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Biomedical subjects

N K Lim

Publications and source records attributed to N K Lim.

12 recordsLinked to original sources
PubMed1999

Crystal structure of the trimeric alpha-helical coiled-coil and the three lectin domains of human lung surfactant protein D.

BACKGROUND: Human lung surfactant protein D (hSP-D) belongs to the collectin family of C-type lectins and participates in the innate immune surveillance against microorganisms in the lung through recognition of carbohydrate ligands present on the surface of pathogens. The involvement of this protein in innate immunity and the allergic response make it the subject of much interest. RESULTS: We have determined the crystal structure of a trimeric fragment of hSP-D at 2.3 A resolution. The structure comprises an alpha-helical coiled-coil and three carbohydrate-recognition domains (CRDs). An interesting deviation from symmetry was found in the projection of a single tyrosine sidechain into the centre of the coiled-coil; the asymmetry of this residue influences the orientation of one of the adjacent CRDs. The cleft between the three CRDs presents a large positively charged surface. CONCLUSIONS: The fold of the CRD of hSP-D is similar to that of the mannan-binding protein (MBP), but its orientation relative to the alpha-helical coiled-coil region differs somewhat to that seen in the MBP structure. The novel central packing of the tyrosine sidechain within the coiled-coil and the resulting asymmetric orientation of the CRDs has unexpected functional implications. The positively charged surface might facilitate binding to negatively charged structures, such as lipopolysaccharides.

Amino Acid Sequence↗
PubMed1991

An in vitro assessment of human fetal pancreatic islets of Langerhans in culture.

The human fetal pancreas is a potential source of islets for transplantation into insulin-dependent diabetic patients. In this study, 35 human fetal pancreas obtained from prostaglandin-induced abortions (12-26 weeks gestation), were placed in culture to determine their capacity to secrete insulin over 30 days. Culture media were sampled twice weekly for insulin and histology was performed serially. Of the 35 pancreases cultured, six were lost due to bacterial contamination, five discarded due to undetectable levels of insulin in culture, nine are still under study, whilst 15 pancreases have been cultured for one month, and insulin studies completed. Three patterns of insulin release were observed: (a) progressive decline (n = 6), indicating non-viable tissue at the onset; (b) delayed decline, indicating significant tissue damage before organ culture (n = 5); and (c) insulin production in vitro over 30 days (n = 4), with viable islets detected histologically. Factors such as gestational age and cold ischaemia time did not correlate with the pattern of insulin secretion observed. This was probably due to a more important variable, not easily assessed, of the period of intrauterine (warm) ischemia. These data suggest: (1) that a small number of fetal pancreases procured from prostaglandin-induced abortuses do yield islets which remain viable in culture over 30 days, and (2) the functional status of islets can be monitored in vivo by measuring insulin secretion, thereby providing a means of identifying tissue suitable for transplantation.

Culture Techniques↗
PubMed1991

Fetal pig pancreas--a preliminary assessment of tissue for transplantation.

Porcine fetal pancreases (PFP) obtained from 4 pregnant sows were pooled, minced into 1 mm3 fragments and studied in organ culture for up to 30 days to determine tissue viability and insulin production in vitro. After 7-9 days in culture, some of these explants were transplanted into euglycemic, N:NIH-nu(s) nude recipient mice, and studied histologically over 69 days following grafting. Using RPMI 1640 supplemented with 5% fetal calf serum as the culture medium in 90% air/10% CO2, it was found that explants were viable with insulin production detected in vitro, which was maximal at day 7 (197 +/- 18.9 mU/L, n = 11), and gradually declined thereafter. By 22 days, insulin levels were less than 60.1 +/- 28.5 mU/L (n = 6). Histology of the explants showed viable tissue with evidence of mitoses present in insulin-positive cells at day 16 in vitro. Beyond this time, tissue viability diminished. Explants transplanted into euglycemic nude mice did not undergo rejection during the observation period of 69 days. Grafts remained viable with evidence of an increase in mitotic activity in the endocrine tissue on immunoperoxidase staining. These preliminary investigations confirm that pancreatic explants from porcine fetuses can be maintained in culture for up to 16 days. Such explants, when transplanted under the kidney capsule of euglycemic, nude mice, did not undergo necrosis, but remained viable, with evidence of mitoses in the islet tissue.

Animals↗
PubMed1991

A study of the xenogeneic response in an isolated liver perfusion circuit--preliminary observations.

An isolated liver perfusion circuit was developed to study the xenogeneic reaction of human blood to porcine liver, and investigate the feasibility of using such a system for liver dialysis in fulminant hepatic failure. Three experimental groups were studied: a control group where pooled porcine blood was perfused through pig liver (n = 12), a xenogeneic group where banked human blood was perfused through porcine liver (n = 23), and a modified xenogeneic group where decomplemented human blood was perfused through porcine liver (n = 4). The following parameters of liver function were assessed: liver function tests, serum electrolytes, bile and ascites production. In addition, liver histology was assessed at the start and completion of each perfusion experiment. Control experiments established that the use of low perfusion pressures (mean inlet pressure of 29.5 +/- 7.4 mmHg), and low haematocrit of 20.5 +/- 8.9% (n = 14), enabled five hour perfusions to be consistently achieved with maintenance of normal acid base and electrolyte balance. Bile production over 5 hours was 18.8 +/- 8.0 ml in controls (n = 5) and 17.0 +/- 6.7 ml in the xenogeneic (human-pig) circuit (n = 11) (NS). Ascites production was 235.8 +/- 157.3 ml/hr in controls (n = 5) and 205 +/- 142.0 ml/hr in the xenogeneic circuit (n = 7) (NS).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗