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1.  Association of BANK1 and TNFSF4 with systemic lupus erythematosus in Hong Kong Chinese 
Genes and Immunity  2009;10(5):414-420.
Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease with complex genetic inheritance. Recently, single nucleotide polymorphisms (SNPs) in BANK1 and TNFSF4 have been shown to be associated with SLE in Caucasian populations, but it is not known whether they are also involved in the disease in other ethnic groups. Recent data from our genome-wide association study (GWAS) for 314 SLE cases and 920 controls collected in Hong Kong identified SNPs in and around BANK1 and TNFSF4 to be associated with SLE risk. On the basis of the results of the reported studies and our GWAS, SNPs were selected for further genotyping in 949 SLE patients (overlapping with the 314 cases in our GWAS) and non-overlapping 1042 healthy controls. We confirmed the associations of BANK1 and TNFSF4 with SLE in Chinese (BANK1, rs3733197, odds ratio (OR)=0.84, P=0.021; BANK1, rs17266594, OR=0.61, P=4.67 × 10−9; TNFSF4, rs844648, OR=1.22, P=2.47 × 10−3; TNFSF4, rs2205960, OR=1.30, P=2.41 × 10−4). Another SNP located in intron 1 of BANK1, rs4522865, was separately replicated by Sequenom in 360 cases and 360 controls and was also confirmed to be associated with SLE (OR=0.725, P=2.93 × 10−3). Logistic regression analysis showed that rs3733197 (A383T in ankyrin domain) and rs17266594 (a branch point-site SNP) from BANK1 had independent contributions towards the disease association (P=0.037 and 6.63 × 10−8, respectively). In TNFSF4, rs2205960 was associated with SLE independently from the effect of rs844648 (P=6.26 × 10−3), but not vice versa (P=0.55). These findings suggest that multiple independent genetic variants may be present within the gene locus, which exert their effects on SLE pathogenesis through different mechanisms.
doi:10.1038/gene.2009.16
PMCID: PMC2834352  PMID: 19357697
SLE; BANK1; TNFSF4; Chinese; genetic association
2.  EpCAM and gpA33 are markers of Barrett's metaplasia 
Journal of Clinical Pathology  2006;59(3):260-263.
Aims
To characterise a specific and sensitive marker of Barrett's metaplasia (BM).
Methods
Cases of normal oesophageal squamous mucosa (11 fresh endoscopic biopsies and 10 formalin fixed, paraffin embedded tissue blocks), BM (11 biopsies and 11 tissue blocks), and normal gastric body mucosa (five biopsies and five tissue blocks) were analysed using reverse transcriptase PCR, Western blotting, and immunohistochemistry for EpCAM, and reverse transcriptase PCR for gpA33.
Results
Strong EpCAM mRNA expression was detected in all the BM cases, in contrast to weak expression in all the normal gastric mucosal samples and no expression in any of the normal oesophageal mucosal samples tested. Strong gpA33 mRNA expression was detected in all the BM cases, in contrast to weak expression in a quarter of the normal gastric mucosal samples and no expression in any of the normal oesophageal mucosal samples tested. Western blotting showed EpCAM protein expression in all the BM cases and in none of the normal gastric or oesophageal mucosal samples tested. Immunohistochemistry showed strong EpCAM protein expression in BM and complete absence of expression in normal oesophageal squamous epithelium. Scattered EpCAM expressing cells were found in the gland bases of normal gastric body mucosa.
Conclusions
EpCAM protein and gpA33 mRNA expressions are specific and sensitive markers of BM.
doi:10.1136/jcp.2005.027474
PMCID: PMC1860330  PMID: 16473928
Barrett's metaplasia; diagnostic marker; EpCAM; gpA33
3.  Purinergic regulation of angiogenesis by human breast carcinoma-secreted nucleoside diphosphate kinase 
British Journal of Cancer  2007;97(10):1372-1380.
MDA-MB-435S human breast cancer cells (435S) secrete nucleoside diphosphate kinase (NDPK) that supports metastases and is inhibited by epigallocatechin gallate (EGCG) and ellagic acid (EA). We hypothesise that 435S cell-secreted NDPK-B supports tumour formation by modulating ATP levels locally to activate endothelial cell (EC) P2Y receptor-mediated angiogenesis. Epigallocatechin gallate (IC50=8–10 μM) and EA (IC50=2–3 μM) suppressed 435S cell growth, but had less effect on human CD31+ EC growth. Epigallocatechin gallate (IC50=11 μM) and EA (IC50=1 μM) also prevented CD31+ EC tubulogenesis on Matrigel™. 435S cell-conditioned media induced tubulogenesis in a cell number, time, and nucleotide-dependent manner. Ellagic acid (1 μM), but not equimolar EGCG, reduced cell number-dependent angiogenesis. P2Y1 receptor activation by NDPK-generated nucleotide (100 μM ATP) or by 10 μM 2-methyl-thio-ATP (2MS-ATP) promoted tubulogenesis on collagen and was blocked by the P2Y1 antagonist MRS2179 (10 μM). Physiological amounts of purified as well as 435S cell-secreted NDPK also promoted angiogenesis that was attenuated by NDPK depletion or 10 μM MRS2179, indicating a P2Y1 receptor-mediated pathway. These results support the notion that secreted NDPK mediates angiogenesis via P2Y receptor signalling and suggests that novel inhibitors of NDPK may be useful as therapeutics.
doi:10.1038/sj.bjc.6604019
PMCID: PMC2360243  PMID: 17940513
breast cancer; tumour angiogenesis; nucleoside diphosphate kinase; polyphenolic compounds; P2Y receptors
5.  SV40 Tag DNA sequences, present in a small proportion of human hepatocellular carcinomas, are associated with reduced survival 
Journal of Clinical Pathology  2003;56(12):904-909.
Aims: To study the association between simian virus 40 (SV40) and human hepatocarcinogenesis.
Methods: Polymerase chain reaction (PCR) to detect SV40 large T antigen (Tag) DNA was performed on: 50 human hepatocellular carcinoma (HCCs) diagnosed between 1978 and 1989 (cohort A); 20 cases of alcoholic liver cirrhosis from the same period; and 20 HCCs diagnosed after 1997 (cohort B). PCR to detect SV40 regulatory sequence and SV40 Tag immunohistochemistry were performed on selected cases from cohorts A and B. Amplified products were directly sequenced. Immunohistochemistry for p53 and pRb and clinicopathological analyses were performed on selected cases from cohorts A and B. Complete survival data were collected for cohort A.
Result: SV40 Tag DNA was found in five cohort A HCCs but not in alcoholic liver cirrhosis cases or cohort B HCCs. Neither SV40 regulatory sequence nor SV40 Tag protein were demonstrated in Tag DNA positive HCCs. No clinicopathological differences existed between Tag DNA positive and negative HCCs, but the presence of Tag DNA was associated with reduced disease specific survival. Relatively fewer Tag DNA positive than negative HCCs expressed p53, but loss of pRb expression was similar in the two groups. Patients with Tag DNA positive HCCs were unlikely to have received SV40 contaminated poliovirus vaccine.
Conclusions: SV40 Tag DNA is present in a small proportion of historical HCCs and may contribute to their pathogenesis and influence their outcome. The source of the virus is uncertain and more recent HCCs show no evidence of SV40.
PMCID: PMC1770131  PMID: 14645347
Simian virus 40; hepatocellular carcinoma; prognosis; p53; retinoblastoma protein
7.  Change in the penetrance of founder BRCA1/2 mutations? A retrospective cohort study 
Journal of Medical Genetics  2002;39(6):407-409.
doi:10.1136/jmg.39.6.407
PMCID: PMC1735145  PMID: 12070248
8.  Glomeruloid microvascular proliferation is associated with p53 expression, germline BRCA1 mutations and an adverse outcome following breast cancer 
British Journal of Cancer  2003;89(6):1031-1034.
doi:10.1038/sj.bjc.6601195
PMCID: PMC2376955  PMID: 12966421
hereditary breast cancer; angiogenesis; factor VIII; chemotheraphy; survival
9.  Nuclear thymidylate synthase expression, p53 expression and 5FU response in colorectal carcinoma 
British Journal of Cancer  2001;85(12):1937-1943.
Thymidylate synthase (TS) is a key enzyme in DNA synthesis and is inhibited by metabolites of the chemotherapeutic agent 5-fluorouracil (5FU). Nuclear expression of TS in human tissue in vivo has not been characterised and its clinicopathological correlates in malignancy are unknown. 52 cases of primary colorectal carcinoma (CRC) and 24 cases of matched metastatic carcinoma were studied immunohistochemically using the monoclonal antibody TS106. The degree of nuclear TS immunostaining correlated closely with levels of TS mRNA expression amongst 10 CRCs studied. Strong nuclear immunostaining was seen in normal basal crypt colonocytes and germinal centre cells, and in a varying proportion of adenocarcinoma cells. Amongst the primary carcinomas, higher TS nuclear expression was associated with prominent extracellular mucin production and right-sided location. Higher TS nuclear expression also showed a significant association with poorer response to protracted venous infusional 5FU therapy. There was no clear association between TS nuclear expression and Ki67 or p53 expression assessed immunohistochemically. There was a strong positive correlation between TS nuclear expression in primary and metastatic CRC but the latter generally showed higher expression than matched primary tumour tissue. These findings confirm the nuclear expression of TS protein in human cells in vivo and provide new insight into how such expression may relate to the behaviour of CRCs. © 2001 Cancer Research Campaign http://www.bjcancer.com
doi:10.1054/bjoc.2001.2175
PMCID: PMC2364004  PMID: 11747337
thymidylate synthase; colorectal carcinoma; immunohistochemistry; 5-fluorouracil; p53 protein; metastases
11.  Measurement of vascularity as a diagnostic and prognostic tool for well differentiated thyroid tumours: comparison of different methods of assessing vascularity. 
Journal of Clinical Pathology  1999;52(8):593-597.
AIMS: To determine whether the measurement of vascularity can be used to differentiate follicular adenomas from follicular carcinomas or to reflect the prognosis of follicular carcinomas and papillary carcinomas of the thyroid gland, and to compare four methods of assessing vascularity. METHODS: Tissue sections from 26 papillary carcinomas, 15 follicular adenomas, and 15 follicular carcinomas were stained with an antibody to CD34. A computerised image analysis system was used to calculate, for each tumour, mean endothelial areas and the mean endothelium to tumour epithelial nucleus area ratio from 10 systematically selected fields across one dimension of the tumour ("systematic field" analysis) or from the three most vascularised fields of the tumour ("hot spot" analysis). A European Organisation for Research on Treatment of Cancer (EORTC) prognostic index was calculated for each papillary carcinoma and follicular carcinoma. RESULTS: Significant differences in vascularity between the three tumour groups could only be shown by comparing mean endothelial area values measured from hot spots. While the hot spot median mean endothelial area of follicular carcinomas was significantly greater than that of follicular adenomas, there was a large overlap between the two groups. For follicular carcinomas, higher hot spot mean endothelial area values were related to worse prognosis as indicated by the EORTC prognostic indices. No association between vascularity and prognosis was found for the papillary carcinomas, regardless of the method of assessing vascularity. CONCLUSIONS: Measuring endothelial area from hot spots using a computerised image analysis system is a sensitive method of assessing the vascularity of thyroid tumours. While vascularity measurement cannot be recommended as a practical tool for differentiating between malignant and benign follicular tumours, the suggestion that vascularity may reflect prognosis for follicular carcinomas deserves further study.
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PMCID: PMC500950  PMID: 10645229
12.  Primary yolk sac tumour of the liver in adulthood. 
Journal of Clinical Pathology  1998;51(12):939-940.
Primary yolk sac tumour of the liver is exceedingly rare. A 28 year old woman presented with a cystic liver mass and a markedly raised serum alpha-fetoprotein concentration. She underwent a partial hepatectomy for a suspected hepatocellular carcinoma but histological examination of the tumour revealed the classical morphological and immunohistochemical features of a yolk sac tumour. There was no evidence of an extrahepatic primary source. Review of this case, together with the six previously reported adult cases of primary yolk sac tumours of the liver, revealed several features of the tumour that may aid differentiation from hepatocellular carcinoma, with potential therapeutic implications.
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PMCID: PMC501033  PMID: 10070339
13.  An analysis of discharge drug prescribing amongst elderly patients with renal impairment. 
Postgraduate Medical Journal  1998;74(873):420-422.
Renal impairment is common amongst elderly patients and increases the risk of drug toxicity. Analysis of the discharge summaries of patients discharged from the geratology wards of an Oxfordshire hospital, showed that renal impairment was not referred to in 42% of patients with calculated creatinine clearances of 10-20 ml/min. A fifth of these patients, and 67% of patients with calculated creatinine clearances < 10 ml/min, had discharge drug prescriptions that contradicted the British National Formulary guidelines for prescribing in the presence of renal impairment. There should be increased awareness that, in elderly patients, normal serum creatinine concentrations do not exclude renal impairment and that several commonly prescribed drugs require dose adjustments or should be avoided in the presence of renal insufficiency.
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PMCID: PMC2360983  PMID: 9799917
14.  Unacceptably high site variability in postmortem blood alcohol analysis. 
Journal of Clinical Pathology  1998;51(3):250-252.
Blood alcohol concentration is a frequently requested test in forensic pathology. The variability of this value was studied by measuring the blood alcohol concentration from six sites in nine subjects at necropsy in whom alcohol was the implicated cause of death. There were small consistent differences in the blood alcohol concentrations between the sites in the nine subjects (p < 0.04). Calculation of the mean blood:vitreous humour alcohol concentration ratio (B:V ratio) showed that vitreous humour alcohol concentration most closely reflected the concentration at the femoral vein (B:V ratio = 0.94, r = 0.98), which is considered the optimal site for blood alcohol measurement. The correlation of left heart blood with femoral blood was lower compared with the other sites. There is a potential for an unacceptably large variation in the postmortem measurement of blood alcohol within each subject.
PMCID: PMC500652  PMID: 9659273
15.  The effect of the I1307K APC polymorphism on the clinicopathological features and natural history of breast cancer 
British Journal of Cancer  1999;81(5):850-854.
The I1307K polymorphism in APC has been found to predispose to colorectal cancer in Ashkenazi Jews, and has recently been associated with an increased risk for breast cancer in the same population. In that study, we genotyped 205 paraffin-embedded breast cancers from Ashkenazi Jewish women diagnosed below the age of 65. We now present an extended analysis, with clinicopathological correlations between carriers of I1307K and non-carriers. Twenty-four of 209 cases (11.5%, 95% confidence interval 7.5–16.6) were found to carry the I1307K polymorphism. When stratifying the data by other relevant clinicopathological variables, we observed no association between the presence of this polymorphism and age at diagnosis (P = 0.52), grade (P = 0.074), tumour size (P = 0.99), lymph node status (P = 0.82), oestrogen receptor status (P = 0.23) or P53 immunoreactivity (P = 0.80). The breast-cancer specific 5-year survival for women with I1307 K polymorphism was 88.9% compared with 81.6% in women without I1307K (P = 0.34). Using microdissected samples and direct sequencing, no somatic mutations were observed in any of the 24 I1307K-positive cases. Single-strand conformation analysis of 158 of the I1307K-negative breast cancers that were available for study revealed no mobility shifts. We conclude that the presence of the I1307K polymorphism does not appear to be associated with any particular clinicopathological feature of breast cancer and importantly, does not affect the prognosis. © 1999 Cancer Research Campaign
doi:10.1038/sj.bjc.6690775
PMCID: PMC2374295  PMID: 10555757
breast cancer; APC; I1307K; polymorphism; survival
16.  Genetic polymorphisms of cytochrome p4502E1 and susceptibility to alcoholic liver disease and hepatocellular carcinoma in a white population: a study and literature review, including meta-analysis 
Molecular Pathology  2000;53(2):88-93.
Aims—To investigate the associations between the Rsa I, Dra I, and Taq I genetic polymorphisms of cytochrome p4502E1 and susceptibility to alcoholic liver disease or to hepatocellular carcinoma.
Methods—DNA samples isolated from 61 patients with alcoholic liver disease, 46 patients with hepatocellular carcinoma, and 375 healthy controls were subjected to polymerase chain reaction amplification followed by digestion with the endonucleases Rsa I, Dra I, or Taq I. Meta-analysis was performed using data from previous studies of Rsa I polymorphism and the risk of alcoholic liver disease.
Results—No association was found between any of the three polymorphisms and susceptibility to hepatocellular carcinoma. The distributions of Rsa I and Dra I alleles among the patients with alcoholic liver disease were not significantly different from those among the control group. Meta-analysis of this data and previous data concerning Rsa I polymorphism and alcoholic liver disease risk failed to demonstrate any significant association between the two. However, the alcoholic liver disease group in this study showed a significantly lower frequency of the less common Taq I allele compared with the healthy control group (odds ratio, 0.33; 95% confidence interval, 0.12 to 0.78).
Conclusions—Possession of the less common Taq I cytochrome p4502E1 allele is associated with reduced susceptibility to alcoholic liver disease. There is no existing evidence that the Taq I polymorphism is directly associated with altered alcohol metabolism, but it might be in linkage disequilibrium with as yet unidentified protective factors.
PMCID: PMC1186911  PMID: 10889908
cytochrome p4502E1; alcoholic liver disease; hepatocellular carcinoma
17.  Lethargy in a patient with cirrhosis. 
Postgraduate Medical Journal  1997;73(857):177-179.
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PMCID: PMC2431269  PMID: 9135840
19.  Small cell variant of Ki-1 lymphoma associated with myelofibrosis and a novel constitutional chromosomal translocation t(3;4) (q13;q12). 
Journal of Clinical Pathology  1996;49(3):259-262.
An unusual case of small cell variant of Ki-1 non-Hodgkin's lymphoma diagnosed one year after an original diagnosis of idiopathic myelofibrosis is reported. On the second occasion, the patient presented with fever, lymphadenopathy and hepatosplenomegaly. A lymph node biopsy specimen confirmed a diagnosis of small cell variant of Ki-1 lymphoma. A repeat bone marrow biopsy specimen showed myelofibrosis with no evidence of lymphomatous infiltration, but cytogenetic studies on blood, bone marrow and skin fibroblasts revealed a novel chromosomal translocation t(3,4)(q13;q12).
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PMCID: PMC500412  PMID: 8675743
20.  Acute respiratory distress secondary to Morgagni diaphragmatic herniation in an adult. 
Postgraduate Medical Journal  1995;71(831):39-41.
Diaphragmatic hernias of the Morgagni type are generally thought to be asymptomatic in adults. This traditional assumption led to a delay in diagnosing a Morgagni hernia as the cause of acute respiratory distress in a chronic schizophrenic man. While Morgagni hernias are usually considered to be long-standing, we present radiological evidence of an acutely expanding hernia. The patient's symptoms were relieved by surgical repair of the hernia. We advise caution before dismissing Morgagni hernias in adults as being long-standing and clinically insignificant.
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PMCID: PMC2397904  PMID: 7708592
21.  Randomly amplified polymorphic DNA typing: a useful tool for rapid epidemiological typing of Klebsiella pneumoniae. 
Epidemiology and Infection  1994;113(3):445-454.
Discriminatory typing methods are invaluable in the investigation of outbreaks of infectious diseases. Single primers were used to generate randomly amplified polymorphic DNA (RAPD) profiles from Klebsiella pneumoniae isolates of various serotype and K. pneumoniae isolates from cases of sepsis at a Malaysian hospital and two English hospitals. RAPD profiles of acceptable reproducibility, a maximum of three minor band variations, were produced using a rapid DNA extraction method. RAPD typing of K. pneumoniae was shown to be as discriminatory as restriction fragment length polymorphism analysis using pulsed field gel electrophoresis yet quicker and less costly. The findings suggest that RAPD typing may be a useful tool for the epidemiological typing of K. pneumoniae.
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PMCID: PMC2271319  PMID: 7995354
22.  Inhibition of expression of major histocompatibility complex class II molecules in macrophages infected with Leishmania donovani occurs at the level of gene transcription via a cyclic AMP-independent mechanism. 
Infection and Immunity  1992;60(5):2115-2120.
Among the important pleiotropic responses to gamma interferon (IFN-gamma) during the activation of macrophages (M phi) is the increased expression of major histocompatibility complex class II genes. In the present study, infection with Leishmania donovani was shown to inhibit in parallel the induction by IFN-gamma of H-2 A beta gene transcription, class II mRNA accumulation, and H-2 Ad protein expression in cells of the murine macrophage cell line P388D1. Treatment of P388D1 cells with either the adenylate cyclase activator cholera toxin or the protein kinase A activator N6-2'-O-dibutyryl cyclic AMP (dibutyryl cAMP) similarly inhibited the induction by IFN-gamma of class II protein expression, and in parallel with Leishmania infection, cholera toxin inhibited the induction of mRNA for the H-2 A alpha and H-2 A beta proteins. Concentrations of intracellular cAMP were significantly increased in cholera toxin-treated cells but not in leishmania-infected cells. These findings indicate that at least one mechanism by which Leishmania infection attenuates the activation of M phi by IFN-gamma involves selective, transcriptional inhibition of major histocompatibility complex class II genes via a cAMP-independent mechanism.
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PMCID: PMC257124  PMID: 1314226
23.  A comparison of Chinese traditional and Western medical approaches for the treatment of mild hypertension. 
We compared the efficacy of Chinese traditional treatment for mild hypertension with that of a standard Western medical regimen in a group of 50 well-matched patients (24 allocated to Western medicine and 26 to Chinese traditional medicine) with mild hypertension (diastolic blood pressure 90-104 mmHg). Those receiving Western therapy were treated in a stepped-care fashion with dihydrochlorothiazide and atenolol. Those in the Chinese traditional therapy group received one of two mixtures of nine herbs and other ingredients, depending on symptoms at initial evaluation. Blood pressure dropped significantly in both groups after only a few days on therapy. After 19 days on treatment, the group receiving Western therapy had a fall in blood pressure from 168.2/96.3 mmHg to 137.3/76.7 mmHg (p less than 0.01), while those on Chinese traditional therapy fell from 168.2/95.9 mmHg to 146.4/80.5 mmHg (p less than 0.01). The fall in blood pressure was significantly greater, however, in those given Western therapy. The relief of existing symptoms or development of possible drug side effects was similar in both groups, except for nocturia, occurring more often in the group treated with Western therapy. We conclude that Western therapy is more effective in reducing blood pressure as compared with Chinese traditional therapy, but effective control of blood pressure in mild hypertensives is possible with either form of treatment.
PMCID: PMC2589443  PMID: 1897264
24.  Liver abscess in the tropics: experience in the University Hospital, Kuala Lumpur. 
Postgraduate Medical Journal  1987;63(741):551-554.
We reviewed 204 cases of liver abscess seen between 1970 and 1985. Ninety were found to be amoebic, 24 pyogenic and one tuberculous. The cause of the abscesses in the remaining 89 patients was not established. The patients were predominantly male, Indians, and in the 30-60 age group. The majority of patients presented with fever and right hypochondrial pain. The most common laboratory findings were leucocytosis, hypoalbuminaemia and an elevated serum alkaline phosphatase. Amoebic abscesses were mainly solitary while pyogenic abscesses were mainly multiple. Complications were few in our patients and included rupture into the pleural and peritoneal cavities and septicaemic shock. An overall mortality of 2.9% was recorded. The difficulty in diagnosing the abscess type is highlighted. The single most important test in helping us diagnose amoebic abscess, presumably the most common type of abscess in the tropics, is the Entamoeba histolytica antibody assay. This test should be used more frequently in the tropics.
PMCID: PMC2428368  PMID: 3658864
25.  Comparison of temporary and permanent catheters for acute peritoneal dialysis. 
Archives of Disease in Childhood  1988;63(7):827-831.
The records of 64 children who underwent acute peritoneal dialysis in the Hospital for Sick Children, Toronto, between 1 January 1982 and 31 December 1985 were reviewed. Rigid Trocath catheters were used in 33 patients and silastic Tenckhoff catheters in 34. The groups were comparable for age, weight, primary disease, duration of dialysis, and outcome. There were no significant differences in the incidences of leakage of dialysate, two way obstruction, bleeding, or peritonitis. Among those who had Trocath catheters there were significantly more episodes of outflow obstruction, and significantly more patients required two or more catheter insertions (12 of 33 compared with one of 34 among those who had Tenckhoff catheters). Furthermore, more catheters were removed because of complications (19 of 33 compared with five of 34 respectively). One patient who had a Tenckhoff catheter died of peritonitis directly related to insertion of the catheter. Overall, the Tenckhoff catheter was associated with fewer complications and is recommended for most patients.
PMCID: PMC1779093  PMID: 3415301

Results 1-25 (32)