Single shot phase contrast imaging using laser-produced Betatron x-ray beams.

Development of x-ray phase contrast imaging applications with a laboratory scale source have been limited by the long exposure time needed to obtain one image. We demonstrate, using the Betatron x-ray radiation produced when electrons are accelerated and wiggled in the laser-wakefield cavity, that a high-quality phase contrast image of a complex object (here, a bee), located in air, can be obtained with a single laser shot. The Betatron x-ray source used in this proof of principle experiment has a source diameter of 1.7 µm and produces a synchrotron spectrum with critical energy E(c)=12.3±2.5 keV and 109 photons per shot in the whole spectrum.

A fast and efficient segmentation scheme for cell microscopic image.

Microscopic cellular image segmentation schemes must be efficient for reliable analysis and fast to process huge quantity of images. Recent studies have focused on improving segmentation quality. Several segmentation schemes have good quality but processing time is too expensive to deal with a great number of images per day. For segmentation schemes based on pixel classification, the classifier design is crucial since it is the one which requires most of the processing time necessary to segment an image. The main contribution of this work is focused on how to reduce the complexity of decision functions produced by support vector machines (SVM) while preserving recognition rate. Vector quantization is used in order to reduce the inherent redundancy present in huge pixel databases (i.e. images with expert pixel segmentation). Hybrid color space design is also used in order to improve data set size reduction rate and recognition rate. A new decision function quality criterion is defined to select good trade-off between recognition rate and processing time of pixel decision function. The first results of this study show that fast and efficient pixel classification with SVM is possible. Moreover posterior class pixel probability estimation is easy to compute with Platt method. Then a new segmentation scheme using probabilistic pixel classification has been developed. This one has several free parameters and an automatic selection must dealt with, but criteria for evaluate segmentation quality are not well adapted for cell segmentation, especially when comparison with expert pixel segmentation must be achieved. Another important contribution in this paper is the definition of a new quality criterion for evaluation of cell segmentation. The results presented here show that the selection of free parameters of the segmentation scheme by optimisation of the new quality cell segmentation criterion produces efficient cell segmentation.

Endothelin-1 in monolayer cultures of articular chondrocytes from young and old rats: regulation by growth factors and cytokines.

The endothelin-1 (ET-1) concentrations were measured by RIA in the media of confluent monolayer cultures of rat articular chondrocyte (RAC) exposed to fetal calf serum (FCS) and several growth factors and cytokines. The cells were obtained from 1- and 18-month-old rats. First passage cells were starved in Dulbecco's modified Eagle's medium (DMEM) containing 0.2% FCS serum for 24 h and then incubated for 48 h in the same fresh medium with each of the following factors: fetal calf serum (FCS), transforming growth factor-beta (TGF-beta), platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin-like growth factor-1 (IGF-1), interleukin-1 beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), lipopolysaccharide (LPS), and NO donor, sodium nitroprusside (SNP). The following was found: the cells from 18-month-old animals accumulated about twice as much ET-1 per microg DNA under basal (low serum) and stimulated conditions as cells from young rats. All, but PDGF and SNP produced concentration-dependent rise in ET-1 levels, the most effective being 10% FCS, IL-1beta, TNF-alpha, EGF, IGF-1 and LPS. TGF-beta caused the smallest stimulation and PDGF was ineffective or slightly inhibitory at high concentrations. SNP caused concentration-dependent decrease of ET-1 concentrations. ET-1-specific mRNA was identified by RT-PCR in cells incubated with the above factors and its concentration paralleled that of the peptide. This suggests that ET-1 found in the culture media of RAC stems, at least in part, from the synthesis. Increased immunoreactive peptide concentration and mRNA expression with the age of the donor rat and its regulation by several growth factors and cytokines suggest the involvement of ET-1 in chondrocytes' physiology and possibly pathology.

Increased endothelin-1 associated with bacterial infection in lung transplant recipients.

BACKGROUND: Endothelin-1 (ET-1) has fibrogenic and inflammatory properties. Its pathogenic role in pulmonary fibrosis and certain inflammatory airway diseases is now well known. Its production is, in part, triggered by infectious processes. Episodes of infection are suspected to be involved in the development of bronchiolitis obliterans syndrome (BOS), which is the main feature of chronic lung rejection and the major factor limiting the long-term survival of transplanted patients. We postulated that ET-1 is upregulated during infectious complications arising from the graft and that this could partly explain the remodeling of airway structures observed in BOS. We, therefore, set up this study to assess ET-1 expression in relation to complications of the graft in human lung transplant recipients. METHODS: ET-1 mRNA was quantified by reverse transcription-competitive polymerase chain reaction in cells from 119 samples of bronchoalveolar lavage (BAL) fluid from 17 lung transplant recipients. ET-1 and big ET-1 proteins were assessed in BAL cell culture supernatants by enzyme immunoassay. Transbronchial biopsies (n=21) were stained immunohistochemically for ET-1 receptors. RESULTS: Episodes of bacterial infection strongly correlated with increased ET-1 mRNA and protein expression. ET-1 receptors were also upregulated during these episodes, especially on endothelial and smooth muscle cells. Five of the seven patients with the highest ET-1 levels subsequently developed BOS. CONCLUSIONS: These results raise the possibility that ET-1, part of whose production is triggered by infectious postgraft complications, might play a role in the development of BOS through its potential effects on airway remodeling.

Indium-111-white blood cell scintigraphy in Crohn's patients with fistulae and sinus tracts.

METHOD: Indium-111-white blood cell (111In-WBC) images of 17 Crohn's patients with fistulae and sinus tracts were reviewed and correlated with radiographic results (n = 16 patients) and surgery (n = 16 patients), to characterize the scintigraphic appearance of fistulization and to determine the role of 111In-WBC scintigraphy in this clinical setting. These were compared to 50 consecutive abnormal 111In-WBC studies obtained in Crohn's patients with suspected active disease but no known fistulae or sinus tracts. RESULTS: Scintigraphic findings which suggested the presence of fistulae were: (1) the presence of concomitant intestinal and extraintestinal lesions and (2) the absence of luminal activity on delayed images when early images detected bowel activity. The extraintestinal lesions were the drainage site of the fistula (n = 7) or an accompanying abscess (n = 6). Absence of luminal activity occurred in seven patients with fistulae and in two without fistulae; two patients had a proximal colostomy, two patients had bowel obstruction and five patients had fistulae to the skin (n = 3) or between the ileum and distal colon (n = 2). The distribution of active bowel disease as assessed scintigraphically was in complete agreement with surgery in 14 of 17 cases (82%) compared to 9 of 15 cases (60%) when correlating radiographic assessment with surgery. All surgically proven abscesses were detected on 111In-WBC images. CONCLUSION: These results indicate that 111In-WBC scintigraphy adds useful information to radiographic studies that is essential for appropriate management of Crohn's patients with fistulae and sinus tracts.

[Memory alterations in non-korsakoff's chronic alcoholics (author's transl)]. / Les altérations de la mémoire chez les alcooliques chroniques non korsakowiens.

There is a memory deficit in non Korsakow chronic alcoholics, mainly in fixation and learning. Retaining less structured and significant elements is particularly altered. Part of the troubles may be due to deterioration of recall mechanisms.

Determination of 1,3-dichloropropanol in soy sauces by automated headspace gas chromatography-mass spectrometry.

There has been recent concern about the levels of carcinogenic chloropropanols in some soy sauces. We have devised and tested a new automated headspace GC-MS technique for the analysis of 1,3-dichloropropan-2-ol (1,3-DCP) in soy sauce and similar products. The method incorporates the use of cryogenic trapping and a deuterium-labelled internal standard. The limit of detection was 0.003 mg kg(-1). After in-house validation testing, the method was applied to soy sauce samples that had previously been analysed for the related contaminant 3-monochloropropanediol (3-MCPD). 1,3-DCP was detected in 10 of 40 sauces, all of which also contained 3-MCPD. The highest level was just >1 mg kg(-1). There was no correlation between the levels of 1,3-DCP and 3-MCPD.

Factors influencing the effectiveness of extracorporeal shock-wave lithotripsy of biliary duct calculi.

The authors examined the effectiveness of extracorporeal shock-wave lithotripsy on patients who had bile-duct stones. Thirty-eight patients were treated with an unmodified HM-3 Dornier lithotripter; 35 (92%) patients either passed the stone fragments spontaneously or had them reduced to a size which allowed removal by percutaneous or endoscopic techniques. As stone size increased so did the number of shocks required for satisfactory fragmentation, the number of treatments and the rate of post-treatment manipulation. The number of stones was not as important a variable as the authors expected. Pre-treatment sphincterotomy was not always necessary and was of no benefit in terms of spontaneous passage of fragments. Complications were minor. Extracorporeal shock-wave lithotripsy of stones throughout the biliary tree is a safe, effective and invaluable adjunct in the management of bile-duct stones.

Pneumatosis coli: is there a relationship with sigmoid colon redundancy?

Pneumatosis coli (PC) is a process characterized by gas-filled cysts in the wall of the large intestine. The barium enema examinations of 14 patients with idiopathic PC were assessed: 93% involved the sigmoid colon, and 84% of patients with sigmoid pneumatosis had sigmoid colon redundancy in comparison to 14% of the control population (p less than 0.001). It is proposed that the sigmoid cysts result in redundancy by affecting the mesentery and colon length. This redundancy may account for the increased risk of sigmoid volvulus in this condition.

Extracorporeal shock wave lithotripsy (ESWL) in the management of complex biliary tract stone disease.

The use of extracorporeal shock wave lithotripsy (ESWL) in the management of ten patients with complex biliary tract stones is described. General or epidural anesthesia was used in all cases, and stone fragmentation was performed, using an unmodified Dornier HM3 waterbath lithotripter (Dornier Medical Systems Inc., Marietta, GA). In all cases, biliary drainage was established before the procedure to allow contrast visualization during and after the procedure, as well as to ensure free drainage of the common bile duct. Indications for ESWL included failure of basket extraction (4 cases), unfavorable anatomy (duodenal diverticulum, previous Billroth II reconstruction, hepatic duct stone, gallbladder stone, cystic duct remnant stone), and immaturity of the T-tube tract (2 cases). Cholangitis was the presenting diagnosis in four cases. Fragmentation of the stones was successful in all patients; in two cases, two ESWL sessions were needed for stone disruption. Morbidity was minimal (there was a minor elevation of LDH and transaminases and asymptomatic hemobilia and hematuria); pancreatitis did not occur. After ESWL, hospital stays ranged from 1 to 13 days (mean of 5.3 days). ESWL can be a valuable adjunct in the management of patients with complex biliary stones.

IL-1 beta inhibits ET-1 production by ATII cells in vitro: evidence for involvement of cyclooxygenase 2 pathway.

The aim of this study was to characterize the effect of alveolar macrophage (AM) secretory products on endothelin (ET)-1 production by rat alveolar type II (ATII) cells in primary culture. We quantified preproendothelin (ppET)-1 mRNA by Northern blot and ET-1 concentrations in cell supernatants by enzyme-linked immunosorbent assay. Conditioned medium (CM) from rat adherent AM decreased the ppET-1 mRNA levels in ATII cells and reduced ET-1 concentrations in cell culture supernatants. This effect was mediated by interleukin (IL)-1 beta as shown by pretreatment of CM with an anti-IL-1 beta neutralizing antiserum. IL-1 beta effect was dependent on protein synthesis, was partially prevented with indomethacin, and was totally prevented with dexamethasone. Specific inhibition of cyclooxygenase 2 activity completely reversed the effect of IL-1 beta. We conclude that rat AM inhibit ET-1 production by rat ATII cells in vitro through IL-1 beta secretion. The IL-1 beta-mediated inhibition is dependent on the cyclooxygenase 2 pathway. Downregulation of ET-1 production by activated AM could limit the intra-alveolar burden of this profibrogenic peptide and thus could prevent fibrosis development.

Extracorporeal shock-wave lithotripsy in the management of bile duct stones.

Extracorporeal shock-wave lithotripsy (ESWL) was performed in 16 patients with bile duct stones. Dornier HM3 water-bath equipment (Dornier Medical Systems, Marietta, GA) was used in all procedures. All patients had either a T-tube or a nasobiliary, cholecystostomy, or transhepatic biliary drainage tube in place at the time of the lithotripsy. In 12 patients, the indication for ESWL was the failure of or anticipated difficulty with basket extraction of the stones, either via a T-tube tract or by means of endoscopic sphincterotomy. In the other four patients, ESWL was performed immediately after retained stones were found on a postoperative cholangiogram, even though no difficulty was anticipated in removing the stones by means of basket extraction. In 15 (94%) of the 16 patients, the stones were successfully fragmented. The fragments passed spontaneously in nine patients but had to be removed by basket in five patients. In one patient, the fragments could not be extracted by basket. The number of ESWL shocks used in a single session ranged from 525 to 3200. Three patients had two ESWL sessions. No significant complications were observed. ESWL is a successful method for the management of patients with bile duct stones when used in conjunction with other nonsurgical techniques. It was also the only treatment required in 56% of our patients.

Endothelin-1 secretion by alveolar macrophages in systemic sclerosis.

Endothelin-1 (ET-1), a potent fibroblast/smooth muscle cells mitogen, has been implicated in the pathogenesis of systemic sclerosis lung disease (SSc). Since monocytes and macrophages are thought to be activated in SSc, we hypothesized that alveolar macrophages (AM) and their precursors blood monocytes from patients with SSc produced more ET-1 than cells from healthy subjects. ET-1 and big ET-1 concentrations were measured in plasma, in bronchoalveolar lavage (BAL) fluids and in cell culture supernatants from monocytes and alveolar macrophages derived from 13 patients with definite SSc with lung involvement and from 10 control subjects. Plasma and BAL fluid ET-1 and big ET-1 levels were similar in both controls and patients with SSc. ET-1 and big ET-1 concentrations in unstimulated alveolar macrophage supernatants were similar in both groups. In contrast, LPS-stimulated alveolar macrophages from patients with SSc secreted higher amounts of ET-1 and big ET-1 than control subjects. ET-1 and big ET-1 concentrations in monocyte supernatants (either LPS-stimulated or not) were not different in patients and controls. These results show that AM from patients with SSc are hyperresponsive to LPS in vitro in terms of ET-1 and big ET-1 production and suggest that AM could participate in vivo in the overproduction of this potentially profibrotic mediator in patients with SSc.

Synergism of interleukin-12 and interleukin-3 on development of hematopoietic progenitors.

The recently cloned cytotoxic lymphocyte maturation factor [CLMF] also called NK cell stimulatory factor [NKSF] or interleukin-12 [IL-12] has been described as a growth factor for mature lymphoid cells. The present study investigated whether purified recombinant human IL-12 could stimulate CFU colony growth. Source of progenitor cells were peripheral blood cells depleted of adherent, CD2- and CD56-positive cells. RhIL-12 was investigated either alone or in combination with rhIL-3, rhIL-6 and rhGM-CSF. RhIL-12 alone did not support colony formation of myeloid or erythroid progenitors. RhIL-12 in combination with rhIL-3 increased the numbers of BFU-E and CFU-GM. No synergism or additive effect was seen with the combination of rhIL-12 and rhGM-CSF or rhIL-12 and rhIL-6. An additive increase in the number of granulocytic colonies was observed when rhIL-3, rhIL-6 and rhGM-CSF were used together with rhIL-12. Our results therefore suggest that, in addition to being a potent lymphopoietic stimulator, IL-12 acts synergistically with IL-3 in enhancing the sensitivity of hemopoietic progenitors to IL-3.

Sensitivity of myeloid progenitors to NK-cell depletion.

To gain additional informations on the role played by Natural Killer cells (NK) in the differentiation of human hematopoietic precursors, we have studied the effect of NK-cell depletion on the in vitro proliferation of hematopoietic cells. NK cells were depleted from blood mononuclear cells by FACS using anti CD3 and anti CD56 monoclonal antibodies. Depletion of NK cells suppressed CFU-GM up to 69% (P < 0.01), while no significant effect on either BFU-E and CFU-Mix growth was observed. To define the threshold of CD56+ cells required to support CFU-GM formation, NK cells were added to NK-depleted cells in a titrated fashion. Enhancement of CFU-GM colony growth was observed at NK/NK- depleted cells ratio of 0.15/1. A dose dependent suppression of CFU growth was observed at ratios ranging from 0.25/1 to 0.5/1. Addition of neutralizing antibodies against IL3 and GM-CSF abrogate the stimulating effect of NK cells. Our results suggest that cells with LGL morphology and NK markers play an important role in differentiation of myeloid precursors and exert a moderate influence on erythroid progenitors. The modulatory effect on hematopoietic progenitors depend on the number of NK cells present in the mixed culture.