bluuebunny/crossref_metadata_2025 huggingface dataset:
Innate immune response to encephalomyocarditis virus infection mediated by CD1d
Authors: Exley Mark A. , Bigley Nancy J. , Cheng Olivia , Shaulov Angela , Tahir Syed Muhammad Ali , Carter Quincy L. , Garcia Jorge , Wang Carren , Patten Kurt , Stills Harold F. , Alt Frederick W. , Snapper Scott B. , Balk Steven P.
Published: November 2003

CD1d‐reactive natural killer T (NKT) cells can rapidly produce T helper type 1 (Th1) and/or Th2 cytokines, can activate antigen‐presenting cell (APC) interleukin‐12 (IL‐12) production, and are implicated in the regulation of adaptive immune responses. The role of the CD1d system was assessed during infection with encephalomyocarditis virus (EMCV‐D), a picornavirus that causes acute diabetes, paralysis and myocarditis. EMCV‐D resistance depends on IL‐12‐mediated interferon‐γ (IFN‐γ) production. CD1d‐deficient mice, which also lack CD1d‐reactive NKT cells, were substantially more sensitive to infection with EMCV‐D. Infected CD1d knockout mice had decreased IL‐12 levels in vitro and in vivo, and indeed were protected by treatment with exogenous IL‐12. IFN‐γ production in CD1d knockout mice was decreased compared with that in wild‐type (WT) mice in response to EMCV‐D in vitro, although differences were not detected in vivo. Treatment with anti‐asialo‐GM1 antibody, to deplete NK cells, caused a marked increase in susceptibility of WT mice to EMCV‐D infection, whereas CD1d knockout mice were little affected, suggesting that NK‐cell‐mediated protection is CD1d‐dependent. Therefore, these data indicate that CD1d is essential for optimal responses to acute picornaviral infection. We propose that CD1d‐reactive T cells respond to early immune signals and function in the innate immune response to a physiological viral infection by rapidly augmenting APC IL‐12 production and activating NK cells.
Insights into the regulation of transcription by scanning force microscopy
Authors: Dame R. T. , Wyman C. , Goosen N.
Published: November 2003

The scanning force microscope (SFM) is a valuable tool for the structural analysis of complexes between protein(s) and DNA. In recent years the application of scanning force microscopy to the field of transcription regulation has been reported in numerous studies. Using this technique, novel insights could be obtained into the architecture and dynamics of complexes, which are relevant to the transcription process and the mechanisms by which this process is regulated. In this article an overview is given of SFM studies addressing, in particular, topics in the field of transcription in prokaryotic organisms.
Encapsulated yeast cells inside Paramecium primaurelia: a model system for protection capability of polyelectrolyte shells
Authors: Krol S. , Cavalleri O. , Ramoino P. , Gliozzi A. , Diaspro A.
Published: November 2003

One of the most promising applications of encapsulated living cells is their use as protected transplanted tissue into the human body. A suitable system for the protection of living cells is the use of nano‐ or microcapsules of polyelectrolytes. These shells can be deposited easily on top of the cells by means of a layer‐by‐layer technique. An interesting feature of the capsules is the possibility to control their properties on a nanometre level, tuning their wall texture via the preparation conditions. Here we introduce a model system to test the protection ability of polyelectrolyte capsules. Common bakery yeast cells were encapsulated. They were coated with a fluorescently labelled shell at conditions known to guarantee cell survival, and the cell interior was stained with DAPI. The protozoan Paramecium primaurelia was incubated with this double‐stained living yeast and visualized by means of two‐photon excitation fluorescence microscopy. Cross‐sections of the dye‐stained material as well as autofluorescence of the fixed protozoan allowed us to follow the digestion of the coated yeast with time. Our investigation reveals that capsules prepared under these deposition conditions are permeable to lysosomal enzymes, leading to degradation of the yeast inside the intact capsules. Our preliminary results indicate the suitability of the introduced model as a test system of this permeability.
Polysaccharide properties probed with atomic force microscopy
Authors: Abu‐Lail N. I. , Camesano T. A.
Published: November 2003

In recent years, polysaccharides have been extensively studied using atomic force microscopy (AFM). Owing to its high lateral and vertical resolutions and ability to measure interaction forces in liquids at pico‐ or nano‐Newton level, the AFM is an excellent tool for characterizing biopolymers. The first imaging studies showed the morphology of polysaccharides, but gradually more quantitative image analysis techniques were developed as the AFM grew easier to use in aqueous liquids and in non‐contact modes. Recently, AFM has been used to stretch polysaccharides and characterize their physicochemical properties by application of appropriate polymer stretching models, using a technique called single‐molecule force spectroscopy. From application of such models as the wormlike chain, freely jointed chain, extensible‐freely jointed chain, etc., properties such as the contour length, persistence length and segment elasticity or spring constant can be calculated for polysaccharides. The adhesion between polysaccharides and surfaces has been quantified with AFM, and this application is particularly useful for studying polysaccharides on microbial and other types of cells, because their adhesion is controlled by biopolymer characteristics. This review presents a synthesis of the theory and techniques currently in use to probe the physicochemical properties of polysaccharides with AFM.
The nacre protein perlucin nucleates growth of calcium carbonate crystals
Authors: Blank S. , Arnoldi M. , Khoshnavaz S. , Treccani L. , Kuntz M. , Mann K. , Grathwohl G. , Fritz M.
Published: November 2003

Atomic force microscopy (AFM) in aqueous solution was used to investigate native nacre of the marine snail Haliotis laevigata on the microscopic scale and the interaction of purified nacre proteins with calcium carbonate crystals on the nanoscopic scale. These investigations were controlled by scanning electron microscopy (SEM), light microscopy (LM) and biochemical methods. For investigations with AFM and SEM, nacre was cleaved parallel to the aragonite tablets in this biogenic polymer/mineral composite. Multilamellar organic sheets consisting of a core of chitin with layers of proteins attached on both sides lay between the aragonite layers consisting of confluent aragonite tablets. Cleavage appeared to occur between the aragonite tablet layer and the protein layer. AFM images revealed a honeycomb‐like structure to the organic material with a diameter of the ‘honeycombs’ equalling that of the aragonite tablets. The walls of the structures consisted of filaments, which were suggested to be collagen. The flat regions of the honeycomb‐like structures exhibited a hole with a diameter of more than 100 nm. When incubated in saturated calcium carbonate solution, aragonite needles with perfect vertical orientation grew on the proteinacous surface. After treatment with proteinase K, no growth of orientated aragonite needles was detected. Direct AFM measurements on dissolving and growing calcite crystals revealed a surface structure with straight steps the number of which decreased with crystal growth. When the purified nacre protein perlucin was added to the growth solution (a super‐saturated calcium carbonate solution) new layers were nucleated and the number of steps increased. Anion exchange chromatography of the water‐soluble proteins revealed a mixture of about 10 different proteins. When this mixture was dialysed against saturated calcium carbonate solution and sodium chloride, calcium carbonate crystals precipitated together with perlucin leaving the other proteins in the supernatant. Thus perlucin was shown to be a protein able to nucleate calcium carbonate layers on calcite surfaces, and in the presence of sodium chloride, is incorporated as an intracrystalline protein into calcium carbonate crystals.
TectoRNA and ‘kissing‐loop’ RNA: atomic force microscopy of self‐assembling RNA structures
Authors: Hansma H. G. , Oroudjev E. , Baudrey S. , Jaeger L.
Published: November 2003

RNA molecules have been much less studied by atomic force microscopy (AFM) than have DNA molecules. In this paper, AFM imaging is presented for two different RNA molecules able to self‐assemble into complex supramolecular architectures. The first one is a molecular dimer of a 230‐nt RNA fragment coming from the RNA genome of a murine leukaemia virus. The monomeric RNA fragment, which appears by AFM as an elongated structure with a mean aspect ratio of 1.4, assembles into a dimer of elongated structures through the formation of a ‘kissing‐loop’ RNA interaction. The second one is a large supramolecular fibre formed of artificial self‐assembling RNA molecular units called tectoRNA. The fibre lengths by AFM suggest that there are 50–70 tectoRNA units per fibre. Some methods and limitations are presented for measuring molecular volumes from AFM images.
A preoperative clinical prognostic model for non‐metastatic renal cell carcinoma
Authors: Cindolo L. , De La Taille A. , Messina G. , Romis L. , Abbou C.C. , Altieri V. , Rodriguez A. , Patard J.J.
Published: November 2003

Authors from Naples, Paris and Rennes describe their efforts to develop a model for the preoperative prediction of outcome for non‐metastatic renal cancer. It is valuable to both urologist and patient to develop such a model, particularly so on preoperative criteria. The results of their study are interesting, leading to possibly helpful findings.

In another article, authors from Iceland estimated the risk of developing prostate and other cancer among relatives of men from that country diagnosed with prostate cancer. They found that a family history is a risk factor for prostate cancer, with the risk potentially higher for relatives of patients who died from the disease.

From the relative dearth of papers on quality of life after radical prostatectomy there are now several, and the authors from Bristol report on the effects of erectile dysfunction on quality of life after this type of treatment. They had a very high response rate (91%) to their questionnaire, and found that erectile dysfunction has a profound effect on quality of life. This finding is not a surprise, but do we need to examine our management of prostate cancer in the light of it?

OBJECTIVE

To develop a model to predict the outcome before surgery for non‐metastatic renal cell carcinoma (RCC).

PATIENTS AND METHODS

The records of 660 patients with non‐metastatic RCC, operated at three European medical institutes, were reviewed. Univariate and multivariate analyses were used to assess the clinical and pathological variables affecting disease‐free survival.

RESULTS

The median (range) follow‐up was 42  (2–180) months; the disease recurred in 110 patients (16%). The 2‐ and 5‐year overall survival was 87% and 54%, respectively. Five variables were significant in the univariate analysis, i.e. clinical presentation, clinical and pathological size, tumour grade and stage (P < 0.05). The preoperative variables, e.g. clinical presentation and clinical tumour size, were retained from the multivariate model. A recurrence risk formula (RRF) was constructed from this model, as (1.28 × presentation (asymptomatic = 0; symptomatic = 1) + (0.13 × clinical size)). Using this equation, the 2‐ and 5‐year disease‐free survival was 96% and 93% for an RRF of ≤ 1.2 and 83% and 68% for an RRF of > 1.2.

CONCLUSION

A formula was developed which, independent of stage, can be used to predict the rate of treatment failure in patients who undergo nephrectomy for non‐metastatic RCC. The RRF might be useful for more accurate sub‐grouping of good‐prognosis patients, and for counselling patients before surgery, their personalized follow‐up or adjuvant treatment once available.
Urodynamic findings in men operated on for an undescended testicle
Authors: Lahdes‐Vasama T.T. , Koskimäki J.E. , Streng T.K. , Fisch R.D. , Nilson E.A. , Santti R.S. , Tammela T.L.J.
Published: November 2003

OBJECTIVE

To test the hypothesis that men with a history of undescended testicle have voiding problems similar to those in rodents exposed to excessive amounts of oestrogens during development, although the role of oestrogen in the failure of the human testicle to descend remains controversial.

PATIENTS AND METHODS

Thirteen men (mean age 45 years) previously operated on for an undescended testicle (testis‐retention, TR group) and 12 age‐matched men operated on for inguinal hernia or appendicitis (control group) participated in a urodynamic examination, transrectal ultrasonography (TRUS) of the prostate, and blood tests for hormones and prostate‐specific protein. They also completed a questionnaire on urinary symptoms.

RESULTS

The free maximum flow rate was significantly lower and the detrusor pressure at maximum flow (PdetQmax) slightly higher in the TR than the control group. Three men in the TR group (and none of the controls) had bladder outlet obstruction (BOO), whereas voiding was not obstructed among 11 control men (and five men in the TR group). The hormone concentrations of the groups did not differ significantly but the prostates were significantly smaller in the TR group. The testosterone concentrations and the ratio between 17β‐oestradiol (E2) and free testosterone (E2/fT) influenced prostate size significantly. An exploratory analysis suggested that E2/fT may influence the maximum detrusor pressure and PdetQmax.

CONCLUSION

Men born with an undescended testicle had smaller prostates but more often had BOO than did the controls. The results suggest that an imbalance between the actions of oestrogen and testosterone may influence the initiation and continuance of BOO among cryptorchid men.
Tubularized incised plate repair of distal hypospadias in toilet‐trained children: should a stent be left?
Authors: El‐Sherbiny M.T.
Published: November 2003

OBJECTIVE

To evaluate the role of stenting in toilet‐trained children undergoing tubularized incised plate (TIP) repair of distal hypospadias, as a stent‐free TIP in boys who are not toilet‐trained has been successful with no unusually prolonged discomfort.

PATIENTS AND METHODS

The study included all toilet‐trained children with distal penile shaft hypospadias who were not suitable candidates for meatal advancement procedures and who underwent TIP repair, by one surgeon, between March and August 2001. Patients were prospectively randomized at the end of surgery to either leaving a stent for 1 week (stented) or no stent (unstented). The study included 64 children (35 stented and 29 unstented; median age 6 years, range 2–17). In the stented group the stent was placed in the bladder for continuous bladder drainage. An adjunct penile block instead of caudal or epidural analgesia was used in all patients, to avoid drug‐induced urinary retention. The early evaluation included an assessment of bladder spasms, dysuria, urinary retention and extravasation. Regular meatal dilatation was provided only to patients with voiding difficulty and an obvious tendency to stenosis.

RESULTS

The median (range) follow‐up was 6 (3–11) months. Voiding was painful in the first week in five (14%) and 13 (45%) of the stented and unstented patients, respectively (P < 0.01); there were bladder spasms in three (8%) and none, respectively (P > 0.05). None of stented patients developed urinary retention or extravasation, compared with seven (24%) and five (17%) in the unstented group, respectively (P < 0.05). Meatal dilatation was required in two (6%) and five (17%) of the stented and unstented patients, respectively (P > 0.05). There were complications requiring re‐operation in nine boys (14%), of whom seven (10%) developed small fistulae and two (4%) had meatal stenosis. Although the re‐operation rate was lower in the stented (9%) than in the unstented group (20%), the difference was not statistically significant (P > 0.05).

CONCLUSIONS

The use of a stent in TIP repair in toilet‐trained children is advantageous; it significantly eliminates the risk of urinary retention and extravasation, and reduces the overall patient discomfort. It is also associated with a lower re‐operation rate.
Prevalence, diagnosis and treatment of prostatitis in Italy: a prospective urology outpatient practice study
Authors: Rizzo M. , Marchetti F. , Travaglini F. , Trinchieri A. , Nickel J.C.
Published: November 2003

OBJECTIVE

To report a prospective, multicentre descriptive study designed to determine the prevalence of the diagnosis of prostatitis in male outpatients examined by urologists in Italy, and to further examine the diagnostic evaluation and treatment of patients identified with a clinical diagnosis of prostatitis.

PATIENTS AND METHODS

Between July 2001 and October 2001, 70 urologists, representing a cross‐section of urological centres in Italy, counted and recorded the overall total of men reported in the clinic and that of patients diagnosed with prostatitis over a 5‐week period. Data on demographics, previous diagnoses, symptoms, physical examination, laboratory data and therapy instituted were collected. Patients with a diagnosis of prostatitis completed questionnaires on symptom frequency and severity, and quality of life.

RESULTS

In all, 8503 patients were included in the primary outcome analysis; 1148 were identified with prostatitis (12.8%; mean age 47.1 years, range 16–83) with all age ranges equally represented, and 68% had had their first symptom within the last year. The most common presenting symptoms were severe, bothersome urinary frequency, obstructive voiding symptoms, perineal, suprapubic and penile pain or discomfort. The self‐administered questionnaire confirmed that the most frequently reported and most severe symptoms at the time of evaluation were irritative voiding symptoms, perineal and suprapubic pain and discomfort. Over three‐quarters of the patients were dissatisfied with their quality of life. While 98% of the patients had a digital rectal examination and expressed prostatic secretion was successfully recovered in 44%, < 3% of the patients had the traditional Meares‐Stamey four‐glass test. The most common treatment prescribed was drug therapy (not antibiotic).

CONCLUSION

The prevalence of a clinical diagnosis of prostatitis in urology outpatient practice in Italy was 12.8%. The prevalence, diagnosis, evaluation and treatment of prostatitis reported in this prospective study was very similar to that reported in other retrospective series from other countries.
The prevalence of male urinary incontinence in four centres: the UREPIK study
Authors: Boyle P. , Robertson C. , Mazzetta C. , Keech M. , Hobbs F.D.R. , Fourcade R. , Kiemeney L. , Lee C. ,   the UrEpik Study Group
Published: November 2003

OBJECTIVES

To report the results of the prevalence and health status associated with male urinary incontinence (UI) in a population‐based, multicentre study in four countries, and to assess the epidemiology of this condition.

SUBJECTS AND METHODS

A standard questionnaire asking about frequency and amount of urine loss, use of pads and stress incontinence, was used to measure the prevalence of UI among men in four countries (Boxmeer, The Netherlands; Auxerre, France; Birmingham, UK; and Seoul, Korea). Self‐reported urinary leakage was also recorded, with details of visits to the doctor for the symptoms. The samples were selected randomly from population registers of men aged 40–79 years, which provided representative samples in each community.

RESULTS

In all, 4979 men responded; from incontinence symptom scores, 7.3% of men aged 40–79 years in Auxerre, 16.3% in Boxmeer, 14.4% in Birmingham and 4.3% in Seoul reported mild to severe UI. On a self‐reported question, 14.8% and 14.5% of men in Birmingham and Boxmeer thought that they had urinary leakage, compared with 7.5% and 7.1% of men in Auxerre and Seoul, but typically the reported incontinence was leaking drops of urine a few times a week. Of men with urinary leakage in the three European centres, 25.9% visited the doctor with this problem, compared with only 9.0% of men in Seoul. Of men in the European centres, 5.9% used pads at least occasionally, compared with only 1.6% of men in Seoul. UI was age‐related and the reporting of it varied among centres, with 14.4% of men in Birmingham and 12.7% of men in Boxmeer aged 40–49 years reporting mild to severe UI, compared with 5.2% in Auxerre and 1.9% in Seoul. Among men aged 60–69 years the corresponding percentages were 13.7% in Birmingham, 22.6% in Boxmeer, 9.2% in Auxerre and 8.0% in Seoul.

CONCLUSIONS

UI is more common in older men and relatively uncommon among younger men. Some men reported no problems on the symptom questionnaire but replied positively to a direct question. Surprisingly many men wear protective pads at least occasionally as a result of their problem. UI appears to be a problem for men and which remains largely untreated; this may partly be a result of cultural differences.
Comparison of multitarget fluorescence in situ hybridization in urine with other noninvasive tests for detecting bladder cancer
Authors: Friedrich M.G. , Toma M.I. , Hellstern A. , Pantel K. , Weisenberger D.J. , Noldus J. , Huland H.
Published: November 2003

OBJECTIVES

To present a single‐centre study investigating aneuploidy at chromosomes 3, 7, 17 and 9p21 (e.g. loss at 9p21) using a multitarget fluorescence in situ hybridization (FISH) system, as identifying genetic alterations in urine specimens is a promising approach for the noninvasive detection of bladder cancer.

PATIENTS AND METHODS

Urine samples from 103 patients were evaluated, including those from 46 with histologically confirmed urothelial carcinoma, two with other urological malignancies, and 55 who acted as controls. The urine samples were taken before any manipulation. The validity of FISH (Urovision, Vysis, Downers Grove, Ill, USA) was compared with other noninvasive urine tests, including the BTA‐Stat test, the nuclear matrix protein (NMP)‐22 test, and immunocytology against 486p3/12 and LewisX. Those evaluating the tests were unaware of the clinical and histopathological data. FISH was considered positive if five or more urinary cells had gains of two or more chromosomes. The threshold for the urine tests were 10 U/mL (NMP‐22), 30% positive cells (486p3/12), or 5% positive cells, respectively (LewisX).

RESULTS

The sensitivity was 69% (FISH), 67% (BTA‐Stat), 69% (486p3/12), 96% (LewisX) and 71% (NMP22), respectively; the respective specificity was 89%, 78%, 76%, 33% and 66%.

CONCLUSION

Multitarget FISH had a better specificity than the other urine markers but because of its inadequate sensitivity it does not seem to be powerful enough to replace endoscopy. Optimizing the marker panel could provide a higher sensitivity.
Eradication of Helicobacter pylori Infection Improves Blood Pressure Values in Patients Affected by Hypertension
Authors: Migneco Alessio , Ojetti Veronica , Specchia Lucia , Franceschi Francesco , Candelli Marcello , Mettimano Marco , Montebelli Rita , Savi Luigi , Gasbarrini Giovanni
Published: November 2003

Background.  Arterial hypertension is a risk factor for atherosclerosis of whose pathogenesis is unknown. Growing evidence underscores the causative role of endothelial dysfunction. A possible association between Helicobacter pylori infection and cardiovascular and autoimmune disorders has been found. The release of cytotoxic substances either of bacterial origin or produced by the host may represent mediators of these systemic sequelae. The aim of our study was to determine the prevalence of H. pylori infection in hypertensive patients and the effects of H. pylori eradication on blood pressure and on digestive symptoms.

Materials and Methods.  Seventy‐two hypertensive patients (34 male and 38 female; mean age 53 ± 12 years) and 70 normotensive controls (35 male and 35 female; mean age 52 ± 10 years) were enrolled. All patients were subjected to a first ambulatory blood pressure monitoring (ABPM) at enrollment, a 13C urea breath test and a test for IgG‐CagA antibodies, and completed the validated dyspepsia questionnaire. H. pylori‐positive patients were treated with triple therapy (amoxicillin, clarithromycin and ranitidine bismute citrate) for 7 days. Control of eradication was assessed by 13C urea breath test, and all patients underwent a second ABPM 6 months after enrollment.

Results.  H. pylori infection was 55% in hypertensive patients, with 90% CagA positivity, and 50% in controls, with 60% CagA positivity. At the first ABPM, blood pressure values were similar in H. pylori‐positive and ‐negative individuals; positive patients showed a significant increase in pyrosis and epigastric pain compared to negative patients. H. pylori was eradicated in 80% of patients and in 85% of controls. At the second ABPM, we found a statistically significant decrease in 24‐hour mean blood pressure values when compared to the first ABPM only in the eradicated hypertensive group.

Conclusions.  Our study demonstrated a significant decrease in blood pressure values, in particular in diastolic blood pressure values, after H. pylori eradication in hypertensive patients. A high prevalence of CagA positivity was found. The association between cardiovascular disease and H. pylori infection seems pronounced only in CagA‐positive patients. The possible links between hypertensive disease and H. pylori infection may involve the activation of the cytokine cascade with the release of vasoactive substances from the primary site of infection, or molecular mimicry between the CagA antigens of H. pylori and some peptides expressed by endothelial cells and smooth muscle cells.
Helicobacter pylori Infection Up‐regulates Gland Mucous Cell‐Type Mucins in Gastric Pyloric Mucosa
Authors: Matsuzwa Masahiro , Ota Hiroyoshi , Hayama Masayoshi , Zhang Mu Xia , Sano Kenji , Honda Takayuki , Ueno Ichiro , Akamatsu Taiji , Nakayama Jun
Published: November 2003

Background.  Two types of mucous cell are present in gastric mucosa: surface mucous cells (SMCs) and gland mucous cells (GMCs), which consist of cardiac gland cells, mucous neck cells, and pyloric gland cells. We have previously reported that the patterns of glycosylation of SMC mucins are reversibly altered by Helicobacter pylori infection. In this study, we evaluated the effects of H. pylori infection on the expression of GMC mucins in pyloric gland cells.

Methods.  Gastric biopsy specimens from the antrums of 30 H. pylori‐infected patients before and after eradication of H. pylori and 10 normal uninfected volunteers were examined by immunostaining for MUC6 (a core protein of GMC mucins), α1,4‐N‐acetyl‐glucosaminyl transferase (α4GnT) (the glycosyltransferase which forms GlcNAcα1‐4Galβ‐R), and GlcNAcα1‐4Galβ‐R (a GMC mucin‐specific glycan).

Results.  MUC6, α4GnT, and HIK1083‐reactive glycan were expressed in the cytoplasm, supranuclear region, and secretory granules in pyloric gland cells, respectively. The immunoreactivity of MUC6 and α4GnT, but not of GlcNAcα1‐4Galβ‐R, in the pyloric gland increased in H. pylori‐associated gastritis, and after the eradication of H. pylori, the increased expression of MUC6 and α4GnT in the gastric mucosa of H. pylori‐infected patients decreased to almost normal levels. This up‐regulation was correlated with the degree of inflammation.

Conclusions.  In addition to the synthesis of GMC mucins increasing reversibly, their metabolism or release may also increase reversibly in H. pylori‐associated gastritis. The up‐regulation of the expression of gastric GMC mucins may be involved in defense against H. pylori infection in the gastric surface mucous gel layer and on the gastric mucosa.
Detection of a Putative Novel Wolinella Species in Patients with Squamous Cell Carcinoma of the Esophagus
Authors: Bohr Ulrich R. M. , Segal Isidor , Primus Anett , Wex Thomas , Hassan Hashiem , Ally Reidwaan , Malfertheiner Peter
Published: November 2003

Background.  Certain regions of South Africa exhibit an extraordinarily high incidence of esophageal carcinoma that develops via an esophagitis–dysplasia–carcinoma sequence. Bacteria belonging to the family Helicobacteraceae are candidates for involvement in the initiation of the esophagitis. We investigated patients with esophageal carcinoma for the occurrence of Helicobacter‐related species.

Methods.  Biopsies from tumor and nonlesional tissue of the esophagus from nine patients with squamous cell carcinoma were investigated for Helicobacteraceae using a PCR‐based method targeting the 16S rRNA gene.

Results.  Four out of nine patients tested negative, while samples from the other five patients revealed an infection by different Helicobacter species. Sequence analysis of the PCR fragments led to the identification of a hitherto unknown bacterium in three of these patients. Phylogenetically, this bacterium was assigned to the genus Wolinella within the family of Helicobacteraceae. Helicobacter pylori was identified in three patients, and one revealed a coinfection with the novel Wolinella species.

Conclusions.  Helicobacteraceae were detected in approximately 50% of South African patients with esophageal carcinoma. Furthermore, a novel bacterium was identified that might be linked to the enhanced incidence of esophagitis and subsequent malignant disease in South Africa.
Chronomics*†: circadian and circaseptan timing of radiotherapy, drugs, calories, perhaps nutriceuticals and beyond
Authors: Halberg Franz , Cornélissen Germaine , Wang Zhengrong , Wan Chaomin , Ulmer Waldemar , Katinas George , Singh Ranjana , Singh R.K. , Singh Rajesh Kumar , Gupta B.D. , Singh R.B. , Kumar Adarsh , Kanabrocki Eugene , Sothern Robert B. , Rao Gundu , Bhatt Madan L.B. , Srivastava Madhu , Rai Gulshan , Singh Subhash , Pati Atanu Kumar , Nath Panchanan , Halberg Francine , Halberg Julia , Schwartzkopff Othild , Bakken Earl ,   His Excellency Governor Shri Vishnu Kant Shastri
Published: November 2003

We suggest a putative benefit from timing nutriceuticals (substances that are both nutrients and pharmaceuticals) such as antioxidants for preventive or curative health care, based on the proven merits of timing nutrients, drugs, and other treatments, as documented, i.a., in India. The necessity of timing melatonin, a major antioxidant, is noted. A protocol to extend the scope of chronoradiotherapy awaits testing. Imaging in time by mapping rhythms and broader time structures, chronomes, for earliest diagnoses, for example detection of vascular disease risk, is recommended. The study of rhythms and broader chronomes leads to a dynamic functional genomics, guided by imaging in time of free radicals and antioxidants, amongst many other variables.
Modulation of antineoplastic activity of cyclophosphamide by Alstonia scholaris in the Ehrlich ascites carcinoma‐bearing mice
Authors: Chandra Jagetia Ganesh , Shrinath Baliga Manjeshwar
Published: November 2003

Alterations in the neoplastic activity of cyclophosphamide (CPA) by the extract of Alstonia scholaris(ASE) were studied in mice transplanted with Ehrlich ascites carcinoma (EAC). The tumor‐bearing animals were injected with various doses of ASE and 25 mg/kg of CPA (1/10th of the LD50 dose). The combination of 120 mg/kg of ASE with 25 mg/kg of CPA was most effective, as it caused the highest tumor regression and enhanced the mean survival time (MST) and the average survival time (AST) up to 42 and 40.7, as against the 29 and 27.5 of CPA alone, respectively. Similarly, when 120 mg/kg of ASE was combined with different doses of CPA (3.125 to 50 mg/kg), a dose‐dependent increase in the anticancer activity was observed up to 25 mg/kg of CPA. However, a further increase in the CPA dose up to 37.5 or 50 mg/kg resulted in toxic side effects and death. The best effect was observed when 120 mg/kg of ASE was combined with 25 mg/kg followed by 12.5 mg/kg of CPA, as evident by the greater tumor remission, when compared with the concurrent doses of either drug alone. The administration of 120 mg/kg of ASE 6 h before the administration of 25 mg/kg of CPA resulted in a drastic decline in the glutathione levels and increased the lipid peroxidation considerably when compared with either drug alone.
In VivoEfficacy of a New Quinolone, DQ-113, against
Streptococcuspneumoniae
in a MouseModel
Authors: Otsu Yoshiko , Yanagihara Katsunori , Fukuda Yuichi , Miyazaki Yoshitsugu , Tsukamoto Kazuhiro , Hirakata Yoichi , Tomono Kazunori , Kadota Jun-ichi , Tashiro Takayoshi , Murata Ikuo , Kohno Shigeru
Published: November 2003

DQ-113
is a new quinolone with potent activity against gram-positive
pathogens. The in vivo activity of DQ-113 against
Streptococcus
pneumoniae
was compared with those of gatifloxacin and
ciprofloxacin in a mouse model. For this purpose, two strains of
S.
pneumoniae
were used: penicillin-susceptible
S.
pneumoniae
(PSSP) and penicillin-resistant
S. pneumoniae
(PRSP). The survival rates of mice infected with PSSP and PRSP at 14
days after infection were 80% in the DQ-113-treated group and 0
to 10% in the other three groups. In murine infections caused by
PSSP, the 50% effective doses (ED
50
s) of DQ-113,
gatifloxacin, and ciprofloxacin were 6.0, 41.3, and 131.6 mg/kg,
respectively. Against PRSP-caused pneumonia in mice, the
ED
50
s of DQ-113, gatifloxacin, and ciprofloxacin were 7.6,
64.7, and 125.9 mg/kg, respectively. Compared with the other drugs,
DQ-113 showed excellent therapeutic efficacy and eradicated viable
bacteria in both PSSP- and PRSP-infected mice. The means ±
standard errors of the means of viable bacterium counts in the lungs of
gatifloxacin-treated, ciprofloxacin-treated, and untreated control mice
infected with PSSP were 2.91 ± 0.34, 3.13 ± 0.48, and
3.86 ± 0.80 log
10
CFU/ml, respectively. The same
counts in mice infected with PRSP treated with the same three agents
were 6.57 ± 0.99, 6.54 ± 0.40, and 7.17 ± 0.43
log
10
CFU/ml, respectively. DQ-113 significantly decreased
the number of viable bacteria in the lungs compared with gatifloxacin
and ciprofloxacin. Of the drugs analyzed, the
pharmacokinetic-pharmacodynamic parameter of area under the
concentration-time curve (AUC)/MIC ratio for DQ-113 was significantly
higher than those for gatifloxacin and ciprofloxacin. Our results
suggest that DQ-113 has potent in vivo efficacy against both PSSP and
PRSP.
Antimalarial 9-Anilinoacridine Compounds Directed atHematin
Authors: Auparakkitanon Saranya , Noonpakdee Wilai , Ralph Raymond K. , Denny William A. , Wilairat Prapon
Published: November 2003

Antimalarial
9-anilinoacridines are potent inhibitors of parasite DNA topoisomerase
II both in vitro and in situ. 3,6-Diamino substitution on the acridine
ring greatly improves parasiticidal activity against
Plasmodium
falciparum
by targeting DNA topoisomerase II. A series of
9-anilinoacridines were investigated for their abilities to inhibitβ
-hematin formation, to form drug-hematin complexes, and to
enhance hematin-induced lysis of red blood cells. Inhibition ofβ
-hematin formation was minimal with 3,6-diamino analogs of
9-anilinoacridine and greatest with analogs with a 3,6-diCl
substitution together with an electron-donating group in the
1′-anilino position. On the other hand, the presence of a
1′-N(CH
3
)
2
group in the anilino ring
produced compounds that strongly inhibited β-hematin formation
but which did not appear to be sensitive to the nature of the
substitutions in the acridine nucleus. The derivatives bound hematin,
and Job's plots of UV-visible absorbance changes in drug-hematin
complexes at various molar ratios indicated a stoichiometric ratio of
1:2. The drugs enhanced hematin-induced red blood cell lysis at low
concentrations (<4 μM). These studies open up the novel
possibility of development of 9-anilinoacridine antimalarials that
target not only DNA topoisomerase II but alsoβ
-hematin formation, which should help delay the rapid onset of
resistance to drugs acting at only a single
site.
In Vitro Evaluation of a New Treatment for UrinaryTract Infections Caused by Nitrate-ReducingBacteria
Authors: Carlsson S. , Govoni M. , Wiklund N. P. , Weitzberg E. , Lundberg J. O.
Published: November 2003

Dietary
and endogenous nitrates are excreted in urine, and during infection
with nitrate-reducing bacteria they are reduced to nitrite. At a low pH
nitrite is converted to a variety of nitrogen oxides that are toxic to
bacteria. We hypothesized that acidification of nitrite-rich infected
urine would result in the killing of the nitrate-reducing bacteria. An
Escherichia coli
control strain and a mutant lacking nitrate
reductase activity were preincubated in urine supplemented with sodium
nitrate (0 to 10 mM) at pH 7.0. Then, the nitrite-containing bacterial
culture was transferred (and diluted 1/10) to slightly acidic urine (pH
5 and 5.5) containing ascorbic acid (10 mM) and growth was monitored.
The control strain produced nitrite in amounts related to the amount of
nitrate added. This strain was killed when the culture was transferred
to acidic urine. In contrast, the mutant that did not produce nitrite
retained full viability. When control bacteria were grown in acidic
urine with nitrate and ascorbic acid present from the start of the
experiment, no inhibition of growth was noted. The MICs and minimal
bactericidal concentrations of sodium nitrite-ascorbic
acid in acidic urine were comparable to those of conventional
antibiotics. Preincubation of nitrate-reducing
E. coli
in
nitrate-rich urine leads to the accumulation of nitrite. Subsequent
acidification of the urine results in generation of nitrogen oxides
that are bactericidal. Killing, however, requires a sequential
procedure in which the bacteria are first allowed to grow in a
nitrate-rich neutral environment, later followed by acidification. We
speculate that ingestion of nitrate followed some hours later by
acidification of urine could be a new therapeutic strategy for the
treatment of urinary tract
infections.
EfrAB, an ABC Multidrug Efflux Pump in
Enterococcusfaecalis
Authors: Lee Eun-Woo , Huda M. Nazmul , Kuroda Teruo , Mizushima Tohru , Tsuchiya Tomofusa
Published: November 2003

A
DNA fragment responsible for resistance to antimicrobial agents was
cloned from the chromosomal DNA of
Enterococcus faecalis
ATCC
29212 by using drug-hypersensitive mutant
Escherichia coli
KAM32 as a host cell. Cells of
E. coli
KAM32 harboring a
recombinant plasmid (pAEF82) carrying the DNA fragment became resistant
to many structurally unrelated antimicrobial agents, such as
norfloxacin, ciprofloxacin, doxycycline, acriflavine,
4′,6-diamidino-2-phenylindole, tetraphenylphosphonium chloride,
daunorubicin, and doxorubicin. Since the sequence of the whole genome
of
E. faecalis
is known, we sequenced several portions of the
DNA insert in plasmid pAEF82 and identified two open reading frames
within the insert. We designated the genes
efrA
and
efrB
. A search of the deduced amino acid sequences of EfrA and
EfrB revealed that they are similar to each other and that they belong
to the ATP-binding cassette (ABC) family of multidrug efflux
transporters. Transformed
E. coli
KAM32 cells harboring
efrAB
showed energy-dependent efflux of acriflavine. The
efflux activity was inhibited by reserpine, verapamil, and
sodium-
o
-vanadate, known inhibitors of ABC efflux
pumps.
FusidicAcid-Resistant Mutants of
Salmonella enterica
SerovarTyphimurium with Low Fitness In Vivo Are Defective in RpoSInduction
Authors: Macvanin Mirjana , Björkman Johanna , Eriksson Sofia , Rhen Mikael , Andersson Dan I. , Hughes Diarmaid
Published: November 2003

Mutants
of
Salmonella enterica
serovar Typhimurium resistant to
fusidic acid (Fus
r
) have mutations in
fusA
, the
gene encoding translation elongation factor G (EF-G). Most
Fus
r
mutants have reduced fitness in vitro and in vivo, in
part explained by mutant EF-G slowing the rate of protein synthesis and
growth. However, some Fus
r
mutants with normal rates of
protein synthesis still suffer from reduced fitness in vivo. As shown
here, Fus
r
mutants could be similarly ranked in their
relative fitness in mouse infection models, in a macrophage infection
model, in their relative hypersensitivity to hydrogen peroxide in vivo
and in vitro, and in the amount of RpoS production induced upon entry
into the stationary phase. We identify a reduced ability to induce
production of RpoS (σ
s
) as a defect associated with
Fus
r
strains. Because RpoS is a regulator of the general
stress response, and an important virulence factor in
Salmonella
, an inability to produce RpoS in appropriate
amounts can explain the low fitness of Fus
r
strains in vivo.
The unfit Fus
r
mutants also produce reduced levels of the
regulatory molecule ppGpp in response to starvation. Because ppGpp is a
positive regulator of RpoS production, we suggest that a possible cause
of the reduced levels of RpoS is the reduction in ppGpp production
associated with mutant EF-G. The low fitness of Fus
r
mutants
in vivo suggests that drugs that can alter the levels of global
regulators of gene expression deserve attention as potential
antimicrobial
agents.
Increased Killing of Staphylococci and Streptococciby Daptomycin Compared with Cefazolin and Vancomycin in an InVitro Peritoneal DialysateModel
Authors: Hermsen Elizabeth D. , Hovde Laurie B. , Hotchkiss John R. , Rotschafer John C.
Published: November 2003

Peritoneal
dialysate fluid (PDF) is a bacteriostatic medium that compromises the
antibacterial activity of cell wall-active agents. By use of an in
vitro static model, methicillin-resistant
Staphylococcus
aureus
(MRSA), methicillin-susceptible
S. aureus
(MSSA),
methicillin-susceptible
Staphylococcus epidermidis
(MSSE), and
Streptococcus sanguis
were exposed to daptomycin
at concentrations of 10, 30, and 100 mg/liter, cefazolin at 125
mg/liter, and vancomycin at 25 mg/liter in cation-adjusted
Mueller-Hinton Broth or Todd Hewitt Broth (for
S. sanguis
) and
PDF at pHs of 5.5 and 7.4. The pH had no effect on antibacterial
activity. Neither cefazolin nor vancomycin produced a bactericidal or a
bacteriostatic effect versus MRSA, MSSA, MSSE, or
S. sanguis
in PDF, while all concentrations of daptomycin were bactericidal
against all organisms in PDF. Daptomycin did not exhibit
concentration-dependent activity in PDF. Daptomycin appears to be a
promising agent for use in peritoneal dialysis-associated peritonitis,
producing bacterial kill to a greater extent and at a higher rate than
cefazolin or vancomycin in
PDF.
Efficacies of Vancomycin, Arbekacin, and GentamicinAlone or in Combination against Methicillin-Resistant
Staphylococcus aureus
in an In Vitro InfectiveEndocarditisModel
Authors: Lee Dong-Gun , Chun Hye-Sun , Yim Dong-Seok , Choi Su-Mi , Choi Jung-Hyun , Yoo Jin-Hong , Shin Wan-Shik , Kang Moon-Won
Published: November 2003

We
adopted an in vitro infective endocarditis model (IVIEM) to compare the
efficacy of vancomycin (VAN), arbekacin (ABK), and gentamicin (GEN)
alone or in combination. Using two strains of clinically isolated
methicillin-resistant
Staphylococcus aureus
, one GEN
susceptible (GS171) and one GEN resistant (GR153), fibrin clots were
prepared and suspended in the IVIEM. Antibiotics were given as boluses
every 6 h (q6h), q12h, or q24h or by continuous infusion with
VAN, q12h or q24h with ABK, and q8h or q24h with GEN. For combination
treatment, VAN q12h plus ABK q24h and VAN q12h plus GEN q24h were
given. Fibrin clots were removed from each model at 0, 8, 24, 32, 48,
and 72 h, and the bacterial densities were determined. The
number of colonies within the fibrin clot was significantly decreased
in all study groups compared with control groups (
P
<
0.001). When VAN and ABK were administered alone, the number of
colonies was significantly lower in GS171 than in GR153 by 8
h after administration (
P
= 0.02) and was lowest in
GS171 when ABK was administered q12h (
P
= 0.01). At
72 h, ABK or VAN alone produced equivalent bacterial
reductions regardless of dosing frequency and GEN resistance. In GR153,
VAN plus ABK showed an additive effect till 24 h, although
VAN plus GEN showed indifference. Our data suggest that ABK could be
used as an alternative to VAN in GEN-resistant staphylococcal
endocarditis. An additive effect was seen when VAN and ABK were used
together in GEN-resistant strains until 24 h; however,
further studies are warranted for the clinical application of this
combination.
Toxicity and Antileishmanial Activity of a New StableLipid Suspension of AmphotericinB
Authors: Larabi Malika , Yardley Vanessa , Loiseau Philippe M. , Appel Martine , Legrand Philippe , Gulik Annette , Bories Christian , Croft Simon L. , Barratt Gillian
Published: November 2003

The
aim of the present study was to evaluate the toxicity and the activity
of a new lipid complex formulation of amphotericin B (AMB) (LC-AMB;
dimyristoyl phosphatidylcholine, dimyristoyl phosphatidylglycerol, and
AMB) that can be produced by a simple process. Like other lipid
formulations, this new complex reduced both the hemolytic activity of
AMB (the concentration causing 50% hemolysis of human
erythrocytes, >100 μg/ml) and its toxicity
toward murine peritoneal macrophages (50% inhibitory
concentration, >100 μg/ml at 24 h). The in
vivo toxicity of the new formulation (50% lethal dose,>
200 mg/kg of body weight for CD1 mice) was similar to those of
other commercial lipid formulations of AMB. The complex was the most
effective formulation against the DD8 strain of
Leishmania
donovani
. It was unable to reverse the resistance of an
AMB-resistant
L. donovani
strain. In vivo LC-AMB was less
efficient than AmBisome against
L.
donovani
.