Effect of maternal undernutrition on vascular expression of micro and messenger RNA in newborn and aging offspring.

The aim of this study was to test the hypothesis that maternal undernutrition (MUN) alters offspring vascular expression of micro-RNAs (miRNAs), which, in turn, could regulate the expression of a host of genes involved with angiogenesis and extracellular matrix remodeling. The expression of miRNA and mRNA in the same aortic specimens in 1-day-old (P1) and 12-mo-old offspring aortas of dams, which had 50% food restriction from gestation day 10 to term, was determined by specific rat miRNA and DNA arrays. MUN significantly downregulated the expression of miRNAs 29c, 183, and 422b in the P1 group and 200a, 129, 215, and 200b in the 12-mo group, and upregulated the expression of miRNA 189 in the P1 group and 337 in the 12-mo group. The predicted target genes of the miRNAs altered in the two age groups fell into the categories of: 1) structural genes, such as collagen, elastin, and enzymes involved in ECM remodeling; and 2) angiogenic factors. MUN primarily altered the expression of mRNAs in the functional category of cell cycle/mitosis in the P1 group and anatomic structure and apoptosis in the 12-mo age group. Several of the predicted target genes of miRNAs altered in response to MUN were identified by the DNA array including integrin-beta(1) in the P1 aortas and stearoyl-CoA desaturase-1 in the 12-mo age groups. These results are consistent with the hypothesis that MUN modulation of offspring gene expression may be mediated in part by a miRNA mechanism.

The role of lipid anchors for small G proteins in membrane trafficking.

Small GTP-binding proteins of the Ras superfamily play diverse roles in intracellular trafficking. In order to perform these functions, the proteins must associate with specific donor vesicles and be recycled after fusion of these vesicles with their acceptor membrane target. Recent results have identified a number of lipid modifications of these proteins, occurring at the N- or C-termini, that contribute to their membrane binding. Recycling appears, in some cases, to be mediated by soluble proteins that bind the lipid-modified tails, removing them from the membrane and allowing their reutilization via the cytosol.

A meta-analysis of 21,178 patients undergoing open or endovascular repair of abdominal aortic aneurysm.

BACKGROUND: Several studies have compared outcomes after elective open and endovascular approaches to abdominal aortic aneurysm (AAA) surgery, with varying results. METHODS: A random-effects meta-analysis was undertaken to compare operative outcomes, postoperative complications, 30-day mortality and long-term patient survival after surgery. Endpoints were compared using odds ratios (ORs), weighted mean differences (WMDs) or log hazard ratios (HRs) as appropriate. RESULTS: Forty-two studies comprising 21,178 patients (10,855 open; 10,323 endovascular) were included. In the elective setting (20,715 patients), the endovascular method was associated with a shorter stay in intensive care (WMD--36 h; P < 0.001) and a shorter total postoperative stay (WMD--5.4 days; P < 0.001). Cardiac (OR 1.76; P = 0.002) and respiratory (OR 4.01; P < 0.001) complications were more common after open surgery. In the endovascular group, 30-day mortality was lower (OR 0.46; P < 0.001). Endovascular surgery was also associated with an improved long-term aneurysm-related mortality (HR 0.39; P < 0.001). For ruptured AAA (463 patients), the less invasive operation was associated with a reduced stay in intensive care (WMD--100.4 h; P = 0.005) and a significantly lower 30-day mortality (OR 0.45; P = 0.005). CONCLUSIONS: The endovascular repair of AAA offers a clear benefit in terms of reduction in postoperative adverse events and 30-day mortality. In the longer term, it is also associated with a reduction in aneurysm-related mortality, but not in all-cause mortality.

Arterial thrombosis associated with malignant disease.

OBJECTIVE: The association between cancer and venous thrombosis is well established, however, that between malignancy and arterial thrombosis is less well described. Isolated cases have been reported and chemotherapy has been implicated as a cause though its significance compared with the malignant disease process itself is not known. This study examines the outcome of patients with malignant disease who present with arterial thrombosis. METHOD: Details of patients with malignant disease who presented with arterial thrombosis were analysed. RESULTS: Twenty patients presented with malignancy and arterial thrombosis, 16 presented in the last four years. The most common malignancy was metastatic breast cancer. Thrombosis involved the leg in 19 cases and the arm in one. Four patients also had venous thromboembolic events and one had a carotid artery thrombosis. Eight patients underwent operative treatment for their thrombosis. Five out of six thromboembolectomies and two out of three bypass procedures failed. Twelve had conservative or palliative treatment. Outcome was generally poor, two patients had major amputations and seventeen died at median follow-up of eight weeks. Survival rate from the time of presentation of arterial thrombosis was 50% at three months and 17% at one year. CONCLUSION: Patients with critical limb ischaemia due to atherosclerotic disease have an expected survival of approximately 80% at one year. The outcome of patients with arterial thrombosis associated with malignant disease is far worse. Arterial thrombosis is an agonal event in many of these patients. Conservative or palliative treatment may be the most appropriate management.

Endovascular and open approaches to non-thrombosed popliteal aneurysm repair: a meta-analysis.

OBJECTIVE: Endovascular repair of popliteal artery aneurysms is a relatively new technique that is still undergoing evaluation. The aim of this study was to compare outcomes following open and endovascular approaches. METHODS: All published studies comparing outcomes following open and endovascular popliteal aneurysm were included. Endpoints included operative duration, length of stay, and postoperative complications including short-term patency rates. Outcomes were combined using a random-effects meta-analytical technique and differences assessed using odds ratios (OR), weighted mean difference (WMD) and log hazards ratio (HR). RESULTS: Three studies comprising 141 patients (37 endovascular; 104 open) were included. No significant differences in patient characteristics were seen. Operative duration was significantly longer for endovascular repair (WMD 120 minutes, p<0.001). Thirty day graft thrombosis (OR 5.05, p=0.06) and reintervention (OR 18.80, p=0.03) were more likely following endovascular repairs. Postoperative length of stay was shorter in the endovascular group (WMD--3.9 days, p<0.001). There was no significant difference in long-term primary patency rates (HR 1.70, p=0.53). CONCLUSIONS: Endovascular repair of popliteal artery aneurysms offers similar medium-term benefits as an open repair. However, short-term graft thrombosis and reintervention rates are significantly greater. With the current technology it is difficult to justify endovascular treatment of popliteal aneurysms.

Prioritisation of vascular outpatient appointments cannot be based on referral letters alone.

BACKGROUND: Vascular referrals include patients with conditions varying from varicose veins of cosmetic nuisance to patients with critical ischaemia, transient ischaemic attacks and abdominal aortic aneurysms. A large number of such referrals are received each week from general practitioners. It is important to prioritise patients with conditions that need to be dealt with quickly. OBJECTIVE: We prospectively reviewed referral letters to two vascular units, one in South Wales and one in the Oxford region to assess whether prioritisation can be made on the basis of referral letters. MATERIAL AND METHODS: All GP referral letters were studied for four months. Only patients with lower limb ischaemic symptoms were included. Degree of urgency requested by the GP was also noted. RESULTS: Of 174 referrals for potential lower limb ischaemia analysed, 145 (83%) proved to be due to peripheral vascular disease. Of these 145 referrals, 72% were referred for claudication. Only 37% and 13% respectively mentioned claudication distance and/or rest pain. Risk factors with reference to diabetes, hypertension, hyperlipidaemia, ischaemic heart disease, atrial fibrillation, cerebrovascular disease and smoking were made in 19%, 43%, 23%, 23%, 10%, 14%, 31% of letters respectively. Clinical signs were poorly documented, with 90% of referrals failing to mention presence or absence of critical ischaemic signs. The GP's own assessment of urgency was not stated in 66% of letters, without which only 3% of patients were seen in the clinic within four weeks, compared with 22% in those in whom urgency was stated. Six per cent of patients when reviewed in outpatients were found to have sufficiently severe symptoms to warrant immediate admission. Regional variation was observed with 57% of Royal Glamorgan Hospital referral letters documenting degree of urgency compared with only 23% of Royal Berkshire Hospital letters (p = < 0.0001). CONCLUSION: Most referral letters were poorly documented with regard to key symptoms, risk factors and clinical signs. The degree of urgency was often not stated. The creation of referral protocols is now being considered.

Management of cardiovascular risk factors by primary care physicians in patients with peripheral arterial disease.

BACKGROUND: Patients with significant coronary artery disease (CAD) are now intensively treated by primary care physicians predominantly because of government pressure and remuneration to prescribe anti-platelet and anti-hyperlipidaemic drugs. Peripheral arterial disease (PAD) with the identical risk factors appeared to us to be less intensively investigated and treated by primary care physicians. OBJECTIVE: To review the treatment of risk factors in all patients referred to two vascular clinics with a diagnosis of suspected PAD. DESIGN: Cross-sectional survey. SETTING: Vascular outpatient clinic in two district general hospitals. PARTICIPANTS: 124 consecutive new patients were studied to determine risk factors and appropriate treatment. RESULTS: Of the 124 patients, 85 (68%) were confirmed to have PAD without evidence of symptomatic CAD. In the PAD alone group, less than 25% received anti-smoking advice (p < 0.0001) and only 36% were prescribed anti-platelet drugs (p = 0.016). Seventy-three per cent of the overall referred patients with hypertension had been treated for this condition and the blood pressure was normal in 71% of the patients with PAD. In patients with hyperlipidaemia, statins had been prescribed in 92% of patients with coexistent symptomatic CAD, but only in 64% of patients with PAD alone (p = 0.009). In the patients with diabetes, only 66% of the PAD alone group had adequate control of their blood sugar (p = 0.185). CONCLUSIONS: It would appear that patients with CAD and PAD are being treated successfully for their risk factors, but patients with PAD alone, sharing the same common risk factors, are being less than optimally treated.

Ventricular arrhythmias after coronary artery bypass graft surgery: incidence, risk factors and long-term prognosis.

The incidence, risk factors and long-term prognosis of complex ventricular arrhythmias after coronary artery bypass graft surgery are not known. Complex ventricular arrhythmias are defined as Lown grades 4a (couplets), 4b (ventricular tachycardia) and 5 (R on T phenomenon). Ninety-two patients with normal left ventricular function who underwent elective coronary artery bypass graft surgery were prospectively evaluated. Ventricular arrhythmias were documented by predischarge 24 hour ambulatory electrocardiographic monitoring; 43% of patients had no or simple ventricular arrhythmias (Lown grades 1 to 3) and 57% had complex ventricular arrhythmias. Risk factors analyzed included age, sex, diabetes, hypertension, smoking, preoperative digoxin or propranolol therapy, cardiopulmonary bypass time, aortic cross-clamp time, number of vessels bypassed, peak creatine kinase (CK) elevation and pericarditis. No risk factor identified patients at higher risk for complex ventricular arrhythmias. Patients were followed up for 6 to 24 months (mean 16). Patients with complex ventricular arrhythmias did not have a higher incidence of sudden death, cardiac death, syncope, angina, myocardial infarction or cerebrovascular accident. It was concluded that: Complex ventricular arrhythmias are common after coronary artery bypass graft surgery. None of the risk factors considered identify high risk patients. Complex ventricular arrhythmias after coronary artery bypass graft surgery do not indicate a poor prognosis in patients with normal left ventricular function.

Results of thoracoscopic sympathectomy for the treatment of axillary and palmar hyperhidrosis with respect to compensatory hyperhidrosis and dry hands.

BACKGROUND: We examined the results of thoracoscopic sympathectomy (TS) for palmar and axillary hyperhidrosis with respect to operative method, symptom control, patient satisfaction and complications. METHODS: We performed a retrospective review of patient records with mail and telephone questionnaire follow-up of 55 patients (15 men) with a median age of 26 years (range, 15-52) who underwent TS between February 1994 and December 2001. RESULTS: There were no differences in complication rates between those having bilateral TS (n = 23) and those having unilateral procedures (n = 20) with a median follow-up of 21 months (range, 2-94). Forty-three patients returned questionnaires (response rate, 78%). Forty patients (93%) were satisfied with the results. Thirty-four patients (79%) noted compensatory hyperhidrosis and 22 (51%) excessively dry hands. CONCLUSION: Despite high rates of compensatory sweating, the majority of patients are very satisfied with the results. The high rate of excessively dry hands is a previously unreported finding and important to discuss when obtaining consent.

MRSA in lower limb amputation and the role of antibiotic prophylaxis.

AIM: Methicillin Resistant Staphylococcus Aureus (MRSA) colonisation is reported in 3-20% of vascular patients. Many develop infective complications. MRSA is associated with poor prognosis. Aim of the study is to assess MRSA in lower limb amputation and efficacy of antibiotic prophylaxis. METHODS: Prospective study of lower limb amputation. MRSA screen and wound swabs were taken at operation. Antibiotic prophylaxis included teicoplanin (400 mg) 1 dose at operation. RESULTS: Twenty-five patients underwent 33 primary amputations. At operation 15 legs (45%) were colonised with MRSA and 18 legs (58%) had active wound infection; MRSA (4) and other (14). Following surgery 3 patients died. Twenty-two legs (76%) had primary healing. Infection developed in 7 stumps (24%), MRSA (5) and Pseudomonas (2). Stump infection increased time to wound healing (p<0.0001). MRSA stump infection increased revision amputation (p=0.009) and duration of hospital stay (p<0.0074). MRSA wound infection at operation increased the risk of MRSA stump infection (p=0.007). Non-MRSA wound infection at operation was not associated with a worse outcome. No patient colonised with MRSA at operation developed postoperative MRSA stump infection. CONCLUSIONS: MRSA is more prevalent that previously reported. MRSA infection has a poor prognosis. Prophylaxis may be effective for patients colonised with MRSA.

Trauma calls: role of the general surgeon and CT scanning.

BACKGROUND AND OBJECTIVE: General surgeons are required in only a minority of trauma call cases to assess for abdominal injuries. Computed tomography (CT) accurately detects blunt abdominal injuries and may have replaced the need for general surgeons at trauma calls. This study evaluated the role of general surgeons at trauma calls and assessed use of CT in cases of suspected abdominal trauma. METHODS: (a) Eighteen month analysis of trauma calls at a district general hospital and (b) three month prospective study of all trauma cases presenting to A&E. RESULTS: (a) There were 73 trauma calls and the mechanism of injury in most cases was a road traffic accident (RTA). Most patients had orthopaedic and/or neurosurgical injuries. The general surgeons assessed 22 trauma call patients. Abdominal injury was excluded in 13 (four by clinical examination and nine following CT). (b) Forty three patients fulfilled the criteria for a trauma call and 14 trauma calls were made. The mechanism of injury in most was RTA and most had orthopaedic and/or neurosurgical injuries. The general surgeons assessed 10/43 potential trauma call patients, and abdominal injury was excluded in five (one by clinical examination and four following CT). CONCLUSION: A&E staff managed most trauma calls. Most patients did not require general surgical intervention. For penetrating injuries, presence of a general surgeon remained crucial. For blunt injuries CT was an important adjunct. These data suggest that general surgeons do not routinely have to attend all trauma calls but can be called if abdominal and/or vascular injuries are specifically suspected.

Comparison of 3.0-T MR vs 3.0-T MR arthrography of the hip for detection of acetabular labral tears and chondral defects in the same patient population.

OBJECTIVE: We report our experience in diagnostic sensitivity of 3.0-T conventional MR vs 3.0-T MR arthrography of the hip for detection of acetabular labral tears and chondral defects in the same patient population. METHODS: 43 consecutive patients had both conventional hip MR and MR arthrography examinations performed. These examinations were reviewed retrospectively by independent reading of two musculoskeletal radiologists who read the MR and MR arthrogram examinations in a randomized fashion (i.e. MR and MR arthrogram examinations were read at separate sittings and in a randomized fashion so as not to bias reviewers). Scans were assessed for acetabular labral tears and chondral defects. All patients went on to arthroscopy. RESULTS: Of these 43 patients, 40 had acetabular labral tears read by Reader 1 and 39 had acetabular labral tears read by Reader 2 on MR arthrogram, 39 had acetabular labral tears read by Reader 1 and 38 had acetabular labral tears read by Reader 2 on conventional MR examination. There were 42 labral tears in 43 patients at arthroscopy. There were four false-negative labral tears compared with arthroscopy on MR and three false negatives on MR arthrography for Reader 1 and five false negatives on MR and four false negatives on MR arthrography for Reader 2. Each reader had one false-positive labral tear compared with arthroscopy on both MR and MR arthrography. There were 32 acetabular chondral defects at arthroscopy. Reader 1 saw 21 acetabular chondral defects on conventional MR and 27 chondral defects at MR arthrography. Reader 2 saw 19 acetabular chondral defects at conventional MR and 25 acetabular chondral defects on MR arthrography. There were no false-positive readings of chondral defects compared with arthroscopy on MR and one false positive for Reader 1 and two false positives for Reader 2 on MR arthrography as compared with arthroscopy. On conventional MR examination, sensitivities and specificities as compared with arthroscopy were as follows: Reader 1 acetabular labral tear (90% sensitivity, 0% specificity) and Reader 2 acetabular labral tear (88% sensitivity, 0% sensitivity). On MR arthrogram, sensitivities and specificities as compared with arthroscopy for Reader 1 were 93%, 0% and for Reader 2 were 90%, 0%, respectively. Sensitivities and specificities for detection of acetabular chondral defects as compared with arthroscopy were Reader 1 conventional MR (65% sensitivity, 100% specificity), Reader 1 MR arthrography (81% sensitivity, 91% specificity), Reader 2 conventional MR (59% sensitivity, 100% specificity) and Reader 2 MR arthrography (71% sensitivity, 82% specificity). CONCLUSION: In this series, 3.0-T MR demonstrated sensitivity for detection of acetabular labral tears that rivals the sensitivity of 3.0-T MR arthrography of the hip. In this series, 3.0-T MR arthrography was more sensitive than conventional 3.0-T MR for detection of acetabular chondral defects. ADVANCES IN KNOWLEDGE: 3.0-T MR and MR arthrography are near equivalent in the diagnosis of acetabular labral tears. This information is useful for pre-operative planning.

Identification and characterization of a novel luciferase-like protein in the human female reproductive tract.

A novel cDNA was cloned from human endometrium, matching a human gene with the interim name KIAA1463. An mRNA identified by 5'-rapid amplification of cDNA ends was found to be 3349 nt in length. PCR analysis also identified another transcript of 6626 nt, with an open reading frame encoding a 900 amino acid protein. A fold recognition program identified similarity to firefly luciferase containing an AMP-binding motif; hence, we refer to the predicted protein as the AMP binding/luciferase-like protein (ALLP). ALLP mRNA and protein were expressed throughout the female reproductive tract with the highest levels found in the ovary and uterus. In situ hybridization and immunohistochemistry showed predominant localization of the ALLP mRNA/protein in endometrial glandular epithelium and within the theca and granulosa cells in the ovary. In the endometrium expression of ALLP, mRNA and protein were higher during d 16-21 of the secretory phase of the cycle. Western blot analysis showed decreased expression of ALLP in the postmenopausal endometrium, and hormone replacement therapy increased the expression of ALLP. Endometrial adenocarcinoma cell lines expressed more ALLP, compared with cultured primary endometrial cells or normal endometrial tissue. The ubiquitous expression of ALLP in reproductive and nonreproductive tissues suggests that this protein, which is probably regulated by ovarian steroids, plays an important metabolic role and may be involved in such processes as implantation and tumorigenesis.

Synthesis and antiinflammatory evaluation of substituted isophthalonitriles, trimesonitriles, benzonitriles, and terephthalonitriles.

In an effort to develop nonacidic, nonsteroidal, antiinflammatory agents without gastrointestinal complications, a series of cyanobenzenes was synthesized for antiinflammatory evaluation. Twenty-seven substituted isophthalonitriles, 19 trimesonitriles, 30 benzonitriles, and 16 terephthalonitriles were tested in the rat utilizing the carrageenan-induced pedal edema assay. Based on the performance of phenylbutazone in this assay (43.8% reduction at 100 mg/kg), six compounds, dosed at 50 mg/kg, produced reductions in inflammation comparable to this standard. However, the LD50 value of each compound dosed at this level was in the range of 40--56 mg/kg in the mouse; therefore, further the LD50 value of each compound dosed at this level was in the range of 40--56 mg/kg in the mouse; therefore, further study was not warranted. Fifteen compounds possessed activity in excess of 20% reduction at 200 mg/kg and also possessed LD50 values greater than 300 mg/kg. Of these cyanobenzenes, trimesonitrile (16), 4-chlorobenzonitrile, 2-chloroterephthalonitrile, and 2-fluoroterephthalonitrile with reductions in edema of 32, 30, 46, and 49%, respectively, represent the best candidates for subsequent study.

Fatty acyl-CoAs inhibit retinoic acid-induced apoptosis in Hep3B cells.

Retinoic acid (RA) induces apoptosis in Hep3B human hepatoma cells. 9-Cis-RA (c-RA) had a similar effect as all-trans-RA (t-RA) in inducing cell death in Hep3B cells. RA-induced Hep3B-cell death was associated with inhibited expression of the hepatocyte nuclear factor 4 (HNF-4) gene. Palmitoyl-CoA ((C16:0)-CoA), the reported HNF-4 ligand, prevented RA-induced apoptosis. The effect of (C16:0)-CoA was specific, since palmitic acid and co-enzyme A had no effect in preventing RA-induced apoptosis. Bovine serum albumin (BSA) also prevented RA-induced apoptosis. However, in contrast to BSA, which induced cell growth, (C16:0)-CoA alone had no effect on cell growth. Investigating the possible role of HNF-4 in apoptosis, the reported HNF-4 antagonist (C18:0)-CoA was employed, and it also prevented RA-induced apoptosis. By transient transfection, overexpression of HNF-4 did not prevent RA-induced apoptosis. The induction and prevention of apoptosis caused by RA and (C16:0)-CoA were associated, respectively with the induction and inhibition of the expression of transforming growth factor beta (TGFbeta), which is known to play a role in apoptosis. Furthermore, RA and (C16:0)-CoA can regulate AP-1, which is a key regulator of the TGFbeta gene. Our data indicate that fatty acyl-CoAs can prevent RA-induced apoptosis and that TGFbeta, rather than HNF-4, may play a role in these regulatory processes. Our data also suggest that (C16:0)-CoA and (C18:0)-CoA are not the agonist and antagonist for HNF4, respectively in the Hep3B cell system.

Gene expression in Peyronie's disease.

Currently, surgical intervention is the only efficacious treatment for Peyronie's disease (PD), a fibromatosis of the tunica albuginea of the penis. Therapies based on the molecular pathways for this disease could provide alternatives to surgical treatment but only recently has the pathophysiology of the Peyronie's disease plaque been investigated at the molecular level. In this review, we examine the current knowledge of gene expression in the PD plaque and the relationship of PD with other fibrotic conditions such as Dupytren's disease. TGFbeta1, along with other growth factors, pro-fibrotic genes, and collagen, are expressed in fibroblasts and myofibroblasts. Myofibroblasts are normally involved in wound contracture and largely eliminated via apoptosis during the late stages of wound remodeling. In the PD plaque, however, these cells persist and may play an important role in the PD plaque fibrosis. The expression levels of TGFbeta1 and pro- and anti-fibrotic gene products, along with the nitric oxide/reactive oxygen species (NO/ROS) ratio in the tunica albuginea, appear to be essential for the formation and progression of the PD plaque and effect the expression of multiple genes. This can be assessed with the recently developed DNA-based chip arrays and results with the PD plaque have been encouraging. OSF-1 (osteoblast recruitment), MCP-1 (macrophage recruitment), procollagenase IV (collagenase degradation), and other fibrotic genes have been identified as being possible candidate regulatory genes. Finally, possible therapeutic avenues for gene-based therapy in the treatment of PD are discussed that may eventually reduce the need for surgical intervention.

Presence of NMDA receptor subunits in the male lower urogenital tract.

Some sexual responses in the male rat, specifically penile erection, are controlled by neural circuits in the brain and spine that are stimulated by the binding of excitatory amino acids (EAAs) to the postsynaptic N-methyl-D-aspartate receptor (NMDAR). In the hypothalamus, EAA/NMDAR interaction triggers the activation of neuronal nitric oxide synthase (nNOS) to produce nitric oxide (NO). The local synthesis of this neurotransmitter in the penile nerve terminals causes corpora cavernosal relaxation and erection. During sexual activity, NO is assumed to participate in seminal emission and ejaculation in the prostate and to inhibit voiding reflexes in the bladder. This study aimed to determine in vitro whether NMDAR is present in these organs and whether it affects the tone of tissue strips through an NO-dependent mechanism. We obtained penile, urinary bladder, and ventral prostate tissues from adult male rats and homologous surgical tissues from human male patients. We detected the NMDAR protein by Western blot and determined the binding of the NMDA antagonist, 3H-CGP. The NMDAR messenger ribonucleic acid (mRNA) was detected by reverse transcription/polymerase chain reaction and identified by cloning and sequencing. The in vitro response to NMDAR antagonists was measured in tissue strips that were precontracted with bethanechol or electrical field stimulation (in rat and human bladder), phenylephrine (in human corpora cavernosa), or norepinephrine (in human prostate). The NMDAR2B protein; ligand-binding activity; and NMDAR1, 2A, and 2B mRNAs were detected in all tissues studied. We found an NMDAR1 variant in rat prostate and penis and in human prostate that is larger than its cerebellar counterpart, but it encodes a 767-amino acid truncated protein (NMDAR1-T). The in vitro contraction of tissue strips was inhibited by NMDAR antagonists against the following sites: polyamine (with ifenprodil); ion channnel high affinity (with dizocilpine); ion channel low affinity (with memantidine, dextrometorphan, and ketamine); and, in an NO-independent, nonadrenergic-noncholinergic pathway that was only partially affected by EAAs. We conclude that, in vitro, all the essential NMDAR subunits are present in the lower urogenital tract, a novel variant of subunit 1 is expressed, the tissues bind an NMDAR ligand, and the NMDAR antagonists induce relaxation of tissue strips. Further work is necessary to determine whether the NMDAR subunits form a fully active receptor and participate in the control of organ tone that is relevant to male sexual activity.

Determination of human immunodeficiency virus-1 protease inhibitors in patient serum using free solution capillary zone electrophoresis.

The goal of this study was to develop a fast, inexpensive and quantitative method for serum determination of the human immunodeficiency virus protease inhibitors Crixivan (C), Viracept (V), Invirase (I) or Fortovase (F), and Norvir (N), using common conditions for isolation and analysis. The best separation procedure developed thus far involves uncoated silica capillary and a buffer containing formic acid and acetonitrile. This procedure allows us to analyze three drugs (C, V and I or F) in 15 min. Norvir requires different analytical conditions. These four drugs are isolated from patient sera with a mixture of ethyl acetate and hexane. Sensitivity of the capillary zone electrophoresis protocols is sufficient for the detection of these pharmacological agents at the lowest clinically relevant concentrations (0.1 microgram/ml).

Theatre delay for general surgical emergencies: a prospective audit.

A prospective audit of emergency theatre use for general surgery has been undertaken. Two month periods were studied before and after the introduction of a fully staffed 24-hour emergency theatre. Data were collected using a proforma documenting the time of the decision to operate, the actual time of the operation and the reason for and duration of any delay. After the introduction of the facility the proportion of procedures performed after midnight fell from 29 cases (21.3 per cent) to 7 (6.3 per cent) (p < 0.05). Emergency operating between 0900-1700 hrs increased from 40 cases (29.4 per cent) to 71 (61.3 per cent) (p < 0.05). There was no significant difference in the causes of delay between the two groups, the commonest being queuing for theatre. However, the length of the delay was significantly reduced. That for an appendicectomy was reduced from a median of 4 hrs 40 mins (range 30 mins-18 hrs 45 mins) to 1 hr 29 mins (0-6 hrs 30 mins) (p < 0.01) and for drainage of abscess from 5 hrs 56 mins (15 mins-20 hrs 30 mins) to 1 hr 51 mins (0-4 hrs 30 mins) (p < 0.01). There was no significant difference in the seniority of the surgeon making the decision to operate. In the first part of the audit we identified problems with regard to delay which were addressed by the introduction of the emergency theatre. The audit cycle has been successfully closed improving the care of general surgical emergencies requiring urgent or emergency operations.

Is a palpable short saphenous vein a useful clinical sign in varicose vein assessment?

The short saphenous vein (SSV) may be palpable in the popliteal fossa in patients with varicose veins. A prospective study has been carried out to determine the significance of this sign in the presence of primary varicose veins. The SSV was assessed by palpation of the popliteal fossa with the knee slightly flexed. Hand-held Doppler insonation (HHD) was also used in the out-patient clinic. All patients had SSV assessment by duplex scanning. One hundred and sixty legs were examined. In 68 the SSV was palpable; 39 (57%) of these had SSV reflux on duplex examination. When the SSV was not palpable (92 legs), only 1% (1 leg) refluxed on duplex scanning. SSV palpability had a 98% sensitivity, 75% specificity, 57% positive predictive value (PPV), and 99% negative predictive value (NPV). In comparison, HHD had 80% sensitivity, 87% specificity, 67% PPV and 93% NPV. The combined tests had 78% sensitivity, 73% specificity, 76% PPV and 100% NPV. Palpation of the SSV is a valuable part of clinical examination. If the SSV is not palpable, it is unlikely to reflux.