Development of a new dynamic method for quantitative evaluation of in vitro hemocompatibility of biomedical materials.

In this study a new dynamic method is introduced allowing the estimation of blood cell adhesion on flat test surfaces by measuring the cell loss in the bulk phase of surface contacting test blood under defined rheological conditions. This was achieved by constructing a novel test chamber permitting the contact of small amounts of blood with a large geometrical test surface. The construction consists of a spiral-shaped flow channel of 0.3 cm width, 0.02 cm height and 78 cm length covered with the biomaterials to be tested from both sides. Laminarity of blood flow in the conduit was confirmed theoretically by the calculation of an equivalent to the Reynolds number for curved systems the so-called Dean number. Furthermore, flow laminarity was proved experimentally finding that the flow rate of blood with different hematocrit values was proportional to the hydrostatic pressure applied. The applicability of the novel 'spiral method' for the estimation of hemocompatibility was demonstrated by evaluation of platelet adhesion onto different polymers in comparison to siliconized and fibrinogen coated glass as reference surfaces. Additionally, it was possible under distinct conditions to determine the adhesion of leucocytes and the detachment of platelet aggregates. Therefore, it was concluded that the spiral method can be used for the assessment of the hemocompatibility of flat biomedical polymers. As main advantages of the new method can be considered the high time efficiency and accuracy without labelling or optical detection of adherent cells.     

Comparative studies of the activity of disaccharidases in the mucosa of the small intestines in dystrophic, formerly hypotrophic-born young children and of patients with flat mucosa of various etiology

The disaccharidase activities of the mucous membrane of the small intestine were determined in formerly hypotrophic children who showing a dystrophy with a morphological normal mucosa (n = 36), patients with a flat mucosa caused by enteral protein intolerances of different genesis (n = 27), patients with a morphologically and functionally normal mucosa (n = 51). In about half of the former small-for-date children were shown activities lower than the simple standard deviation of the normal value, for lactase n = 17, for sucrase n = 14, for maltase n = 12. Some children showed pathologically reduced activities even below the double standard deviation for the normal value: lactase n = 8, sucrase n = 5, maltase n = 3.     

Ambulatory 24-hour esophageal pH monitoring

If 24-hour esophageal pH monitoring is to be a useful diagnostic tool, it must reliably discriminate gastroesophageal reflux patients despite daily variations in distal esophageal acid exposure. To address this issue, we studied 53 subjects (14 healthy normals, 14 esophagitis patients, and 25 patients with atypical symptoms) with two ambulatory pH tests performed within 10 days of each other. Intrasubject reproducibility of 12 pH parameters to discriminate the presence of abnormal acid reflux was determined. As a group, the parameters of percent time with pH<4 (total, upright, recumbent) were most reproducible (80%). Therefore, a subject was defined as having gastroesophageal reflux disease if at least one of these three values were abnormal. Intrasubject reproducibility for the diagnosis of reflux disease was 89% for the entire sample. Among subsets, the reproducibility was 93% for the normals and esophagitis patients and 84% for the atypical symptom patients. Total percent time with pH<4 was the single most discriminate pH parameter (85%) and nearly equaled that of the three combined parameters (89%). The intrasubject variability of this parameter was determined by the mean ±2sd of the relative differences between the two test results for all 53 subjects. Total percent time with pH<4 may vary between tests by a factor of 3.2-fold or less (218% higher to 69% lower). We conclude: (1) ambulatory 24-hr esophageal monitoring is a reproducible test for the diagnosis of gastroesophageal reflux disease; and (2) the large intrastudy variability in 24-hr total acid exposure may limit this test's usefulness as a measurement of therapeutic improvement.Supported, in part, by Public Health Services Grant AM 34200-01A1 from NIADDIK.     

Suppression of canine antral gastrin secretion by urine

Distention of the gastric antrum with an alkaline fluid normally results in the secretion of gastrin. Following gastrocystoplasty in humans, however, hypergastrinemia has not been observed. We explored the possibility that a component of urine may suppress antral gastrin activity in the dog. Partial cystectomy and antral transposition to the bladder (ATB) was performed in five animals and antral transposition to the colon (ATC) was performed in five other dogs to serve as a hypergastrinemic controls. At four and eight weeks after surgery the mean serum gastrin levels in the ATC dogs were significantly greater than the mean preoperative levels (p less than 0.05). In contrast, at four and eight weeks after surgery the mean serum gastrin levels in the ATB animals were significantly less than the mean preoperative levels (p less than 0.05). The antral G-cell density as determined by immunohistochemical study at eight weeks after surgery was greater than normal in the ATC dogs but less than normal in the ATB dogs; but the differences did not achieve statistical significance. In another series of experiments using four other dogs a 4% aqueous peptone solution and a 4% peptone solution in concentrated dog urine were instilled into exteriorized antral pouches. The mean serum gastrin levels at 60 and 90 minutes after instillation of the former were significantly increased (p less than 0.05), but there was little or no change after instillation of the latter. Urine, or a component of urine, appears to suppress canine antral gastrin secretion and may explain the absence of hypergastrinemia following gastrocystoplasty in humans.     

Histological comparison of implanted cadaveric and porcine dermal matrix grafts.

OBJECTIVES: Histological comparison of human-based (AlloDerm) and porcine-based (ENDURAGen) dermal matrices regarding tissue incorporation and neovascularization as potential soft tissue augmentation materials. STUDY DESIGN: In vivo, rat model. METHODS: Subcutaneous implantation of 1-mm thick, 1 cm x 1 cm pieces of AlloDerm, ENDURAGen, and meshed ENDURAGen was performed in 24 Sprague Dawley rats. Implant materials were harvested at 4 (n = 12) and 8 weeks (n = 12). Histological quantification of soft tissue ingrowth and microvascular density was performed following hematoxylin-eosin staining and CD34 immunohistochemistry, respectively. RESULTS: AlloDerm showed significantly greater soft tissue ingrowth and microvascular density compared with both ENDURAGen and meshed ENDURAGen at 4 and 8 weeks (P < 0.001). CONCLUSIONS: Although these results may differ in human host tissues, AlloDerm seems to be a more suitable dermal matrix implant than ENDURAGen for cases in which tissue incorporation and neovascularization are sought for the optimal outcome based on this animal model.     

Assessment of renal artery stenosis by phase-contrast magnetic resonance angiography

Three-dimensional (3D) phase-contrast magnetic resonance angiography (MRA) and velocity-encoded cine magnetic resonance (VEC-MR) imaging were performed in 23 subjects to assess the severity of renal artery stenosis. MRA was used for detection of stenosis, demonstrating a sensitivity of 100% and a specificity of 80%; the severity of stenosis was overestimated in 33%. VEC-MR was used to quantify the renal flow oattern and was successful in 11 subjects. Mean blood flow of normal renal arteries (420 +- 107 ml/min) was significantly higher (P < 0.01) than mean blood flow of stenotic arteries (131 +- 46ml/min). The flow profile displayed both systolic and diastolic peaks in 75% of the normal arteries, while the flow in stenotic arteries showed only a single systolic peak in all cases. The systolic peak in stenotic arteries occurred significantly later (32 +- 3% of the period of one cardiac cycle) than in normal subjects (21 +- 7%) (P < 0.05). Phase-contrast MR is likely to gain considerable importance in the noninvasive aetection and quantification of renal artery stenosis. Correspondence to: C. S. Richter     

Intestinal first-pass metabolism of nitrosamines. 2. Metabolism of N-nitrosodibutylamine in isolated perfused rat small intestinal segments

In a previous study a high first-pass metabolism of N-nitrosodi-[1-¹⁴C]butylamine(NDBA, O.32–730p.µM) has been shown in isolatedperfused rat small intestinal segments. In the present workthe identification and quantitation of metabolitesin samplesof perfusate and absorbed fluid (absorbate) is reported. AfterHPLC enrichment and purification five metabolites could be identifiedby GLC-MS: N-nitrosobutyl-(3-hydroxybutyl)amine (NB3HBA), N-nitrosobutyl-(4-hydroxybutyl)amine(NB4HBA), N-nitrosobutyl-(3-carboxypropyl)amine (NB3CPA), N-nitrosobutyl-(2-hydroxy-3-carboxypropyl)amine(NB2H3CPA) and N-nitrosobutylcarboxymethylamine (NBCMA), representing> 95% of total metabolites. -hydroxylatlon leading to NB4HBAand NB3CPA accounted for 75–95% of total metabolites.The formation of their follow-up products NB2H3CPA and NBCMAwas too small for quantitative analysis. In absorbate NB3CPAwas by far the most important metabolite. The hydroxylationof NB4HBA to NB3CPA was more efficient in jejunal as comparedto ileal segments.-1-hydroxylation to NB3HBA and, as reportedpreviously, -hydroxylation were only minor metabolic pathways.In conclusion, a high first-pass metabolism of NDBA to the proximatebladder carcinogen NB3CPA takes place during its absorptionin rat small intestine. This is in contrast to the observationin rat liver preparations, where - and -1-hydroxylation arethe predominant pathways. The intestinal first-pass metabolismmay play a key role explaining the increased incidence of bladdertumors in rats after low oral doses of NDBA.     

Impaired tactile pattern recognition in the early stage of primary degenerative dementia compared with normal aging

Tactile pattern recognition of ten different patterns which were engraved upon 4 x 4 cm plastic plaquettes with one hand and thereafter drawing with the other or the same hand was tested in 21 patients with a mild stage primary degenerative presenile onset dementia of Alzheimer type (DAT) and in 14 patients with a questionable dementia on the background of a chronic cardiovascular disease (CVD) in comparison to 15 healthy subjects of the same age (51 years old, elder controls, (EC)) and 16 younger subjects (22 years old, younger controls, (YC)). Patients made about four times more errors than the EC group. Duration from the begin of touching to the end of drawing was significantly longer in patients than in controls. In a second task the subjects had to recognize four subsequent patterns, then to perform a standard multiplication task and afterwards to draw all four patterns in their correct sequence. Patients made about ten times more errors in pattern recognition and series reproduction than age-matched controls.