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Biomedical subjects

M Pack

Publications and source records attributed to M Pack.

At least 19 recordsLinked to original sources
PubMed2001

Genetic analysis of digestive physiology using fluorescent phospholipid reporters.

Zebrafish are a valuable model for mammalian lipid metabolism; larvae process lipids similarly through the intestine and hepatobiliary system and respond to drugs that block cholesterol synthesis in humans. After ingestion of fluorescently quenched phospholipids, endogenous lipase activity and rapid transport of cleavage products results in intense gall bladder fluorescence. Genetic screening identifies zebrafish mutants, such as fat free, that show normal digestive organ morphology but severely reduced phospholipid and cholesterol processing. Thus, fluorescent lipids provide a sensitive readout of lipid metabolism and are a powerful tool for identifying genes that mediate vertebrate digestive physiology.

Animals↗
PubMed2000

A monoclonal antibody to the DEC-205 endocytosis receptor on human dendritic cells.

DEC-205 is a multilectin receptor for adsorptive endocytosis, expressed in mouse dendritic cells (DC) and some epithelia. DEC-205 is homologous to the macrophage mannose receptor (MMR). A cDNA for murine DEC-205 was used to identify 3 overlapping human DEC-205 clones from a lymphocyte library. The human homologue is a transmembrane protein of 1722 aminoacids with 10 externally disposed C-type lectin domains having 77% identity to the mouse counterpart. The NH(2) terminal cysteine-rich and fibronectin type II domains were expressed and used to immunize mice. A hybridoma, MG38, which specifically recognized the immunogen was obtained from a DEC-205 knockout mouse. The antibody precipitated a 205 kD protein from metabolically labeled, monocyte-derived DCs. MG38 labeled mature monocyte-derived DCs but showed weak or no labeling of other peripheral blood mononuclear cells. In tissue sections, MG38 identified DEC-205 on thymic cortical epithelium and DCs in the thymic medulla and tonsillar T cell areas. In contrast, an anti-MMR antibody stained DEC-205 negative, macrophages in the thymus cortex, the trabeculae of the thymus and tonsil, as well as efferent lymphatics in the tonsil. Therefore, the MG38 anti-DEC-205 antibody is useful for identifying DCs and reveals clear differences in sites where MMR and DEC-205 are expressed in lymphoid tissues.

Animals↗
PubMed1999

Experimental computer-assisted alloplastic sandwich augmentation of the atrophic mandible.

PURPOSE: This study evaluated the effectiveness of a technique that combined computer-aided surgery with alloplastic augmentation and implant-borne prosthodontic rehabilitation of the atrophic mandible. MATERIALS AND METHODS: Computed tomographic (CT) data from an atrophic cadaver mandible were transferred to a computer-aided design (CAD) system that prepared an anterior sandwich osteotomy. The cranial segment was moved upward and backward to provide an ideal alveolar relationship, and the geometry of the intermediate space was used to design a titanium implant. Furthermore, a surgical template was derived for the osteotomies, and insertion of dental implants was planned to stabilize both the transposed bone and the intermediate implant on the bony base. An identical implant for augmentation was also fabricated from poly-D,L-lactide in a mold as a possible resorbable carrier for osteoinductive proteins. RESULTS: The experimental surgery was successfully performed with maximum precision on the dried mandible. The fabrication of an implant made out of poly-D,L-lactide for the same purpose was also possible. CONCLUSIONS: This preliminary experiment showed that it is possible to use CAD/computer-aided manufacturing (CAM) technology to prepare a prefabricated template and a corresponding titanium implant for mandibular augmentation with a high degree of exactness. Dental implants could be planned and integrated in this procedure as well. The fabrication of a mold using this method also provided the opportunity to give a complex shape to possible carriers of osteoinductive substances.

Alveolar Bone Loss↗
PubMed1999

Ideal amino acid profile and dietary lysine specification for broiler chickens of 20 to 40 days of age.

1. The aim of the study was to determine the ideal ratio of the essential amino acids lysine (Lys), methionine (Met), threonine (Thr), tryptophan (Trp), arginine (Arg), valine (Val) and isoleucine (Ile) and to assess the required dietary lysine concentration for optimum performance in broiler chickens of 2 commercial strains from 20 to 40 d of age. 2. An identical basal diet was used throughout all experiments. It consisted mainly of maize and soyabean meal and contained 172 g crude protein and 13.2 MJ AME(N) per kg. For each experiment, the basal diet was adequately supplemented with all essential amino acids except the one to be tested, which was supplemented in 6 graded concentrations in exchange for maize starch. One (Met, Trp, Arg, Val, Ile) or 2 (Lys, Thr) growth trials were conducted per amino acid tested and the response in weight gain, food: gain ratio, breast meat yield and abdominal fat were examined. 3. The ideal amino acid ratio relative to Lys (expressed as a percentage) was calculated to be 75% Met+Cys, 63% Thr, 19% Trp, 112% Arg, 71% Ile and 81% Val on a true faecal digestible basis when the data were subjected to broken-line regression analysis. From both lysine studies the concentration for optimum food: gain ratio was calculated, by exponential regression analysis, to be 11.5 g true faecal digestible lys per kg diet.

Amino Acids, Essential↗
PubMed1999

Estimates of essential amino acid requirements from dose-response studies with rainbow trout and broiler chicken: effect of mathematical model.

A total of 37 dose-response experiments with essential amino acids performed with rainbow trout and broiler chicken were re-evaluated with different mathematical approaches: an exponential model, a four-parameter logistic function, the saturation kinetics model and the broken line approach. The different approaches were compared both with regard to the goodness of fit (r2 and sy.x) and with regard to the allowances which were derived regarding the optimal amino acid level in the diet. The experimental design, particularly the chosen range in dietary amino acid concentration was found to be important for the comparison of models. Amongst the non-linear models, the four-parameter logistic function and the saturation kinetics model appeared superior to the exponential approach, when the range in dietary amino acid concentration was very wide and included both a severely deficient basal level and a level that exceeded the needs of the animal by approximately the factor 2. In these cases, allowances derived from individual experiments were considerably different depending on the model. The allowances based on the exponential and the saturation kinetics approach were 27.7 and 20.7 g lysine/kg DM and 8.0 and 6.3 g methionine/kg DM, respectively, for rainbow trout. For other amino acids studied in rainbow trout the difference due to model was less. Consequently, the predicted 'ideal protein' for rainbow trout was considerably different depending on the model used. The maximum deviation found in different experiments with broiler chicken for the exponential vs. the saturation kinetics approach was 13.0 and 9.7 g lysine/kg and 11.4 and 8.2 g sulfur-containing amino acids/kg, respectively. However, the more restricted the range in dietary concentration was, the lesser became the differences between the different non-linear models. No definite recommendation can therefore be extracted regarding the most suitable, generally applicable mathematical model.

Amino Acids, Essential↗
PubMed1999

Effect of enzyme supplementation of broiler diets based on corn and soybeans.

Digestibility of diets based on corn and soybean meal or soybeans treated by roasting or extrusion, with or without an enzyme supplementation, was measured by "true" (Sibbald) methods, by analysis of excreta, and by analysis of ileal digesta. Only analysis of ileal digesta was able to consistently measure differences between soybean and enzyme treatments in the digestibility of CP, starch, fat, and ME. The amino acid (AA) digestibility of the diets was measured by analysis of the ileal contents. Whereas enzyme supplementation improved overall CP digestibility by 2.9%, this improvement was not equal for all AA. Of the AA most important for broilers fed corn-soybean diets, the digestibilities of Lys, Met, and Arg were not improved or not improved significantly by the enzyme supplementation; however, that of Val was improved by 2.3% and that of Thr was improved by 3.0%. A performance trial demonstrated that enzyme supplementation with equal diet formulation improved BW and the feed conversion ratio by 1.9 and 2.2%, respectively. A second performance trial compared standard diet formulations with formulations using enzyme supplementation and energy levels that were reduced by the amount of improvement provided by the inclusion of enzyme in the first performance trial. No difference was seen between treatments, showing that the improvement of nutrient utilization brought about by enzyme supplementation completely compensated for the reduced energy content. Whereas enzyme supplementation should allow a reduction in CP formulation as well, individual AA were not improved equally by supplementation and should also be balanced.

Adipose Tissue↗
PubMed1998

Efficient presentation of phagocytosed cellular fragments on the major histocompatibility complex class II products of dendritic cells.

Cells from the bone marrow can present peptides that are derived from tumors, transplants, and self-tissues. Here we describe how dendritic cells (DCs) process phagocytosed cell fragments onto major histocompatibility complex (MHC) class II products with unusual efficacy. This was monitored with the Y-Ae monoclonal antibody that is specific for complexes of I-Ab MHC class II presenting a peptide derived from I-Ealpha. When immature DCs from I-Ab mice were cultured for 5-20 h with activated I-E+ B blasts, either necrotic or apoptotic, the DCs produced the epitope recognized by the Y-Ae monoclonal antibody and stimulated T cells reactive with the same MHC-peptide complex. Antigen transfer was also observed with human cells, where human histocompatibility leukocyte antigen (HLA)-DRalpha includes the same peptide sequence as mouse I-Ealpha. Antigen transfer was preceded by uptake of B cell fragments into MHC class II-rich compartments. Quantitation of the amount of I-E protein in the B cell fragments revealed that phagocytosed I-E was 1-10 thousand times more efficient in generating MHC-peptide complexes than preprocessed I-E peptide. When we injected different I-E- bearing cells into C57BL/6 mice to look for a similar phenomenon in vivo, we found that short-lived migrating DCs could be processed by most of the recipient DCs in the lymph node. The consequence of antigen transfer from migratory DCs to lymph node DCs is not yet known, but we suggest that in the steady state, i.e., in the absence of stimuli for DC maturation, this transfer leads to peripheral tolerance of the T cell repertoire to self.

Animals↗
PubMed1997

High levels of a major histocompatibility complex II-self peptide complex on dendritic cells from the T cell areas of lymph nodes.

T lymphocytes recirculate continually through the T cell areas of peripheral lymph nodes. During each passage, the T cells survey the surface of large dendritic cells (DCs), also known as interdigitating cells. However, these DCs have been difficult to release from the lymph node. By emphasizing the use of calcium-free media, as shown by Vremec et al. (Vremec, D., M. Zorbas, R. Scollay, D.J. Saunders, C.F. Ardavin, L. Wu, and K. Shortman. 1992. J. Exp. Med. 176:47-58.), we have been able to release and enrich DCs from the T cell areas. The DCs express the CD11c leukocyte integrin, the DEC-205 multilectin receptor for antigen presentation, the intracellular granule antigens which are recognized by monoclonal antibodies M342, 2A1, and MIDC-8, very high levels of MHC I and MHC II, and abundant accessory molecules such as CD40, CD54, and CD86. When examined with the Y-Ae monoclonal which recognizes complexes formed between I-Ab and a peptide derived from I-Ealpha, the T cell area DCs expressed the highest levels. The enriched DCs also stimulated a T-T hybridoma specific for this MHC II-peptide complex, and the hybridoma underwent apoptosis. Therefore DCs within the T cell areas can be isolated. Because they present very high levels of self peptides, these DCs should be considered in the regulation of self reactivity in the periphery.

Animals↗
PubMed1997

Response of rainbow trout (Oncorhynchus mykiss) to supplements of individual essential amino acids in a semipurified diet, including an estimate of the maintenance requirement for essential amino acids.

We studied the effects of increasing dietary concentrations of each of the following amino acids on growth, feed intake, feed conversion ratio and composition of gain in rainbow trout in six dose-response experiments: L-lysine, L-tryptophan, L-histidine, L-valine, L-leucine and L-isoleucine. Semipurified diets containing 20.1 MJ digestible energy/kg dry matter, with wheat gluten and crystalline amino acids as sole sources of amino acids, were fed to rainbow trout [initial mean body weight (BW) 40-51 g, depending on the amino acid studied]. In one series of 24 diets, lysine concentration ranged from 4.5 to 58.0 g/kg dry matter; in five further series of 12 diets each, concentrations ranged from (in g/kg dry matter): tryptophan, 1.3 to 5.6; histidine, 2.6 to 13.5; valine, 6.2 to 34.2; leucine, 10.0 to 42.0 and isoleucine, 5.0 to 15.3. Each diet was fed to a group of 20 fish for 53-64 d, depending on the amino acid studied. Dry matter intake, weight gain, feed conversion ratio, protein concentration of gain and total protein deposition followed exponential response functions. To achieve 95% of the maximum protein deposition, dietary concentrations of 27.7 g lysine, 2.0 g tryptophan, 5.8 g histidine, 15.7 g valine, 13.6 g leucine and 13.7 g isoleucine/kg dry matter were required. Maintenance requirements, estimated from exponential functions for protein deposition, were [in mg/(100 g BW.d)]: lysine, 1.93; tryptophan, 1.05; histidine, 1.07; valine, 2.92; leucine, 8.26 and isoleucine, 0.91. This corresponds to 4% of the requirement for protein deposition for lysine and isoleucine but 32% for leucine, with the other amino acids being intermediate. Therefore, different dietary amino acid requirement patterns were derived from protein deposition data depending on the chosen level of performance.

Amino Acids, Essential↗
PubMed1997

Replacement value of betaine for DL-methionine in male broiler chicks.

The effect of DL-methionine and betaine supplementation on growth performance of 2,400 male broilers in the age period of 1 to 38 d, and on carcass composition of a subsample of 384 birds was examined. Three dose levels of DL-methionine (0, 0.05, and 0.10%) and two doses of betaine (0 and 0.04%) were supplemented in different combinations to methioninedeficient diets. Two types of diets were fed as starters and growers: either corn-soybean diets or practical diets typical for the Dutch broiler industry. All diets were fortified with 220 ppm choline in order to avoid a deficiency in methyl groups. Increasing DL-methionine supplementation significantly improved daily weight gain and feed conversion efficiency. Supplemental betaine did not affect bird growth. Betaine slightly improved feed conversion in diets without supplemental DL-methionine, but did not affect this parameter in diets with added DL-methionine. Breast meat yield was significantly increased by about 1.5 percentage points by the addition of 0.05% DL-methionine, whereas 0.04% betaine only tended to increase breast meat yield in the range of 0.3 to 0.6 percentage points. The type of diet did not have any effect on the responses obtained. In summary, there was no evidence for betaine to spare DL-methionine as an essential amino acid supplement in broiler diets.

Analysis of Variance↗
PubMed1997

Effects of dietary threonine in starting, growing, and finishing turkey toms.

The present study investigates the threonine requirements of British United Turkey (BUT) Big 6 turkey toms from 0 to 4, 8 to 12, and 16 to 20 wk of age. Growth, feed conversion, and carcass quality responses to increasing dietary threonine levels were measured. From 0 to 4 wk of age (57 g to 1.1 kg live weight), a dietary threonine level of about 0.95% (3.3 g/Mcal ME) was found to be adequate to obtain optimum growth and feed conversion. From 8 to 12 wk of age (4.0 to 8.6 kg live weight), there was no significant response to dietary threonine beyond the basal level of 0.69% (2.2 g/ Mcal ME). From 16 to 20 wk of age (13.1 to 18.7 kg live weight), a threonine level of about 0.58% (1.8 g/Mcal ME) appeared to be adequate to obtain optimum growth. For optimum breast meat deposition, a higher level in the range of 0.64% (2.0 g/Mcal ME) dietary threonine was required. The present findings may point to the importance of threonine as potentially third limiting amino acid after methionine and lysine in cereal-based turkey diets.

Aging↗
PubMed1997

Dendritic cells in the T-cell areas of lymphoid organs.

Substantial numbers of dendritic cells (DCs) are found in the T-cell areas of peripheral lymphoid organs such as the spleen, lymph node and Peyer's patch. By electron microscopy these DCs (also called interdigitating cells) form a network through which T-cells continually recirculate. The cytological features of DCs in the T-cell areas, as well as a number of markers detected with monoclonal antibodies, are similar to mature DCs that develop from other sites such as skin and bone marrow. Some markers that are expressed in abundance are: MHC II and the associated invariant chain, accessory molecules such as CD40 and CD86, a multilectin receptor for antigen presentation called DEC-205, the integrin CD11c, several antigens within the endocytic system that are detected by monoclonal antibodies but are as yet uncharacterized at the molecular level, and, in the human system, molecules termed S100b, CD83 and p55. DCs in the periphery can pick up antigens and migrate to the T-cell areas to initiate immunity. However, there are new observations that DCs within the T-cell areas also express high levels of self-antigens and functional fas-ligand capable of inducing CD4+ T-cell death. We speculate that there are at least 2 sets of DCs in the T-cell areas, a migratory myeloid pathway that brings in antigens from the periphery and induces immunity, and a more resident lymphoid pathway that presents self-antigens and maintains tolerance.

Animals↗
PubMed1996

Effects of dietary protein content, addition of nonessential amino acids and dietary methionine to cysteine balance on responses to dietary sulphur-containing amino acids in broilers.

1. Two experiments were conducted with male broiler chicks from 2 to 5 weeks of age to determine the effect of dietary protein content and amino acid balance on the response to dietary sulphur-containing amino acids (SAA) in terms of performance and carcase quality. 2. In experiment 1, 5 graded amounts of a DL-methionine and L-cysteine (1:1 by weight) mixture were added to basal diets containing 197 or 233 g crude protein/kg. The diets containing 197 g protein/kg were fed with or without the further addition of 36 g crude protein/kg from nonessential amino acids. The amino acid balance of all diets was kept constant for all essential amino acids except the SAA. In experiment 2, 5 graded amounts of SAA from either a crystalline source (DL-methionine or a mixture of DL-methionine and L-cysteine) or from intact proteins were added to a diet containing 208 g protein/kg. 3. At each protein concentration there were significant responses to the SAA addition in weight gain, food conversion efficiency, and carcase quality. Non-linear exponential regression analyses were used to describe bird responses to SAA concentration. The broiler chick's requirement for SAA increased with increasing dietary protein concentrations ranging from 197 to 259 g protein/kg. 4. The utilisation of SAA differed also with differences in origin (crystalline or peptide-bound), and methionine:cysteine balances. Compared to DL-methionine, a 1:1 mixture of DL-methionine and L-cysteine was only 81% or 86% as effective in supporting growth or food conversion, respectively. SAA from added protein was even less effectively utilised. 5. The addition of nonessential amino acids tended to decrease food intake without affecting SAA utilisation. 6. Slaughter yield and breast meat yield were clearly increased while fat deposition was clearly decreased, by SAA addition. The response in breast meat yield suggested an important economic benefit for further meat processing. Nitrogen retention was significantly enhanced by SAA supplementation from crystalline sources, and this led to reductions of up to 30% in the amount of nitrogen excreted per kg weight gain.

Animal Feed↗
PubMed1996

Mutations affecting development of zebrafish digestive organs.

The zebrafish gastrointestinal system matures in a manner akin to higher vertebrates. We describe nine mutations that perturb development of these organs. Normally, by the fourth day postfertilization the digestive organs are formed, the epithelial cells of the intestine are polarized and express digestive enzymes, the hepatocytes secrete bile, and the pancreatic islets and acini generate immunoreactive insulin and carboxypeptidase A, respectively. Seven mutations cause arrest of intestinal epithelial development after formation of the tube but before cell polarization is completed. These perturb different regions of the intestine. Six preferentially affect foregut, and one the hindgut. In one of the foregut mutations the esophagus does not form. Two mutations cause hepatic degeneration. The pancreas is affected in four mutants, all of which also perturb anterior intestine. The pancreatic exocrine cells are selectively affected in these four mutations. Exocrine precursor cells appear, as identified by GATA-5 expression, but do not differentiate and acini do not form. The pancreatic islets are spared, and endocrine cells mature and synthesize insulin. These gastrointestinal mutations may be informative with regard to patterning and crucial lineage decisions during organogenesis, and may be relevant to diabetes, congenital dysmorphogenesis and disorders of cell proliferation.

Animals↗