Comparative phenotypic analysis of "Clostridium neonatale" and Clostridium butyricum isolates from neonates.

"Clostridium neonatale" was recently described as a new species within the Cluster I of the Clostridium genus sensu stricto. In this study, we characterized "C. neonatale" isolates (n = 42) and compared their phenotypic properties with those of Clostridium butyricum (n = 26), a close related species. Strains isolated from fecal samples of healthy neonates were tested for different phenotypic characteristics. Compared to C. butyricum, "C. neonatale" showed a significant higher surface hydrophobicity (p = 0.0047), exopolysaccharide production (p = 0.0069), aero-tolerance (p = 0.0222) and viability at 30 °C (p = 0.0006). A lower swimming ability (p = 0.0146) and tolerance against bile (0.3%) (p = 0.0494), acid (pH 4.5) (p < 0.0001), osmolarity (NaCl 5%, p = 0.0188) and temperature at 50 °C (p = 0.0013) characterized "C. neonatale" strains. Our results showed that "C. neonatale" behaves very differently from C. butyricum and suggests specific responses to environmental changes. Besides it is the first study on clinical isolates for these two anaerobic members of the newborns' gut microbiota and broadens our knowledge about their phenotypic traits.

Carcinoma de células claras variedad no hialinizante de parótida: comunicación de dos casos con diferente grado de diferenciación / Non-hyalinizing clear cell carcinoma of the parotid: report of two cases with different grade of differentiation

El carcinoma de células claras es una neoplasia rara de bajo grado que se desarrolla casi exclusivamente en glándulas salivales menores humanas. El objetivo de esta comunicación es analizar las características estructurales, histoquímicas e inmunohistoquímicas (IHQ) de dos casos de carcinoma de células claras de parótida y realizar el diagnóstico diferencial con otros tumores salivales primarios y metastásicos que presentan células claras. Cortes de ambos tumores fueron procesados para H/E, tricrómicos de Masson y Dane, Azul de toluidina, Azul alciano, PAS y PAS/diastasa; marcaciones IHQ para citoqueratinas de bajo y alto peso molecular, ki67, HMB45, p63 y proteína S-100. El patrón estructural de estos tumores estaba determinado por nidos y cordones de células claras delimitados por un estroma no hialinizado. En un tumor se observó una neoplasia maligna a células claras con manifiesta anaplasia. La expresión de ki67 fue importante. El otro tumor estaba constituido por células claras monomorfas sin signos manifiestos de atipia y casi nula expresión de ki67. Con PAS se demostró la presencia de glucógeno y no se observó un desarrollo importante del estroma colágeno en ambas neoplasias. En ambos casos resultó francamente positiva la inmunomarcación para citoqueratinas de bajo y alto peso molecular. Por el contrario resultó negativa para HMB45 y p63. La proteína S-100 tuvo su expresión en células aisladas. Se concluye que los casos presentados son de localización poco común (parótida) según lo descripto en la bibliografía; corresponden a la variedad no hialinizante ya que no está presente un importante estroma colágeno hialinizado, y ambos tumores son primarios de glándulas salivales, de origen epitelial, no melanocítico ni hematopoyético (linfomas) como lo demuestran las marcaciones IHQ.

The clear cell carcinoma is a rare neoplam of low level grade that develops almost exclusively in human minor salivary glands. The objective of this present work is to analyze the structural, histochemical and inmunohistochemical (IHQ) characteristics of two cases of parotid clear cell carcinoma and make a differential diagnosis with other primary and metastatic salivary tumors that clear cells show. Serial cuts of both tumors were processed for H/E, Masson and Dane trichromes, Toluidine blue, Alcian blue, PAS and PAS/diastase; IHQ marking of both high-and low-molecular weight cytokeratins, ki67, HMB45, p63 and S-100 protein. The structural pattern of these tumors were determined by nests and cords of clear cells delimited by a stroma non hyaline. In a tumor a malign neoplasia was observed in clear cells with anaplasia. The ki67 expression was important. The other tumor was constitued of clear monomorph cells without manifestation signs of atypia and almost no ki67 expression. With PAS was confirmed the presence of glycogen and not observed an important development of the collagen stroma in both neoplasms. Both cases resulted very positive the immunostaining of both high and low molecular weight cytokeratins. It resulted negative for the HMB45 and p63. The protein S-100 has it´s expression in isolated cells. We conclude that: the presented cases are very uncommon localization (parotid) as described by the literature; it matches a non hyalinizing variety because a very important hyalinized collagen stroma is not present, both primary tumors of salivary glands, of epithelial origin, non melanocytic, renal or hematopoietic lymphomas which shows immunostaining IHQ.

The role of MRI and CT in diagnosis and treatment planning of cherubism: a 13-year follow-up case report.

AIM: Cherubism is characterised by mesenchymal alterations during the development of the jaws secondary to perivascular fibrosis. According to the ALARA (As Low As Reasonably Achievable) principle, it is important to avoid conditions where the amount of radiation used is more than that needed for the procedure, because there is no benefit from unnecessary radiation. However, the use of MRI has been poorly studied in cherubism. MATERIALS AND METHODS: The patient underwent head and neck MRI and 3D CT for imaging assessment. RESULTS: MRI is necessary to evaluate the extension of dysplastic tissue and the cystic part of the lesions. Bone window CT only allows evaluation of strong densitometric alterations of cherubism lesions. Moreover, on radiographic film it is not always possible to distinguish fibrous tissue from mucous pseudocystic tissue. By contrast, these differences are readily evident on MRI. CONCLUSION: MRI, in addition to other traditional radiographs and CT, could be useful in helping the clinician in the diagnosis and treatment of cherubism.

Reinforcement of lumbosacral instrumentation using S1-pedicle screws combined with S2-alar screws.

OBJECTIVE: Increasing construct stability of lumbosacral instrumentations using S2-ala screws as an alternate to iliac screws. INDICATIONS: Revision surgery after failed lumbosacral fusion; long instrumentations to the sacrum; L5-S1 fusion without anterior support. CONTRAINDICATIONS: Lack of sacral bone stock. SURGICAL TECHNIQUE: Midline approach. The entry point for S2-ala screws is caudal to the posterior S1 foramen and close to the lateral sacral crest. Screw tract preparation for S2-ala screws necessitates 30-45° angulation in the axial plane. Biplanar fluoroscopy with inlet and outlet views ensure screw accuracy. With S2-ala screws, bicortical fixation is the goal. POSTOPERATIVE MANAGEMENT: Patients are mobilized under the surveillance of physiotherapists on day 1 and released from the hospital after 10 days. Clinical and radiographic controls are performed at 6, 12 and 24 months. RESULTS: Retrospective review of 80 patients undergoing S2-ala screw fixation. Main diagnosis was degenerative lumbar instability, adult scoliosis, high-grade listhesis, and nonidiopathic scoliosis. In 66% of patients, the instrumentation using S2-ala screws was part of a major lumbosacral revision surgery. Follow-up averaged 26 months. There were no deaths or major neurovascular complications. First time fusion rate at L5-S1 was greater than 90%. Eight patients (10%) experienced a complication which could be related to the S2-ala screws. Out of 160 S2-ala screws, 16 screws were judged to cause focal irritation and were removed, indicating a survival rate of 90% for the S2-ala screw.

[Restoration of sagittal balance in treatment of thoracic and lumbar vertebral fractures]. / Wiederherstellung der sagittalen Balance bei der Versorgung thorakaler und lumbaler Wirbelkörperfrakturen.

Posttraumatic kyphosis (PTK) is a possible consequence of a missed fracture, a wrong indication for conservative therapy or an inadequate surgical technique but PTK can also be a complication after adequate surgery. Avoidance of PTK is of importance because subsequent surgical therapy can be extensive. A thorough planning as well as surgical experience with anterior and posterior revision cases is necessary. The various types of osteotomy which allow correction of local as well as global deformities should also be mastered. Knowledge of the principles of sagittal balance and spinopelvic parameters are indispensable in the treatment of PTK. Our experience and results from the literature show that a good long-term outcome with limited complications can only be achieved when considering the biomechanical principles as well as restoration of sagittal balance.

High-throughput mapping of protein occupancy identifies functional elements without the restriction of a candidate factor approach.

There are a variety of in vivo and in vitro methods to determine the genome-wide specificity of a particular trans-acting factor. However there is an inherent limitation to these candidate approaches. Most biological studies focus on the regulation of particular genes, which are bound by numerous unknown trans-acting factors. Therefore, most biological inquiries would be better addressed by a method that maps all trans-acting factors that bind particular regions rather than identifying all regions bound by a particular trans-acting factor. Here, we present a high-throughput binding assay that returns thousands of unbiased measurements of complex formation on nucleic acid. We applied this method to identify transcriptional complexes that form on DNA regions upstream of genes involved in pluripotency in embryonic stem cells (ES cells) before and after differentiation. The raw binding scores, motif analysis and expression data are used to computationally reconstruct remodeling events returning the identity of the transcription factor(s) most likely to comprise the complex. The most significant remodeling event during ES cell differentiation occurred upstream of the REST gene, a transcriptional repressor that blocks neurogenesis. We also demonstrate how this method can be used to discover RNA elements and discuss applications of screening polymorphisms for allelic differences in binding.

Is anterior release effective to increase flexibility in idiopathic thoracic scoliosis? Assessment by traction films.

With the advent of thoracoscopy, anterior release procedures in adolescent idiopathic scoliosis (AIS) have come into more frequent use, however, the indication criteria for an anterior release in thoracic AIS are still controversial in the literature. To date, few studies have assessed the influence on spinal flexibility and no study has so far been able to show a beneficial effect on the correction rate as compared to a single posterior procedure. The objective of this study was to evaluate the influence of thoracic disc excision on coronal spinal flexibility. Six patients (5 females, 1 male) with AIS and a mean age of 15.6 years (range 13-20 years) underwent an open anterior thoracic release prior to posterior instrumentation. Cotrel dynamic traction along with radiographs of the whole spine including traction films were conducted pre- and postoperatively and were evaluated retrospectively. The mean preoperative thoracic curve was 89.7 degrees +/- 15.4 degrees (range 65 degrees-110 degrees). The flexibility rate in Cotrel traction was 22.8 +/- 8.1%. After performance of the anterior release the thoracic curve showed a mean increase of coronal correction by 5.5 degrees +/- 5.0 degrees as assessed by traction radiographs. The flexibility index changed by 6.2 +/- 5.6%. After posterior instrumentation the thoracic curve was corrected to a mean of 36.5 degrees +/- 10.1 degrees (correction rate 59.6%). Disc excision in idiopathic thoracic scoliosis only slightly increased spinal flexibility as assessed by traction films. In our view a posterior release with osteotomy of the concave ribs (concave thoracoplasty, CTP) is more effective in increasing spinal flexibility. According to our clinical experience, an anterior release prior to posterior instrumentation in AIS should only be considered in hyperkyphosis, coronal imbalance or massive curves.

Management and outcomes of hepatic cirrhosis: Findings from the RING study.

BACKGROUND/AIM: Hepatic cirrhosis is a frequent reason for ordinary hospital admission (OA). The RING study collected hospital discharge files (HDF) from Italian hospital gastroenterology units (IGU). This caselist provides a broad picture of the patients admitted for this pathology. MATERIAL/METHODS: More than 50,000 HDF for OA were collected between 2001 and 2004 from 26 IGU. RESULTS: Eight thousand four hundred and eighty-seven HDF (16%) had a diagnosis of hepatic cirrhosis; Child-Pugh classes were 20.2% A, 34.8% B and 45.0% C. Patients' mean age was 63.7+/-12.1 years and 62.5% were male. A 61.1% of the cirrhosis cases had ascites, 29.9% portal-systemic encephalopathy, 29.2% hepatocellular carcinoma (HCC), 10% bleeding varices, 3.0% hepatorenal syndrome (HRS). Mortality for OA for cirrhosis was 5.7% versus 2.6% for other diagnoses. The proportion varied with the severity of the cirrhosis: 0% for Child A, 1.1% B, 10.5% C. Mortality was significantly associated with: Child-Pugh at admission (odds ratio: OR 9.2), HRS (OR 11.7), bleeding varices (OR 2.2), HCC (OR 1.8). CONCLUSIONS: Hepatic cirrhosis was found in 16% of the OA to IGU and mortality was double the rate for all the other pathologies in the same wards. Child-Pugh is a useful prognostic tool, higher classes implying a greater risk of death. HRS and bleeding varices were the complications with most influence on in-hospital mortality.

Seasonal patterns of hospital treatment for inflammatory bowel disease in Italy.

AIM: It is still debated whether clinical flare-ups of chronic inflammatory bowel disease follow a seasonal pattern, and the various reports are based on general practitioners' records or hospital discharge charts. There are, however, no specific figures for treatment in hospital gastroenterology units, which serve as a reference point for these disorders. This study was therefore designed to investigate whether there is a seasonal pattern in admissions for inflammatory intestinal disease in Italy, differing from what is generally known about gastrointestinal pathologies, since there are no nation-wide figures on the subject. METHODS: The RING (Ricerca Informatizzata in Gastroenterologia) project is an observational study collecting hospital discharge forms from 22 centers in Italy. RESULTS: From winter 2000 to autumn 2003, the 22 gastroenterology units participating in the RING project discharged 32,357 patients following ordinary hospital admissions. Of these, 2,856 (8.8%) had a main diagnosis of inflammatory bowel disease: 1,541 Crohn's disease, and 1,315 ulcerative colitis. No seasonal patterns were detected for either category, or when the analysis was done by age, sex and site of disease. CONCLUSIONS: The most serious flare-ups of inflammatory bowel disease, i.e. those requiring routine hospital treatment, do not appear to follow any seasonal pattern, regardless of the site of the disease or the patient's age or sex.

Tumores neuro-endocrinos del intestino / Neuroendocrine tumors of the small intestine

Los tumores carcinoides son neoplasias poco frecuentes en el tubo digetivo y que se desarrollan a partir de las células que forman el sistema APUD o neuroendocrinas. El objetivo es analizar la nueva nomenclatura, la incidencia en el aparato digetivo, la presentación clínica, diagnóstico y tratamiento quirúrgico

Tumores neuro-endocrinos del intestino / Neuroendocrine tumors of the small intestine

Los tumores carcinoides son neoplasias poco frecuentes en el tubo digetivo y que se desarrollan a partir de las células que forman el sistema APUD o neuroendocrinas. El objetivo es analizar la nueva nomenclatura, la incidencia en el aparato digetivo, la presentación clínica, diagnóstico y tratamiento quirúrgico

Liquid chromatographic determination of triasulfuron in soil.

Two extraction methods were developed for the determination of triasulfuron in soil. Method I included extraction with methanol-phosphate buffer at pH 7 (2 + 1, v/v), liquid-liquid partition with dichloromethane, and cleanup on a liquid chromatographic Si adsorption solid-phase extraction tube. In Method II, Extrelut was added and the sample was then extracted with acetonitrile. In both cases, the extracts were analyzed by liquid chromatography (LC) with UV detection and the LC peak was confirmed by LC/mass spectrometry (MS). The 2 methods were tested on 3 soils having different physicochemical characteristics. Method I gave 83% average recovery and a determination limit of 0.4 microg/kg soil. Method II gave 67% average recovery and a determination limit of 2 microg/kg soil. Examples of application of Method I to field samples are reported.

The natural history of ulcerative proctitis: a multicenter, retrospective study. Gruppo di Studio per le Malattie Infiammatorie Intestinali (GSMII).

OBJECTIVE: The aim of this study was to evaluate the clinical features and the long term evolution of patients with a well defined initial diagnosis of ulcerative proctitis. METHODS: Patients with an original diagnosis of ulcerative proctitis who had been seen at any of 13 institutions from 1989 to 1994 were identified. Data on disease onset and subsequent evolution were recorded. In addition, 575 patients with more extensive disease, treated in the same centers, were used as controls. RESULTS: A total of 341 patients satisfied the inclusion criteria. The percentage of smokers in these patients was slightly lower than in controls; no differences were found in the other clinical/demographic variables evaluated. A total of 273 patients entered long term follow-up (mean, 52 months). Proximal extension of the disease occurred in 74 of them (27.1%). The cumulative rate of proximal extension and of extension beyond the splenic flexure was 20% and 4% at 5 yr and 54% and 10% at 10 yr, respectively. The risk of proximal extension was higher in nonsmokers, in patients with >3 relapses/yr, and in patients needing systemic steroid or immunosuppressive treatment. Refractory disease was confirmed as an independent prognostic factor at multivariate analysis. CONCLUSIONS: Proximal extension of ulcerative proctitis is frequent and may occur even late after the original diagnosis. However, the risk of extension beyond the splenic flexure appears to be quite low. Smoking seems to be a protective factor against proximal extension, whereas refractoriness is a risk factor for proximal extension of the disease.

Are we correctly using the inhibitors of gastric acid secretion and cytoprotective drugs? Results of a multicentre study.

AIM: To evaluate indications and frequency of prescriptions of pH-modifying and cytoprotective drugs in hospital departments. DESIGN OF THE STUDY: Multicentre cohort transverse relevation. MATERIALS AND METHODS: On a randomly selected day the clinical charts of 3685 inpatients were reviewed by a gastroenterologist and data were collected concerning the drug used, its dosage and indications for the prescription. RESULTS: 26.8% of hospitalized patients were under treatment on the day of the study. Ranitidine was the most frequently prescribed (73.9%). The purpose of treatment was cure of acute disease or symptoms in 27% of cases and prevention of peptic damage in 73%. Fear of development of stress mucosal lesions (28%) and therapy with other drugs (56%) motivated prophylactic treatment in the majority of cases. The examination of clinical charts failed to demonstrate any need for treatment in 51.4% of the patients. Disparities in the frequency of prescription between different departments and hospitals were found. CONCLUSION: Criteria used for prescription of drugs for peptic diseases in hospitalized patients should be strictly monitored to avoid excessive and unmotivated use also in general practice.

Eosinophils within the healthy or inflamed human intestine produce substance P and vasoactive intestinal peptide.

The purpose of this study was to show if inflammatory cells within healthy or diseased human intestinal mucosa produce some regulatory neuropeptides. First, inflammatory cells were isolated from the intestinal lamina propria of 11 patients with ulcerative colitis or Crohn's disease. Also collected were cells from anatomically normal intestine derived from five patients requiring bowel resection for diseases not related to inflammatory bowel disease. Extracts of these isolated cells contained authentic substance P (SP) and vasoactive intestinal peptide (VIP) as shown by RIA and their elution profiles on HPLC. Immunostaining of cells from nine of 13 additional patients localized immunoreactive SP and VIP to secretory granules within most mucosal eosinophils. No other cell types stained positive. Messenger RNA encoding SP and VIP was localized to lamina propria eosinophils by in situ hybridization. Mucosa inflammatory cells, from eight of nine more patients, cultured in vitro, released detectable amounts of VIP, but not SP. It is concluded that intestinal eosinophils produce SP and VIP. Since the eosinophils store and release more VIP than SP, it is possible that VIP is the preferred secretory product.

Acetyl-L-carnitine affects aged brain receptorial system in rodents.

Acetyl-L-carnitine (ALCAR), the acetyl ester of carnitine, is regarded as a compound of considerable interest because of its capacity to counteract several physiological and pathological modifications typical of brain ageing processes. In particular, it has been demonstrated that ALCAR can counteract the age-dependent reduction of several receptors in the central nervous system of rodents, such as the NMDA receptorial system, the Nerve Growth Factor (NGF) receptors, those of glucocorticoids, neurotransmitters and others, thereby enhancing the efficiency of synaptic transmission, which is considerably slowed down by ageing. The present review thus postulates the importance of ALCAR administration in preserving and/or facilitating the functionality of carnitines, the concentrations of which are diminished in the brain of old animals.

Intestinal epithelial cells contribute to the enhanced generation of platelet activating factor in ulcerative colitis.

Generation of platelet activating factor by intestinal mucosal epithelial cells and lamina propria mononuclear cells was evaluated to elucidate the possible role of this mediator in the pathogenesis of inflammatory bowel disease. Epithelial and lamina propria mononuclear cells were isolated from surgical specimens from control, Crohn's disease, and ulcerative colitis patients. Platelet activating factor was extracted from highly purified cell preparations with 80% ethanol after stimulation with and without 0.2 uM calcium ionophore A23187 and was measured by platelet aggregation assay. Both cell types generated platelet activating factor activity and this was generally comparable for epithelial and lamina propria cells. Basal and stimulated platelet activating factor activity of epithelial and lamina propria cells from ulcerative colitis but not Crohn's disease patients was appreciably higher than that of control. Stimulation with calcium ionophore increased appreciably platelet activating factor activity in lamina propria cells from all groups. In contrast, only epithelial cells from ulcerative colitis showed an appreciable increase after calcium ionophore induction. These results suggest that epithelial cells are important contributors to intestinal platelet activating factor generation under normal and inflammatory conditions and that epithelial cells actively play a part in the pathogenesis of ulcerative colitis.

[Unilateral gynecomastia induced by the use of anabolic steroids. A clinical case report]. / Ginecomastia unilaterale indotta dall'uso di steroidi anabolizzanti. Descrizione di un caso clinico.

The authors studied a case of left unilateral gynecomastia occurring in a 26-year-old body-builder after anabolic steroids assumption. The mammal features were found by echography and mammography and they showed the presence of a 3 cm diameter retroareolar area of gland tissue surrounded by fibrotic tissue. After excluding every likely cause giving gynecomastia through laboratory tests, it was clear a relationship with the assumption of high doses of anabolic hormones, with particular regard to methyltestosterone. Therefore, also this substance which on its own is not aromatized by extrasplanchnic tissues, can be responsible for man's mammal hypertrophy, if joined to a slight iatrogenic liver cytolysis.