Mitsuda negative and seropositive, with antibody production varying from low levels, as seen in tuberculoid patients, to much higher levels. Indeterminate leprosy patients included 62% Mitsuda-positive subjects and 54% seropositive subjects with a large dispersion of antibody levels. Comparing the results of the Mitsuda test to those of the ELISA by factorial analysis allowed us to define several subgroups among this population: some (25%) showed a "lepromatous-like" immune status (Mitsuda negative, seropositive); others (54%) exhibited "tuberculoid-like" profiles (Mitsuda positive without antibodies or with low antibody levels). "Lepromatous-like" cases were significantly older than "tuberculoid-like" patients. A group of subjects (17%) was Mitsuda negative and seronegative, thus displaying a true "indeterminate" immune profile, which had not been seen in other forms of the disease and had been observed in only 2 out of 51 healthy controls. A large majority of contacts was Mitsuda positive, with 33% of them being seropositive, indicating that the prevalence of M. leprae infection greatly exceeds that of overt leprosy in this population.(ABSTRACT TRUNCATED AT 250 WORDS)

Primary and secondary dapsone resistance were studied among lepromatous patients living in Martinique, Guadeloupe, New Caledonia, Tahiti, Senegal, and Paris. Four hundred fifteen biopsies were taken from clinically active and bacteriologically positive (bacterial index greater than 2) patients in the 6-year period of 1980-1985. Among these, 280 biopsies that contained 5 x 10(4) acid-fast bacilli per ml with a morphological index of at least 0.10 were inoculated into the mouse foot pad, and 229 harbored infective Mycobacterium leprae. Among the 129 infective M. leprae isolated from new cases, 54% had some degree of dapsone resistance, a low degree being prominent in all cases. Among the 100 infective M. leprae isolated from relapsed cases, 79% had a high or an intermediate degree of dapsone resistance. The annual incidence of secondary dapsone resistance was estimated to be about 0.55% in Guadeloupe.

A seroepidemiological surveillance of a contact population was started in 1984 in French Polynesia. The ELISA test was used to measure IgM anti-ND-O-BSA in the sera. Specific antibody levels were higher in healthy Polynesians than in normal individuals living in a nonendemic country. The positive threshold of the reaction was fixed according to this background activity in healthy Polynesians. Under these conditions, 100% of the multibacillary patients were detected as seropositive as compared to 5% of the paucibacillary group. In the population of 724 household contacts tested and observed for 2 years: 93 (12.8%) were seropositive, with 8 (1.1%) showing activity equivalent to multibacillary patients (1 of these 8 individuals developed a lepromatous form of leprosy); 631 (87%) were seronegative and 3 developed a paucibacillary form of the disease (2 BT, 1 I) without any antibody increase. Among those four contacts who developed leprosy, three were related to a multibacillary index case. These data suggest that this test may be useful for the prediction of multibacillary leprosy. A long-term surveillance of this high-risk population will be able to evaluate the diagnostic and prognostic value of the serological assay.

The authors assess of three decades of struggle against Bancroftian filariasis in French Polynesia. Wuchereria bancrofti var. pacifica, aperiodic filaria, and Aedes polynesiensis, mosquito with high parasitologic output, set up a cycle very well adapted to the Polynesian environment; after numerous tests, the chemoprophylaxis with diethylcarbamazine (3 mg/kg/half-year) of all the exposed population has been decided, in association with methods of vector-control (use of predatory crustaceans). However, the achievement of this strategy is impeded by economical contingencies and the endemic comes up again particularly in areas close to eradication. Entomological clues show a high transmission over the greater part of French Polynesia.