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Biomedical subjects

N Lucas

Publications and source records attributed to N Lucas.

13 recordsLinked to original sources
PubMed2002

Comparison of bacterial colonization rates of antiseptic impregnated and pure polymer central venous catheters in the critically ill.

A study was performed on critically ill patients to evaluate the rate of colonization and catheter-related sepsis using antiseptic bonded (Arrowguard Arrow International) versus smooth pure polymer (Infectguard MedexMedical Ltd) central venous catheters. Two hundred and thirty-two catheters were inserted into 181 patients. Indications for removal included local or systemic infection, the catheter was no longer required and patient death. No statistical difference in colonization rate was found between the two types of catheter.

Anti-Infective Agents, Local↗
PubMed2000

Transplantation of fetal liver tissue suspension into the spleens of adult syngenic rats: effects of different cytotoxins on cytochrome P450 isoforms expression and on glycogen content.

Syngenic fetal liver tissue suspensions were transplanted into the spleens of adult male Fisher 344 inbred rats. Four months after surgery, transplant recipients and age matched control rats were treated with different cytotoxins (allyl alcohol [AAL], bromobenzene [BBZ], carbon tetrachloride [CCl4], or thioacetamide [TAA]) or the respective solvents 24 or 48 hours before sacrifice. Effects of the cytotoxins on the expression of three cytochrome P450 (P450) isoforms, 1A1, 2B1 and 3A2, within spleens and livers were assessed by immunohistochemistry. Additionally, effects on glycogen content within the hepatocytes were examined. In the livers AAL caused small lesions and fatty degeneration of hepatocytes only in some periportal areas. BBZ led to a perivenous necrosis of single cells only, whereas CCl4 and TAA caused complete necrosis of the centrilobular parenchyma. Treatment with each of the four cytotoxins led to necrosis and fatty degeneration of single or groups of hepatocytes within the intrasplenic transplants. This effect was most pronounced with CCl4 and TAA. The orthotopic livers of both solvent treated transplant recipients and control rats displayed only in few lobules a slight P450 1A1, but in all lobules a strong P450 2B1 and 3A2 expression, all mainly located in the hepatocytes around the central veins. AAL administration led to an increase in the P450 2B1 expression in the perivenous hepatocytes, whereas the staining for P450 1A1 was not affected and that for P450 3A2 in the periportal areas was even decreased. BBZ administration caused a P450 1A1 expression in the periportal hepatocytes but a decrease in this staining of the perivenous cells. The number of hepatocytes positively stained for P450 2B1 and 3A2 in the perivenous and intermediate zones was diminished in comparison to the livers of solvent treated rats. TAA and, more pronounced, CCl4 administration caused a strong reduction in the expression of all three P450 isoforms. Spleens of control rats displayed almost no P450 isoforms expression, independent of the treatment with the cytotoxins. Similar to adult liver, the hepatocytes in the transplant containing spleens showed nearly no P450 1A1, but a noticeable P450 2B1 and 3A2 expression. No staining was observed within the bile duct cells of the intrasplenic transplants. AAL administration slightly reduced the P450 2B1 and 3A2 expression in the transplants. BBZ and, much more pronounced, CCl4 and TAA treatment diminished the staining for all three P450 isoforms. AAL administration led to a marked decrease in the glycogen content of the hepatocytes of the periportal zones of the liver lobules, whereas after BBZ, CCl4 and TAA treatment a strong perivenous reduction in the glycogen content was seen. Similarly, within the intrasplenic transplants a remarkable decline in the glycogen content of the hepatocytes was caused by the treatment with each of the four cytotoxins. Especially after AAL and BBZ treatment the glycogen depletion within both livers and transplants was much more pronounced than the effects on morphology or P450 isoforms expression. It can be concluded that the effects of cytotoxins like AAL, BBZ, CCl4 or TAA seen in normal orthotopic liver are exerted in a similar way also in intrasplenic liver cell transplants.

Animals↗
PubMed1999

Transplantation of fetal liver tissue suspension into the spleens of adult syngenic rats: effects of various mitogens and cytotoxins on cytochrome P450 (P450) isoforms expression and on P450 mediated monooxygenase functions.

Syngenic fetal liver tissue suspensions were transplanted into the spleens of adult male Fisher 344 inbred rats. Four months after surgery, transplant recipients and age matched control rats were treated with various mitogens (fluorene [FEN], fluorenone [FON] and 2-acetylaminofluorene [AAF]) or cytotoxins (allyl alcohol [AAL], bromobenzene [BBZ] and carbon tetrachloride [CCl4]) or the respective solvents 24 or 48 hours before sacrifice. The expression of three cytochrome P450 (P450) isoforms, 1A1, 2B1 and 3A2, within spleens and livers was assessed by immunohistochemistry and P450 mediated monooxygenase functions in spleen and liver 9000 g supernatants by the model reactions ethoxyresorufin O-deethylation (EROD), ethoxycoumarin O-deethylation (ECOD), and ethylmorphine N-demethylation (EMND). The orthotopic livers of both solvent treated transplant recipients and control rats displayed only in few lobules a slight P450 1A1, but in all lobules a moderate P450 2B1 and 3A2 expression, all mainly located in the hepatocytes around the central veins. Correspondingly, regular EROD, ECOD and EMND activities were observed. Each of the three mitogens increased the P450 1A1 expression in the hepatocytes of the perivenous zones of the liver lobules. FON administration caused an additional P450 1A1 immunostaining in the periportal areas, and AAF treatment a P450 1A1 expression in bile duct epithelia. Also the staining for P450 2B1 and 3A2 in the hepatocytes of the perivenous and intermediate zones of the liver lobules was intensified after treatment with any of the mitogens. The three model reactions were significantly increased within the livers after FEN and FON administration, whereas after AAF treatment only ECOD was enhanced, EROD remained unaffected and EMND was decreased. The cytotoxin AAL caused small lesions and fatty degeneration of hepatocytes only in some periportal areas. BBZ only produced a perivenous necrosis of single cells, whereas CCl4 caused complete necrosis of the centrilobular parenchyma. Immunohistochemically, AAL administration led to an increase in the P450 2B1 expression in the perivenous hepatocytes, whereas the staining for P450 1A1 was not affected and that for P450 3A2 was even decreased in the periportal areas. Due to AAL treatment EROD and EMND activities were not affected and ECOD activity was increased. BBZ administration caused a P450 1A1 expression in the periportal hepatocytes but a decrease in this staining of the perivenous cells. The number of hepatocytes positively stained for P450 2B1 and 3A2 in the perivenous and intermediate zones was diminished in comparison to the livers of solvent treated rats. After BBZ treatment, EROD and EMND activities were decreased, ECOD activity was not affected. CCl4 administration caused a strong reduction in the expression of all three P450 isoforms and in the activity of all three model reactions. Spleens of control rats displayed almost no P450 isoforms expression and P450 mediated monooxygenase functions, without as well as after treatment with the mitogens or cytotoxins. Similar to adult liver, the hepatocytes in the transplant containing spleens showed nearly no P450 1A1, but a noticeable P450 2B1 and 3A2 expression. No staining was observed within the bile duct cells of the intrasplenic transplants.

2-Acetylaminofluorene↗
PubMed1998

Developmental expression of cytochrome P450 isoforms after transplantation of fetal liver tissue suspension into the spleens of adult syngenic rats.

In the present study, the developmental expression of three cytochrome P450 (P450) isoforms, 1A1, 2B1 and 3A2, and the ability to store glycogen was investigated in intrasplenic liver cell explants in comparison to adult and fetal liver. Fetal liver tissue suspensions were transplanted into the spleens of adult male syngenic Fisher inbred rats. Animals were sacrificed at 3 days, 1, 2, 4 weeks, 2, 4, 6 months and 1 year after transplantation. Spleens and livers of transplant recipients were compared to those of sham operated and control rats. Three days after transplantation little bulks of hepatocytes and only few bile ducts were seen in the red pulp of the transplant containing spleens. A massive hypertrophy and proliferation of bile ducts and also an augmentation in the number of hepatocytes were observed 4 weeks after transplantation. One month later, however, the bile ducts had become more and more atrophic, while instead the number of hepatocytes continuously increased. One year after surgery large masses of hepatocytes with apparent cord structure and only few but well preserved bile ducts were seen. Within the livers of adult rats, P450 1A1 was only slightly expressed by some hepatocytes around the central veins. P450 2B1 and 3A2 isoforms expression was much stronger, but also predominantly located in the hepatocytes of the central zone of the liver lobule. Hepatocytes of fetal livers displayed a moderate P450 1A1 expression. In some cells also a very mild staining for P450 2B1 and 3A2 was observed. Within the hepatocytes of the intrasplenic liver cell explants P450 1A1 was still expressed 3 days after transplantation, disappeared at 1 week after surgery, but reappeared at 4 weeks after transplantation. After 2, 4 and 6 months no staining for P450 1A1 was detectable any more. One year after transplantation again a slight P450 1A1 expression appeared. With P450 2B1 and 3A2 a mild to moderate expression was seen already at 3 days after transplantation. Four weeks after surgery nearly all of the hepatocytes were stained for P450 2B1 and 3A2, but there were marked differences between the individual cells in the extent of the expression of these two P450 subtypes, like it was also the case with normal adult liver. Within hepatocytes of the fetal livers strongly stained glycogen granules were seen, which, in comparison to adult livers, were rather coarse-grained. Three days after transplantation the glycogen granules in the transplanted hepatocytes were still coarse-grained, but from 1 week after transplantation on, they became more and more fine-grained. As it was also the case with normal adult liver cells, there were marked differences between the individual transplanted hepatocytes in their glycogen content. These results demonstrate that transplanted liver cells originating from syngenic fetal liver tissue suspensions can survive in host organs like the spleen for at least 1 year. They proliferate, differentiate, are able to store glycogen, and express different P450 isoforms, like normal adult liver cells.

Animals↗
PubMed1998

Transplantation of fetal liver tissue suspension into the spleens of adult syngenic rats: effects of beta-naphthoflavone, phenobarbital and dexamethasone on cytochrome P450 isoforms expression and on glycogen storage.

In the present study, the effect of beta-naphthoflavone (BNF), phenobarbital (PB) and dexamethasone (DEX) on the expression of three cytochrome P450 (P450) isoforms, 1A1, 2B1 and 3A2, and on glycogen storage was investigated in intrasplenic liver cell explants in comparison to adult liver. Fetal liver tissue suspensions were transplanted into the spleens of adult male syngenic Fisher inbred rats. Four months after surgery, transplant recipients and age matched controls were orally treated with BNF (1 x 50 mg/kg body weight (b.wt.)), PB (1 x 50 mg/kg b.wt.), DEX (for 3 days 4 mg/kg b.wt. per day), or the respective solvents (dimethylsulfoxide or 0.9% NaCl). The animals were sacrificed 24 (BNF, DEX) or 48 (PB) hours after the last treatment. The livers of both solvent treated transplant recipients and control rats displayed only in few liver lobules a slight P450 1A1, but in all lobules a strong P450 2B1 and 3A2 expression, which was all mainly located in the hepatocytes around the central veins (zone III, according to Rappaport). After BNF administration a P450 1A1 expression was induced in the hepatocytes of the peripheral regions of the liver lobules (zone I, according to Rappaport), whereas the staining of the hepatocytes around the central veins disappeared. Also the staining for P450 2B1 in the hepatocytes of zone III became slightly more pronounced. Following PB treatment the P450 1A1 expression in the hepatocytes of the central regions (zone III), as seen in few lobules after solvent treatment only, was reduced, whereas the staining for P450 2B1 and 3A2 was more pronounced in the hepatocytes of the intermedial and central regions of the liver lobules (zone II and III). DEX treatment diminished P450 1A1 and 2B1 expression within the livers of both transplant recipients and control rats. In contrast, the staining for P450 3A2 was enhanced in all regions of the liver lobules. Transplantation of fetal liver tissue suspensions into the spleens did not influence the inducibility of P450 isoforms expression within the respective livers of the animals. Spleens of control rats displayed no P450 isoforms expression without as well as with induction. In the explant containing spleens, however, similar to normal liver, the transplanted hepatocytes displayed nearly no P450 1A1, but a strong P450 2B1 and 3A2 expression. After BNF treatment a staining for P450 1A1 was induced and also the P450 2B1 expression was slightly more pronounced. PB treatment caused an increase in the staining for P450 2B1 and 3A2 and DEX administration for P450 3A2 within the transplanted hepatocytes. Additionally, after DEX treatment some bile ducts of the explants displayed a slight staining for P450 1A1, 2B1 and 3A2. All hepatocytes within the livers of both solvent treated transplant recipients and control rats displayed a slightly PAS-positive cytoplasma and, in most cases, homogeneously distributed, fine-grained, strongly PAS-stained granules indicating glycogen storage. No regional variance in the glycogen content of the hepatocytes was seen within the liver lobules, but there was a marked difference between the individual hepatocytes of the same lobular region in the extent of glycogen accumulation. The hepatocytes within the explants displayed the same type of glycogen storage as did the adult liver cells. BNF treatment did not display any effect on the glycogen accumulation in livers and intrasplenic liver cell explants. After PB administration, only in livers, but not in the transplants, the glycogen content in the hepatocytes around the central veins was slightly reduced. DEX treatment lead to an excessive storage of fat within the hepatocytes of both livers and spleens. Thus, the glycogen was displaced, leading to a "spoke-wheel" like pattern of glycogen storage. Additionally, within the hepatocytes of both livers and liver cell explants a higher amount of glycogen seemed to be stored and the granules appeared to be more coarse-grained. (ABSTRACT

Animals↗
PubMed1996

When should the "infected" subcutaneous infusion reservoir be removed?

Subcutaneous central venous infusion reservoirs (central venous catheters) are one of the primary devices for administration of intravenous chemotherapy. Usually these devices have few problems, and they provide dependable long term central venous access. Infection of these catheters is a significant problem that usually requires removal. When infection is suspected, it is often difficult to make this determination without actually removing the catheter. Thorough preoperative evaluation may help the surgeon decide which catheters are infected and should be removed. A total of 817 subcutaneous infusion reservoirs were placed at our institution from January 1, 1990 through November 1, 1994. During the same time period, 143 catheters were removed, 63 for suspected infection. The charts of these 63 patients were reviewed to determine to what extent available preoperative information could be used to predict which catheters were infected, thus avoiding unnecessary removal. Twenty-three preoperative parameters were assessed, including physical exam, body temperature, leukocyte count, platelet count, blood cultures from the catheter and peripheral blood, time from placement to removal, whether or not the catheter was functional, and whether it was currently in use. Forty catheters (65%) removed for suspected infection were infected, as demonstrated by a positive culture from the catheter or the wound. Staphylococcus was the most common microorganism. Physical exam (local erythema, tenderness, or swelling) correlated significantly with catheter infection (P = 0.0238). In contrast, blood culture data and the other clinical and laboratory parameters showed no significant association with catheter infection. We conclude that physical exam is the best indicator of catheter infection. Commonly used parameters such as fever, leukocytosis, and positive blood cultures are nonspecific, may not be due to catheter infection, and were not significant in our study. Removal and subsequent restoration of long term intravenous access is associated with significant morbidity and expense. Clinical decision making should not be based on isolated laboratory findings, but must be individualized in each patient with suspected catheter infection.

Adult↗
PubMed1994

Protein purification, gene cloning and sequencing of an acidic endoprotease from Myxococcus xanthus DK101.

An acidic endoprotease (MAEP) secreted during vegetative growth by Myxococcus xanthus DK101 was purified to homogeneity by a series of chromatographic procedures. The endoprotease cleaved the Phe-Met bond of kappa-casein under acidic conditions (pH 5.9). Its apparent molecular mass and its isoelectric point have been estimated to be 12 kDa and 4.5, respectively. From the N-terminal amino acid sequence, a set of two primers for polymerase chain reaction have been designed. Amplification of the corresponding DNA fragment (84 bp) generated a probe, then used to screen an expression DNA library of M. xanthus and to isolate a recombinant plasmid which contained a 2127-bp insert. The nucleotide sequence included an open reading frame (ORF) of 585 nucleotides, encoding 195 amino acids, that exhibited a high degree of similarity with the N-terminal amino acid sequence of the purified MAEP. The polypeptide sequence inferred from this ORF revealed that the mature enzyme should contain 131 amino acids arising from a 195-amino-acid precursor protein.

Amino Acid Sequence↗
PubMed1974

A new method for the preparation of uniformly 14C-labelled compounds by using Anacystis nidulans.

1. Growth conditions were devised which enabled Anacystis nidulans to be grown on high specific radioactivity (14)CO(2) (at over 98% isotopic abundance of (14)C) in a closed system with greater than 85% substrate utilization. 2. The DNA and RNA content of A. nidulans was estimated by various methods and compared with that of Chlorella pyrenoidosa. 3. A procedure was developed for the quantitative extraction and separation of the DNA and RNA from A. nidulans. 4. The fractionation and analysis of the DNA and RNA includes methods for the preparation of the uniformly (14)C-labelled deoxyribonucleoside monophosphates and the uniformly (14)C-labelled ribonucleoside monophosphates. 5. The base 6-methylaminopurine was identified as a component of the DNA. 6. The preparation and total yield of uniformly (14)C-labelled amino acids from A. nidulans are also briefly described. The results are discussed in relation to the use of other micro-organisms for the preparation of compounds labelled with (14)C.

Adenosine Monophosphate↗