bluuebunny/crossref_metadata_2025 huggingface dataset:
Establishment of a testicular carcinoma cell line producing α‐fetoprotein
Authors: Ueno M. , Nakashima J. , Ohigashi T. , Deguchi N. , Ban S. , Akita M. , Murai M.
Published: March 2003

Objective To characterize a newly established human testicular carcinoma cell line that continuously produces α‐fetoprotein (AFP), and to investigate the effects of retinoic acid on AFP production.

Materials and methods A 24‐year‐old man underwent a radical orchidectomy for a right testicular tumour and was found to have two separate metastatic lesions in the lungs, both of which were removed surgically. The cancer cells were isolated from one of the tumours, which was composed of undifferentiated germ cells and produced AFP; the cells were cultured in a monolayer. This cell line was designated as KU‐MT.

Results The cell line was successfully maintained both in athymic nude mice and in culture. Histological examination showed that the xenografted tumours were composed of cells in the reticular, solid and glandular patterns of a yolk sac tumour, and of embryonal carcinoma cells. These cells immunostained positively for AFP. On electron microscopy, the extracellular deposition of a basement lamina‐like substance, a typical feature of yolk sac tumour, was detected. The AFP production in mice correlated well with the tumour weight of the xenograft. The cultured KU‐MT cells were oval to polygonal in morphology and grew exponentially, with a population doubling time of ≈ 2 days. Chromosomal analysis showed a modal number of 57 with consistent structural abnormalities of +add(1)(p13), del(1)(q32), del(2)(q31), add(6) (q21), +add(9)(p22), add(11)(p15), and add(14)(p11). Reverse‐transcription polymerase chain reaction analysis showed that the retinoic acid receptors (RAR)‐α, RAR‐γ, and retinoid X receptor‐α were present in the cells. The expression of AFP mRNA was up‐regulated in response to all‐trans‐retinoic acid; treatment with this agent caused morphological changes and induced apoptosis in the cells.

Conclusions This newly established cell line provides a reproducible model system that should offer a good insight into the differentiation of testicular carcinoma.
The urethral plate – does it grow into the genital tubercle or within it?
Authors: Penington E.C. , Hutson J.M.
Published: March 2003

Objective To determine whether the urethral plate grows into the genital tubercle or develops within it.

Materials and methods Sprague‐Dawley rat embryos were examined at 11–21 days of gestation and compared with available human embryo sections. Rat embryos were examined by a combination of light microscopy, dissection, serial histological section and immunohistochemistry.

Results In rat embryos the urethral plate extended to the tip of the genital tubercle from the earliest stages of genital tubercle development. The plate formed from adherence in the sagittal plane of the inner walls of the primitive cloacal cavity. The urethral plate continued to extend to the tip of the genital tubercle throughout penile development in the male and developed a lumen ahead of the advancing perineal mesenchyme. In the female the perineal side of the urethral plate disintegrated by massive apoptosis. This loss of perineal substance allowed the genital tubercle to collapse on itself and left the urogenital opening at the base of the genital tubercle. Human embryos also showed the presence of the urethral plate at the tip of the genital tubercle throughout genital tubercle development.

Conclusion The urethral plate forms ab initio within the genital tubercle and extends to the tip; it does not grow into the genital tubercle as is often described. The contribution of ectodermal ingrowth to urethral development at the tip of the penis is minimal.
Prognostic value of the Gleason score in prostate cancer
Authors: Egevad L. , Granfors T. , Karlberg L. , Bergh A. , Stattin P.
Published: March 2003

Objective To investigate the prognostic value of the Gleason score in prostate cancer.

Patients and methods A consecutive series of 305 men with prostate cancer diagnosed at transurethral resection (1975–1990) and with no curative treatment was analysed. There was no assessment of prostate‐specific antigen level during this period. The mean (range) age at diagnosis was 73.7 (52–95) years and the mean follow‐up was 6.4 (0–22) years. The influence of Gleason score and the percentage of the specimen area with tumour (% cancer) on disease‐specific survival were assessed using Kaplan–Meier analyses.

Results Of 305 cancers, 22% had a Gleason score of 4–5, 29% of 6, 18% of 7 and 32% of 8–10. At the follow‐up, 89% of the men had died, of whom 42% had died from prostate cancer. The disease‐specific 10‐year survival was 56%. The disease‐specific mean survival (DSMS) for Gleason score 4–5, 6, 7 and 8–10 was 20, 16, 10 and 5 years, respectively (P < 0.001). The DSMS did not differ significantly between Gleason 4 and 5 or between 8–10. There was a trend towards shorter survival for Gleason 4 + 3=7 (DSMS 9 years) than GS 3 + 4=7 (DSMS 13 years; P = 0.16). Gleason score and % cancer were independent predictors of DSMS (P < 0.001).

Conclusion The long‐term prognosis of prostate cancer on deferred treatment is predicted well by the Gleason score. Four prognostic categories of prostate cancer are suggested, i.e. Gleason score 4–5, 6, 7 and 8–10.
Paediatric penile trauma
Authors: El‐Bahnasawy M.S. , El‐Sherbiny M.T.
Published: March 2003

Objective  To report our experience with paediatric penile trauma in a retrospectively evaluated series.

Patients and methods

 The records of 64 boys (mean age 7 years,
sd 4) who were hospitalized over the last 20 years because of penile trauma were reviewed. The cause of trauma was circumcision in 43 (67%), a human hair‐tie strangulation injury in 10 (16%), an animal attack in four (6%), a bicycle accident in four (6%), a zipper injury in two (3%) and electrical injury in one (2%). Patients were managed according to the severity of the injury. Eight (12%) with minimal skin loss or meatal injury underwent primary skin closure or meatoplasty; 40 (62%) with urethrocutaneous fistulae underwent repair and five (8%) with a glans hanging on a thin pedicle had the glans and the urethra reconstructed. Patients with partial or complete amputation of the glans (10) underwent primary haemostasis and meatoplasty; the penis was lengthened in one. One child with complete avulsion of the penis underwent perineal urethrostomy.

Results

 Fifty‐four patients (84%) were followed for a mean (
sd ) of 5.7 (4) years; there were good cosmetic and functional results in 45 (83%). Fifteen patients are now adults; 13 (86%) reported normal sexual function. Of the 40 patients assessed with circumcision‐related injuries, six (15%) had functional disability (short penis in one and fistulae in five). Of the 10 patients with a hair‐tie injury, none lost their glans. Of the four injuries caused by animal attacks, three had poor results (emasculation in one, short penis in one and severe curvature in the remaining patient). There was no functional disability in the remaining forms of trauma.

Conclusions  In our region, ritual circumcision and hair‐tie strangulation injuries are the most common causes of penile trauma in children. Good functional and cosmetic results are possible in most cases. However, animal attacks are associated with the highest rate of long‐term functional and cosmetic disability.
Polyphenols in red wine inhibit the proliferation and induce apoptosis of LNCaP cells
Authors: Romero I. , Páez A. , Ferruelo A. , Luján M. , Berenguer A.
Published: March 2003

Objective To assess the effect of five polyphenol constituents of red wine (quercetin, morin, rutin, gallic acid and tannic acid) on the proliferation of LNCaP cells, and to quantify the extent of apoptosis with each polyphenol.

Materials and methods LNCaP cells (500) were cultured in microtitre plates and treated with gallic acid, tannic acid, quercetin (1, 5 and 10 µmol/L), rutin and morin (25, 50 and 75 µmol/L). A colorimetric immunoassay was then used to determine the extent of proliferation at 24, 48, 72 and 96 h, and a cell‐death detection assay to assess apoptosis at 24, 48 and 72 h.

Results Gallic and tannic acid (5 and 10 µmol/L), morin (50 and 75 µmol/L), quercetin (5 and 10 µmol/L) and rutin (50 and 75 µmol/L) all significantly inhibited (P<0.05) cell proliferation compared with the control. Apoptotic indexes were significantly greater (P<0.01) in the presence of gallic (5 and 10 µmol/L) and tannic acid (5 and 10 µmol/L), and rutin (75 µmol/L, P<0.05) than in the control. The apoptotic effect of morin (75 µmol/L), although significant (P<0.01), only appeared at 72 h. Conversely, while significant (P<0.05) quercetin (5 and 10 µmol/L) had a transient (first 48 h) apoptotic effect compared with the control.

Conclusion Quercetin, rutin, morin, gallic acid and tannic acid inhibited the growth of LNCaP cells at different concentrations, and induced apoptosis. The results provide a strong rationale for studying the in vivo effects of these compounds.
Does delaying pyeloplasty affect renal function in children with a prenatal diagnosis of pelvi‐ureteric junction obstruction?
Authors: Chertin B. , Rolle U. , Farkas A. , Puri P.
Published: March 2003

Objective  To evaluate retrospectively children with an antenatal diagnosis of pelvi‐ureteric junction (PUJ) obstruction who required pyeloplasty for deteriorating renal function during the follow‐up, specifically assessing the recovery of function after surgery.

Patients and methods

 Between 1988 and 2000, 44 consecutive patients with a prenatal diagnosis of PUJ obstruction who were initially treated by observation underwent pyeloplasty because their renal function deteriorated during the follow‐up. Patients with bilateral disease, a single kidney or vesico‐ureteric reflux were excluded from the study. The mean (
sd
) age of the patients was 13.3 (4.5) months and their mean (range) initial renal function 39.7 (30.4–45)%. In all patients the diagnostic criteria and indications for surgery were identical. The main indication for surgery was deteriorating renal function of >5%, confirmed by renal scintigraphy. Of 44 patients, 35 (77%) had severe dilatation of the renal pelvis by >3 cm. The mean (
sd
) deterioration in renal function during the follow‐up, just before surgery, was 8.2 (2.4)%, and the mean time between the last acceptable renogram and the study showing deterioration was 8.3 (1.3) months; the mean follow‐up was 5.1 (1.6) years. The results were assessed statistically using the unpaired nonparametric Mann–Whitney
U ‐test.

Results  An improvement in hydronephrosis was confirmed in all patients, with renal function returning to the initial levels in 36 of 44 (81%) patients 6–12 months after surgery, although in two patients renal function did not improve after surgery. There was no significant difference between the preoperative characteristics and the degree of hydronephrosis, renal function or patient age with the level of renal functional improvement after surgery.

Conclusion  In patients with an antenatal diagnosis of PUJ obstruction, expectant management is recommended and spares the children unnecessary surgery. Even if renal function deteriorates, delayed pyeloplasty recovers the initial functional level.
Gap junctions and connexin expression in human suburothelial interstitial cells
Authors: Sui G.P. , Rothery S. , Dupont E. , Fry C.H. , Severs N.J.
Published: March 2003

Objective  To determine whether suburothelial interstitial cells of the human bladder express gap junctions, and if so, to establish their extent and composition, using immunocytochemistry, confocal microscopy and electron microscopy.

Materials and methods  Bladder tissue was obtained at cystectomy; the tissue was: (i) frozen for cryosectioning and Northern blot analysis; (ii) fixed and embedded for standard thin‐section electron microscopy; and (iii) processed using low‐denaturation conditions in Lowicryl for immunogold‐label electron microscopy. Cryosections were immunofluorescently labelled using antibodies against connexins 43, 40 and 45, vimentin, desmin and c‐Kit ligand, and examined by confocal microscopy. Double labelling was used to determine the spatial relationship of labelling for connexin43 with that of vimentin and desmin. Thin‐section electron microscopy was used to investigate interstitial cell ultrastructure and permit unequivocal identification of gap junctions, and immunogold labelling of Lowicryl sections was applied to localize connexin43.

Results  Immunoconfocal microscopy showed prominent labelling for the gap junction protein, connexin43, in a suburothelial band of cells that was also strongly positive for vimentin. The connexin43/vimentin‐positive cells showed only weak labelling for desmin and c‐Kit ligand, and were immunonegative for connexins 40 and 45. Northern blotting showed a corresponding abundance of connexin43 transcript in the mucosal layer but not the detrusor layer of the bladder wall. Electron microscopy revealed abundant gap junctions, recognized by their pentalaminar structure, between the cell processes of interstitial cells in the suburothelial zone. That these interstitial cell gap junctions were the source of the connexin43 immunolabelling observed by immunoconfocal microscopy was confirmed by immunogold labelling in sections of Lowicryl‐embedded tissue examined by electron microscopy.

Conclusion  A network of interstitial cells, extensively linked by connexin43‐containing gap junctions, is located beneath the urothelium in human bladder. As gap junctions provide pathways for direct cell‐to‐cell communication, the interstitial cellular network may operate as a functional syncytium, integrating signals and responses in the bladder wall.
Thermo‐expandable intraprostatic stents in bladder outlet obstruction: an 8‐year study
Authors: Perry M.J.A. , Roodhouse A.J. , Gidlow A.B. , Spicer T.G. , Ellis B.W.
Published: March 2003

Objective To assess the use of a thermo‐expandable intraprostatic stent (Memokath®, Engineers and Doctors A/S, Copenhagen, Denmark) for bladder outlet obstruction in men unable to undergo transurethral resection of the prostate (TURP), assessing symptoms, complications and duration of stent life.

Patients and methods The Memokath stent is a coil of a nickel‐titanium alloy which has ‘shape memory’, the lower end expanding when heated to 55 °C. Risks associated with inserting the stent with a flexible cystoscope under local anaesthesia are minimal. Men were selected who were either permanently or temporarily unfit for TURP. Indications included severe respiratory and cardiovascular disease. Exclusion criteria included bladder carcinoma, calculi or detrusor failure; in all, 211 men were fitted with 217 intraprostatic stents over 8 years.

Results There were 1511 TURPs during the study period; the mean age of men receiving a stent was 80.2 years, compared with 70.2 years for those undergoing TURP. The International Prostate Symptom Score decreased from a mean of 20.3 to 8.2 (P < 0.001) in the first 3 months after stent placement; there was virtually no change over 7 years. During the follow‐up, 38% of men died with their stents in situ, 34% remain alive, 23% have had their stents removed for failure and 4% were removed as they were no longer required. There was a 13% migration rate and 16% repositioning rate. There were few side‐effects (pain 3%, haematuria 3%, incontinence 6% and infection 6%). These frail men were more likely to die than have their stent fail.

Conclusion The Memokath intraprostatic stent is a valuable addition to the armamentarium of the urologist treating elderly or frail men with advanced bladder outlet obstruction and complements existing technologies.
Surgical treatment of disseminated peritoneal metastases from urological cancer: results from a prospective study
Authors: Otto T. , Suhr J. , Krege S. , Goebell P.J. , Wishahi M. , Bier U.W. , Kimmig R. , Rübben H.
Published: March 2003

Objective  To report a prospective phase II study of patients with disseminated peritoneal carcinomatosis and symptomatic disease, in whom the peritoneal metastases were resected.

Patients and methods  From 1995 to 1999, 32 patients (20 men and 12 women, median age 56 years, range 32–75) with peritoneal carcinomatosis were enrolled in the trial. Pain and ascites were determined according to the National Cancer Institute score/criteria, and performance scored according to the World Health Organisation criteria.

Results  All patients had intraperitoneal disseminated malignancies with clinically evident ascites, and presented with abdominal pain. The median (range) operative duration was 2.9 (1–5.5) h and the hospital stay 25 (10–44) days, with no deaths at 30 days. The ascites was completely cured in 25 of the 32 patients, pain relieved in 28 and the performance score improved in 25. The median survival time was 1 year; the 1‐ and 2‐year survival rates were 45% and 38%, respectively. Patients with residual metastases after incomplete resection had a significantly worse prognosis, but the prognosis was significantly better in those with a low tumour burden.

Conclusions  Peritoneal carcinomatosis is treatable; radical peritonectomy improves the performance score in selected patients with cancer‐related ascites and/or pain, and is now the standard approach in the authors’ Cancer Centre.
A randomized, double‐blind crossover study of tamsulosin and controlled‐release doxazosin in patients with benign prostatic hyperplasia
Authors: Kirby R.S.
Published: March 2003

OBJECTIVE

To compare the effects of the doxazosin gastrointestinal therapeutic system, extended‐release (doxazosin‐GITS) formulation, and tamsulosin, another α1‐antagonist, on total International Prostate Symptom Score (IPSS) and maximum urinary flow rate (Qmax) in treating patients with benign prostatic hyperplasia (BPH).

PATIENTS AND METHODS

Data were analysed from a prospective, randomized, double‐blind, crossover study of men aged 50–80 years with concomitant BPH and hypertension as inclusion criteria. Fifty‐two men were treated in four phases: phase I, placebo run‐in for 2 weeks; phase II, first study drug doxazosin‐GITS or tamsulosin for 8 weeks; phase III, washout with placebo for 2 weeks; and phase IV, second study drug tamsulosin or doxazosin‐GITS for 8 weeks. Doxazosin‐GITS was started at 4 mg/day and tamsulosin at 0.4 mg/day, and then titrated to 8 mg/day and 0.8 mg/day, respectively, after 4 weeks of therapy if the increase in Qmax was < 3 mL/s or the reduction in total IPSS was < 30%. Efficacy assessments included the IPSS and Qmax. Changes in blood pressure were not analysed, as most patients were actually not hypertensive. Endpoint efficacy data were analysed using an analysis of covariance model, with terms for sequence, phase, patients and sequence within patients, in addition to the baseline as covariate. Forty‐seven men were treated in both efficacy arms of the study and were evaluable for analysis.

RESULTS

Doxazosin‐GITS and tamsulosin significantly relieved lower urinary tract symptoms and significantly increased Qmax from baseline (P = 0.001). Doxazosin‐GITS produced significantly greater improvements than tamsulosin in total IPSS (P = 0.019) and obstructive subscores (P = 0.004) at the last treatment visit. The difference between doxazosin‐GITS and tamsulosin in improving Qmax approached significance in favour of the former (mean change from baseline 2.6 vs 1.7 mL/s, respectively; between‐group difference P = 0.089). Both treatments were well tolerated.

CONCLUSIONS

Treatment with doxazosin‐GITS was significantly more effective than tamsulosin in relieving lower urinary tract symptoms.
The role of neuromodulation in the management of urinary urge incontinence
Authors: Abrams P. , Blaivas J.G. , Fowler C.J. , Fourcroy J.L. , Macdiarmid S.A. , Siegel S.W. , Van Kerrebroeck P.
Published: March 2003

Neuromodulation is increasingly becoming an important part of the treatment strategy for bladder dysfunction. In this issue a group of urologists have analysed outcome measures from all patients in pivotal clinical trials who have had this treatment, and have suggested that sacral nerve stimulation has an vital role in the management of refractory overactive bladder and retention problems.

In a carefully performed study, authors from Mansoura and Scottsdale have evaluated high‐energy TUMT and found that although symptomatic improvement occurred in 82.5% of patients, and that the peak flow rate improved from a median of 9.2 to 15 mL/s, pressure‐flow improvement occurred in just 50% of the group. However, they found that younger patient age and higher grade of obstruction, are good predictors of urodynamic and symptomatic success respectively. In another study, Marshall's group from Adelaide emphasize the value of urodynamics and other factors in patients having TURP. The value of trans‐abdominal ultrasound is described by Foo and his colleagues from Singapore, who state that intravesical protrusion of the prostate is an important indicator of bladder outlet obstruction.

OBJECTIVE

To examine the benefit‐risk profile of neuromodulation in treating refractory urinary urge incontinence and other voiding disorders.

PATIENTS AND METHODS

The outcome measures from all patients in pivotal clinical trials who had undergone sacral nerve stimulation were analysed retrospectively.

RESULTS

Neuromodulation was effective in several clinical studies; the response is durable and the benefit‐risk profile good.

CONCLUSION

Sacral nerve stimulation is becoming the standard of care for refractory overactive bladder and retention problems. The potential benefit of neuromodulation should be included in female urology and gynaecology training programmes.
The evolution of diabetes information systems
Authors: Young R. J. , Khong C. K. , Vaughan N. J. A. , New J. , Roxburgh M.
Published: March 2003

Diabetes information systems have already evolved rapidly in recent years along a developmental pathway initiated by the St Vincent Declaration, fuelled by the rapid pace of IT development in the 1990s and now endorsed by the emerging NHS information strategy. They will be central to the delivery of ‘patient‐centred’ care and essential to supporting and monitoring the diabetes national service framework implementation. Widespread experience has identified three key principles. Firstly the need for a core data set that supports both service delivery and quality development. Secondly, because of the multiprofessional, multisector nature of diabetes care, there is a need to reconcile information from many diverse sources into unitary diabetes care records. Thirdly the crucial importance of making data collection a by‐product of every day care delivery (i.e. no duplicate data entry). The work of many local innovators, allied to the increasing experience of the Diabetes UK sponsored UKDIABS project has generated substantial expertise. With the aid of new extraction/analysis tools such as QUIDS and a consistent approach to assessment, this work has hopefully laid secure foundations for monitoring the implementation of the national service framework. Furthermore, parallel developments under the aegis of the National electronic Library for Health (NeLH) should enable those involved with diabetes care to access relevant knowledge and information with ease. Increasingly user friendly ways by which patients can interact with their electronic records and linked knowledge sources will create many new opportunities. Diabetes information systems are likely to be at the forefront of diabetes care delivery in the future, providing patients and professionals with timely and accurate data for the organization and delivery of care.
Reversal of corticosteroid‐induced diabetes mellitis with supplemental chromium
Authors: Ravina A. , Slezak L. , Mirsky N. , Bryden N. A. , Anderson R. A.
Published: March 2003

Aims To determine if the stress of corticosteroid treatment increases chromium (Cr) losses and if corticosteroid‐induced diabetes (steroid diabetes) can be reversed by supplemental chromium.

Methods The effects of corticosteroid treatment on chromium losses of 13 patients 2 days prior to steroid administration and the first 3 days following treatment were determined. Since steroid‐induced diabetes was associated with increased chromium losses and insufficient dietary chromium is associated with glucose intolerance and diabetes, we treated three patients with steroid‐induced diabetes with 600 μg per day of chromium as chromium picolinate.

Results Urinary chromium losses following corticosteroid treatment increased from 155 ± 28 ng/d before corticosteroid treatment to 244 ± 33 ng/d in the first 3 days following treatment. Chromium supplementation of patients with steroid‐induced diabetes resulted in decreases in fasting blood glucose values from greater than 13.9 mmol/l (250 mg/dl) to less than 8.3 mmol/l (150 mg/dl). Hypoglycaemic drugs were also reduced 50% in all patients when given supplemental chromium.

Conclusions These data demonstrate that corticosteroid treatment increases chromium losses and that steroid‐induced diabetes can be reversed by chromium supplementation. Follow‐up, double‐blind studies are needed to confirm these observations.

Diabet. Med. 16, 164–167 (1999)
Painful diabetic neuropathy
Authors: Benbow S. J. , Cossins L. , MacFarlane I. A.
Published: March 2003

Chronic painful diabetic neuropathy causes symptoms that can last for many years and severely impair the quality of life of affected patients. This review describes the epidemiology, pathophysiology and treatment of chronic neuropathic pain. Particular emphasis is placed on a comprehensive review of the management of painful symptoms through a detailed review of the published literature using a variety of databases particularly Medline and EMBASE.
Increased nerve growth factor mRNA in lateral calf skin biopsies from diabetic patients
Authors: Diemel L. T. , Cai F. , Anand P. , Warner G. , Kopelman P. G. , Fernyhough P. , Tomlinson D. R.
Published: March 2003

Aims This study set out to establish a novel procedure for the measurement of human nerve growth factor (NGF) messenger ribonucleic acid (mRNA) and to use this method to measure NGF expression in skin biopsies from control subjects and from patients with early neuropathies. NGF mRNA levels were related to functional measures of the competence of NGF‐responsive nerves.

Methods mRNA levels were measured by competitive reverse transcription with polymerase chain reaction amplification (cRT‐PCR). Functional correlates of this observation were assessed by indices of thermal sensitivity – mediated by C‐fibres, whose phenotype is regulated by NGF.

Results NGF mRNA was increased in skin biopsies from 19 diabetic patients (5.12 ± 3.88 (
sd)) compared with samples from eight controls (1.57 ± 0.95; P = 0.001). Diabetic patients showed significantly (P < 0.001) diminished detection of cool and warm stimuli compared to age matched control group (n = 24), but there were no differences in detection of heat as pain, or correlation with NGF mRNA levels.

Conclusions These findings suggest abnormally increased expression of NGF in diabetic neuropathy, which may represent a compensatory mechanism for impaired phenotype in NGF‐responsive neurones.
Identification of new mutations in the hepatocyte nuclear factor 4a gene among families with early onset Type 2 diabetes mellitus
Authors: Malecki M. T. , Yang Y. , Antonellis A. , Curtis S. , Warram J. H. , Krolewski A. S.
Published: March 2003

Aims Mutations in hepatocyte nuclear factor (HNF)‐4α gene located on chromosome 20q have been found to be responsible for the development of early onset Type 2 diabetes mellitus (DM). Through a national campaign, 53 families with autosomal dominant, early onset Type 2 DM (n = 654) were assembled to determine the frequency of mutations in the HNF‐4α gene and their contribution to the development of diabetes.

Methods Twelve exons and the promoter region of the HNF‐4α gene were screened in probands of the families by a double gradient, denaturing gradient gel electrophoresis (DG‐DGGE) protocol combined with automated bi‐directional sequencing of the PCR products of all heterozygous individuals.

Results We detected two new mutations in the HNF‐4α gene that changed the amino‐acid sequence. The first mutation was a Gly→Ser substitution in codon 115 within a highly conserved DNA binding domain, and all six carriers of this mutation had diabetes and low insulin secretion. The second mutation was an Ile→Val substitution in codon 454 within the transactivation domain. It was carried by four family members, two of whom also carried a mutation in the HNF‐1α gene. Of those having only the mutation in HNF‐4α one had diabetes and the other had normal glucose tolerance and both were obese and hyperinsulinaemic. Thus, it is uncertain that this mutation is responsible for any of the diabetes in this family.

Conclusion We have found that mutations in the HNF‐4α gene account for a small proportion, about 2–4%, of families with early onset, autosomal dominant, Type 2 DM in US Caucasians.

Diabet. Med. 16, 193–200 (1999)
Twinning project: Israel and Georgia – the birth of a Diabetes‐in‐Pregnancy Centre in Georgia
Authors: Hod M. , Asatiani N. , Elphick A. , Kurashvili R. , Natsvlishvili M. , Chanturia T. , Bar J. , Peled Y.
Published: March 2003

Aims To determine the effectiveness of a joint Israeli–Georgian twinning project aimed at introducing modern methods of perinatal care to Georgian women with pregestational diabetes mellitus.

Methods A Diabetes‐in‐Pregnancy Centre was established in Georgia (in the former USSR). Thirty‐two women with Type 1 diabetes mellitus participated in the first stage of the study (January 1997–June 1998). All were maintained under strict metabolic surveillance starting at least three months prior to conception and were given organized instruction in methods of self‐monitoring of glucose levels, insulin dose adjustment, dietary management and close fetal surveillance throughout pregnancy. A second stage of the project was started in January 1998 and aimed at the diagnosis of gestational diabetes mellitus (GDM).

Results Of the 32 women, 20 had had a total of 44 prior pregnancies without proper perinatal care (1990–96); only five (11.3%) ended in the birth of a healthy infant. On entry to the study, all 32 patients had unsatisfactory metabolic indices. HbA1c levels decreased significantly from the pre‐conception period (P < 0.001) and were maintained at the lower level throughout pregnancy; insulin doses decreased significantly until the third trimester (P < 0.01) and then increased (P < 0.001). Since January 1997, 21 women have become pregnant. Eighteen have given birth, 12 (67%) by Caesarean section and six (33%) by vaginal delivery. Gestational age at birth was 36–39 weeks, and birth weight ranged from 2300 to 4100 g. The only neonatal complications were mild respiratory distress syndrome and hypoglycaemia. There were no significant maternal complications in the 236 women screened, eight were diagnosed as having GDM and 12 impaired glucose tolerance. They were actively managed to a successful outcome.

Conclusions The establishment of the Diabetes‐in‐Pregnancy Centre in the Republic of Georgia has significantly reduced the prior high pre‐programme perinatal morbidity and mortality as well as the incidence of maternal complications in pre‐GDM as well as in GDM.
The Charcot foot
Authors: Jeffcoate W. , Lima J. , Nobrega L.
Published: March 2003

Aims To review the clinical manifestations of the Charcot foot in diabetes mellitus, with particular reference to theories concerning aetiology.

Methods Systematic review of the published literature, searching for the keywords ‘Charcot’, ‘foot and diabetes’ and ‘neuropathy’ on Medline, as well as by examination of the references in recent published reviews.

Conclusions The Charcot foot of diabetes mellitus is a common problem, and yet is not widely recognized by non‐specialists. The failure of professionals to identify the condition in its early phases is probably largely responsible for the gross deformity which follows continued weight‐bearing. The condition is confined to those with severe peripheral neuropathy. It is thought to result from three factors: motor neuropathy leading to the development of abnormal forces within the foot, subsequent disorganization of the foot as a result of associated osteopenia and progressive destruction from continued weight‐bearing, enabled by reduced pain sensation. The cause of the osteopenia is not known, but it is associated with increased bone blood flow, which may be mainly the result of loss of sympathetic innervation. The importance of increased limb blood flow in the pathogenesis of the Charcot foot has been recognized for over a century. Paradoxically, the increased flow is associated with evidence of macrovascular disease, in that the prevalence of vascular calcification of pedal vessels approaches 90%. After an interval of many months, the condition tends to evolve: the increased blood flow lessens, the osteopenia is reduced and the disorganized bones become sclerotic. This tendency for the condition to evolve remains unexplained, since it would not be expected if the condition was caused solely by progressive denervation. As a result, it is suggested that another factor may be involved in the pathogenesis of the Charcot foot: an abnormal vasomotor reflex, analogous to reflex sympathetic dystrophy, occurring against a background of severe peripheral neuropathy. The resolution of the condition occurs because it is the reflex component of the hyperaemia which proves self‐limiting.
Iohexol as a marker of glomerular filtration rate in patients with diabetes: comparison of multiple and simplified sampling protocols
Authors: Pucci L. , Bandinelli S. , Pilo M. , Nannipieri M. , Navalesi R. , Penno G.
Published: March 2003

SUMMARY

Aims  To report on the reproducibility of iohexol glomerular filtration rate (GFR) estimation, to compare the plasma clearance of iohexol with that of[51Cr]EDTA and to evaluate the reliability of reduced sampling schedules in estimating GFR in Type 1 and Type 2 diabetes mellitus.

Methods  Agreement was assessed in 15 Type 1 and 26 Type 2 diabetics with creatinine ranging from 53 to 564 μmol/l.

Results  The regression between multiple‐sample iohexol and[51Cr]EDTA clearances was 0.999 in Type 1 and 0.987 in Type 2 diabetes (P < 0.0001 for both). A seven‐sample design and the three‐sample approach by Brøchner–Mortensen were validated by comparison with the full‐sample schedule in 87 patients (51 Type 1, 36 Type 2). Full‐sample GFR was 80.3 ± 43.8, seven‐sample 79.5 ± 43.9 (r = 0.990) and three‐sample 79.8 ± 45.2 ml.min−1.1.73 m−2 (r = 0.972). The coefficients of variation of GFR were 2.7 ± 1.4% and 3.8 ± 1.9% for the full‐sample and the seven‐sample approaches, respectively, and significantly higher for the three‐sample design (6.9 ± 3.4%, P = 0.0001).

Conclusions  After iohexol injection, the Brøchner–Mortensen schedule does not provide an accurate estimate of GFR. The seven‐sample approach gives acceptable errors and allows a good estimate of GFR throughout a wide range of renal function.
Variability of glomerular filtration rate estimation using single injection methods in Type 1 diabetic patients
Authors: Siebenhofer A. , Meinitzer A. , Brandmair J. , Plank J. , Pieber T. R.
Published: March 2003

Aim  Single‐injection inulin clearance (SIIC) methods without urine collection have often been compared with classical constant‐infusion inulin clearance methods. Therefore, we used repeated SIIC measurements to prove the hypothesis of a possible overestimation or variability of inulin clearance in Type 1 diabetic patients.

Methods  Two SIIC periods, glomerular filtration rate 1 and 2 (GFR 1, GFR 2) were performed consecutively on the same day. In diabetic patients, GFR 3 was measured several days later at the same time of day as the GFR 1 had been done. The calculation used a two‐compartment modelling system (TCM) which showed excellent agreement to ratio of dose over area under the curve (D/AUC) calculation.

Results  Twelve normotensive Type 1 diabetic patients (mean and sd), age 39 (10.7) years, diabetes duration 20 (10.7) years, HbA1c 8.5% (0.82%), albuminuria 13.8 (15.7) mg/24 h and six controls, age 26 (2.6) years were examined. Healthy controls showed an excellent repeatability between GFR 1 111.8 (11.44) and GFR 2 110.8 (11.41) ml.min−1.1.73 m2−1. However, in diabetic patients there was a highly significant decrease between GFR 1 129.4 (11.86) and GFR 2 118.1 (13.05) ml.min−1.1.73 m2−1. GFR 1 with 129.4 (11.86) ml.min−1.1.73 m2−1 did not differ significantly from GFR 3 with 129.9 (12.40) ml.min−1.1.73 m2−1 in diabetic patients (P < 0.839).

Conclusions  In contrast to normal subjects, in Type 1 diabetic patients single‐injection inulin clearance technique showed a clinically relevant decrease in GFR on two repeated measurements on the same day. Overestimation of GFR by the first inulin clearance may be caused by incomplete inulin distribution in the slow compartment.

Diabet. Med. 18, 464–468 (2001)
The influence of isolated small nerve fibre dysfunction on microvascular control in patients with diabetes mellitus
Authors: Pfützner A. , Forst T. , Engelbach M. , Margin T. , Goitom K. , Löbig M. , Beyer J. , Kunt T.
Published: March 2003

Aim The aim of the study was to investigate the influence of isolated small nerve fibre dysfunction on microvascular skin blood flow and transcutaneous oxygen tension in patients with diabetes mellitus.

Methods  Small nerve fibre dysfunction was assessed by the measurement of thermal and pain perception thresholds. Patients with evidence of large fibre disturbances as evaluated by means of vibration perception threshold were excluded from the study. Microvascular blood flow was investigated with laser‐Doppler‐fluxmetry (LDF) following stimulation with acetylcholine and mild thermal injury.

Results  Diabetic patients with small nerve fibre injury showed a significantly reduced increase in the laser‐Doppler‐flux signal following the application of acetylcholine compared with patients without neuropathy or healthy control subjects (2.8 arbitrary units (AU) (1.3–5.5) vs. 7.2 AU (4.1–25.5); P = 0.007 and vs. 8.5 AU (3.0–17.0), P = 0.02, respectively). The increase in LDF following thermal injury was also diminished in patients with small nerve fibre dysfunction compared with patients without neuropathy or the control group (29.8 AU (17.2–46.5) vs. 51.2 AU (29.5–93.5); P = 0.02 and vs. 54.6 AU (39.7–97.7); P = 0.004, respectively). In addition, they showed a significantly reduced transcutaneous oxygen tension compared with the other groups (42.9 mmHg (41.6–55.5) vs. 56.1 mmHg (49.2–60.8); P = 0.04 and vs. 59.0 mmHg (54.6–80.3), P = 0.03, respectively).

Conclusions  Our study confirms an association between small nerve fibre injury and skin microvascular dysfunction. It further underlines the concept of neurovascular disturbances in the pathogenesis of neurotrophic foot ulceration.

Diabet. Med. 18, 489–494 (2001)
Audit of influenza and pneumococcal vaccination uptake in diabetic patients attending secondary care in the Northern Region
Authors: Wahid S. T. , Nag S. , Bilous R. W. , Marshall S. M. , Robinson A. C. J.
Published: March 2003

Aims  To document uptake of influenza and pneumococcal vaccination in diabetic patients attending secondary care in the Northern Region, and to explore influencing factors.

Methods  Diabetic patients attending out‐patients in Middlesbrough, Gateshead and Newcastle were questioned from October 1999 to March 2000. Physicians enquired about influenza and pneumococcal vaccination status using a standardized questionnaire. Data collected included age, year of diagnosis, duration of diabetes, type of diabetes, and the presence of other recognized indications for vaccination.

Results  Two hundred and sixty‐eight diabetic patients, 42% (113/268) with Type 1 diabetes, 34% (91/268) with ischaemic heart disease, 10% (26/268) with chronic pulmonary disease (CPD) and 10% (27/268) with chronic renal disease, were questioned. Thirty‐five percent (93/268) of patients received both influenza and pneumococcal vaccines, 24% (64/268) received only influenza vaccine, and none received pneumococcus vaccine alone. Most vaccinees received advice about influenza and pneumococcal vaccination from their general practitioner (90% (142/157) and 87% (81/93), respectively). A large number of non‐vaccinees were unaware of the need for influenza and pneumococcal vaccination (69% (76/111) and 91% (159/175), respectively). Using multiple logistic regression co‐existing CPD increased the odds of receiving influenza (odds ratio (OR) (95% confidence interval (CI)) = 1.99 (1.07–14.12)) or pneumococcal (OR = 3.77 (1.69–21.76)) vaccination. Furthermore, each 1‐year increase in age increased the chance of receiving influenza or pneumococcal vaccination by 22% (OR = 1.22 (1.09–1.67) and 29% (OR = 1.29 (1.07–1.72)), respectively.

Conclusions  Vaccination rates in these diabetic patients are unsatisfactory. Secondary care health professionals might increase rates by raising the topic in consultations.

Diabet. Med. 18, 599–603 (2001)
Deep tissue biopsy vs. superficial swab culture monitoring in the microbiological assessment of limb‐threatening diabetic foot infection
Authors: Pellizzer G. , Strazzabosco M. , Presi S. , Furlan F. , Lora L. , Benedetti P. , Bonato M. , Erle G. , De Lalla F.
Published: March 2003

Aims  The results of ulcer swabbing vs. deep tissue biopsy have been compared prospectively in 29 diabetic patients with limb‐threatening foot infection, to investigate the effectiveness and reliability of each method, and to evaluate whether any of the two could be more suitable for the microbiological follow‐up of severe lesions.

Methods  Microbiological samples were collected by using both methods at fixed intervals after therapy commencement (i.e. at day 0, 7, 14, and 30). Statistical comparison was performed between the results of each sampling procedure after the end of follow‐up.

Results  At enrolment, the mean number of isolates per patient was 2.34 by swabbing and 2.07 by tissue biopsy sampling; the rate of isolation for anaerobes with the two methods was 35% and 25%, respectively; no statistical differences could be observed between the two procedures in terms of either species or frequency of isolation. Anaerobic species were never detected after the first 2 weeks of appropriate treatment, and those ulcers which were still active at day 30 yielded almost exclusively Gram‐positive bacteria. At the end of follow‐up, deep tissue cultures appeared to exhibit a higher diagnostic sensitivity with respect to swabs.

Conclusions  Swabbing and deep tissue cultures appear to be equally reliable for the initial monitoring of antimicrobial treatment in severe diabetic foot infection. However, our experience seems to suggest that deep tissue might be more sensitive than swabbing for monitoring those isolates that have been selected for antibiotic resistance, i.e. those from ulcers that are still active after 30 days of treatment.

Diabet. Med. 18, 822–827 (2001)
Assessing diabetic control — reliability of methods available in resource poor settings
Authors: Rotchford A. P. , Rotchford K. M. , Machattie T. , Gill G. V.
Published: March 2003

Aims and methods To examine the reliability of random venous or capillary blood glucose testing, random urine glucose testing, and a current symptom history in predicting a high HbA1c in Type 2 diabetic patients taking oral hypoglycaemic agents in a poorly controlled rural African population.

Results For a cut‐off point for HbA1c of ≥ 8%, for random venous plasma glucose of ≥ 14 mmol/L (present in 47.2% of subjects), specificity was 97.1% (95% CI 85.1–99.9), sensitivity 56.8% (48.8–64.5) and positive predictive value (PPV) 98.9% (94.2–99.9). HbA1c≥ 8% is predicted by a random capillary blood glucose of 17 mmol/L (present in 28.4% of subjects) with specificity 100% (90.0–100.0), PPV 100% (93.7–100.0) and sensitivity of 34.3% (27.2–42.1). HbA1c≥ 8% is predicted by the presence of heavy glycosuria (≥ 55 mmol/L) (present in 35.6%) with specificity 94.1% (80.3–99.3), sensitivity of 41.9% (34.1–49.9) and PPV 97.1% (89.9–99.6). Polyuria/nocturia (present in 31.3%) was the only symptom found to be associated with poor control, with a specificity for predicting HbA1c of ≥ 8% of 81.5% (61.9–93.7), PPV 89.1% (76.4–96.4) and sensitivity 30.6% (22.9–39.1).

Conclusions Where resources are short, random glucose testing can be used to detect a significant proportion of those with the worst control with a high degree of specificity enabling primary care staff to modify treatment safely. Where facilities are limited capillary blood or urine testing with reagent strips, may be substituted for venous plasma testing in the laboratory. A symptom history was insufficient to replace biochemical testing, but where this is unavailable, urinary symptoms may be helpful.

Diabet. Med. 19, 195–200 (2002)
Cystic fibrosis‐related diabetes: the role of peripheral insulin resistance and β‐cell dysfunction
Authors: Yung B. , Noormohamed F. H. , Kemp M. , Hooper J. , Lant A. F. , Hodson M. E.
Published: March 2003

Aims The goal of this study was to identify the glycaemic status and investigate the roles of peripheral insulin resistance (IR) and pancreatic β‐cell dysfunction in the pathogenesis of cystic fibrosis‐related diabetes (CFRD) in adult cystic fibrosis (CF) patients with no previous history of glycaemic disturbances.

Methods The glucose tolerance status of 68 CF patients was determined using 2‐h oral glucose tolerance tests (OGTTs). Peripheral IR was measured using the homeostasis model assessment for insulin resistance (HOMA‐IR) in the CF group and 46 normal healthy control subjects. Pancreatic β‐cell function, calculated as the ratio between the 30‐min increment in plasma insulin and the corresponding 30‐min post‐OGTT plasma glucose concentration, was also measured in a subset of 30 CF patients and 16 normal healthy controls. Extended 180‐min OGTTs, with frequent plasma glucose and insulin sampling, were also undertaken in 24 CF patients and eight normal healthy controls to determine glucose‐induced insulin response.

Results Of the 68 CF patients studied, 41, 18 and nine were found to have normal, impaired and diabetic glucose tolerances, respectively. The mean HOMA‐IR values (mU/mmol) in the CF patients, as a whole, were not significantly different compared with the normal healthy controls (CF 2.2 ± 1.1 vs. control 1.8 ± 0.9; NS). Within the CF group, glycaemic status had no impact on HOMA‐IR (mU/mmol): 2.2 ± 1.2 (normal glucose tolerance); 2.0 ± 1.0 (impaired glucose tolerance); and 2.3 ± 1.1 (diabetic glucose tolerance). β‐cell function (mU/mmol) was not only significantly lower in the CF group (CF 1.65 ± 1.8; P < 0.001) but also in the CF group with normal glucose tolerance (2.25 ± 2.10; P < 0.01) compared with healthy control (4.98 ± 2.38). Mean plasma glucose concentrations were generally higher and mean plasma insulin concentrations lower in the CF group as a whole when compared with normal healthy controls. Within the CF group, there was a progressive decline in glucose‐induced insulin release with worsening glycaemic status.

Conclusions A lack of difference in peripheral IR, measured using HOMA‐IR, in the CF group and healthy controls or within the CF group with differing glycaemic status suggests that IR does not have a significant role in the pathogenesis of CFRD. Pancreatic β‐cell function, already subnormal in CF patients with OGTT‐defined normal glucose tolerance status, deteriorated further with worsening glycaemic status. This suggests that insulinopenia plays a prominent role in the pathogenesis of glucose intolerance and subsequent development of CFRD.