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Summary We review the imaging appearances of hepatic angiomyolipomas in patients with and without tuberous sclerosis. Sporadic hepatic angiomyolipomas have a varied appearance because of the inconstant proportion of fat, making confident imaging diagnosis difficult and necessitating biopsy in many cases. In patients with tuberous sclerosis, hepatic angiomyolipomas have a more consistent imaging appearance and, together with other features of the syndrome, can be more easily diagnosed. Preoperative diagnosis helps obviate unnecessary surgery.

A study comparing the pre- and post-radiotherapy computed tomographic scans of patients treated for nasopharyngeal carcinoma revealed that the incidence of major mucosal abnormality was significantly increased and approximately doubled after radiotherapy. The most significant factor predicting major mucosal abnormality after radiotherapy was the presence of tumour in the sinus before treatment. In the maxillary sinus there was significant association of major mucosal abnormality before and after radiotherapy although this was not so for the other sinuses. The mucosal changes observed were evident as early as six months after radiotherapy.

Objective: Severe arterial bleeding from the gastrointestinal (GI) wall usually arises from submucosal or subserosal arteries. Endosonography (EUS) is a method which can image small structures like GI wall vessels and, thus, has potential for improving the diagnosis and directing the therapy of vascular disorders of the β tract. Furthermore, ultrasonography is increasingly used in open and laparoscopic surgery to detect arteries, thereby possibly reducing bleeding complications. The aim of this study was to evaluate the ability of two different ultrasound (US) systems to image small arteries in the submucosa, subserosa and mesentery in vitro. Methods: A special system was used which allowed the ultrasound images to be precisely compared with histology of the same area of tissue. US images were made with an 8.5 MHz linear array and a 20 MHz single crystal US probe. Fifty measurements were obtained from 48 porcine and human GI arteries. All arteries were localized in the submucosa, subserosa or mesentery. Two of these arteries were also partly located in the mucosa and muscularis propria, respectively. Results: The arteries had an internal diameter between 0.04 and 2.20 mm (median 0.25 mm) on histology. Both US systems could display arteries with a diameter as small as 0.1 mm, but arteries less than 0.20 mm in diameter were better resolved with the 20 MHz system than by the 8.5 MHz system. Arteries with diameters less than 0.1 mm could not be reliably distinguished from artifacts. Arterial lumens were anechoic, thus giving a good contrast to the echogenic submucosa and subserosal fat. The two arteries in the mucosa and muscularis propria could not be seen on US images due to their similar hypoechoic appearance. Conclusions: These findings indicate that small gastrointestinal arteries can be defined in vitro using ultrasound frequencies between 8.5 MHz and 20 MHz. Further studies with ultrasonography done in vivo are needed to define their potential clinical role.

Forty-three vascularized iliac crest grafts in the femoral neck region in 38 patients were assessed with marrow scintigraphy. Twenty grafting procedures had been performed for treatment of fractures, 16 for nontraumatic osteonecrosis, and seven for focal bone lesions. Scintiscans were obtained preoperatively and in the early and late postoperative period. Anterior images of the hips were obtained 30 minutes after intravenous injection of 4 mCi (148 MBq) of technetium-99m tin colloid by using a gamma camera with a low-energy, general-purpose collimator. The viable graft was detected as an area of tracer uptake corresponding to the configuration of the graft on radiographs. Marrow scintigraphy at 12 weeks showed that 24 grafts were viable, 11 were nonviable, and eight were indeterminate. Early and late results concurred in 36 of 43 (84%) grafts. Nonviable grafts occurred most frequently in fractures in the elderly. In seven patients who subsequently required total hip replacement, six grafts were nonviable while one was indeterminate at 12 weeks. Results indicate that marrow scintigraphy is useful in the treatment of patients with vascularized iliac crest bone grafts.

SUMMARYDaily and hourly variations in microbial indicators densities in the beach-waters of Hong Kong have been described. The levels of Escherichia coli at a number of beaches was observed to be influenced by tide, and for staphylococci, by bather numbers. The tidal influence was most obvious during spring tides; and for the effect of bathers, during neap tides. Both organisms are present in high densities in external sources of faecal pollution of bathing beaches, with the average staphylococci to E. coli ratios being 0·04–3. Staphylococci may serve as an indicator of bather density and the risk of cross-infection amongst bathers (rather than as another indicator of faecal contamination) when the average staphylococci to E. coli ratio for a bathing beach is considerably higher than 3. The variability of microbial indicator densities means the routine sampling of bathing beaches should be carried out on weekend days with maximum numbers of swimmers exposed to the water, and spread throughout the bathing season.

SUMMARYThe standard membrane filtration method of the UK has been modified in order to improve its specificity for enumeratingEscherichia coliin the subtropical waters of Hong Kong. This involves incorporating into the membrane lauryl sulphate (mLS) method either anin situurease test (the mLS-UA method), or anin situβ-glucuronidase test (the mLS-GUD method). The false-positive errors of the mLS-UA and mLS-GUD methods are low, ranging from 3–5%. A comparison between the membrane filtration (mLS-UA) method and the multiple tube technique in testingE. coliin subtropical beach-waters has demonstrated that the former can give much more precise counts, and is the method of choice for such a purpose. The mLS-GUD method, for which automated counting ofE. colicolonies is possible, is a good alternative to mLS-UA in routine enumeration of this bacterial indicator in environmental waters.

A prospective epidemiological study was undertaken in Hong Kong in 1987, in which 18,741 usable responses were obtained. It showed bathing in the coastal beaches of Hong Kong poses an increased risk of developing gastrointestinal, ear, eye, skin, respiratory and total illness. Swimmers immersed in the more polluted beach waters are exposed to a significantly higher risk of contracting swimming-associated gastrointestinal, skin, respiratory and total illness. E. coli was found to be the best indicator for swimming-associated gastroenteritis and skin symptoms amongst the bathers, and a linear relationship could be established. Staphylococci was a good indicator for ear, respiratory and total illness, and should be used in complementary to E. coli. Beach water quality objectives for both E. coli and staphylococci have been proposed. A 4-tier classification system (rather than a single acceptability criterion) based on swimming-associated health risks has been developed for the beaches of Hong Kong. Information on the bacterial water quality and health risk levels of individual beaches is reported to the public both annually and fortnightly, so that beach-goers can choose where to go for swimming based on health effects data.

Organ donor shortage is now the greatest limitation to the success of organ transplantation. To assess and compare the role of a regional trauma center in identifying and referring cadaveric organ donors to that of a community hospital, we retrospectively reviewed records of potential cadaveric organ donors during a comparable 5-year period in each. The trauma center (TC) contributed 44 donors, while the community hospital (CH) supplied seven. Five hundred sixty-five possible donors were not harvested at the TC, compared to 126 at the CH. While most "non-donors" from both institutions were not suitable candidates, 8% could have served as good organ donors. Inability to obtain consent was the major obstacle in obtaining good candidates (70% at CH, 83% at TC). Failure to recognize and refer possible donors occurred in 2% of the cases at CH, compared to 0.4% at the TC. We conclude that regional trauma centers can play an important role in alleviating organ donor shortage.

The effects of intracoronary ultrapure human Interleukin-1 on the regional distribution of coronary blood flow (radioactive microspheres), contractile function (subendocardial ultrasonic length gauges) and systemic hemodynamics were studied in open-chest, anesthetized dogs (n = 7). Bolus doses of Interleukin-1 (10, 20, and 30 u) administered directly into the left anterior descending coronary artery increased coronary blood flow from 43 to 71, 80 and 87 ml/min, respectively. The increase in blood flow produced by Interleukin-1 was distributed uniformly to the subendocardium, midmyocardium, and subepicardium of the left ventricular free wall without effect on regional function or systemic hemodynamics. Indomethacin (1 mg/kg i.v.) attenuated the increase in blood flow, especially to the subepicardium. Due to the selective diminution of the Interleukin-1-mediated increase in subepicardial blood flow by indomethacin, the subendocardial to subepicardial perfusion ratio was increased by Interleukin-1 in the presence of indomethacin. The present results demonstrate that Interleukin-1 has direct coronary vasodilator actions, a portion of which is mediated by a product of cyclooxygenase metabolism.

One hundred and two liver transplants have been performed since the program's inception in Hawai'i in 1993. Viral hepatitis continues to be the primary indication for liver transplant, though hepatocellular cancer is involved in 23.5% of cases. One, 3 and 5-year patient survival rates have been 88%, 79%, and 74% respectively which is comparable to mainland centers and national data.

The well-known gap between organ-donor supply and demand in the United States is particularly acute for Asian Americans. Lower participation in organ donation programs by Asian Americans has been hypothesized as one explanation for this observation. This study finds that, relative to European Americans, Asian Americans hold more negative attitudes toward and participate less frequently in a large, urban organ-donor program. The study also hypothesizes and test possible reasons for subcultural differences in attitudes toward donation. Two cultural belief constructs hypothesized to more strongly predict Asian American attitudes and behaviors appear to impact both groups equally. Reasons for these results along with public policy implications and future research directions are discussed.

Aims: To review 25 cases of breast hamartoma and discuss the pathological criteria, and the usefulness of imaging modalities, fine needle aspiration cytology (FNAC), and needle core biopsy in the diagnosis. Methods: The hamartomas were assessed for interlobular fibrotic stroma, stromal adipose tissue content, pseudo-angiomatous stromal hyperplasia, and epithelial changes (hyperplasia, adenosis or apocrine metaplasia, and cyst formation). All imagings, previous FNACs, and biopsies were also reviewed. Results: Imaging (mammography, ultrasound, and magnetic resonance imaging) was performed in 18 cases, and mostly showed encapsulated masses with a heterogeneous appearance. Microscopically, all hamartomas demonstrated good demarcation with fibrous tissue condensation. Adipose tissue was noted in all cases (5–90%; mean, 31%), and interlobular fibrosis in 21 cases. Benign epithelial hyperplasia occurred in 10 cases, and pseudo-angiomatous stromal hyperplasia or cystic ducts in eight cases each. Apocrine metaplasia, calcification, stromal giant cells, and adenosis occurred in four cases or less. Two cases showed coexisting ductal carcinoma in situ limited to within the hamartoma. Needle core biopsies (four cases) and FNAC (14 cases) were largely insufficient, inconclusive, or non-specific. Conclusions: Hamartomas do not possess specific diagnostic histological features. The role of FNAC and needle core biopsy in making the diagnosis is limited, and requires clinical and radiological correlation to avoid underdiagnosis.

We report a rare epithelioid haemangioendothelioma in the frontal lobe. The CT and MRI findings are characteristic and correspond to histological features intermediate between those a cavernous haemangioma and an aggressive vascular tumour.

Synovial fluid basic calcium phosphate (BCP) crystals are markers of severe joint degeneration in osteoarthritis. These crystals are mitogenic and induce protooncogene expression and matrix metalloproteinase (MMP) synthesis and secretion in human fibroblasts, effects that are specifically blocked by phosphocitrate (PC). We have recently determined that crystals transduce signals to the nucleus via the activation of the p42 and p44 mitogen-activated protein (MAP) kinases (Nair et al., 1997, J Biol Chem 272:18920-18925). Treatment of human fibroblasts (HF) with BCP induces phosphorylation of p42/44 MAPK, which is inhibited by PC in a dose-dependent manner. Blocking of p42/44 MAPK signal transduction with an inhibitor (PD98059) of MEK1, an upstream activator of MAPKs, reduces crystal-induced p42/44 MAPK activation and significantly inhibits crystal-induced cell proliferation. Based on these findings, we sought to determine the role of the p42/44 MAPK signal transduction pathway in crystal-induced expression of matrix MMPs. We demonstrate suppression of crystal-induced MMPs via the utilization of two different MEK inhibitors: PD98059 and the recently described U0126, a novel inhibitor of MEK1 and MEK2. Treatment of HF with PD98059 blocks the induction of crystal-stimulated collagenase 1 (MMP-1) and stromelysin (MMP-3) expression. PD98059 and PC reduced the level of crystal-induced MMP-1 and MMP-3 mRNA expression to that observed in nonstimulated cells. Likewise, PD98059 treatment of HF blocked the epidermal growth factor (EGF)- and crystal-induced increases in MMP-1 and MMP-3 protein expression and secretion as demonstrated by Western blotting and zymography. Treatment of HF with U0126 inhibits EGF-induced phosphorylation of p42/44 MAPK as well as crystal- and EGF-induced upregulation of MMP-1 mRNA. Additionally, we demonstrate that treatment of HF with BCP, EGF, or PD98059 does not significantly alter levels of gelatinase A (MMP-2) mRNA and protein expression.

To investigate cellular signaling mechanisms that influence chondrocyte production of inorganic pyrophosphate (PPi), which promotes calcium pyrophosphate dihydrate (CPPD) crystal deposition.Articular chondrocyte and cartilage cultures were stimulated with protein kinase C (PKC) activator and adenyl cyclase activator. Generation of extracellular PPi was measured.Adenyl cyclase activation resulted in diminished pyrophosphate generation. PKC activation stimulated pyrophosphate elaboration.Two signaling pathways, cAMP and PKC, modulate generation of extracellular pyrophosphate by cartilage and chondrocytes. They are novel targets for potentially diminishing extracellular pyrophosphate elaboration that leads to CPPD crystal deposition.

Synovial fluid basic calcium phosphate (BCP) crystals are markers of severe joint degeneration in osteoarthritis. BCP crystals cause mitogenesis of articular cells and stimulate matrix metalloprotease production, thus promoting degradation of articular tissues. Previous work suggested that BCP crystal-induced cell activation required intracellular crystal dissolution, induction of proto-oncogene expression, and activation of signal transduction pathways involving protein kinase C and mitogen-activated protein kinases. Here we further elucidate the mechanisms of BCP crystal-induced cell activation as BCP crystals activate transcription factors nuclear factor kappaB and activator protein 1 in human fibroblasts. We confirm the role of protein kinase C in BCP crystal-induced mitogenesis in human fibroblasts. In contrast, we demonstrate that BCP crystals do not activate signal transduction pathways involving protein tyrosine kinases or phosphatidylinositol 3-kinase. These data further define the mechanism of cell activation by BCP crystals and confirm its selectivity, an observation that may have therapeutic implications.

Varicella-zoster virus (VZV), one of the human herpes viruses, is a well known virus that can infect and induce both chicken pox (varicella) and shingles (zoster). General vaccination against VZV is now seriously considered mainly for economical reasons. A live attenuated varicella vaccine was developed using Oka strain by a cell culture method (1). This varicella vaccine was approved for use in Japan, Korea, and USA (Biken), and use for healthy children is under consideration in some countries in Europe (2). However, so far it has been difficult to obtain high yield of infectious cell-free VZV for vaccine from cell culture. As an attempt to develop a cell-free VZV production process, we established a human embryonic lung cell culture, and VZV strains were isolated from patients and attenuated.

To prospectively compare magnetic resonance (MR) cholangiography with endoscopic retrograde cholangiography (ERC) in the diagnosis of choledocholithiasis.Forty-seven patients with suspected choledocholithiasis underwent non-breath-hold, heavily T2-weighted, respiratory-triggered turbo spin-echo MR cholangiography. They then underwent ERC within 5 hours. The results of the two procedures were compared in 45 patients.The absence of ductal dilatation was shown in 16 patients at MR cholangiography and at ERC. MR cholangiography showed common duct dilatation in 28 of the 29 patients with dilatation shown at ERC. MR cholangiography helped correctly identify 18 of the 19 patients with choledocholithiasis and 22 of the 26 patients without choledocholithiasis. Sensitivity with MR cholangiography was 95%, specificity was 85%, positive predictive value was 82%, and negative predictive value was 96%. Two of the false-positive findings were due to pneumobilia.Non-breath-hold MR cholangiography is as accurate for the evaluation of choledocholithiasis as ERC.

A retrospective study of computed tomography scans of the paranasal sinuses of 131 control subjects in Hong Kong revealed minor mucosal abnormalities in more than half of the ethmoid sinuses. Major abnormalities were present in seven per cent of maxillary, five per cent of anterior ethmoid and four per cent of posterior ethmoid sinus. In 85 patients with nasopharyngeal carcinoma the prevalence of minor mucosal abnormalities in the sinuses was similar to that of the control group but major mucosal abnormalities were significantly more common in the anterior and posterior ethmoids at 15 per cent and 21 per cent of the respective sinuses (p

Combined neutral endopeptidase (NEP) and angiotensin-converting enzyme (ACE) inhibition produces greater acute hemodynamic effects than either treatment alone. We investigated whether BMS-182657 (BMS), which bears inhibitory activities against both NEP and ACE, elicited similar enhanced effects. BMS inhibited NEP and ACE, in vitro (IC50 = 6 and 12 nM, respectively) and the pressor response to Ang I in rats. In deoxycorticosterone acetate (DOCA)-salt hypertensive rats sensitive to NEP inhibition but not to ACE inhibition, BMS at 100 mumol/kg i.v. lowered mean arterial pressure (MAP) from 180 +/- 6 to 151 +/- 5 mm Hg. In sodium-depleted, spontaneously hypertensive rats (SHR) sensitive to ACE inhibition but not to NEP inhibition, BMS at 100 mumol/kg p.o. lowered MAP from 151 +/- 4 to 123 +/- 5 mm Hg. Cardiomyopathic hamsters with heart failure were administered vehicle or one of the following (30 mumol/kg i.v.): the ACE inhibitor enalaprilat; the NEP inhibitor SQ-28603; or BMS. Enalaprilat and SQ-28603 had minimal hemodynamic effects. BMS decreased left ventricular end-diastolic pressure by 12 +/- 2 and 10 +/- 1 mm Hg and left ventricular systolic pressure by 27 +/- 2 and 23 +/- 3 mm Hg at 30 and 60 min, respectively (P.05 vs. each other group). These changes were associated with a 40% increase in cardiac output, a 47% decrease in peripheral vascular resistance and a lowering of MAP by 21 +/- 3 mm Hg at 60 min (P.05 vs. each other group). There were no significant differences in the changes in heart rate or left ventricular stroke work index among the four groups. Hence, BMS-182657 is a dual inhibitor of NEP and ACE, is antihypertensive irrespective of the activity of the renin-angiotensin system and has acute hemodynamic effects in hamsters with heart failure greater than those produced by selective inhibition of NEP or ACE. The NEP and ACE inhibitory activities of BMS-182657 act synergistically and mimic the interaction resulting from combining selective inhibitors of these enzymes.

Catheter infection is a major complication of continuous ambulatory peritoneal dialysis (CAPD) therapy for end-stage renal disease. Catheter exist-site infections were treated with a new surgical technique consisting of dissection and removal of the existing catheter in the subcutaneous layer, insertion of a catheter connector and new catheter piece, and creation of a new subcutaneous tunnel. The new surgery can be performed on an outpatient basis and allows for the continuation of CAPD, thereby avoiding the cost associated with inpatient admission and interim hemodialysis.

A means to monitor intestinal allografts will be crucial for the future success of small bowel transplantation. We have previously demonstrated the ability of high-frequency ultrasound (US) to diagnose porcine intestinal ischemia in vitro. The aim of this study was to compare the histologic appearance of normal porcine small bowel versus bowel undergoing acute rejection, using 8.5-MHz US images. We allowed porcine heterotopic small bowel allograft transplants to reject and then removed, at scheduled intervals from postoperative day 0 to 12, specimens of both the transplanted bowel and the native bowel. We examined the tissues in vitro with an 8.5-MHz linear array US system then studied them histologically. Histologically, the earliest changes of rejection occurred at days 4 to 5, with mild submucosal edema, endotheliitis, and vasculitis affecting the small vessels; the mucosa remained normal. By days 7 to 8, the submucosal endotheliitis became more prominent, with focal thrombosed small vessels; the mucosa now appeared abnormal with flattened villi, erosions, and necrosis. By days 10 to 12, marked submucosal edema, vasculitis, endotheliitis, and necrotic mucosa were present. Ultrasonically, normal intestinal wall has five wall layers, corresponding to mucosa, submucosa, muscularis propria, and subserosal fat. The US criteria for abnormality were loss of folds, decreased numbers of echo layers, discontinuity of layers, and a homogeneous appearance. Using these US criteria, blinded observers differentiated normal from abnormal bowel wall after transplantation with a sensitivity of 84% and a specificity of 81%. Most of the errors occurred with the day 4 and 5 specimens, which appeared nearly normal on US. This study demonstrates that US can potentially differentiate normal from abnormal changes after small bowel transplantation in vitro, and may be useful in monitoring the transplanted intestine. Abnormal US findings might then warrant a biopsy procedure. Further studies are necessary to fully evaluate the potential clinical use of US in intestinal transplantation.

Endoscopic ultrasound imaging of the gastrointestinal wall can be performed through intraluminal fluid or by direct transducer contact with the wall. We tested the hypothesis that the ultrasound appearance of the gastrointestinal wall is influenced by the amount of pressure applied when the transducer is in contact with the tissue. Fresh autopsy specimens from the porcine gastrointestinal tract were examined in vitro using an 8.5-MHz linear array ultrasound system. As transducer pressure against the wall was increased from 0 to 10 KPa, changes were seen on the images in wall thickness, tissue echogenicity, and the number of layers. The stomach and rectum were more resistant to compression than the esophagus, duodenum, and colon. Wall echogenicity increased with increasing degrees of applied pressure and some layers were obliterated by this pressure. The second ultrasound layer, or deep mucosa, appeared to be the most susceptible to compression. Endoscopic ultrasound imaging artifacts should be reduced by limiting the amount of pressure applied to the wall with the transducer.

Cheung AHS, Wang KY, Jiranek GC, Odegaard S, Kimmey MB, Silverstein FE. Evaluation of a 20-MHz ultrasound transducer used in diagnosing porcine small bowel ischemia. The authors have previously demonstrated the ability of an 8.5-MHz linear array to detect moderate or severe intestinal ischemia in a porcine model. This study compares the ability of the 8.5-MHz linear array with a prototype miniature 20-MHz ultrasound (US) imaging probe in detecting small bowel ischemia. Five piglets were studied in which vascular clamps were applied to isolated jejunal pedicles, then released sequentially at hourly intervals to induce ischemia from 0 to 6 hours. After 24 hours of reperfusion, the tissue was removed and examined with both the 8.5-MHz linear array and the 20-MHz probe. A histologic examination also was done. The acoustical criteria used for interpretation were presence or absence of folds, number of echo layers, relative thickness of layers and homogeneity and continuity of layers. The 8.5-MHz system predicted the duration of ischemia with a kappa value of 0.66 ± 0.03, whereas the 20-MHz system had a kappa value of 0.49 ± 0.03. Both systems were able to distinguish normal or mild ischemia from moderate or severe ischemia with sensitivity and specificity rates of at least 94%. Both 8.5− and 20-MHz US systems detected intestinal ischemia in vitro. Further studies are indicated to determine the ideal frequency and design for a US system that can be used clinically.

To evaluate the use of ultrasound (US) to detect intestinal wall ischemia, we isolated segments of jejunum on single vascular pedicles in five piglets. We sequentially clamped these segments in intervals of 0 to 6 h, reperfused them for 24 h, and then examined them in vitro histologically and with an 8.5 MHz US scan. All segments were grossly viable except those with 6 h of ischemia. Histologically, mild submucosal edema developed after 1 to 2 h of ischemia; after 3 to 4 h, mucosal necrosis, loss of folds, worsening submucosal edema, and prominent neutrophilic infiltration occurred; after 5 to 6 h, severe mucosal necrosis with hemorrhage and submucosal edema was present. Ultrasonically, we saw five wall layers in the control group corresponding to mucosa, submucosa, muscularis propria, and subserosal fat. After mild (1 to 2 h) ischemia, all layers were present except for a discontinuity in layer 3. After moderate (3 to 4 h) ischemia, the five layers persisted but with a markedly thickened submucosal layer, reduced echogenicity, and flattened mucosal folds. With severe (5 to 6 h) ischemia, we observed a loss of all normal layers with no discernible architecture. Using these US criteria, blinded observers were able to differentiate normal/mild from moderate/severe ischemia with a sensitivity and specificity of 100%. These data suggest that US can differentiate, in vitro, normal from moderate and severe degrees of intestinal wall ischemia that correlates well with the histological appearance. Endoscopic US or surgically implantable US probes can potentially help diagnose clinical intestinal wall ischemia.

An 82-year-old man developed periarticular roentgenographic calcifications of the knee joint. Fragments of meniscus and peritendinous tissue were collected during total knee arthroplasty. Crystals were released from tissues by collagenase digestion. Calcium pyrophosphate dihydrate crystals were identified in the meniscus, and basic calcium phosphate crystals were identified in the peritendinous tissue. Methods of crystal identification included compensated polarized light microscopy, scanning electron microscopy with X-ray energy dispersive analysis, and X-ray diffraction.

SUMMARYProspective epidemiological studies of beach water pollution were conducted in Hong Kong in the summers of 1986 and 1987. For the main study in 1987, a total of 18741 usable responses were obtained from beachgoers on nine beaches at weekends. The study indicated the overall perceived symptom rates for gastrointestinal, ear, eye, skin, respiratory, fever and total illness were significantly higher for swimmers than non-swimmers; and the swimming-associated symptom rates for gastrointestinal, skin, respiratory and total illness were higher at ‘barely acceptable’ beaches than at ‘relatively unpolluted’ ones.Escherichia coliwas found to be the best indicator of the health effects associated with swimming in the beaches of Hong Kong. It showed the highest correlation with combined swimming-associated gastroenteritis and skin symptom rates when compared with other microbial indicators. A linear relationship betweenE. coliand the combined symptom rates was established. Staphylococci were correlated with ear, respiratory and total illness, but could not be used for predicting swimming-associated health risks. They should be used to complementE. coli. The setting of health-related bathing-water quality standards based on such a study is discussed.

The disordered production of extracellular inorganic pyrophosphate (PPi) by cartilage contributes to calcium pyrophosphate dihydrate crystal formation and associated diseases. We have previously shown that a factor(s) derived from human platelets markedly stimulates the accumulation of PPi in the media of porcine articular cartilage in organ culture. This is the first known physiologic modifier of PPi production by cartilage. We report herein that platelet derived growth factor (PDGF) is not the platelet factor responsible for PPi stimulation and that the active factor is not mitogenic for chondrocytes. PDGF added to the media of articular cartilage explants in the presence of 0.5% (platelet-poor) plasma (PPP) produces 3.92 +/- 1.6 mumol/L PPi compared with 2.85 +/- 0.7 mumol/L PPi in cartilage exposed to PPP alone. The platelet extract (PE) and PDGF are mitogenic for adult articular chondrocytes in high-density monolayer cultures. Anti-PDGF antibodies block the mitogenic effects of PDGF and PE. The uptake of thymidine labeled with tritium is 157% of control in cells exposed to PE, PPP, and polyclonal goat immunoglobulin G (IgG); 114% of control in cells exposed to PE, PPP, and anti-PDGF antibody; 148% of control in cells treated with PDGF, PPP, and goat IgG; and 98% of control in cells treated with PDGF, PPP, and anti-PDGF antibody. However, anti-PDGF antibody has no effect on PPi accumulation. PPi levels are 17.22 +/- 1.6 mumol/L in media from cartilage treated with PPP, goat IgG, and PE and 17.62 +/- 2.2 mumol/L in cartilage exposed to PE, PPP, and anti-PDGF antibody. We have further characterized the platelet factor responsible for the stimulation of PPi by cartilage. It is not mitogenic for chondrocytes, and it is not PDGF.

Saccharomyces cerevisiae was immobilised by adsorption to untreated sugar cane bagasse in a packed bed reactor. Complete conversion of glucose to ethanol was obtained at a dilution rate of 0.19 h−1. Continuous ethanol production was maintained for up to 57 days. Reactor productivity increased with increasing packing density of the bagasse. Plugging of void spaces due to cell overgrowth led to channelling of the feed and decreased reactor productivity. Increasing the average column temperature alleviated plugging and restored column performance over a short period; however prolonged exposure to the high temperature resulted in decreased ethanol production rates. Bagasse has advantages as a support material for ethanol production from sugar cane or beet, including negligible cost, ready availability and the capacity to support a high yeast population.

Captopril is a remarkably effective new antihypertensive drug designed and developed as a potent and specific inhibitor of angiotensin-converting enzyme, a zinc metallopeptidase that participates in the synthesis of a hypertensive peptide, angiotensin II, and in the degradation of a hypotensive peptide, bradykinin. Earlier studies with a snake venom peptide (teprotride or SQ 20881) that could be administered only by injection demonstrated that specific inhibitors of angiotensin-converting enzyme could be highly effective as antihypertensive drugs, and helped to clarify the specificity and mechanism of action of the enzyme. A hypothetical model of the active center of angiotensin-converting enzyme based on its presumed analogy to the well characterized zinc metallopeptidase carboxypeptidase A was used to guide logical sequential improvements of a weakly active prototype inhibitor that led eventually to the highly optimized structure of captopril. The hypothetical working model of the active site of angiotensin-converting enzyme used to develop captopril continues to provide a firm basis for development of new types of specific inhibitors of this biologically important enzyme.

Synovial tissue hyperplasia in basic calcium phosphate deposition disease has been suggested to develop through the stimulation of cell growth by basic calcium phosphate (BCP) crystals deposited in joints. These crystals have been used in vitro to stimulate DNA synthesis in quiescent fibroblasts to experimentally study this proliferative disease. The stimulation of DNA synthesis, in density-arrested Balb/c 3T3 cells, by BCP crystals was inhibited after down-regulating protein kinase C activity with 12-O-tetradecanoyl-phorbol 13-acetate (TPA). No effect on platelet-derived growth factor (PDGF)-stimulated DNA synthesis was observed under the same conditions. The expression of c-myc and c-fos increased in response to BCP stimulation in a manner similar to the increase produced by stimulation with PDGF. The BCP stimulation of c-fos and c-myc messages was inhibited 60 and 90%, respectively, in TPA-pretreated, protein kinase C-down-regulated cells. The induction of these transcripts by PDGF was unaffected in cells pretreated with TPA. TPA was unable to stimulate c-fos and c-myc expression or DNA synthesis following protein kinase C down-regulation. Both PDGF and TPA stimulated phosphorylation of an 80-kDa protein, whereas BCP crystals had no effect on phosphorylation of this protein. The exposure of density-arrested Balb/c 3T3 cells to BCP crystals had no effect on high affinity epidermal growth factor receptor binding under conditions in which PDGF and TPA reduced epidermal growth factor binding. The data suggest that PDGF can act to stimulate c-fos and c-myc expression as well as DNA synthesis through a protein kinase C-independent pathway, whereas BCP crystals require at least endogenous levels of protein kinase C to stimulate these events.

Basic calcium phosphate (BCP) crystals control the traverse of cells from the G0-G1 to S-phase of the cell cycle and initiate proliferation by rendering fibroblasts competent to respond to insulin-like growth factors in plasma. The present study examines whether BCP crystals induce transcription of the protooncogenes c-fos and c-myc and the effect of beta-interferon (IFN-beta) on protooncogene transcription as well as BCP crystal-induced DNA synthesis. Stimulation of density-arrested BALB/c-3T3 cells with either BCP crystal or platelet-derived growth factor (PDGF) results in maximal accumulation of c-fos mRNA at 30 min after stimulation. Induction of c-myc transcription by BCP crystal or PDGF occurs within 1 h and is maximal at around 3 h after stimulation. Simultaneous addition of IFN-beta with either BCP crystals or PDGF had little effect on c-fos induction but delayed both c-myc message accumulation and entry into S phase. The delay in c-myc message induction after IFN-beta treatment cannot account for the observed delay in the onset of DNA synthesis, since IFN-beta can be added at up to 6 h after stimulation with either PDGF or BCP crystals, and a similar delay in the onset of DNA synthesis is still observed.

From available clinical, radiographic, and synovial fluid findings, coupled with in vivo radiolabelled crystal turnover data and in vitro experimental data, a hypothesis has been formulated relative to the pathogenesis of BCP crystal deposition diseases (Fig. 2). Synovial lining cells phagocytose BCP crystals and particulate collagens in the joint fluid. During and/or after internalization these cells are stimulated in a variety of ways: 1) protease synthesis and secretion is relentlessly stimulated, which may damage joint tissues producing clinically evident loss of collagenous tissues including cartilage, bone, and tendon, and which may release additional amounts of crystals and particulate collagens into the synovial fluid, completing a vicious cycle; 2) PGE2 production is greatly augmented; 3) DNA synthesis is stimulated as a result of increased inositol phospholipid turnover and intracellular crystal dissolution. The increased number of synovial cells also augments the total local generation of proteases and prostenoids. Mechanical factors such as trauma or joint overuse also contribute to the pathogenesis of joint destruction as discussed in the article on the clinical aspects of BCP crystal deposition.

An in vitro model of ADA deficiency is selectively toxic to cartilage from immature rabbits with a greater effect on growth plate than articular cartilage. The selective toxicity observed appears to be the consequence of ATP depletion. These results support the hypothesis that the chondro-osseous dysplasia observed in patients with ADA deficiency is caused by the disordered metabolism that results from the enzyme deficiency.

To examine the repair collagens produced by cells in injured cartilage, the femoral articular surfaces of three groups of New Zealand white rabbits were injured by making both superficial and deep lacerations and drill holes. Eight weeks after surgery, the rabbits were killed and slices of injured articular cartilage were harvested. The types of collagen being synthesized at the site of these lesions were identified by labeling the recovered specimens in vitro with 3H-proline and by characterizing the collagen using sodium dodecyl sulphate electrophoresis, carboxymethyl cellulose chromatography, and cyanogen bromide peptide analysis. In all cases, tissue-specific type II ([alpha1 (II)]3) cartilage collagen was synthesized. Histological examination using toluidine blue showed that the chondrocytes bordering the cartilage defect produced by deep lacerations and drill holes responded by increased cellular activity, as shown by cell cloning and increased matrix staining. The drilled holes were completely filled by tissue with staining and morphological characteristics similar to those of hyaline cartilage.

The healing of surgically induced defects in mature rabbits' hyaline cartilage was examined histologically and biochemically. The subchondral bone underneath the lesion was the source of repair. At the end of the first month, mushroom-shaped chondroid buds were seen sprouting from the subchondral bone. By the tenth week, these buds fused to a cartilagenous plug filling the lesion. The repaired cartilage synthesized Type II collagen.

Collagenase has been implicated in the pathogenesis of several arthropathies. Arteparon [glycos-aminoglycan (GAG) polysulfate] is a proteinase inhibitor that is being investigated as a therapeutic agent in osteoarthritis. We found that cultures of calcified lapine synovium release collagenase, one tenth of which is already activated. Normal synovium produces no active collagenase. GAG polysulfate suppresses the amount of active collagenase by one half. GAG polysulfate may interfere with the activation process of collagenase.

Angiotensin-converting enzyme is one of the enzyme components of the renin-angiotensin system (Figure 1), the products of which play physiologically important roles in maintenance of cardiovascular homeostasis and contribute to the elevation of arterial blood pressure in various hypertensive disease states.1–4 The immediate product of the action of angiotensin-converting enzyme is the octapeptide, angiotensin II, the most potent naturally occurring pressor substance known. Angiotensin III, a hepta-peptide derived from a further enzymatic cleavage of angiotensin II, is a potent stimulator of secretion of aldosterone by the adrenal cortex.5 Angiotensin-converting enzyme also plays a biologically important role in the inactivation of the potent vasodepressor peptide bradykinin.6

The similarity of the biologically important enzyme angiotensin-converting enzyme to the structurally characterized digestive enzyme carboxypeptidase A has led us to develop a hypothetical model of the mechanism of binding of substrates to its active site. In this model, a positively charged group on the enzyme forms an ionic bond with the negatively charged carboxyl group of the substrate; a hydrogen bonding group of the enzyme binds with the terminal peptide bond of the substrate, and the tightly bound zinc ion of the enzyme binds to the penultimate (scissile) peptide bond of the substrate. Succinyl- l -proline (SQ 13,745) was synthesized as a potential inhibitor of angiotensin-converting enzyme by analogy to d -2-benzylsuccinic acid, an inhibitor of carboxypeptidase A; it was a moderately potent but specific inhibitor of the enzyme. Structure-activity studies carried out using the hypothetical model as a guide led to the synthesis of d -2-methyl-succinyl- l -proline (SQ 13,-297) and d -2-methylglutaryl- l -proline (SQ 14,-102), more potent inhibitors of the enzyme that were shown to be orally active in rats. Attempts to replace the zinc-binding carboxyl group of these compounds with groups with greater affinity for zinc have led to the synthesis of extremely potent inhibitors such as 3-mercapto-propanoyl- l -proline (SQ 13,863) and d -3-mercapto-2-methylpropanoyl- l -proline (SQ 14,225). The most active compound, SQ 14,225, is a purely competitive inhibitor of angiotensin-converting enzyme with an enzyme-inhibitor dissociation constant (Ki) of 1.7 × 10−9M. It is an extremely potent and specific inhibitor of angiotensin-converting enzyme and appears to have great potential for the treatment of hypertensive disease.

Hydroxyapatite, like other calcium-containing crystals previously studied by us, is mitogenic for cultured human fibroblasts. This mitogenic effect is not a result of increased ambient calcium concentration due to extracellular crystal dissolution. Synthetic crystals labelled uniformly with calcium 45 (45Ca) undergo endocytosis when incubated with cells and are solubilized. Such solubilization is inhibited by chloroquine or ammonium chloride in concentrations that significantly block the mitogenic effect of crystals but not that of serum. The data suggest that mitogenesis and intracellular crystal dissolution are related phenomena.

Nachweis mit Analogen der im Gifte vonBothrops jararaca gefundenen Peptide, dass die Inhihibition des Angiotensins «converting enzyme» von zwei eindeutigen Teilsequenzen dieser Peptide abhangig ist. Die hohe spezifische kompetitive Inhibition, hervorgerufen durch die Peptide vonBothrops jararaca, wird der Bindung ihrer Tripeptidreste vom Carboxyterminus mit dem aktiven Zentrum des Enzyms zugeschrieben, die in gleicher Weise wie die Peptidsubstrate mit dem Enzym gebunden werden. Die Wirksamkeit der Giftpeptide hangt von der Bindung eines zweiten Teiles der Peptide mit dem Enzym ab.