Phytoextraction efficiency of Pteris vittata grown on a naturally As-rich soil and characterization of As-resistant rhizosphere bacteria.

This study evaluated the phytoextraction capacity of the fern Pteris vittata grown on a natural arsenic-rich soil of volcanic-origin from the Viterbo area in central Italy. This calcareous soil is characterized by an average arsenic concentration of 750 mg kg-1, of which 28% is bioavailable. By means of micro-energy dispersive X-ray fluorescence spectrometry (µ-XRF) we detected As in P. vittata fronds after just 10 days of growth, while a high As concentrations in fronds (5,000 mg kg-1), determined by Inductively coupled plasma-optical emission spectrometry (ICP-OES), was reached after 5.5 months. Sixteen arsenate-tolerant bacterial strains were isolated from the P. vittata rhizosphere, a majority of which belong to the Bacillus genus, and of this majority only two have been previously associated with As. Six bacterial isolates were highly As-resistant (> 100 mM) two of which, homologous to Paenarthrobacter ureafaciens and Beijerinckia fluminensis, produced a high amount of IAA and siderophores and have never been isolated from P. vittata roots. Furthermore, five isolates contained the arsenate reductase gene (arsC). We conclude that P. vittata can efficiently phytoextract As when grown on this natural As-rich soil and a consortium of bacteria, largely different from that usually found in As-polluted soils, has been found in P. vittata rhizosphere.

Anterior trans-frontal endoscopic management of colloid cyst: an effective, safe, and elegant way of treatment. Case series and technical note from a multicenter prospective study.

Different management options are available for the treatment of colloid cysts. Goals of those procedures are to achieve a complete resection avoiding potential long-term recurrence along with CSF pathways restoration with minimal morbidity and mortality. The two main surgical options are endoscopic resection or direct removal by either transfrontal or transcallosal approach. The efficacy of endoscopic technique to achieve gross total colloid cyst excision has been well documented. In the present study, authors describe a series of 29 patients who underwent surgery by a variation of the standard worldwide implemented endoscopic technique. Using a more anterior approach, it is easier to reach the roof of the cyst, its possible adherences with the tela choroidea, plexus, and the internal cerebral veins. The described approach has shown to be safe, quick, and very effective with a total cyst removal rate of 86.2%.

Diagnostic value of PRND gene expression profiles in astrocytomas: relationship to tumor grades of malignancy.

Doppel (PRND) is a paralogue of the mammalian prion (PRNP) gene. It is abundant in testis and, unlike PRNP, it is expressed at low levels in the adult central nervous system (CNS). Besides, doppel overexpression correlates with some prion-disease pathological features, such as ataxia and death of cerebellar neurons. Recently, ectopic expression of doppel was found in two different tumor types, specifically in glial and haematological cancers. In order to address clinical important issues, PRND mRNA expression was investigated in a panel of 111 astrocytoma tissue samples, histologically classified according to the World Health Organization (WHO) criteria (6 grade I pilocytic astrocytomas, 15 grade II low-grade astrocytomas, 26 grade III anaplastic astrocytomas and 64 grade IV glioblastoma multiforme). Real-time PRND gene expression profiling, after normalisation with GAPDH, revealed large differences between low (WHO I and II) and high grade (III and IV) of malignancy (P<0.001). Extensive differences in PRND gene expression were also found within each grade of malignancy, suggesting that PRND mRNA quantitation might be useful to distinguish astrocytoma subtypes, and important in disease stratification and in the assessment of specific treatment strategies.

[Intestinal intussusception: easy diagnosis in the 21st Century? Outcome of five years experiences]. / Invaginazione intestinale: una diagnosi facile nel 21 degrees secolo? Outcome degli ultimi 5 anni.

UNLABELLED: Intussusception is an entity well codify in pediatric surgery, with guidelines well defined since years in terms of imaging and procedures. MATERIAL: From January '99 to Dec. '03, 25 children were admitted to the Department of Pediatric Surgery of the Children's Hospital of Milan. All the aspects related to the pathology have been evaluated in the study. RESULTS: 25 patients have been studied. 23/25 presented severe pain, 19/25 normal bowel movements in the last 3 hours and in 9/25 a mass were detected at clinical examination.19/25 were submitted to laparotomy after failure of reduction through barium enema, and in 17/19 a manual reduction were performed. Only 2 patients required a bowel resection with primary anastomosis. The analysis of the delay of diagnosis, the delta-T between beginning of the symptoms and first medical evaluation was of 16,23 h. (3-72), and the delta-T between the first evaluation and diagnosis was 18,7 h. The interval between diagnosis and surgery was of 4,3. CONCLUSION: Authors believe that intussusception still represent a challenge for medical and surgical emergency team, supported by the data in literature. Imaging procedures must not be constricted in case of minimal suspicions of intestinal intussusception.

Effects of exogenous propylene on softening, glycosidase, and pectinmethylesterase activity during postharvest ripening of apricots.

Apricots (Prunus armeniaca L. cv. Boccuccia spinosa) picked at the commercial ripening stage [soluble solids content (SSC) 12.6%] were left to reach full ripening in continuously humidified air at 20 degrees C. Changes in the rate of ethylene production, firmness, soluble solids concentration, and titratable acidity were measured. The alpha-D- and beta-D-glucosidases, alpha-L-arabinofuranosidase, alpha-D- and beta-D-galactosidases, beta-D-xylosidase, and alpha-D-mannosidase activities were assayed. To evaluate the influence of ethylene on glycosidase activity, propylene (500 microL x L(-1)) was applied to apricots for 24 and 48 h. In apricots ripened in air, ethylene production increased on the first day and exhibited a typical climacteric pattern. Good edible quality was reached in 5 days when SSC was at least 14% and acidity was between 1.1 and 1.2% (% malic acid). During postharvest ripening, alpha-L-arabinofuranosidase activity increased from 1.9 to 11.6 nkat until day 7. alpha-D-Galactosidase, alpha-D-mannosidase, and beta-D-galactosidase activity increased continuously but at a lower rate. beta-D-Xylosidase activity also increased, but the level of activity was lower than the other glycosidases assayed. Pectinmethylesterase (PME) decreased during the postharvest ripening, and propylene enhanced this pattern, by stimulating ethylene production. Even the activities of alpha-L-arabinofuranosidase, beta-D-xylosidase, alpha-D-mannosidase, and beta-D-galactosidase were greatly stimulated by the propylene treatment, which consequently induced rapid softening of the fruits.

Experience with spiral computed tomography as the sole diagnostic method for traumatic aortic rupture.

BACKGROUND: Spiral computed tomographic (CT) scan is an excellent screen for aortic trauma. Traditionally, aortography is performed when injury is suspected to confirm the diagnosis. We hypothesized that it is safe and expeditious to forgo aortography when the spiral CT demonstrates aortic injury. METHODS: Retrospective review of 54 patients undergoing aortic repair from July 1994 to December 1999. Spiral CT was the initial diagnostic study in 52 patients. Pseudoaneurysm or aortic wall defect in the presence of mediastinal hematoma was considered diagnostic. Angiography, initially routine, was later performed only when requested by the surgeon, and for all "nonnegative" studies (periaortic hematoma without detectable aortic injury). RESULTS: Twenty-six patients underwent angiography before operation (group 1). Nineteen group 1 spiral CTs were unequivocally diagnostic; 7 were nonnegative and angiography was required. Twenty-eight other patients underwent repair based on spiral CT alone (group 2). There was one false-positive result in both groups. There were no unexpected operative findings. Mean time from admission to diagnosis was 5.7+/-3.4 hours for group 1 and 1.7+/-1.7 hours for group 2 (p < 0.01). CONCLUSIONS: Operating on the basis of a diagnostic spiral CT is safe and expeditious. Aortography may be reserved for those with equivocal studies.

Heparinless partial cardiopulmonary bypass for the repair of aortic trauma.

OBJECTIVE: We hypothesized that partial cardiopulmonary bypass with a heparin-bonded system would be a technically simple, effective adjunct for reducing paraplegia during repair of traumatic aortic rupture. It avoids the risk of heparin, but, unlike left atrial-arterial bypass, it can heat, cool, oxygenate, and rapidly infuse volume if needed. METHODS: A retrospective review was conducted of patients admitted for aortic trauma from July 1994 to December 1999. Bypass consisted of femoral venous (right atrial) cannulation, a centrifugal pump, and an oxygenator-heater/cooler. Arterial return was to the femoral artery or distal aorta. The entire system was heparin-bonded and no systemic heparin was given. RESULTS: Heparin-bonded partial bypass was established in 50 patients (mean age 43 +/- 17 years). Crossclamp time was 32 +/- 11 minutes (range 14-70 minutes), mean flow 3.0 +/- 0.8 L/min, and bypass time 64 +/- 43 minutes. During repair, 58% of patients received volume through the system (mean 1.1 +/- 1.9 L). Core temperature rose slightly (35.9 degrees C +/- 0.7 degrees C to 36.3 degrees C +/- 0.8 degrees C). Three of the 15 patients who underwent percutaneous femoral arterial and venous cannulation concomitant with their angiograms had vessel injury, with one limb loss, and this procedure was discontinued. Thirty-five patients underwent percutaneous femoral vein and direct distal aortic cannulation without event. The mortality rate for patients supported by bypass was 10%, and all deaths were due to other injuries. There were no new cases of paraplegia and no worsening of intracranial or pulmonary injuries. CONCLUSIONS: Heparin-bonded bypass is technically simple to use and avoids the risk of anticoagulation. Paraplegia was avoided. The ability to correct hypothermia, oxygenate, and rapidly infuse volume may simplify management and improve outcomes.

[Sentinel lymph nodes in skin melanoma]. / Il linfonodo sentinella nel melanoma cutaneo.

BACKGROUND: Early node dissection offers increased survival in patients with node metastases only. The study of sentinel node (SN) using blue dye and radiolocalization permits to identify patients who could undergo lymphadenectomy. METHODS: At Department of Surgery of Macerata General Hospital 22 patients with melanoma of trunk or limbs at I and II stage were submitted to SN biopsy. RESULTS: SN was identified in all cases by combined approach. Dynamic lymphoscintigraphy permitted identification of SL when it was not the node nearest the tumor. Nodal metastases were found in 2 cases (9.1%) and the patients underwent regional lymphadenectomy. In 1 patient the SN was the only node with metastases. Both patients had high thickness melanomas. The measurement of radioactive exposition for operating room personnel and pathologist demonstrated that the technique is safe and without risks. CONCLUSIONS: Biopsy of SN is effective for identification of occult nodal metastases from cutaneous melanoma. Combined technique allows to localize SN in 100% of cases. There is not radio-exposition for operators.

Reversible pulmonary trunk banding with a balloon catheter: assessment of rapid pulmonary ventricular hypertrophy.

OBJECTIVE: We sought to assess the rapid hypertrophy of the right ventricle of young goats submitted to progressive pressure load by a balloon catheter. METHODS: The hearts of 6 young goats were assessed by means of echocardiography and cell morphology during and after right ventricular hypertrophy had been produced by a balloon catheter. Myocardial samples of the right ventricular outflow tract were harvested for microscopic studies. The external diameter of longitudinally sectioned myocytes was measured at the nucleus level. The volume density of mitochondria was also determined. A balloon catheter was then placed through the right ventricular outflow tract in the pulmonary trunk and progressively inflated every 2 days. Postoperative serial echocardiography was performed at intervals of 1 to 2 days. The animals were killed after 2 to 3 weeks of right ventricular training for morphologic analysis. RESULTS: Under optical microscopy, there was a 20.5% increase in the mean diameter of the myocyte of the trained right ventricle. However, under electron microscopy, there was no significant change in the mean volume density of mitochondria from the trained right ventricle. Serial echocardiography showed equalization of the ventricular thickness over a short interval of 6 to 10 days of progressive balloon inflation. CONCLUSIONS: The balloon catheter permits the manipulation of the pressure load over the right ventricle, causing rapid hypertrophy in a 6- to 10-day period. This study suggests that nonsurgical preparation of the "pulmonary ventricle" in patients with transposition of great arteries with intact ventricular septum beyond the neonatal period could probably be accomplished within a very few days.

Prognostic relevance of altered Fas (CD95)-system in human breast cancer.

The Fas ligand (FasL) and its receptor Fas (APO-1 or CD95) are members, respectively, of the tumor necrosis factor family that, upon interaction with each other, play a key role in the initiation of one apoptotic pathway. Faulty regulation of the Fas system has been described in a variety of human tumors with different histogenetic origin. Here, we describe the expression and distribution of Fas receptor and ligand pair antigens in surgical samples collected from a cohort of 186 patients bearing breast neoplasms (45 benign and 141 malignant lesions). Immunoperoxidase staining of formalin-fixed tissues showed that 91.1% of benign lesions expressed Fas, which was present in only 56.7% of malignant tumors. On the other hand, FasL was found positive in 22.2% of benign neoplasms and up-regulated in in situ as well as invasive carcinomas (53.9%). Moreover, in malignant tumors, the expression of receptor and ligand antigens appeared to be inversely related. When these findings were correlated with pathological parameters of prognostic relevance, a significant association was observed between FasL and the presence of metastatic lymph nodes and larger tumor size while Fas expression correlated to node-negative status and smaller tumor size. Patients with Fas positive tumors exhibited longer disease-free survival than those with Fas-negative carcinoma while FasL did not influence patient outcome. These relationships indicate that benign and malignant mammary lesions are characterized by differential cellular expression of Fas and FasL and suggest that a neoplastic Fas negative/FasL positive phenotype may be linked to breast cancer progression.

The effect of angiotensin II on mitogen-activated protein kinase in human cardiomyocytes.

The role of angiotensin II (Ang II)-receptors on mitogen-activated protein kinase (MAPK) activation in cardiomyocytes remains controversial. Therefore, the current study was designed to investigate the actions of AT(1)- and AT(2)-receptors on Ang II-induced extracellular signal-regulated kinase (ERK), p38 and the c-Jun N-terminal kinase (JNK) MAPK activities in human cardiomyocytes. Human cardiac tissue was obtained from open-heart surgery (n=6). The cardiac tissue was minced and incubated in the special tissue culture system for 24 hours in the absence or presence of Ang II (10(-7) M). These studies were repeated with the AT(1)-receptor antagonist losartan (10(-6) M) or the AT(2)-receptor antagonist PD-123319 (10(-6) M). Immunohistochemical staining and Western blot analysis with phospho-antibodies were performed to determine ERK, JNK and p38 activities. Ang II increased ERK and p38 activities in human cardiomyocytes. The effects of Ang II were abolished by losartan and enhanced by PD-123319. Co-incubation with both losartan and PD-123319 resulted in a decrease of ERK and p38 activities in cardiomyocytes. The immunohistochemical staining of JNK showed no significant differences between Ang II alone, Ang II plus losartan and Ang II plus PD-123319 groups. In conclusion, Ang II has a potent effect on ERK and p38 MAPK activities in cardiomyocytes, by acting through AT(1)-receptors. This effect of Ang II is modified by AT(2)-receptors. Therefore, Ang II, via AT(1)- and AT(2)-receptor stimulation, has a distinct effect on MAPK activity in cardiomyocytes.

Traumatic aortic rupture: recent outcome with regard to neurologic deficit.

BACKGROUND: Traumatic aortic rupture is highly lethal, and its surgical treatment is complicated by a high rate of paraplegia. METHODS: The charts of 263 patients with traumatic aortic rupture from vehicular accidents treated between 1971 and 1998 were reviewed. Patients were grouped according to four periods: group 1, 1971 to 1975, (n = 31); group 2, 1976 to 1985, (n = 83); group 3, 1986 to 1994, (n = 82); and group 4, 1994 to 1998 (n = 67). Seventy-one patients died of exsanguination before definitive care. One hundred-ninety two patients had surgical repair with the following techniques: clamp and sew, 6 in group 1, 22 in group 2, 54 in group 3, none in group 4; shunt, 23 in group 1, 39 in group 2, 2 in group 3; cardiopulmonary bypass, 2 in group 1, 1 in group 3. Forty-three patients had partial bypass with the centrifugal pump and heparin-coated circuits in group 4. RESULTS: Operative mortality was 6 of 31 (19%) in group 1, 22 of 61 (36%) in group 2, 15 of 57 (26%) in group 3, and 7 of 43 (16%) in group 4. There was one case of paraplegia in group 1 (4%), ten in group 2 (18%), 11 in group 3 (26%), and none in group 4. This difference of paraplegia between the groups was significant (p<0.002). Significant factors for paraplegia were intraoperative hypotension (p<0.000002), cross-clamp time longer than 30 minutes (p<0.008), pump versus no pump (p<0.008), and younger age group (28+/-11 versus 39+/-17 years) (p<0.03). CONCLUSIONS: There were no statistically significant improvements in mortality rate over the four periods, although, the mortality rate was lowest in the last period when partial bypass with the centrifugal pump was used exclusively. Further, the use of the centrifugal pump with heparin-coated circuits, with femoral vein cannulation into the right atrium and distal aortic perfusion, reduced paraplegia significantly.

Alternative surgical management of post-infarction septal rupture: a case report.

Weaning a patient from cardiopulmonary bypass after repair of a postinfarction ventricular septal defect in the face of severe right ventricular failure is likely to be fatal. The use of a ventricular assist device may seem to be the only available option. We present an alternative surgical approach that we carried out in a 72-year-old woman. The right ventricular preload was decreased by adding a bidirectional cavopulmonary anastomosis to the septal rupture repair.

Antimitotic effects of usnic acid on different biological systems.

Usnic acid is a biosynthesis product characteristic of several epiphytic lichens such as Evernia, Cladonia and Parmelia. Usnic acid has several interesting biological properties. It is an antibiotic and it also seems to exert an antimitotic action. It has even been postulated that usnic acid can play a role as an environmental indicator, since its concentration varies according to the presence of toxic agents. A series of tests have been run on different biological systems such as fungi, yeasts, plant cells and neoplastic human cell cultures in order to make a general evaluation of the properties of usnic acid and to highlight any analogy between its effects on phylogenetically distant organisms. The results obtained confirm some of the already known properties of usnic acid and identify concentration ranges that are active against cells from different organisms. Furthermore, at low concentrations, the acid displays a capacity to stimulate cell metabolism in some of the biological systems tested.

Oligogalacturonides Prevent Rhizogenesis in rolB-Transformed Tobacco Explants by Inhibiting Auxin-Induced Expression of the rolB Gene.

Oligogalacturonides elicit several defense responses and regulate different aspects of growth and development in plants. Many of the development-related effects of oligogalacturonides appear to be amenable to an auxin antagonist activity of these oligosaccharins. To clarify the role of oligogalacturonides in antagonizing auxin, we analyzed their effect on root formation in leaf explants of tobacco harboring the plant oncogene rolB. We show here that oligogalacturonides are capable of inhibiting root morphogenesis driven by rolB in transgenic leaf explants when this process requires exogenous auxin. Because rolB expression is induced by auxin and dramatically alters the response to this hormone in transformed plant cells, the inhibiting effect of oligogalacturonides could be exerted on the induction of rolB and/or at some other auxin-requiring step(s) in rhizogenesis. We show that oligogalacturonides antagonize auxin primarily because they strongly inhibit auxin-regulated transcriptional activation of a rolB-[beta]-glucuronidase gene fusion in both leaf explants and cultured leaf protoplasts. In contrast, oligogalacturonides do not inhibit rhizogenesis when rolB transcriptional activation is made independent of auxin, as shown by the lack of inhibition of root formation in leaf explants containing rolB driven by a tetracycline-inducible promoter.

Efficacy of leukocyte filters in the bypass circuit for infants undergoing cardiac operations.

BACKGROUND: Leukocyte filters have been used in cardiopulmonary bypass circuits to decrease potentially deleterious consequences of white blood cell product release in adults undergoing cardiac operations. Studies in animal models have also demonstrated benefits of white blood cell depletion during cardiopulmonary bypass. We sought to examine the efficacy of a leukocyte filter placed in the bypass circuit of infants undergoing operation for correction of congenital heart disease. METHODS: We conducted a retrospective study of infants undergoing correction of congenital heart defects (all had left-to-right shunts) between July 1993 and May 1994. Six infants (group I) had a Leukoguard-3 leukocyte filter (Pall Biomedical Products) placed in the arterial limb of the bypass circuit, and 6 infants (group II) had no filter. We analyzed for differences between groups in intraoperative variables (total pump time, aortic cross-clamp time, deep hypothermic cardiac arrest time, and time for rewarming), postoperative clinical variables (fever in the first 24 hours, days of mechanical ventilation, length of intensive care unit stay, and total hospital stay) and laboratory variables (change in white blood cell and platelet counts preoperatively to postoperatively, and arterial oxygen tension/inspired oxygen fraction ratio). RESULTS: Comparisons between groups were made using Student's t test or chi 2 where appropriate, and results were considered significant at p less than 0.05. No differences between groups were seen in the demographic data or in any of the intraoperative variables. Fever was the only parameter (clinical or laboratory) showing statistical significance between the groups, affecting 1/6 patients in group I and 5/6 patients in group II (chi 2 = 5.33; p < 0.05). CONCLUSIONS: Although these data suggest that the leukocyte filter offers little clinical benefit, a prospective, randomized study should be undertaken to further elucidate the efficacy of leukocyte depletion for infants undergoing cardiopulmonary bypass for correction of congenital heart disease.

rol genes of Agrobacterium rhizogenes cucumopine strain: sequence, effects and pattern of expression.

By sequencing the central region of the cucumopine-type T-DNA of Agrobacterium rhizogenes strain 2659, we identified three open reading frames homologous, to different extents, to ORFs 10, 11 and 12 (rolA, B and C) of the agropine-type (1855) T-DNA. Recombinant Agrobacterium strains encompassing the ORFs of 2659 T-DNA--which we refer to as rol alpha, beta and gamma--were utilized to infect carrot discs and to obtain transgenic tobacco plants, in order to compare the morphogenetic capabilities to those of the 1855 rol genes. Moreover, a long segment of the 5' non-coding region of rol alpha and rol beta was fused to the GUS reporter gene and the pattern of expression and the responsiveness to auxin of the constructs was analysed in transgenic tobacco. Differences in the auxin requirement for root induction between the 2659 rol genes and their respective 1855 counterparts were pinpointed. These differences are not due to gene regulation and presumably reflect functional differences in the proteins encoded. Differences were also observed in the pattern of expression of rol beta in roots of transgenic plants, as compared to rolB. In addition, the pattern of expression of rol alpha-GUS construct in roots was found to be analogous to that observed for a construct driven by two of the five regulatory domains of the rolB promoter.

Expression in different populations of cells of the root meristem is controlled by different domains of the rolB promoter.

Selective gene expression in different populations of cells of the root apex of transgenic tobacco could be evidenced by means of GUS constructs with deletions of the rolB promoter and fusions with the CaMV 35S minimal promoter. Five regulatory regions have been broadly identified in the rolB 5' non-coding region. The presence of all five domains (A to E) directs gene expression in the root cap, in the protoderm and in the different tissues within the root meristematic region: the dermatocalyptrogen, the cortex and the vascular cylinder. Deletion of domain A (-623 to -471) selectively suppresses expression in non-meristematic cells, i.e. the root cap and the protoderm. Deletion of either domain B (-341 to -306) or E (80 bp around the TATA box) causes loss of expression in all cells of the root apex: constructs C + D + E, B + C + D, B + C are inactive. Domain D (70 bp around the CAAT box) is necessary for gene expression in the dermatogen and in meristematic cells of the cortex but not in the innermost meristematic layer: construct B + C + E is active only in vascular meristematic cells. Domain C (-216 to -158) seems to have a double regulatory role as construct B + E is no longer expressed in meristematic cells of the vascular cylinder but is very active in the protoderm. Constructs allowing gene expression in meristematic cells are also inducible by auxin in leaf protoplasts, while activation of the regulatory elements necessary for gene expression in the non-meristematic cells of the root apex do not seem to depend upon the hormone. The connection between auxin induction and meristematic expression is discussed.

Bacterial plant oncogenes: the rol genes' saga.

The rol genes are part of the T-DNA which is transferred by Agrobacterium rhizogenes in plant cells, causing neoplastic growth and differentiation. Each of these bacterial oncogenes deeply influences plant development and is finely regulated once transferred into the plant host. Both from the study of the effects and biochemical function of the rol genes and from the analysis of their regulation, important insight in plant development can be derived. Some of the most intriguing aspects of past, current and future research on this gene system are highlighted and discussed.