Expression of anti-inflammatory macrophage genes within skeletal muscle correlates with insulin sensitivity in human obesity and type 2 diabetes.

AIMS/HYPOTHESIS: Low-grade systemic inflammation and adipose tissue inflammatory macrophages are frequently detected in patients with obesity and type 2 diabetes. Whether inflammatory macrophages also increase in skeletal muscle of individuals with metabolic disorders remains controversial. Here, we assess whether macrophage polarisation markers in skeletal muscle of humans correlate with insulin sensitivity in obesity and type 2 diabetes. METHODS: Skeletal muscle biopsies were obtained from individuals of normal weight and with normal glucose tolerance (NGT), and overweight/obese individuals with or without type 2 diabetes. Insulin sensitivity was determined by euglycaemic-hyperinsulinaemic clamps. Expression of macrophage genes was analysed by quantitative RT-PCR. RESULTS: Gene expression of the inflammatory macrophage phenotype marker cluster of differentiation (CD)11c was higher in muscle of type 2 diabetes patients (p = 0.0069), and correlated with HbA1c (p = 0.0139, ρ = 0.48) and fasting plasma glucose (p = 0.0284, ρ = 0.43), but not after correction for age. Expression of TGFB1, encoding the anti-inflammatory marker TGF-ß1, correlated inversely with HbA1c (p = 0.0095, ρ = -0.50; p = 0.0484, ρ = -0.50) and fasting plasma glucose (p = 0.0471, ρ = -0.39; p = 0.0374, ρ = -0.52) in two cohorts, as did HbA1c with gene expression of macrophage galactose-binding lectin (MGL) (p = 0.0425, ρ = -0.51). TGFB1 expression was higher in NGT individuals than in individuals with type 2 diabetes (p = 0.0303), and correlated with low fasting plasma insulin (p = 0.0310, ρ = -0.42). In exercised overweight/obese individuals, expression of genes for three anti-inflammatory macrophage markers, MGL (p = 0.0031, ρ = 0.71), CD163 (p = 0.0268, ρ = 0.57) and mannose receptor (p = 0.0125, ρ = 0.63), correlated with high glucose-disposal rate. CONCLUSIONS/INTERPRETATION: Muscle expression of macrophage genes reveals a link between inflammatory macrophage markers, age and high glycaemia, whereas anti-inflammatory markers correlate with low glycaemia and high glucose-disposal rate.

Effect of selected modified atmosphere packaging on Campylobacter survival in raw poultry.

Most current research on Campylobacter has focused on preharvest or processing plant cross-contamination. Little is known about the effect of storage environment on the survival of Campylobacter on raw poultry. We evaluated the effects of modified storage atmosphere and freezing on the survival of naturally occurring Campylobacter on raw poultry. Broiler carcasses (n = 560) were collected as they exited the chiller in 2 commercial processing plants and were sampled for the detection of Campylobacter, Escherichia coli, psychrophiles, and total aerobes at 0 and 14 d of refrigerated (2°C) storage. Gases evaluated were air, 100% O(2), 100% CO(2), and a standard poultry modified atmosphere packaging mixture (5% O(2) + 10% CO(2) + 85% N). Freezing was included as a control group. All carcasses were sampled by the whole-carcass rinse method. The rinse fluid was recovered and pooled from 5 individual rinses, and serial dilutions were made for examination of Campylobacter (42°C, 48 h), E. coli (37°C, 24 h), psychrophiles (plate count agar, 4°C, 7 d), and total aerobic bacterial populations (plate count agar, 37°C, 24 h). Campylobacter counts for all treatments were reduced during the 14-d storage period but the 100% O(2) treatment caused a significantly (P < 0.05) greater reduction than the other gas treatments. For the psychrophiles, storage in air resulted in the greatest growth after 14 d, with reduced psychrophilic growth allowed by either O(2) or the modified atmosphere packaging mixture (not different from each other). Of the treatments evaluated, CO(2) allowed the least growth of psychrophiles. Proliferation of E. coli and aerobes was the greatest when packaged in air after 14 d, whereas CO(2) packaging resulted in the least growth. These data suggest that storage under O(2) may reduce Campylobacter recovery and slow psychrophile and aerobe recovery following storage.

Lipid oxidation stability of omega-3- and conjugated linoleic acid-enriched sous vide chicken meat.

Lipid oxidation is known to occur rather rapidly in cooked chicken meat containing relatively high amounts of polyunsaturated fatty acids. To assess the lipid oxidation stability of sous vide chicken meat enriched with n-3 and conjugated linoleic acid (CLA) fatty acids, 624 Cobb × Ross broilers were raised during a 6-wk feeding period. The birds were fed diets containing CLA (50% cis-9, trans-11 and 50% trans-10, cis-12 isomers), flaxseed oil (FSO), or menhaden fish oil (MFO), each supplemented with 42 or 200 mg/kg of vitamin E (dl-α-tocopheryl acetate). Breast or thigh meat was vacuum-packed, cooked (74°C), cooled in ice water, and stored at 4.4°C for 0, 5, 10, 15, and 30 d. The lipid oxidation development of the meat was estimated by quantification of malonaldehyde (MDA) values, using the 2-thiobarbituric acid reactive substances analysis. Fatty acid, nonheme iron, moisture, and fat analyses were performed as well. Results showed that dietary CLA induced deposition of cis-9, trans-11 and trans-10, cis-12 CLA isomers, increased the proportion of saturated fatty acids, and decreased the proportions of monounsaturated and polyunsaturated fatty acids. Flaxseed oil induced higher deposition of C18:1, C18:2, C18:3, and C20:4 fatty acids, whereas MFO induced higher deposition of n-3 fatty acids, eicosapentaenoic acid (C20:5), and docosahexaenoic acid (C22:6; P < 0.05). Meat lipid oxidation stability was affected by the interaction of either dietary oil or vitamin E with storage day. Lower (P < 0.05) MDA values were found in the CLA treatment than in the MFO and FSO treatments. Lower (P < 0.05) MDA values were detected in meat samples from the 200 mg/kg of vitamin E than in meat samples from the 42 mg/kg of vitamin E. Nonheme iron values did not affect (P > 0.05) lipid oxidation development. In conclusion, dietary CLA, FSO, and MFO influenced the fatty acid composition of chicken muscle and the lipid oxidation stability of meat over the storage time. Supranutritional supplementation of vitamin E enhanced the lipid oxidation stability of sous vide chicken meat.

Dietary lipid source and vitamin E effect on lipid oxidation stability of refrigerated fresh and cooked chicken meat.

The fatty acid composition of chicken muscle may affect the lipid oxidation stability of the meat, particularly when subjecting the meat to thermal processing and storage. The objective of this study was to evaluate the diet effect on lipid oxidation stability of fresh and cooked chicken meat. Six hundred broilers were raised for a 6-wk feeding period and were assigned to 8 treatments with 3 repetitions. Broilers were fed a basal corn-soybean meal diet, including 5% of either animal-vegetable, lard, palm kernel, or soybean (SB) oil, each supplemented with a low (33 mg/kg) or high (200 to 400 mg/kg) level of vitamin E. Fresh breast and thigh meat and skin were packaged and refrigerated (4°C) for 15 d. Breast and thigh meat were frozen (-20°C) and stored for ~6 mo and then thawed, deboned, ground, and formed into patties of 150 g each. Patties were cooked (74°C), cooled, packaged, and stored in refrigeration for 6 d. The lipid oxidation development of the products was determined using the TBA reactive substances analysis. The results showed that the lipid oxidation development, in both fresh chicken parts and cooked meat patties, was influenced by the interaction of either dietary lipid source or vitamin E level with storage time. Fresh breast meat showed no susceptibility to lipid oxidation, but thigh meat and skin presented higher (P < 0.05) malonaldehyde values in the SB oil treatment, starting at d 10 of storage. In cooked patties, during the entire storage time, the SB oil showed the highest (P < 0.05) lipid oxidation development compared with the other treatments. Regarding vitamin E, in both fresh parts and cooked meat patties, in most sampling days the high supplemented level showed lower (P < 0.05) malonaldehyde values than the control treatment. In conclusion, the lipid oxidation stability of chicken meat is influenced by the lipid source and vitamin E level included in the diet upon storage time and processing of the meat.

Soybean, palm kernel, and animal-vegetable oils and vitamin E supplementation effect on lipid oxidation stability of sous vide chicken meat.

There is an increasing demand in precooked chicken meat products for restaurants and catering services. Because cooked chicken meat develops lipid oxidation relatively fast, sous vide chicken meat was studied to assess its shelf-life. Six hundred Cobb x Ross broilers were fed for 6 wk with a basal corn-soybean meal diet including soybean, palm kernel, or animal-vegetable oil, each supplemented with 33 or 200 mg/kg of dl-alpha-tocopheryl acetate. Broilers were randomly assigned into 6 treatments and 4 repetitions with 25 birds each. Boneless breast or thigh muscle pieces were dissected into 5 x 5 x 5 cm cubes, vacuum-packed, cooked in water bath (until 74 degrees C internal temperature), chilled, and stored at 4 degrees C for 1, 5, 10, 25, and 40 d. For each storage day, each pouch contained 3 pieces of meat, either breast or thigh. Thiobarbituric acid reactive substances analysis, to quantify malonaldehyde (MDA) values, was conducted to estimate the lipid oxidation development. Nonheme iron values of cooked meat were analyzed. Fatty acid methyl esters analysis was performed in chicken muscle to determine its fatty acid composition. There was no interaction between dietary fat and vitamin E level in all of the variables studied except in nonheme iron. Dietary fat significantly influenced the fatty acid composition of the muscle (P < 0.01), but it did not affect the MDA values, regardless of differences in the muscle fatty acid composition between treatments. Supplementation of the high level of vitamin E significantly reduced the MDA values in both breast and thigh meat (P < 0.01). The maximum MDA values were observed at d 40 of storage in thigh and breast meat in animal-vegetable and soybean oil treatments with the low levels of vitamin E, 0.91 and 0.70 mg/kg, respectively. Nonheme iron values in thigh meat differed between treatments at 1 or 25 d of storage but not in breast meat. In conclusion, refrigerated sous vide chicken meat has a prolonged shelf-life, which is enhanced by dietary supranutritional supplementation of vitamin E.

Research developments in pale, soft, and exudative turkey meat in North America.

Pale, soft, and exudative (PSE) refers to meat that is pale in color, forms soft gels, and has poor water-holding ability. Most frequently used in reference to pork, this defective meat is being seen with increasing frequency in turkey and broiler processing plants. It has been estimated that this PSE-type meat represents 5 to 40% of meat that is produced in the poultry industry. With the increased production of further-processed products, this PSE problem has become more apparent in the turkey industry. It has been estimated that due to the high incidence, a single turkey processing plant could be losing $2 to 4 million per year, resulting in a loss in excess of $200 million dollars by the turkey industry alone.

Factors influencing the variation in tenderness of seven major beef muscles from three Angus and Brahman breed crosses.

Beef carcasses (n=30) from 3/4 Angus (A)×1/4 Brahman (B), 1/4A×3/4B, and 1/2A×1/2B F(1) crosses were used to evaluate breed type, electrical stimulation, and postmortem aging on the M. semimembranosus (SM), M. semitendinosus (ST), M. biceps femoris (BF), M. vastus lateralis (VL), M. gluteus medius (GM), M. longissimus dorsi lumborum (LD), and M. triceps brachii (TB). Shear force values decreased with increased postmortem aging to a greater extent in steaks from 3/4A×1/4B than steaks from the other breed types. Shear force values for steaks from the round (SM, ST, BF, VL) were higher than steaks from the loin (LD, GM) and chuck (TB) for both electrically stimulated and non-electrically stimulated muscles. In the LD muscle, calpastatin activities were similar among breed types. Muscle type played the greatest role in determining tenderness.

Comparison of flavor changes in cooked-refrigerated beef, pork and chicken meat patties.

Beef and pork longissimus dorsi (LD) and semimembranosus (SM) and chicken breast (B) and thigh (T) muscles excised 24 h postmortem were ground by muscle/species group, formed into patties, pan-fried, refrigerated for 0, 3 or 6 days, and evaluated by a trained sensory panel for intensity of specific flavors. The rate of decline in species-specific natural meat flavor intensity and the rate of increase in "cardboard" (CBD) flavor intensity during the first half of the 6-day storage were fastest for beef, while such decline and increase during the entire storage period were slowest for chicken B. Overall trends of natural meat flavor and CBD intensity changes for chicken T appeared more like those for the red meats than chicken B. It was concluded that, while flavor deterioration can occur in cooked-stored meats from all the species, quantitative or the magnitude of differences between species would depend on muscle types and sensory terms/method used.

Skin pigmentation evaluation in broilers fed natural and synthetic pigments.

Broiler carcass skin color is important in the United States and Mexico. This study evaluated the use of natural and synthetic pigments in broiler diets at commercial levels. Birds were fed natural or synthetic pigments at low or high levels, simulating US and Mexican commercial practices. Skin color was measured during live production (3 to 7 wk of age) and after slaughter and chilling. The natural pigments had consistently greater skin b* values (yellowness) than the synthetic pigments. The high levels produced greater skin b* values than the low levels, regardless of source. The synthetic pigments had a slower increase in skin b* but reached the same level as the natural low by 7 wk. There was no difference in skin a* values (redness) due to pigment source or level or the age of the bird. By 7 wk, all pigment sources approached plateau levels in the blood, but the synthetic pigment diet produced higher blood levels of yellow and red pigments than the natural pigment diets. Processing intensified skin yellowness and reduced skin redness. These data suggest that although synthetic pigments might have been absorbed better than natural ones, natural pigments were more efficient at increasing skin yellowness and there were only small differences between high and low levels for each pigment source. This finding may allow reduction in pigment use and feed cost to achieve the same skin acceptance by the consumer.

Research note: Functionality of electrically stimulated broiler breast meat.

Postmortem electrical stimulation (ES) tenderizes meat by acceleration of adenosine triphosphate (ATP) depletion, pH decline, and physical disruption of muscle fibers. It has been demonstrated that rigor development at elevated temperatures, as with slow chilling, can cause meat to develop pale color and poor water-holding capacity. The objective of this study was to compare the functionality of broiler breast meat from control and electrically stimulated carcasses with and without normal rapid chilling. Broilers were either electrically stimulated (450 mA, 450 V, 2 s on, 2 s off for 7 pulses) immediately after bleeding or used as nonstimulated controls. The ES birds were either chilled immediately (ES2) or had chilling delayed for 2 h at room temperature (ESD2). All ES breast fillets were harvested at 2 h postmortem. The control carcasses were chilled immediately and had fillets harvested at 2 h postmortem (C2) or at 8 h postmortem (C8). Electrical stimulation accelerated pH decline and prevented toughening when breast meat was deboned at 2 h postmortem, regardless of chilling rate. The water released from the gels during cooking was higher for the ESD2 than the ES2 group, which was not different from the C2 group, suggesting that ES followed by slow chilling reduced water-holding capacity compared with the ES2 and C2 groups. There were no differences in expressible moisture, gel strength, or lightness among the ES2, ESD2, and C2 treatments. These results indicated that high voltage ES followed by normal chilling did not impair protein functionality or cause pale, soft, exudative meat. However, there was some evidence that slow chilling after ES may negatively affect some water-holding properties of the meat.

Early postmortem carcass trim effects on the tenderness of broiler breast fillets.

Broiler carcasses are often trimmed during evisceration to remove damaged areas of the carcass. Because deboning before rigor mortis development can toughen meat, trimming during evisceration may toughen the meat. This study evaluated the effects of trimming on the tenderness of broiler breast meat. To evaluate the effect of wing removal on tenderness, breast halves from 2 flocks were collected after chilling at a commercial plant. One-third were untrimmed controls, one-third had small amounts of breast meat removed with wing (WMin), and one-third had large amounts of breast meat removed with wing (Wmax). Salvage fillets from the 2 flocks were also collected from the salvage table of the plant. Carcasses were also processed to evaluate the effect of breast blister trimming that removed a superficial amount of muscle tissue, half of which had breast blister trims, and half did not. All front halves or carcasses were aged until 24 h postmortem and deboned. Salvage fillets were held refrigerated until 24 h postmortem. Fillets were cooked and then sheared in 2 locations on the fillet, upper and lower, to determine if tenderness was more affected at areas close to the trim. Carcasses with wing trims had significantly higher shear values compared with the control, and shear values from the upper portion of the fillets from the WMax and WMin (nearer the trim) were significantly greater than for the lower portion. Location, however, did not affect shear values in the control carcasses. This finding indicated that tenderness of the areas nearest the trim might be affected more by the trim process. Salvage table deboning significantly increased shear values throughout the fillet. There was no significant difference in shear value due to breast blister trimming. The results of this study suggest that trimming carcasses by wing or breast fillet removal results in decreased meat tenderness.

CGRP receptors mediating CGRP-, adrenomedullin- and amylin-induced relaxation in porcine coronary arteries. Characterization with 'Compound 1' (WO98/11128), a non-peptide antagonist.

1. Calcitonin gene-related peptide (CGRP), amylin and adrenomedullin (AM) belong to the same family of peptides. Accumulating evidence indicate that the calcitonin (CT) receptor, the CT receptor-like receptor (CRLR) and receptor-activity-modifying proteins (RAMPs) form the basis of all the receptors in this family of peptides. 2. Using reverse transcriptase - polymerase chain reaction the presence of mRNA sequences encoding the CRLR, RAMP1 and RAMP2 were demonstrated in porcine left anterior descending (LAD) coronary arteries, whereas porcine calcitonin (CT) receptor mRNA was not present. The partial porcine mRNA sequences shared 82 - 92% nucleotide identity with human sequences. 3. The human peptides alphaCGRP, betaCGRP, AM and amylin induced relaxation with pEC(50) values of 8.1, 8.1, 6.7 and 6.1 M respectively. 4. The antagonistic properties of a novel non-peptide CGRP antagonist 'Compound 1' (WO98/11128), betaCGRP(8 - 37) and the proposed AM receptor antagonist AM(22 - 52) were compared to the well-known CGRP(1) receptor antagonist alphaCGRP(8 - 37). 5. The alphaCGRP(8 - 37) and betaCGRP(8 - 37) induced concentration-dependent (10(-7) - 10(-5) M) rightward shift of both the alphaCGRP and betaCGRP concentration-response curves. betaCGRP(8 - 37) (10(-6) M) had the same effect as alphaCGRP(8 - 37) (10(-6) M), but with less potent rightward shift of the concentration-response curves for alphaCGRP, AM and amylin. 6. Preincubation with 'Compound 1' (10(-7) - 10(-5) M) and AM(22 - 52) (10(-6) M) had no significant antagonistic effect. 7. In conclusion, the building blocks forming CGRP and AM receptors were present in the porcine LAD, whereas those of the amylin receptor were not. alphaCGRP, betaCGRP, AM and amylin mediated vasorelaxation via the CGRP receptors. No functional response was detected to adrenomedullin via the adrenomedullin receptor.

Potassium-induced cardioplegia. Additive protection against ischemic myocardial injury during coronary revascularization.

Potassium-induced cardioplegia during anoxic arrest was utilized in a study of 190 consecutive patients undergoing revascularization (average 2.8 grafts per patient) from August, 1975, through August, 1976. Surgical technique, moderate systemic hypothermia with intermittent anoxic arrest, and the surgeon were the same for all patients. One hundred thirty-five patients (KC1-treated) received a bolus (150 ml.) of potassium solution injected into the proximal aortic root whenever the aortic cross-clamp was applied; 55 others served as control subjects. The mortality rate was 2.2% (three of 135) in the KCl-treated group and one of 55 in the control group. New Q waves appeared in 5.9% (eight of 135) of the KCl-treated patients and 11% (6 of 55) of control subjects (p = N.S.). Catecholamine drips were required after bypass in 4.4% (six of 135) of patients given potassium and 18% (10 of 55) of control patients (p less than 0.05). Profound myocardial relaxation was of added technical value with potassium. It is our impression that hearts treated with potassium exhibited more prompt cardioversion, separated from cardiopulmonary bypass with less need for inotropic support, and exhibited less myocardial injury during the revascularization procedure.

Equipotent in vitro actions of alpha- and beta-CGRP on guinea pig basilar artery are likely to be mediated via CRLR derived CGRP receptors.

The aim of the present study was to investigate and compare specific in vitro pharmacological actions of human alpha- and beta-CGRP applied as single concentrations to prostaglandin F2alpha precontracted segments of guinea pig basilar artery. To support the suggestion of a possible link between the pharmacological actions of alpha- and beta-CGRP and a specific receptor, we wished to determine whether mRNAs required for the expression of calcitonin receptor-like receptor (CRLR) derived CGRP receptors were present in the guinea pig basilar artery. In the pharmacological experiments we demonstrated an increase in the cAMP content by 2.5-fold and a concomitant significant vasorelaxation of the precontracted basilar artery segments following 1 min of stimulation by 10(-7) M alpha- or beta-CGRP. In another set of experiments, the time course of alpha- and beta-CGRP induced vasodilatation was investigated and concentration dependent responses of the two peptides were demonstrated. No significant differences between the actions of alpha- and beta-CGRP regarding induction of cAMP formation, amount of vasodilatation, time course of vasodilatation and mode of inhibition by the CGRP receptor antagonist, human alpha-CGRP(8-37), could be detected. The presence of mRNA encoding the guinea pig CRLR and the guinea pig CGRP receptor component protein (RCP) in the guinea pig basilar artery was demonstrated by RT-PCR methods. Furthermore, a partial sequence of mRNA encoding the guinea pig CRLR was determined. The expression in this tissue of a CRLR derived CGRP receptor and a functional RCP is therefore likely, and the equipotent pharmacological actions of alpha- and beta-CGRP might be mediated via CRLR derived CGRP receptors.

Characterisation of the effects of a non-peptide CGRP receptor antagonist in SK-N-MC cells and isolated human cerebral arteries.

The cerebral circulation is innervated by calcitonin gene-related peptide (CGRP) containing fibers originating in the trigeminal ganglion. During a migraine attack, there is a release of CGRP in conjunction with the head pain, and triptan administration abolishes both the CGRP release and the pain at the same time. In the search for a novel treatment of migraine, a non-peptide CGRP antagonist has long been sought. Here, we present data on a human cell line and human and guinea-pig isolated cranial arteries for such an antagonist, Compound 1 (4-(2-Oxo-2,3-dihydro-benzoimidazol-1-yl)-piperidine-1-carboxylic acid [1-(3,5-dibromo-4-hydroxy-benzyl)-2-oxo-2-(4-phenyl-piperazin-1-yl)-ethyl]-amide). On SK-N-MC cell membranes, radiolabelled CGRP binding was displaced by both CGRP-(8-37) and Compound 1, yielding pK(i) values of 8.9 and 7.8, respectively. Functional studies with SK-N-MC cells showed that CGRP-induced cAMP production was antagonised by both CGRP-(8-37) and Compound 1 with pA(2) values of 7.8 and 7.7, respectively. Isolated human and guinea pig cerebral arteries were studied with a sensitive myograph technique. CGRP induced a concentration-dependent relaxation in human cerebral arteries which was antagonized by both CGRP-(8-37) and Compound 1 in a competitive manner. In guinea pig basilar arteries, CGRP-(8-37) antagonised the CGRP-induced relaxation while Compound 1 had a weak blocking effect. The clinical studies of non-peptide CGRP antagonists are awaited with great interest.

CGRP and adrenomedullin receptor populations in human cerebral arteries: in vitro pharmacological and molecular investigations in different artery sizes.

The aim of the present study was to determine functional and molecular characteristics of receptors for calcitonin gene-related peptide (CGRP) and adrenomedullin in three different diameter groups of lenticulostriate arteries. Furthermore, the presence of perivascular neuronal sources of CGRP was evaluated in these arteries. In the functional studies, in vitro pharmacological experiments demonstrated that both CGRP and adrenomedullin induce alpha-CGRP-(8-37) sensitive vasodilation in artery segments of various diameters. The maximal amounts of vasodilation induced by CGRP and adrenomedullin were not different, whereas the potency of CGRP exceeded that of adrenomedullin by 2 orders of magnitude. Significant negative correlations between artery diameters and maximal responses were demonstrated for CGRP and adrenomedullin. In addition, the potency of both peptides tended to increase in decreasing artery diameter. In the molecular experiments, levels of mRNAs encoding CGRP receptors and receptor subunits were compared using reverse transcriptase polymerase chain reactions (RT-PCR). The larger the artery, the more mRNA encoding receptor activity-modifying proteins 1 and 2 (RAMP1 and RAMP2) was detected relative to the amount of mRNA encoding the calcitonin receptor-like receptor. By immunohistochemistry, perivascular CGRP containing nerve fibres were demonstrated in all the investigated artery sizes. In conclusion, both CGRP and adrenomedullin induced vasodilation via CGRP receptors in human lenticulostriate artery of various diameter. The artery responsiveness to the CGRP receptor agonists increased with smaller artery diameter, whereas the receptor-phenotype determining mRNA ratios tended to decrease. No evidence for CGRP and adrenomedullin receptor heterogeneity was present in lenticulostriate arteries of different diameters.

Expression of calcitonin receptor-like receptor and receptor-activity-modifying proteins in human cranial arteries.

Recently a new type of proteins modulating the pharmacological profile of the calcitonin receptor-like receptor (CRLR) were identified. The receptor-activity-modifying proteins (RAMPs) were shown to be essential for the expression of a functional CRLR and furthermore the RAMPs seemed to modify ligand selectivity of CRLR: coexpression of CRLR and RAMP1 resulted in a CGRP1 type of receptor while an adrenomedullin receptor resulted when CRLR and RAMP2 were coexpressed. In the present study significant molecular expression of CRLR concomitant with RAMP1, 2 and 3 were demonstrated in human meningeal, cerebral and temporal arteries by use of reverse transcriptase polymerase chain reactions (RT-PCR). These findings support previous studies demonstrating functional CGRP1 receptors in human cranial arteries. Furthermore the present study suggests the potential for functional adrenomedullin receptors in human cranial arteries.

Communication of the ulnaris lateralis bursa with the equine elbow joint and evaluation of caudal arthrocentesis.

Elbows from cadaver limbs were evaluated to determine the presence of a communication between the ulnaris lateralis bursa (ULB) and the joint and the extent of the bursa. Thirty-two pairs of joints were studied: 12 pairs were frozen, then transversely sectioned and 20 pairs were injected with methyl methacrylate. The 12 frozen-section pairs revealed a communication between the ULB and the elbow joint in 9/24 joints (37.5%) and a true bursa (absence of communication) in 15/24 joints (62.5%). The mean bursal length in adult horses was 3.8 cm. There was no significant difference in the length of the bursa or presence of bursa-joint communication between the right and left limbs. In the acrylic specimens a communication between the ULB and the joint was found in 19/40 specimens (47.5%). There was no significant correlation between age or sex and frequency of communication. There was a significantly greater prevalence of communications present in Quarter Horse than in non-Quarter Horse (P < 0.05) joints. A communication between the ULB and the joint is not always present, and therefore injection of the elbow joint via the ULB may be unreliable.

In vitro comparison of three fixation methods for humeral fracture repair in adult horses.

The stiffness, load to failure, and bending moments of adult intact equine humeri and humeri repaired with 3 fixation techniques were determined in vitro. Bones were tested in axial compression (30 pairs), mediolateral 3-point bending (15 pairs), and caudocranial 3-point bending (15 pairs). An oblique osteotomy of 1 humerus of each pair was performed to simulate the long spiral oblique fractures that occur clinically in horses. Bones were repaired in 3 ways: group 1--nylon band cerclage fixation (20 bones); group 2--multiple intramedullary pinning (20 bones); and group 3--nylon band cerclage fixation and multiple intramedullary pinning (20 bones). Intact bones were significantly (P less than 0.05) stronger than repaired bones in each testing mode. Bones repaired with bands only were significantly less stiff in bending than were bones repaired with pins only or with pins and bands. In compression, only specimens repaired with pins and bands were significantly stiffer than were bones repaired with bands only. Bones repaired with bands only required significantly less load to failure in compression and in caudocranial bending than did bones repaired with pins only or with pins and bands. Bones repaired with pins only deformed through the full displacement of the actuator (5 cm), and pins deformed plastically. Bones repaired with pins and bands were stiffer and had higher bending moments than did bones repaired with pins only, but the differences were not significant.

Temporal matrix synthesis and histologic features of a chondrocyte-laden porous collagen cartilage analogue.

Cartilage resurfacing by chondrocyte transplantation, using porous collagen matrices as a vehicle to secure the cells in cartilage defects, has been used experimentally in animals. This in vitro study evaluated the temporal morphologic features and proteoglycan synthesis of chondrocyte-laden collagen matrices. Forty-two porous collagen disks were implanted with a minimum of 6 x 10(6) viable chondrocytes, covered by a polymerized collagen gel layer, and 6 disks were harvested after 0, 3, 7, 10, 14, 18, or 22 days of incubation in supplemented Ham's F12 medium at 37 C and 5% CO2. Histologic and histochemical evaluation of formalin-fixed segments of the cultured disks indicated that the chondrocytes proliferated in the implant, producing small groups and linear segments of cells by day 14. The collagen framework remained intact over the course of the study with thick areas attributable to depositions of matrix material after day 10. Alcian blue-stained matrix was evident in the pericellular region of chondrocytes in sections of disks harvested on days 14, 18, and 22. Glycosaminoglycan (GAG) assay by dimethylmethylene blue dye binding after papain digestion of the disk segments revealed negligible amounts of GAG at day 0. Significant (P < or = 0.0001) increase in total GAG content was observed by day 3 (0.329 micrograms/mg of disk) and further increases were observed until a plateau in GAG quantity was seen on day 14. Mean peak GAG content was 0.553 +/- 0.062 micrograms/mg. Secondary treatment of the papain-digested implants with keratanase and chondroitinase ABC yielded similar trends in chondroitin sulfate (CS) and keratan sulfate (KS) concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)