bluuebunny/crossref_metadata_2025 huggingface dataset:
rRNA probe‐based cell fishing of bacteria
Authors: Stoffels Marion , Ludwig Wolfgang , Schleifer Karl Heinz
Published: March 2003

We have developed a new, cultivation‐independent, fast and flexible method for the rRNA‐targeted probe‐based enrichment of bacteria. The target cells were labelled by in situ hybridization with biotinylated polyribonucleotide probes. These probes were generated by in vitro transcription of amplified rDNA of a variable region in domain III of the 23S rRNA molecules. The probes were about 300 nucleotides in length and were labelled by incorporation of biotin‐UTP during the transcription. Probes were hybridized with bacterial cells and incubated with paramagnetic streptavidin‐coated particles. The labelled target cells can be separated in a column filled with steel wool inserted into the field of a permanent magnet. Unlabelled, non‐target cells pass through the column, whereas labelled cells are retained. They were eluted from the column after removal of the magnetic field. Up to now, the method has been tested with mixtures of different pure cultures. For the first time, transcript probes have been used for the labelling of the target cells and for their specific separation. The enrichment of the target cells can be monitored by a streptavidin–fluorescein staining of the biotinylated target cells and/or by a subsequent in situ hybridization with fluorescently labelled oligonucleotide probes. Enrichment rates of up to 90‐fold, depending on the original abundance of the cells of interest, could be determined. To demonstrate that the sorted cells were amenable to molecular analysis, we amplified and sequenced a part of the tuf gene of enriched Acinetobacter calcoaceticus cells.
Online and in situ monitoring of environmental pollutants: electrochemical biosensing of cadmium
Authors: Biran Israel , Babai Reuven , Levcov Klimentiy , Rishpon Judith , Ron Eliora Z.
Published: March 2003

Online sensitive monitoring of gene expression is essential for understanding microbial life and microbial communities, especially under stress‐inducing conditions, such as the presence of environmental pollutants. We describe here a novel use of promoter‐based electrochemical biosensing for online and in situ monitoring of gene expression in response to pollutants. As a model system, we used a cadmium‐responsive promoter from Escherichia coli fused to a promoterless lacZ gene, which was monitored using an electrochemical assay of β‐galactosidase activity. This whole‐cell biosensor could detect, within minutes, nanomolar concentrations of cadmium in water, sea water and soil samples, and it can be used for continuous online and in situ monitoring.
Remarkably low temperature optima for extracellular enzyme activity from Arctic bacteria and sea ice
Authors: Huston Adrienne L. , Krieger‐Brockett Barbara B. , Deming Jody W.
Published: March 2003

Extracellular degradative enzymes released by psychrophilic marine bacteria (growing optimally at or below 15°C and maximally at 20°C) typically express activity optima at temperatures well above the upper growth limit of the producing strain. In the present study, we investigated whether or not near‐zero Arctic environments contain extracellular enzymes with activity optimized to temperatures lower than previously reported. By applying fluorescently tagged substrate analogues to measure leucine‐aminopeptidase and chitobiase activity, the occurrence of extracellular enzymatic activity (EEA) with remarkably low temperature optima (15°C) was documented in sea‐ice samples. An extremely psychrophilic bacterial isolate, strain 34H, yielded an extract of cell‐free protease with activity optimized at 20°C, the lowest optimum yet reported for cell‐free EEA from a pure culture. The use of zymogram gels revealed the presence of three proteolytic bands (between 37 and 45 kDa) in the extract and the release of the greatest quantities of the proteases when the strain was grown at −1°C, suggesting a bacterial strategy for counteracting the effects of very cold temperatures on the catalytic efficiency of released enzymes. The detection of unusually cold‐adapted EEA in environmental samples has ramifications not only to polar ecosystems and carbon cycling but also to protein evolution, biotechnology and bioremediation.
Phylogenetic characterization of epibiotic bacteria in the accessory nidamental gland and egg capsules of the squid Loligo pealei (Cephalopoda: Loliginidae)
Authors: Barbieri Elena , Paster Bruce J. , Hughes Deborah , Zurek Ludek , Moser Duane P. , Teske Andreas , Sogin Mitchell L.
Published: March 2003

Sexually mature female squid Loligo pealei harbour dense bacterial communities in their accessory nidamental glands (ANGs) and in their egg capsules (ECs). This study describes a molecular approach using the 16S rRNA gene (rDNA) to identify bacterial populations within the ANG and the ECs of the North Atlantic squid species L. pealei. Fluorescent in situ hybridization (FISH) and 16S rDNA analysis showed that predominantly alpha‐ and, to a lesser extent, gamma‐proteobacteria were the predominant components of the ANG and EC bacterial communities. Sequencing results showed the presence of alpha‐proteobacterial populations affiliated with the Roseobacter group and additional deep‐branching alphaproteobacterial lineages. In contrast, isolates from the ANG and ECs contained only a few alpha‐proteobacteria of the Roseobacter group compared with several gamma‐proteobacterial isolates, mostly Shewanella and Pseudoalteromonas species. Most of the ANG‐associated bacterial populations were also found within the ECs of L. pealei. The molecular approach allowed the visualization of alpha‐proteobacteria as major constituents of a bacterial symbiosis within the reproductive system of the Loliginidae.
Bacterial community associated with Pfiesteria‐like dinoflagellate cultures
Authors: Alavi Mohammad , Miller Todd , Erlandson Karn , Schneider Rachel , Belas Robert
Published: March 2003

Dinoflagellates (Eukaryota; Alveolata; Dinophyceae) are single‐cell eukaryotic microorganisms implicated in many toxic outbreaks in the marine and estuarine environment. Co‐existing with dinoflagellate communities are bacterial assemblages that undergo changes in species composition, compete for nutrients and produce bioactive compounds, including toxins. As part of an investigation to understand the role of the bacteria in dinoflagellate physiology and toxigenesis, we have characterized the bacterial community associated with laboratory cultures of four ‘Pfiesteria‐like’ dinoflagellates isolated from 1997 fish killing events in Chesapeake Bay. A polymerase chain reaction with oligonucleotide primers specific to prokaryotic 16S rDNA gene sequences was used to characterize the total bacterial population, including culturable and non‐culturable species, as well as possible endosymbiotic bacteria. The results indicate a diverse group of over 30 bacteria species co‐existing in the dinoflagellate cultures. The broad phylogenetic types of dinoflagellate‐associated bacteria were generally similar, although not identical, to those bacterial types found in association with other harmful algal species. Dinoflagellates were made axenic, and the culturable bacteria were added back to determine the contribution of the bacteria to dinoflagellate growth. Confocal scanning laser fluorescence microscopy with 16S rDNA probes was used to demonstrate a physical association of a subset of the bacteria and the dinoflagellate cells. These data point to a key component in the bacterial community being species in the marine alpha‐proteobacteria group, most closely associated with the α‐3 or SAR83 cluster.
Hungry bacteria – definition and properties of a nutritional state
Authors: Ferenci Thomas
Published: March 2003

Bacteria are sometimes neither starving nor under nutrient‐excess conditions. When growing with suboptimal levels of nutrients, hungry bacteria express appropriate cellular responses. This review discusses approaches to defining the hunger response in both molecular and growth kinetic terms. The gene expression changes unique to hunger conditions are described, using Escherichia coli as the primary example. Metabolite changes with hunger and starvation and the differing role of the stationary phase regulator RpoS also lead to the hypothesis proposed in this review that bacteria undertake distinct approaches to hunger and starvation. Indeed, an understanding of the difference between hunger and starvation and the incompatibility between hunger and starvation responses explains some of the paradoxical mutational adaptations, such as rpoS inactivation, found in natural populations.
Antibiotic resistance in bacteria from magpies (Pica pica) and rabbits (Oryctolagus cuniculus) from west Wales
Authors: Livermore David M. , Warner Marina , Hall Lucinda M. C. , Enne Virve I. , Projan Stephen J. , Dunman Paul M. , Wooster Sarah L. , Harrison Graham
Published: March 2003

The prevalence of antibiotic‐resistant bacteria in wild animal and bird populations is largely unknown, with little consistency among the few published reports. We therefore examined intestinal bacteria from magpies (Pica pica) and rabbits (Oryctolagus cuniculus) collected in rural west Wales. Escherichia coli isolates resistant to multiple antibiotics were grown from eight of 20 magpies trapped in spring, 1999 and one of 17 in spring, 2000; the most prevalent resistance trait among these isolates was to tetracycline, but resistances to ampicillin, chloramphenicol, kanamycin, sulphonamide, tetracycline and trimethoprim were also found. Tetracycline‐resistant Enterococcus spp. were found in one of 20 magpies in 1999 and three of 17 in 2000. Only one resistant E. coli isolate was detected among gut bacteria from 13 rabbits, and this strain was resistant only to tetracycline. Differences in the prevalence of resistance between bacteria from rabbits and magpies may reflect differences in diet: rabbits graze field edges, whereas magpies are omnivorous and opportunistic. The resistance genes found in E. coli isolates from magpies mostly corresponded to those common among human isolates, but those conferring tetracycline resistance were unique.
Prevalence and microdiversity of Alteromonas macleodii‐like microorganisms in different oceanic regions
Authors: García‐Martínez Jesús , Acinas Silvia G. , Massana Ramon , Rodríguez‐Valera Francisco
Published: March 2003

The presence, prevalence and variability of microorganisms related to the species Alteromonas macleodii, a well known culturable γ‐Proteobacterium, has been studied in different seawater samples from diverse geographical locations, in both the Northern and Southern hemispheres, and tested with two molecular techniques (rRNA hybridization and gene cloning and sequencing). Results show that A. macleodii‐like microorganisms are present in high proportions in North Atlantic and, especially, Mediterranean waters, being higher at deep samples and particle‐associated fractions, in agreement with previous findings. In contrast, Southern samples (all from very cold areas near Antarctica) presented no significant hybridization signals. The analysis of the ribosomal ITS (16S‐23S internal transcribed spacers) revealed that A. macleodii‐like microorganisms from Mediterranean, North Atlantic, Caribbean and Red Sea waters differed in both size and sequence, mostly depending on their geographical origin, with Mediterranean and North Atlantic clones clustering into two main groups whereas Caribbean and Red Sea clones appeared separated.
The LysR‐type regulator SftR is involved in soil survival and sulphate ester metabolism in Pseudomonas putida
Authors: Kahnert Antje , Mirleau Pascal , Wait Robin , Kertesz Michael A.
Published: March 2003

Sulphate esters make up a large proportion of the available sulphur in agricultural soils, and many pseudomonads can desulphurize a range of aryl‐ and alkylsulphate esters to provide sulphur for growth. After miniTn5 transposon mutagenesis of Pseudomonas putida S‐313, we isolated 19 mutants that were defective in cleavage of the chromogenic sulphate ester 5‐bromo‐4‐chloro‐3‐indoxylsulphate (X‐sulphate). Analysis of these strains revealed that they carried independent insertions in a gene cluster that comprised genes for a sulphate ester/sulphonate transporter (atsRBC) a LysR‐type regulator (sftR), an oxygenolytic alkylsulphatase (atsK), an arylsulphotransferase (astA) and a putative TonB‐dependent receptor (sftP). The SftP protein was localized in the outer membrane, and the arylsulfphotransferase was identified as an intracellular enzyme. Expression of sftR was repressed in the presence of inorganic sulphate, and the sftR gene was required for the expression of atsBC, atsRK and sftP–astA. An sftR mutant was unable to grow with aryl‐ or alkylsulphate esters in laboratory media and showed significantly reduced survival compared with the parent strain during incubation in Danish agricultural and grassland soils. This effect suggests that sulphate esters are an important sulphur source for microbes in aerobic soils and highlights the importance of the microbial population in the soil sulphur cycle.
Development of a novel, bioluminescence‐based, fungal bioassay for toxicity testing
Authors: Weitz Hedda J. , Campbell Colin D. , Killham Ken
Published: March 2003

Naturally bioluminescent fungi, Armillaria mellea and Mycena citricolor, were used to develop a novel, bioluminescence‐based bioassay for toxicity testing. Bioassays were carried out to assess the toxicity of 3,5‐dichlorophenol (3,5‐DCP), pentachlorophenol (PCP), copper and zinc. The results suggested that 60 min was a suitable exposure time for the bioassay. Light reduction was observed in response to 3,5‐DCP, PCP and Cu for both A. mellea and M. citricolor, but to Zn only for A. mellea. Armillaria mellea was significantly less sensitive to 3,5‐DCP and PCP than M. citricolor. The EC50 values for A. mellea and M. citricolor were similar to EC50 values for 3,5‐DCP, PCP and Cu (but not Zn) of bioluminescence‐based bacterial biosensors. They were also similar to EC50 values for Cu and Zn of a bioluminescence‐based yeast biosensor. The results highlighted the importance of using both prokaryotic and eukaryotic biosensors. The novel bioassay provides a rapid and sensitive method to assess bioavailability of pollutants as well as a method to determine their toxicity to filamentous fungi. It also expands the range of organisms that can be used for bioluminescence‐based toxicity testing by complementing existing biosensors.
Pseudomonas aeruginosa displays an epidemic population structure
Authors: Pirnay Jean‐Paul , De Vos Daniel , Cochez Christel , Bilocq Florence , Vanderkelen Alain , Zizi Martin , Ghysels Bart , Cornelis Pierre
Published: March 2003

Bacteria can have population structures ranging from the fully sexual to the highly clonal. Despite numerous studies, the population structure of Pseudomonas aeruginosa is still somewhat contentious. We used a polyphasic approach in order to shed new light on this issue. A data set consisting of three outer membrane (lipo)protein gene sequences (oprI, oprL and oprD), a DNA‐based fingerprint (amplified fragment length polymorphism), serotype and pyoverdine type of 73 P. aeruginosa clinical and environmental isolates, collected across the world, was analysed using biological data analysis software. We observed a clear mosaicism in the results, non‐congruence between results of different typing methods and a microscale mosaic structure in the oprD gene. Hence, in this network, we also observed some clonal complexes characterized by an almost identical data set. The most recent clones exhibited serotypes O1, 6, 11 and 12. No obvious correlation was observed between these dominant clones and habitat or, with the exception of some recent clones, geographical origin. Our results are consistent with, and even clarify, some seemingly contradictory results in earlier epidemiological studies. Therefore, we suggest an epidemic population structure for P. aeruginosa, comparable with that of Neisseria meningitidis, a superficially clonal structure with frequent recombinations, in which occasionally highly successful epidemic clones arise.
Microbial diversity in ikaite tufa columns: an alkaline, cold ecological niche in Greenland
Authors: Stougaard Peter , Jørgensen Flemming , Johnsen Mads G. , Hansen Ole C.
Published: March 2003

Ikaite tufa columns from the Ikka Fjord in south‐western Greenland constitute a natural, stable environment at low temperature and with a pH ranging from neutral at the exterior to very alkaline (pH 10.4) at the interior of the column. Phylogenetic analysis of culturable organisms revealed ten different isolates representing three of the major bacterial divisions. Nine of the isolates showed 94–99% similarity to known sequences, whereas one isolate displayed a low degree of similarity (less than 90%) to a Cyclobacterium species. Seven of the isolates were shown to be cold active alkaliphiles, whereas three isolates showed optimal growth at neutral pH. Phylogenetic analysis of DNA isolated directly from the ikaite material demonstrated the presence of a microbial flora more diverse than the culturable isolates. Whereas approximately half of the phylotypes showed 90–99% similarity to known meso‐ or thermophilic alkaliphiles, the rest of the sequences displayed less than 90% similarity when compared to known 16S rRNA gene sequences in databases. Thus, in the present paper, we demonstrate that ikaite columns that host a specialized macroscopic flora and fauna also contain a unique, cold active, alkaliphilic microflora.
Stable augmentation of activated sludge with foreign catabolic genes harboured by an indigenous dominant bacterium
Authors: Watanabe Kazuya , Teramoto Maki , Harayama Shigeaki
Published: March 2003

Comamonas sp. rN7 is a phenol‐degrading bacterium that represents the dominant catabolic population in activated sludge. The present study examined the utility of this bacterium for establishing foreign catabolic genes in phenol‐digesting activated sludge. The phc genes coding for phenol hydroxylase and its transcriptional regulators of C. testosteroni R5 were integrated into the chromosome of strain rN7. The specific phenol‐oxygenating activity of a resultant transformant designated rN7(R503) was three times higher than the activity of strain rN7, and the phc genes were stably inherited by rN7(R503) grown in a non‐selective laboratory medium. Inoculation of phenol‐acclimatized activated sludge with rN7(R503) resulted in a high phenol‐oxygenating activity and improved resistance to phenol‐shock loading compared to sludge inoculated with either no cells, rN7 or R5. Quantitative competitive polymerase chain reaction (PCR) showed that the phc genes were retained in the rN7(R503)‐inoculated sludge at a density of more than 108 copies per ml of mixed liquor for more than 35 days, whereas those in the R5‐inoculated sludge disappeared rapidly. No transfer of the phc genes to other indigenous populations was apparent in the rN7(R503)‐harbouring sludge. From these results, we concluded that the phenol treatment of the activated sludge was enhanced by the phc genes harboured by the rN7(R503) population. This study suggests a possible bioaugmentation strategy for stably utilizing foreign catabolic genes in natural ecosystems.
Decline in diversity and abundance of endophytic fungi in twigs of Fagus sylvatica L. after experimental long‐term exposure to sodium dodecylbenzene sulphonate (SDBS) aerosol
Authors: Danti Roberto , Sieber Thomas N. , Sanguineti Giovanni , Raddi Paolo , Di Lonardo Vincenzo
Published: March 2003

Sodium dodecylbenzene sulphonate (SDBS) is an anionic synthetic detergent found in polluted sea aerosol and is known for its harmful effects on leaf surface ultrastructure on conifers and broadleaved trees. Four‐year‐old saplings of European beech were sprayed weekly for three consecutive growing seasons with either a 50 mg l−1 solution of SDBS in deionized water or with pure deionized water (control). Two‐ to three‐ year‐old twigs were collected from SDBS‐treated and control plants during the growing season one year after the last treatment to isolate endophytic fungi. The frequency of colonization by endophytic fungi was significantly lower on SDBS‐treated plants (63.8%) than on control plants (85.4%). Multiple colonization of twigs occurred more frequently and diversity of endophyte species was higher in control plants than in SDBS‐treated plants. Thirty‐six fungal species were isolated from 360 twigs. Cladosporium cladosporioides, Coryneum compactum, Phialocephala dimorphospora, and a species each of Mycosphaerella and Phomopsis were the most abundant endophytes with frequencies of colonization of more than 5%. The abundance of the Phomopsis species proved to be significantly reduced by the SDBS treatment. Within the limits of the indoor experimental conditions, the obtained results suggest that long‐term exposure of aerial parts of beech to SDBS can affect the amount and composition of endophytic fungal communities of lignified twigs. Degradation of the leaf epicuticular wax layer and changes of the assimilation capacity and leaf water content (transpiration) of the crowns are presumed to be responsible for the reduction of endophytic fungi detected in twigs of SDBS‐treated plants.
Detection of multiple extracytoplasmic function (ECF) sigma factors in the genome of Pseudomonas putida KT2440 and their counterparts in Pseudomonas aeruginosa PA01
Authors: Martínez‐Bueno Manuel A. , Tobes Raquel , Rey Manuel , Ramos Juan‐Luis
Published: March 2003

Pseudomonas putida KT2440 is highly successful in colonizing a variety habitats, including aquatic and edaphic niches. In accordance with this ability and with the need to adapt to changing environmental conditions, P. putida has developed sophisticated mechanisms of transcriptional regulation. We analysed, at the genome level, the repertoire of sigma factors in P. putida KT2440 and identified 24 sigma factors, 19 of which corresponded to the subfamily of extracytoplasmic function (ECF) sigma factors. We detected 13 ECF sigma factors that showed similarity to the Escherichia coli FecI sigma factor, which is involved in iron acquisition. In 11 cases, a fecR‐like gene was found adjacent to the fecI‐like gene and, in 10 cases, a gene encoding an iron receptor lies in the vicinity of the fecI/fecR cluster. This may explain the ability of P. putida KT2440 to grow under low iron availability conditions. Five fecI/fecR/iron receptor gene clusters from P. putida were also identified in the human pathogen Pseudomonas aeruginosa.
Microbial community and physicochemical analysis of an industrial waste gas biofilter and design of 16S rRNA‐targeting oligonucleotide probes
Authors: Friedrich Udo , Van Langenhove Herman , Altendorf Karlheinz , Lipski André
Published: March 2003

A study was conducted to investigate the microbial community structure, the physicochemical properties, and the relationships between these parameters of a full‐scale industrial biofilter used for waste gas abatement in an animal‐rendering plant. Fluorescencein situhybridization (FISH) was successfully combined with digital image analysis to study the composition of the microbial community. Several new nucleic acid probes were designed and established based on published 16S rDNA sequences and on ones retrieved from the biomass of the biofilter under investigation. Bacterial detection rates varied greatly over time and filterbed depth between 27.2% and 88.1% relative to DAPI counts. Overall, members of theBetaproteobacteriafollowed byActinobacteria,Alphaproteobacteria,Cytophaga‐Flavobacteria,FirmicutesandGammaproteobacteriawere the most abundant groups. Among the groups below phylum level, members of theAlcaligenes/Bordetellalineage were on average the most abundant group accounting for up to 8.5% of DAPI‐stained cells. Whereas the community composition generally showed no vertical gradient, the lower 50 cm of the biofilter proved to be the most active part for the degradation of aldehydes such as 2‐ and 3‐methylbutanal, 2‐methylpropanal, and hexanal. This zone of the filterbed being operated in up‐flow direction degraded about 80% of these compounds. Dimethyldisulphide was the most common reduced sulphur compound. Statistical analysis of microbial versus waste gas parameters generally revealed only weak or non‐significant correlations between the two. Possible explanations for this finding are discussed.
Emergency subtotal colectomy as treatment of choice in obstructing carcinomas of the left colon
Authors: Pérez‐Díaz  , Turégano‐Fuentes  , Calvo‐Serrano  , Tomás‐Palacios J. De , Fuenmayor‐Valera M. L. De , Hernández E. Del Valle , Jiménez F. Muñoz , Quintans‐Rodríguez
Published: March 2003

Purpose

To assess the morbidity, mortality and function following emergency subtotal colectomy (ESC) with primary anastomosis in obstructing carcinomas of the left colon, to determine whether this technique is warranted as the current treatment of choice for this condition.

Methods

Retrospective review of a series of 70 patients with obstructing carcinomas of the left colon who underwent an ESC with primary anastomosis between September 1989 and December 1996. Morbidity, mortality, hospital stay and functional results were analysed, as well as the percentage of patients submitted to this technique.

Results

The use of ESC has steadily increased at our institution over the last few years, from 22% in 1990 to 84% of all the procedures performed for obstructing carcinomas of the left colon in 1995. The mortality rate of the series was 4.2%. Anastomotic leak rate was 10%, with only three major leaks. A prolonged obstructive ileus was a frequent and distressing complication (26%), and the main determinant of hospital stay. Average daily stool frequency on discharge from hospital was of 2 (range 1–6), with many patients having discontinued anti‐diarrhoeal medications.

Conclusion

Emergency SC with primary anastomosis has evolved into the treatment of choice for most patients with acutely obstructing malignancies of the left colon at our institution. It can be performed with low mortality, a reasonable morbidity and good functional results.
Immediate reconstruction of the anal sphincter after fistulectomy in the management of complex anal fistulas
Authors: Roig  , Garcia‐Armengol  , Jordán  , Alos  , Solana
Published: March 2003

An analysis was made of the indications and outcome of immediate reconstruction of the anal sphincter after fistulectomy in the management of a selected group of patients who were at risk of post‐operative incontinence. A total of 31 patients underwent surgery, with 25 high trans‐sphincteric fistulas (80.6%), four low trans‐sphincteric fistulas (12.9%), and two suprasphincteric fistulas (6.5%). The median post‐operative stay was 7.0 days, with a median follow up of 24.0 months. We describe one case (3.2%) of post‐operative infection and dehiscence of the muscle suture. The fistula recurred in three patients (9.7%). At the end of follow up of 25 patients with full preoperative continence, five patients (20.0%) presented with perianal soiling and one (4.0%) had incontinence to flatus. Sphincter reconstruction after fistulectomy constitutes a management option that should be considered in the treatment of certain fistulas. It allows both rapid recovery and the preservation of anal function.
Medical negligence in coloproctology
Authors: Schofield
Published: March 2003

There has been a significant increase of litigation in the UK in recent years and for this reason it is appropriate to analyse the reasons for civil litigation. Alleged medical negligence in the colorectal field comes from the activity of general surgeons and gynaecologists as well as colorectal surgeons. This communication deals with problems in the coloproctology area irrespective of the surgeon involved.

Two hundred and forty‐five cases are reviewed, consisting of 49 anal problems, 160 abdominal problems with 24 laparoscopic and 12 colonoscopic injuries. Thirty‐two of 49 anal cases had incontinence and these were often difficult to defend, especially the 20 cases due to childbirth injury because of poor documentation. The abdominal problems were due to delay or complications of surgery. The cases based on delay in diagnosis of cancer were often unsuccessful, but the cases of delay in diagnosis of intra‐abdominal infection were difficult to defend. Problems occurring in the operative or early post‐operative period produced allegations which were often justified, but the late complications of surgery were rarely due to poor standard of care.

The laparoscopic injuries included bowel perforation, bleeding and major vascular damage which were not recognized at the time. The colonoscopic cases were chiefly due to perforation, which is easily defended provided the endoscopist is experienced and the risk is recorded in the consent.

The cases have yielded a number of lessons which are discussed and compared with the American experience. It is concluded that litigation is traumatic to both plaintiff and surgeon and that better ways should be found to resolve complaints of inadequate care.
Anal plug for faecal incontinence
Authors: Norton C. , Kamm M. A.
Published: March 2003

Objective

Despite the availability of pharmacological, behavioural and surgical treatments for faecal incontinence, many patients remain symptomatic. There are few devices designed specifically to cope with this, and perineal pads are inefficient and unacceptable for many patients. This study aimed to evaluate a new device in patients with intractable faecal incontinence.

Method

Two sizes of a purpose‐designed anal plug were evaluated in 20 patients with intractable faecal incontinence for solid or liquid stool. Each plug size was tested for 2 weeks, with patients completing a structured questionnaire after each size.

Results

The majority [14/20] could not tolerate a plug due to discomfort. Four patients (20%) wished to continue to use a plug on a regular basis after the study, and two others on an occasional basis. However, for this minority who could tolerate a plug, it was highly successful at controlling faecal incontinence. There was no clear preference for the smaller or larger plug. There was no association between comfort in using the plug and anorectal sensitivity as measured by electrophysiological tests. It was not possible to predict which patients would benefit from plug use.

Conclusion

The anal plug is effective in controlling faecal incontinence and is well tolerated in a minority of patients. Evaluation quickly reveals whether the patient will find it an effective and acceptable option. This device therefore offers a further management option for patients with faecal incontinence.
Using quality of life scores to help determine treatment: is restoring bowel continuity better than an ostomy?
Authors: Ko C. Y. , Rusin L. C. , Schoetz D. J. , Coller J. A. , Murray J. J. , Roberts P. L. , Moreau L.
Published: March 2003

Objective

In colorectal surgery, evaluation of heath‐related quality of life (HRQL) has been relatively minimal when compared to other medical specialties. Would the performance of such HRQL evaluations change our decision‐making in patient care? In familial adenomatous polyposis (FAP), procedures that restore bowel continuity (i.e. Ileorectal anastomosis or ileal pouch anal anastomosis) are routinely preferred to ileostomy because of the perceived, but unproven, better HRQL. This study evaluates FAP patients who underwent prophylactic colectomy with either permanent ileostomy or ‘restored bowel continuity’ reconstruction. The functional outcomes of both groups are reported, and the HRQL assessments are compared.

Methods

All FAP patients who underwent (procto) colectomy resection with reconstruction, either restored bowel continuity (BC) or permanent ileostomy (OST), between 1980 and 1998 were studied. Functional data were obtained by questionnaire and medical record review. HRQL was assessed by 2 validated instruments – the SF‐36 Physical and Mental Health Summary Scales and the SF‐36 Health Survey – which measure physical summary (PSF) and mental summary functioning (MSF) as well as eight separate health quality dimensions including health perception (HP), physical (PF) and social functioning (SF), physical (PR) and emotional role limitations (ER), mental health (MH), bodily pain (BP), and energy level (E).

Results

Results were obtained in 54 patients; bowel continuity (44), ileostomy (10). Mean patient age was 39 years, mean follow up time was 10.5 years. Mean patient age at operation was 28 years. Functional results for BC included number of bowel movements/day (6.7), leakage (30%), having to wear a pad (11%), perianal skin problems (25%), food avoidance (68%), and inability to distinguish gas (27%). Functional results for OST were routinely excellent. Results of the HRQL surveys reveal no significant differences for BC vs OST (HP: 67 ± 28 vs 79 ± 39; PF: 91 ± 14 vs 90 ± 17; SF: 86 ± 23 vs 97 ± 5; PR: 79 ± 34 vs 83 ± 40; ER: 86 ± 28 vs 88 ± 27; MH: 77 ± 19 vs 82 ± 14; BP: 78 ± 24 vs 71 ± 32; E 60 ± 21 vs 58 ± 18, respectively).

Conclusion

Although the perceived quality of life for ileostomy patients is generally worse than the ‘restored bowel continuity’ group, the measured HRQL is the same for both groups. These results suggest that a permanent ileostomy should be included as a viable and appropriate first line treatment option for FAP patients after resection. This study also suggests that HRQL should play a greater role in the evaluation of care and treatment in colorectal surgery.
A prospective study to assess the implementation of a fast‐track system to meet the two‐week target for colorectal cancer in Somerset
Authors: Davies R. J. , Ewings P. , Welbourn R. , Collins C. , Kennedy R. , Royle C.
Published: March 2003

Objective

This prospective study assesses the introduction of a fast‐track referral system for patients with suspected colorectal cancer.

Patients and methods

The referral system was initiated in Yeovil District Hospital and Taunton and Somerset Hospital using six screening criteria to select high‐risk patients. Data on all high‐risk patients from 1 November 1999 to 30 April 2000 was recorded prospectively. Patients with proven colorectal cancer diagnosed between 1 November 1998 and 30 April 1999 have been identified for comparison.

Results

There were 158 new cases of colorectal cancer in total (111 elective, 47 emergency). 59 cases of colorectal cancer were diagnosed from 433 fast‐track referrals (53% of total elective cases). Median time from referral to diagnosis in the fast‐track group was 11 days vs. 32.5 days for nonfast‐track elective patients (P < 0.001). Median time from referral to diagnosis for all elective cases was 17 days vs. 38.5 days for patients presenting one year earlier (P < 0.001). 75% of fast‐track patients were diagnosed with colorectal cancer within two weeks, compared with 17% of nonfast‐track elective patients (P < 0.001). The proportion of emergency admissions was reduced from 40% to 30% (P=0.07) following the introduction of this system.

Conclusion

Data from the first six months reveal that over half of the elective cases of colorectal cancer were diagnosed within the fast‐track system. The median time from referral to diagnosis in these patients was within two weeks and there has been a significant reduction in the time to diagnosis associated with the introduction of this service.
Altered serotonin immunoreactivities in the left colon of patients with colonic inertia
Authors: Zhao R. H. , Baig M. K. , Mack J. , Abramson S. , Woodhouse S. , Wexner S. D.
Published: March 2003

Background

Serotonin is an important positive regulator of colonic motility and transit. Its quantity and distribution in the left colon could be abnormal in patients with colonic inertia (CI) and contribute to the disease.

Aim

To evaluate serotonin positive cells and immuno‐reactivities in the mucosa, submucosa and muscularis propria of the left colon from patients with CI was compared to a control group.

Patients and methods

Nineteen patients who underwent subtotal colecotomy for CI were assessed. The control group consisted of 15 patients who underwent left hemicolectomy for colonic cancer; histologically normal tissue specimens were used. Immunohistochemical staining for serotonin was performed. In the mucosa, the average number of serotonin positive cells per microscopic field (200 × ) was determined. The positively stained area (square pixels) in the mucosa, submucosa and muscularis propria per microscopic field (200 × ) was calculated utilizing a computer image analysis program.

Results

In the mucosa, both the number of serotonin positive cells and positively stained area were significantly higher in the patient group than in controls (P < 0.05). The difference in serotonin positive area in the submucosa in the CI group compared to the control group was not statistically significant. There was a very significant correlation between the serotonin positive area in the submucosa and muscularis propria in controls (r=0.65, p < 0.01), but not in patients with CI.

Conclusion

The increased serotonin level in patients with CI may contribute to the disease or be an adaptive response to some other pathology. The lack of a positive correlation in serotonin levels between the submucosa and muscularis propria in CI patients suggests that the coordinated distribution of serotonin may be disrupted in CI.
Diverticular colitis – therapeutic and aetiological considerations
Authors: Evans J. P. , Cooper J. , Roediger W. E. W.
Published: March 2003

Objective

Treatment of diverticulitis may change when associated with mucosal inflammation of either ulcerative‐like or Crohns‐like colitis. To determine effective treatment of diverticular colitis, four cases were analysed. Mechanisms to account for the colitis associated with diverticulitis are put forward.

Patients and treatment

Four cases had colitis and active diverticulitis established by clinical, colonoscopic or CT evidence of active inflammation. Biopsies confirmed mucosal inflammation: two with ulcerative colitis‐like and two with granulomata suggestive of Crohns‐like colitis. Treatment for colitis in all cases included sulphasalazine and steroids in two cases.

Results

Colitis subsided in three cases but one had continuing inflammatory polyps and one case did not resolve. Segmental resection was performed in two cases one with continuing colitis and one with inflammatory polyps. No further attacks of colitis have occurred since the initial observations were made or operations performed.

Conclusion

An association of colitis with diverticulitis has been confirmed by present observations and case reports of others. Colitis requires medical treatment and if inflammation fails to resolve, segmental resection is indicated. Diverticular colitis, either ulcerative colitis‐like or Crohns‐like, is part of the spectrum of acute diverticulitis.
Prospective analysis of quality of life after reversal of a defunctioning loop ileostomy
Authors: Camilleri‐Brennan J. , Steele R. J. C.
Published: March 2003

Objective

To measure changes in patients' quality of life following reversal of a defunctioning ileostomy in patients who had a curative anterior resection for rectal cancer.

Patients and methods

Twenty consecutive patients undergoing reversal of a loop ileostomy following low anterior resection for primary rectal cancer participated. They answered three quality of life questionnaires: the European Organization for the Research and Treatment of Cancer (EORTC) QLQ‐C30 and QLQ‐CR38, and the SF‐36. Questionnaires were answered before reversal, on discharge home, and at 3 and 6 months postoperatively.

Results

Reversal of the defunctioning ileostomy resulted in significant improvements in global quality of life, physical function, social function, role‐physical and energy‐vitality scores. Post‐operative diarrhoea and frequency improved over six months, but one third of patients had mild incontinence of faeces which did not improve with time. Emotional function, mental health, future perspective, perception of body image and general health perception did not change significantly after reversal. There was a significant negative correlation between defaecation‐related problems, global quality of life and body image.

Conclusion

Reversal of a defunctioning loop ileostomy improves most patients' overall quality of life. Therefore, once it is considered safe to reverse a stoma, an early operation may give the patient a longer time with an improved quality of life.
Hyperplastic polyposis of the colon and rectum
Authors: Renaut A. J. , Douglas P. R. , Newstead G. L.
Published: March 2003

The significance of multiple hyperplastic polyps in relation to the risk of colon cancer is unknown although recent investigation suggests a causative link. We have prospectively identified a small but distinct group of patients that also suggests an association. These patients have either numerous (usually more than twenty, in sites other than the rectosigmoid alone) or large (greater than 1 cm) hyperplastic polyps, in association with either adenomatous polyps, polyps of mixed pathology or carcinoma of the colon and rectum. Additionally, there is frequently a first or second degree family history of colorectal carcinoma.
The contemporary management of haemorrhoids
Authors: Beattie G. C. , Wilson R. G. , Loudon M. A.
Published: March 2003

Background  New concepts in the management of haemorrhoidal disease have recently rekindled interest in this common pathology. General and subspecialist colorectal surgeons were surveyed to assess their impact on the current management of haemorrhoids.

Methods  A questionnaire was sent to all members of the Association of Coloproctology of Great Britain and Ireland (ACPGBI) and the Association of Surgeons of Great Britain and Ireland (ASGBI). Regarding indications for surgery, surgical techniques, day case haemorrhoidectomy (DCH) and postoperative treatment regimens. ASGBI members were asked to state their subspecialist interest and estimated time devoted to colorectal practice.

Results  There were 406 (71%) ACPGBI respondents and 483 (68%) ASGBI respondents. Eighty‐four (12%) ASGBI respondents performed no elective colorectal surgery. One hundred and ninety‐nine (35%) of ACPGBI respondents saw between 6 and 10 new haemorrhoid patients per week whereas three hundred (42%) of ASGBI respondents saw between 1 and 5 per week. Non‐operative management included routine advice on fluid and diet by the majority of surgeons, with banding carried out in 79% (ACPGBI) and 75% (ASGBI) and injection sclerotherapy in 61% (ACPGBI) and 56% (ASGBI). The Milligan Morgan haemorrhoidectomy was performed in 265 (46%; ACPGBI) and 336 (47%; ASGBI). ACPGBI members used Submucosal diathermy (148, 26%vs 67, 9%; ASGBI (P < 0.01; χ2 test with Yates correction)) and stapled anoplasty (61, 11%vs 14, 2%; ASGBI (P < 0.01; χ2 test with Yates correction)) more often. DCH was performed in 117 (20%; ACPGBI) and in 48 (7%; ASGBI)(P < 0.01; χ2 test with Yates correction).

Conclusions  In this sample of surgeons, operative management varies according to specialist interest. There was a trend towards day case haemorrhoidectomy. Whilst more surgeons have accepted the use of postoperative techniques to reduce pain, only a small minority have, as yet, adopted new surgical techniques such as stapling.
A novel indicator for surveillance colonoscopy following colorectal cancer resection
Authors: Skaife P. , Seow‐Choen F. , Eu K. W. , Tang C. L.
Published: March 2003

Introduction  Current surveillance for recurrent intraluminal or metachronous colorectal cancer following resection is largely undertaken by colonoscopic examination of the remaining colon. The burden on colonoscopic services is high and the procedure is expensive. Immunological faecal occult blood testing (FOBT) is a sensitive and specific test for detecting colorectal cancer, and may fine tune the need for timely surveillance colonoscopy.

Methods  Consecutive patients due for surveillance colonoscopy following colonic resection for cancer were prospectively studied. Each patient had a single faecal sample obtained at per rectal examination on a gloved examining finger. This was subjected to immunological FOBT in the clinic, and patients were categorized as FOBT positive or negative, according to the result. Colonoscopy as well as ultrasound or CT of the liver were performed within eight weeks of FOBT.

Results  Six hundred and eleven patients had both FOBT and colonoscopy. Fifty‐nine (13.6%) were categorized as FOBT‐positive. Of these, nine had biopsy‐proven recurrent or metachronous cancer, 12 patients had one, or more adenomatous polyps, one patient had radiation proctitis and two patients had pan‐colonic mucositis following chemotherapy. In the remaining 552 FOBT‐negative patients, no cancers were found. Thirty‐eight patients had polyps that were removed. The sensitivity and specificity for detecting cancer by immunological FOBT was 100% sensitivity for detecting adenomatous polyps was 24% but specificity was 93%.

Conclusion  The immunological faecal occult blood test provides sensitive detection of metachronous and recurrent cancer in postoperative surveillance. Routine application may be used to reduce the frequency of colonoscopic surveillance, as a negative FOBT may be taken as a sign that colonoscopy may be deferred safely.
Effects of niacin‐bound chromium supplementation on body composition in overweight African‐American women
Authors: Crawford V. , Scheckenbach R. , Preuss H. G.
Published: March 2003

Aim: This pilot study was designed to determine whether 600 μg niacin‐bound chromium ingested daily over 2 months by African‐American women undergoing a modest dietary and exercise regimen influences weight loss and body composition.

Methods: Twenty overweight African‐American women, engaged in a modest diet–exercise regimen, participated in a randomized, double‐blinded, placebo‐controlled, crossover study. They received placebo three times a day (t.i.d.) during the control period and niacin‐bound chromium, 200 μg t.i.d., during the verum period. Control and verum periods were each 2 months in duration. One‐half received placebo first (group 1), the other half received chromium first (group 2). Body weights (b.w.) and blood chemistries were measured by routine clinical methodology. Fat and nonfat body masses were estimated using bioelectrical impedance (electrolipography).

Results: In the first group of 10 women receiving niacin‐bound chromium after the placebo period (group 1), b.w. loss was essentially the same, but fat loss was significantly greater and non‐fat body mass loss significantly less with chromium intake. In contrast to the previous findings, there was a significantly greater loss of fat in the placebo compared to the verum period in the second group of eight women who received chromium first (group 2). Blood chemistries were not affected by intake of chromium for 2 months.

Conclusions: Niacin‐bound chromium given to modestly dieting‐exercising African‐American women caused a significant loss of fat and sparing of muscle compared to placebo. Once chromium was given at these dose levels, there was a ‘carry‐over’ effect. Blood chemistries revealed no significant adverse effects from the ingestion of 600 μg of niacin‐bound chromium daily over 2 months.
Pathogenesis of vascular disease
Authors: Colwell John
Published: March 2003

Only recently are we beginning to understand the complex interplay of factors involved in vascular disease and diabetes. Insulin resistance provides a starting point to explain the many factors that lead to the more severe vascular disease characteristic of diabetes. Insulin resistance syndrome comprises insulin resistance and compensatory hyperinsulinaemia as well as hypertension, dyslipidaemia, macrovascular disease, and increased plasminogen activator inhibitor‐1 activity. The development of type 2 diabetes may be viewed as the inability of the pancreas to continue to overcome insulin resistance, even with excessive insulin production.