Longitudinal changes in vitamin D and its metabolites in pregnant South Africans.

BACKGROUND: Vitamin D deficiency (VDD) has been associated with adverse maternal and foetal outcomes and is determined by measuring 25 hydroxyvitamin D (25(OH)D). The 25(OH)D is catabolized to 24, 25-(OH) 2D and the ratio of 25(OH) D to 24, 25-(OH)2D, the vitamin D metabolite ratio (VMR), is thought to be a superior marker of VDD, being elevated in such states. The aims of this study were to assess the longitudinal vitamin D status of pregnant women by measuring cholecalciferol, 25(OH)D, 24, 25-(OH)2D and VMR at two time points and also to determine any association of vitamin D and metabolites with gestational age at birth, birth length and weight. METHODS: We recruited 400 pregnant black African women in their first trimester (V1) and measured weights and heights. Ultrasound scans were performed for gestational age. Blood was drawn at V1 and at about 26 weeks (V2) of gestation for cholecalciferol, 25(OH)D, 24, 25-(OH)2D, VMR and parathyroid hormone (PTH). An OGTT was performed at V2 where fasting glucose, insulin and 30-minute glucose were measured. At birth, we measured birth weight, length and gestational age. Maternal insulin, PTH and vitamin D binding protein (VDBP) were measured by immunoassay. Maternal albumin was measured colorimetrically. Maternal cholecalciferol, 25(OH)D and 24, 25-(OH)2D, were measured by mass spectrometry and free and bioavailable vitamin D were calculated. Initial gestation was determined by ultrasound. We compared analytes by visit as well as by 25(OH)D status. Vitamin D deficiency (<30 nmol/L) was defined according to the National Academy of Medicine guidelines. Linear regression analysis was used to determine associations of vitamin D molecules with maternal blood pressure, fasting and 30-minute insulin and blood glucose and neonatal parameters. RESULTS: Results are presented for participants for whom we had complete data (n = 330-346 depending on variable). The prevalence of vitamin D deficiency (VDD) was 35.8 % at V1 and 32.4 % at V2. Levels of 25(OH)D did not change significantly between visits. Levels of 24, 25(OH)2D dropped from the first to the second visit (17.64 ± 12.64 to 9.39 ± 9.07 nmol/L; p < 0.0001) while VMR increased ((3.15 (1.31; 7.67) to 7.90 (2.44; 25.98); p < 0.0001). The proportion of women with the lowest cholecalciferol concentrations increased at V2 compared to the V1 (36.1-42.8 %; p = 0.02). In multivariable regression models 25(OH)D was negatively associated with 30-minute glucose concentrations (p = 0.038) whilst 24, 25-(OH)2D was positively associated with fasting insulin (p = 0.017) and HOM A-I R (p = 0.023). There was no correlation of 25(OH)D or metabolites with infant birth weight, birth length or gestational age. CONCLUSIONS: Maternal VDD is common in pregnant black South African women. Decreased VMR suggest that catabolism of 25(OH)D is reduced in pregnancy to maintain adequate free vitamin D levels.

In vitro marginal and internal adaptation of metal-ceramic crowns with cobalt-chrome and titanium framework fabricated with CAD/CAM and casting technique.

AIM: The purpose of this study was to compare the marginal and internal fit of Co-Cr and titanium ceramic crowns fabricated with 2 different techniques: CAD/CAM milling and traditional casting (TC). MATERIALS AND METHODS: Ten traditional casting of Co-Cr (TCC), 10 CAD/CAM milling of Co-Cr (MC), 10 traditional casting of titanium (TCT), 10 CAD/CAM milling of titanium ceramic crowns (MT) were fabricated. Silicone replicas were obtained to measure internal gap volume, marginal gap and internal adaptation that was evaluated at 3 regions: axial wall, axio-occlusal angle, and occlusal surface. Measurements were made with a X-ray micro computerized tomography (micro-CT) and analyzed with Bonferroni and Dunnet T3 post-hoc tests (α = 0.05). RESULTS: One-way ANOVA revealed no statistically significant differences among the groups for measurements at the marginal gap (P > 0.05). At axial wall region the mean values of TCT group were higher than those of groups but only statistically not significant for TCC group (P < 0.05). TCC group statistically gives better results than MC group in axio-occlusal and occlusal regions (P < 0.05). The mean measurement of internal gap volume were 20.59 ± 0.83 mm3 for TCC, 22.73 ± 0.82 mm3 for MC, 22.83 ± 1.11 mm3 for TCT and 20.51 ± 1.16 mm3 for MT. Mean internal gap volume values MT group were smaller than those of groups but only statistically not significant for TCC group (P > 0.05). CONCLUSION: All groups performed similar marginal adaptation. The cement film thickness at axio-occlusal angle point and occlusal region were higher for MC crowns.

Effects of sandblasting and silicoating on bond strength between titanium and porcelain.

PURPOSE: The aim of this study was to evaluate the effects of the different sized alumina particles (50 and 150 µm) and tribochemical silica-modified alumina particles (110 µm) on titanium (Ti) surface to identify the most effective method of increasing the bond strength between porcelain and Ti. MATERIALS AND METHODS: Thirty rectangular plates (15 mm × 50 mm × 1 mm) of commercially pure Ti (Cp Ti) Grade 5 (GC Dental Industrial Corporation, Tokyo, Japan) were divided into three groups for different surface modification procedures (n = 10). Ti bonder porcelain, opaque, and dentin layers were fired separately on Ti plates. All specimens were placed in a bending jig for four-point bending test. The load and crosshead displacement data were collected to calculate the strain energy release rate as a G value. RESULTS: Lowest mean G values in J/m2 were in the group sandblasted with 150 µm Al2O3particles (Group 2) (18.6 ± 5), followed by the group sandblasted with 50 µm Al2O3particles (Group 3) (20.8 ± 6.1) and the group sandblasted with 110 µm silicoated Al2O3particles (Group 1) (24.5 ± 4.1). The one-way ANOVA and Bonferroni post hoc tests indicated that there was a statistically significant difference between Group 1 and Group 2 (P < 0.05). There were also no statistically significant differences between Group 1 and Group 3 and Group 2 and Group 3 (P > 0.05). CONCLUSION: The size of alumina particles is not a factor that is directly effective in enhancing the bond strength of Ti-porcelain systems. The bond strength of Ti-porcelain systems can be extremely improved by the application of sandblasting with silica-coated alumina particles.

Visceral adiposity is a predictor of parathyroid hormone levels in healthy adults.

PURPOSE: Parathyroid hormone (PTH) has been shown to correlate positively with fat mass, however there are no studies that have investigated whether this association is a result of, or is modified by, body fat distribution. The aim of this study was to investigate the association of PTH with several body composition indices, namely visceral (VAT) and subcutaneous adiposity (SCAT) as well as with lean mass and with serum leptin, which has been reported to increase PTH. METHODS: This was a cross-sectional study in which PTH was measured by chemiluminescent assay; body fat and lean mass by dual-energy X-ray absorptiometry (DXA) and abdominal fat by ultrasonography in 714 healthy adults aged 18-65 years. Serum leptin was measured by ELISA. RESULTS: In a multivariate linear regression model that included height, age, gender, ethnicity, serum 25 hydroxyvitamin D, leptin levels, calcium, magnesium and phosphate concentrations, glomerular filtration rate, smoking status, and calcium and vitamin D supplementation as independent variables and PTH as the dependent variable, VAT (ß = 0.094, p = 0.035) correlated significantly with PTH levels but SCAT (ß = -0.045, p = 0.386) and body fat mass (ß = 0.098, p = 0.126) did not. Leptin did not correlate with PTH (ß = 0.013, p = 0.832) in this regression model. CONCLUSIONS: Plasma PTH is significantly associated with VAT in healthy adults. In view of the association of PTH with increased cardiovascular mortality, it is important to investigate this association further.

Effect of chlorhexidine on bond strength between glass-fiber post and root canal dentine after six month of water storage.

Aim of this study was to assess the influence of chlorhexidine digluconate (CHX) application on bond strength of glass fibre reinforced composite (FRC) posts to root dentine using adhesive luting systems. Forty extracted human mandibular premolars were endodontically treated and root canals were prepared. Teeth were divided into 2 groups according to luting system as SuperBond C&B (etch-and-rinse/chemical cure) and FuturaBond DC (self-etch/ dual cure). Each group was further divided in 2 subgroups (n=10) according to whether CHX was applied or not. Group Futura/CHX: FuturaBond DC + CHX; group Futura: FuturaBond DC; group Super/CHX: Super Bond C&B + CHX; group Super: SuperBond C&B + CHX. Mean and standard deviation (in parenthesis) values of bond strength in MPa were: group Futura/CHX: 8.86 (1.96), group Futura: 7.65 (1.01), group Super/CHX: 17.47(2.93), group Super: 12.41 (3.83). Bond strength values were affected by the type of luting agent and CHX irrigation (p=0.001, two-way ANOVA). There were statistically significant differences among the groups according to one-way ANOVA (p<0.001). Significant differences were observed in bond strength between groups Super/CHX and Super (p=0.023), between groups Futura/CHX and Super/CHX (p<0.001). Application of CHX before luting procedure with etch-and-rinse/chemical cure luting agent of glass FRC post improved long-term bond strength between glass FRC and root dentine.

Characterization of CD34⁺ thymocytes in newborn dogs.

Using two-color flow cytometry, we characterized CD34(+) cells in the newborn canine thymus. CD34(+) thymic cells comprised approximately 5% of cells recovered by thymus tissue teasing and both large and small thymocytes have been present in this population, the former being 7-12 times more frequent. All CD34(+) cells expressed the pan-leukocyte antigen CD45. The expression of CD44 profile on the large and small CD34(+) thymocytes differed: almost all large CD34(+) cells were CD44(+), while only 75% of small CD34(+) thymocytes co-expressed the CD44 antigen. We have previously described that CD172α is present on the surface of CD34(+) bone marrow cells in dogs. In the thymus, CD172α was expressed on 5-10% and less than 5% of large and small CD34(+) cells, respectively. Some CD34(+) thymocytes also co-expressed T-lineage-specific markers like CD3, CD4, CD8, TCR1 and TCR2. Their expression increased during the large-to-small thymocyte transition. Based on our findings we suggest that thymocyte progenitors enter their primary differentiation center as large CD34(+), CD44(+), CD45(+) and CD172α(+) cells. T-cell specific markers appear on their surface at early stages of differentiation. As the size of progenitors decreases with terminal primary differentiation, the CD34, CD44, and CD172α surface markers are down-regulated.

The evaluation of displacement resistance of glass FRC posts to root dentine using a thin slice push-out test.

AIM: To investigate and compare the displacement resistance of glass fibre reinforced composite (FRC) posts to root dentine after luting with different adhesive systems. METHODOLOGY: A total of 32 noncarious extracted human mandibular premolars were prepared for post-cementation using the FRC Postec system (Ivoclar Vivadent, Schaan, Liechtenstein) and divided into four groups (n = 8). The posts in each group were luted with one or other of the following materials. Group 1: Variolink II/Excite DSC (etch-and-rinse, dual-curing), group 2: Clearfil Esthetic Cement/ED Primer II (self-etch, dual-curing), group 3: Multilink/Multilink Primer (self-etch, chemical-curing) and group 4: Multilink Sprint (self-adhesive, dual-curing). Specimens were sectioned to obtain slices with the post in the centre and with the root dentine overlaid by the autopolymerizing acrylic resin on each side. The displacement resistance was measured using a Universal Testing Machine at a crosshead speed of 0.5 mm min(-1). The displacement resistance of the specimens were calculated and expressed in MPa. Data were analysed with one-way ANOVA and post hoc Tukey's test (P < 0.05). RESULTS: Mean (SD) values of displacement resistance data in MPa are as follows: group 1, 12.08 (2.13); group 2, 12.39 (2); group 3, 11.3 (1.23); group 4, 14.29 (1.84). There were statistically significant differences amongst the groups (P = 0.021). A statistically significant difference was observed for the displacement resistance values between groups 3 and 4 (P = 0.015), that is between Multilink/Multilink Primer and Multilink Sprint. CONCLUSIONS: Glass FRC posts luted with self-adhesive luting system exhibited higher displacement resistance than when luted with chemical-curing self-etch luting system.

Abnormal cation exchange in insulin-resistant patients with essential hypertension.

OBJECTIVES: To identify important factors that may contribute to abnormal glucose tolerance in elderly patients with treated hypertension with primary reference to changes in the following parameters: calculated insulin resistance, endogenous insulin processing and secretion; platelet cation concentration and membrane ATPase activity. DESIGN: Thirty-nine patients receiving antihypertensive therapy (including low-dose thiazide treatment) were compared to 13 normotensive, normoglycaemic control subjects. Total platelet cation concentration and membrane ATPase activity were measured and, following a 75-g oral glucose test, serum insulin, proinsulin and 31-32 des-proinsulin responses were measured in prospectively defined hypertensive patients with normal glucose tolerance (NG), impaired glucose tolerance (IGT) and diabetes mellitus (DM). RESULTS: Of the total patient cohort, seven patients manifested newly diagnosed DM, 18 had IGT and 14 NG. Among the three groups, no difference in duration of drug use (thiazides and beta-blockers) was noted; BMI and waist-to-hip ratio increased progressively from NG to IGT to overt DM. Compared to NG patients, serum insulin responses were significantly greater in the IGT (all time points) and DM (two-hour measurements) subjects. Proinsulin and 31-32 des-proinsulin serum responses were likewise significantly higher in the IGT and DM groups. The derived measure of insulin resistance in the hypertensive patients showed a significant increase in the progression from NG to IGT and DM. Mean total platelet potassium concentration was reduced in the DM compared to the IGT and the control groups, while platelet sodium, calcium and magnesium concentrations showed no significant differences. Platelet membrane magnesium ATPase activity was significantly higher in the normotensive control versus the hypertensive group. Sodium, potassium and calcium ATPase activity showed no significant differences among the subgroups. CONCLUSION: Our findings support the strong link between essential hypertension, insulin resistance/hyperinsulinaemia and regional adiposity. Beta-cell dysfunction (hypersecretion and abnormal insulin processing) is manifest in the progression from normality to overt diabetes. The use of antihypertensive therapy (low-dose thiazides and cardioselective beta-blockers) possibly added diabetogenic effect(s). The reduction in platelet total potassium concentration paralleled the diabetic state while a reduced membrane magnesium ATPase activity correlated with the hypertensive state.

Abnormal Cation Exchange in Insulin-Resistant Patients with Essential Hypertension : Cardiovascular Topics

Objectives: To identify important factors that may contribute to abnormal glucose tolerance in elderly patients with treated hypertension with primary reference to changes in the following parameters: calculated insulin resistance; endogenous insulin processing and secretion; platelet cation concentration and membrane ATPase activity. Design: Thirty-nine patients receiving antihypertensive therapy (including low-dose thiazide treatment) were compared to 13 normotensive; normoglycaemic control subjects. Total platelet cation concentration and membrane ATPase activity were measured and; following a 75-g oral glucose test; serum insulin; proinsulin and 31-32 des-proinsulin responses were measured in prospectively defined hypertensive patients with normal glucose tolerance (NG); impaired glucose tolerance (iGT) and diabetes mellitus (DM). Results: of the total patient cohort; seven patients manifested newly diagnosed DM; 18 had iGT and 14 NG. Among the three groups; no difference in duration of drug use (thiazides and beta-blockers) was noted; BMi and waist-to-hip ratio increased progressively from NG to iGT to overt DM. Compared to NG patients; serum insulin responses were significantly greater in the iGT (all time points) and DM (two-hour measurements) subjects. Proinsulin and 31-32 des-proinsulin serum responses were likewise significantly higher in the iGT and DM groups. The derived measure of insulin resistance in the hypertensive patients showed a significant increase in the progression from NG to iGT and DM. Mean total platelet potassium concentration was reduced in the DM compared to the iGT and the control groups; while platelet sodium; calcium and magnesium concentrations showed no Significant differences. Platelet membrane magnesium ATPase activity was significantly higher in the normotensive control versus the hypertensive group. Sodium; potassium and calcium ATPase activity showed no significant differences among the subgroups. Conclusion: our findings support the strong link between essential hypertension; insulin resistance / hyperinsulinaemia and regional adiposity. Beta-cell dysfunction (hypersecretion and abnormal insulin processing) is manifest in the progression from normality to overt diabetes. The use of antihypertensive therapy (low-dose thiazides and cardioselective beta-blockers) possibly added diabetogenic effect(s). The reduction in platelet total potassium concentration paralleled the diabetic state while a reduced membrane magnesium ATPase activity correlated with the hypertensive state

Systemic and local antibody responses after experimental infection with Actinobacillus pleuropneumoniae in piglets with passive or active immunity.

The objectives of the present study was to describe different dynamics of humoral immune responses to experimental infection in piglets of different stages of infection and immunity. Two groups of piglets originating from non-immune (group 1) and immune (group 2) sows at the age of 3 weeks were subdivided as follows: a half of each group of piglets was exposed to a low-dose infection with Actinobacillus pleuropneumoniae (APP) strain 9. At the age of 8 weeks, all four groups of piglets were challenged with a high infection dose of APP of the same strain. Isotype characterization of the specific antibodies in sera and in bronchoalveolar lavage fluids (BALF) to a lipopolysaccharide was carried out, besides monitoring clinical signs and post-mortem examinations. A typical primary immune response was observed in specific antibody-free piglets infected with a challenge infection. Colostrum-derived immunoglobulin-G (IgG) antibodies persisted in sera and BALF of piglets up to the age of 8 weeks. However, they did not prevent induction of specific-primary antibody response, either in 8 or 4 weeks of age, when levels of specific colostrum-derived antibodies were still high. It was demonstrated by the increase of specific IgM antibodies in sera. The infection induced an increase in the levels of IgA antibodies in BALF regardless the severity of infection and presence of specific colostrum-derived antibodies. The specific antibodies of IgG isotype increased only in BALF from piglets without colostrum-derived antibodies.

Experimental Actinobacillus pleuropneumoniae infection in piglets with different types and levels of specific protection: immunophenotypic analysis of lymphocyte subsets in the circulation and respiratory mucosal lymphoid tissue.

Actinobacillus pleuropneumoniae (APP) infection in piglets results in severe and fatal fibrinous hemorrhagic necrotizing pneumoniae. The aim of our study was to analyze changes in lymphocyte subset distribution in peripheral blood, bronchoalveolar lavage fluid (BALF) and tracheobronchal lymph nodes (TLN) in non-immune piglets upon a challenge with a high dose of APP and to compare the quality of such changes in unprotected piglets with counterparts exhibiting specific immunity mediated by high titers of colostrum-derived APP-specific antibodies and/or a low dose APP infection in the early postnatal period. Challenge with APP resulted in a massive increase in CD8-negative gammadelta T-cells in parallel with a reduction in numbers of CD3-CD8low cells in BALF independent of the type and level of immunity and this seems to be a general phenomenon associated with experimental infection. An increase in B-lymphocyte numbers in TLN was another characteristic feature accompanying APP infection in all experimental groups. In piglets with colostrum-derived APP-specific antibodies, this was associated with higher relative numbers of IgM+CD2+ lymphocytes in TLN, while B-cells with the CD2- surface phenotype apparently expanded in the absence of passive humoral immunity.

Effect of luting agents and reconstruction techniques on the fracture resistance of pre-fabricated post systems.

Fracture resistance and fracture modes of endodontically treated maxillary central incisors restored with different post-and-core systems covered with all-ceramic copings were evaluated. Ten samples were prepared for each group. Groups 1, 2 and 3 consisted of tooth-coloured post-and-core, zirconia post (Cosmopost) with a composite core (Tetric Ceram), zirconia post (Cosmopost) with a custom made ceramic core (Cosmo Ingot), glass fibre-reinforced post (FRC Postec) with a composite core (Tetric Ceram), respectively. Group 4 consisted of a titanium post (ERpost) with a composite core (Tetric Ceram). The control group (group 5) consisted of root-filled incisors without posts. Tooth-coloured posts were cemented in the roots using Variolink-2, while titanium posts were cemented in the roots using Harvard cement. The all-ceramic copings were cemented using Variolink-2. Static load was applied to 2 mm below the incisal edge on the palatinal surface of each sample until they were fractured. Fracture data were obtained and statistically analysed with One-way anova and a Tukey's test. The results of the means and standard deviations of the fracture resistance during static loading were: 497.5 +/- 61.94 (1), 474.61 +/- 96.84 (2), 494.61 +/- 104.67 (3), 581.34 +/- 105.36 (4), 420.42 +/- 127.48 (5). There were statistically significant differences between groups 4 and 5. Glass fibre-reinforced posts and composite cores (group 3) showed the most catastrophic failure. Consequently, zirconia ceramic posts can be used in clinical practice.

Lymphatic organ development in dogs: major lymphocyte subsets and activity.

In the present study, we have characterized lymphocyte subsets and activity in peripheral blood, spleen, mesenteric and popliteal lymph nodes in pups from birth till the age of one month and compared the results with the situation in the group of three adult dogs. In neonatal pups, lower numbers of CD3(+) T-cells were detected in both the spleen and peripheral blood than in lymph nodes. In contrast to the other compartments, CD21(+) B-cells prevailed in the spleen, which resulted in low values (<1) of the CD3(+)/CD21(+) ratio. Low numbers of CD8(+) lymphocytes were characteristic in all compartments immediately after birth; consequently a high CD4(+)/CD8(+) ratio has been calculated. Postnatal development was characterized by an increasing frequency of CD8(+) lymphocytes in all organs studied. Another typical feature of the early period of life was a relative decrease of B-cell numbers, which was compensated by an increasing proportion of T-lymphocytes, particularly in the peripheral blood and spleen. DNA synthesis in newborn pups' cells as measured by in vitro thymidine incorporation was surprisingly high in non-stimulated control samples, notably in the spleen. Further development of lymphocyte activity was characterized by the decline in spontaneous activity in all organs. Stimulation indices upon mitogen-induced proliferation increased proportionally to the decrease in spontaneous activity. Based on our experimental data, we have concluded that pups are born with a relatively competent immune system the structure of which, however, markedly develops during a few postnatal weeks.

The intrauterine environment is a strong determinant of glucose tolerance during the neonatal period, even in prematurity.

The aim of this study was to determine the contribution of birth weight and gestational age to glucose tolerance in premature neonates. The study group consisted of 100 premature and/or small-for-gestational age infants. Anthropometric measurements were performed both at birth and at the time of a standardized milk feed carried out at 19.6 +/- 12.1 d (range, 1-65 d) after birth. Fasting and postprandial glucose and insulin levels were measured. Birth weight, as a proxy mirror of the intrauterine environment, was found to influence the glucose concentration following a standardized milk feed (beta = -0.46; P = 0.01 for birth weight z-score with 60-min glucose level), whereas gestational age did not. Small-for-gestational age neonates had higher 60-min insulin levels than appropriate-for-gestational age neonates (115.4 +/- 9.5 vs. 68.4 +/- 14.2; P < 0.05) despite similar glucose levels. Neonates born of mothers who were on antihypertensive treatment were smaller and had a higher insulin secretory response than neonates from normotensive mothers. Postnatal growth velocity (kilograms per day) correlated with birth weight (beta = -0.65; P < 0.0001) and insulin resistance (beta = -0.31; P = 0.0004), independently of each other. This study shows that glucose tolerance of the neonate is determined by weight attained at birth irrespective of gestational age and that maternal blood pressure may influence insulin sensitivity of the newborn. Furthermore, catch-up growth in neonates is determined by birth weight and insulin sensitivity.

Postnatal development of leukocyte subset composition and activity in dogs.

The aim of the presentation is to summarise our data on the counts and activity of circulating canine leukocytes at birth and on their changes in the first 3 months of life. On day 1, neutrophil counts were almost three times higher than lymphocyte counts. During the first week of life, a decrease of neutrophil and an increase of lymphocyte counts, resulting in a predominance of lymphocytes, were observed. Neutrophil counts reached values comparable with those in adults in 1 month. Lymphocyte counts were higher than those in adults during the first 3 months. From birth to the age of 3 months, the phagocytic activity of neutrophils was nonsignificantly higher than in young adults. When compared with adults, the peripheral blood of new-born pups contained a lower proportion of T lymphocytes (detected by CD3 and CD5 markers), with a very low percentage of CD8(+) cells and a higher proportion of CD21(+) B lymphocytes. The counts of individual subsets levelled out during the first 3 months of life, although the proportion of CD21(+) B cells remained higher all the time. Lymphocytes of new-born pups were able to respond to nonspecific mitogen stimulation. Spontaneous proliferation in vitro was higher during the first week of life. Although in vitro stimulation of lymphocytes with Concanavalin A in some pups was comparable with that of adult dogs, mean activity was weaker. Pups with zero or very low levels of maternal antibodies were able to develop specific immune responses to a parvovirus antigen as early as at 2 weeks of age. On the basis of these data, we assume that pups are born with an immune system that can respond to external stimuli. Nevertheless its development continues in the postnatal period and some parameters differ from adult values for at least 3 months after birth.

Lymphocyte subsets in peripheral blood of dogs--a flow cytometric study.

Slight differences in the results of papers describing lymphocyte subsets distribution in the peripheral blood of healthy dogs may be explained by differences in monoclonal antibody clones and sources, breed and age of animals examined, methods of sample treatment, or methods of result analysis. In this paper, we described the effect of sample processing and of sample storage as well as the effect of age, breed, and gender of dogs on lymphocyte subset distribution. No significant differences were found between samples processed following a whole-blood lysis method and samples processed after density gradient separation. Furthermore, no significant differences were found between samples processed within 2h after collection and those stored at 4 degrees C for 12-16 h before processing. Age-related changes were evident in lymphocyte subset distribution in the peripheral blood of 38 Beagles divided according to their age into the six groups: (1) 5-6 days; (2) 2 months; (3) 6 months; (4) 1-2 years; (5) 3-5 years; and (6) >5 years. The percentage of B-lymphocytes (CD21-like positive cells) in the peripheral blood of newborn pups was 39.5+/-5.7 and decreased with advancing age. The percentage of CD8+ lymphocytes was 7.7+/-3.4 after birth and increased with advancing age. No age-related changes were observed in the percentages of CD4+ lymphocytes. The CD4+:CD8+ ratio decreased with advancing age. No significant age-related change was observed for lymphocytes bearing the gammadelta-TCR. Some breed differences were evident. Adult (1-5-year-old) Beagles, German Shepherds, Dalmatians, and Dachshunds were examined. The percentages of lymphocytes were higher in Beagles and Dachshunds than in Dalmatians and German Shepherds. The highest and the lowest absolute lymphocyte counts were found in Beagles and German Shepherds, respectively. As a consequence, German Shepherds showed the lowest absolute counts of the individual lymphocyte subpopulations and the widest neutrophil:lymphocyte ratio. Dalmatians showed the lowest percentage of CD3+ cells, the highest percentage of CD21+ cells, and the lowest CD4+:CD8+ ratio. German Shepherds showed the lowest percentage of CD21+ cells and the highest CD4+:CD8+ ratio. Females in Beagles and Dachshuns had nonsignificantly higher percentages of total lymphocytes, CD3+, CD4+, and nonsignificantly lower percentages of CD21+ lymphocytes. We concluded that there are age-, breed-, and perhaps also gender-related differences in lymphocyte subset distribution in the peripheral blood of dogs. Therefore, there is need to use appropriate control group in the experimental protocols. Among-breed differences could explain, at least partly, breed predisposition for some diseases.

Weight-related differences in glucose metabolism and free fatty acid production in two South African population groups.

OBJECTIVE: The effects of free fatty acids (FFA), leptin, tumour necrosis factor (TNF) alpha and body fat distribution on in vivo oxidation of a glucose load were studied in two South African ethnic groups. DESIGN AND MEASUREMENTS: Anthropometric and various metabolic indices were measured at fasting and during a 7 h oral glucose tolerance test (OGTT). Body composition was measured using bioelectrical impedance analysis and subcutaneous and visceral fat mass was assessed using a five- and two-level CT-scan respectively. Glucose oxidation was evaluated by measuring the ratio of (13)CO(2) to (12)CO(2) in breath following ingestion of 1-(13)C-labelled glucose. SUBJECTS: Ten lean black women (LBW), ten obese black women (OBW), nine lean white women (LWW) and nine obese white women (OWW) were investigated after an overnight fast. RESULTS: Visceral fat levels were significantly higher (P<0.01) in obese white than black women, despite similar body mass indexes (BMIs). There were no ethnic differences in glucose oxidation however; in the lean subjects of both ethnic groups the area under the curve (AUC) was higher than in obese subjects (P<0.05 for both) and was found to correlate negatively with weight (r=-0.69, P<0.01) after correcting for age. Basal TNF alpha concentrations were similar in all groups. Percentage suppression of FFAs at 30 min of the OGTT was 24+/-12% in OWW and -38+/-23% (P<0.05) in OBW, ie the 30 min FFA level was higher than the fasting level in the latter group. AUC for FFAs during the late postprandial period (120--420 min) was significantly higher in OWW than OBW (P<0.01) and LWW (P<0.01) and correlated positively with visceral fat mass independent of age (r=0.78, P<0.05) in the OWW only. Leptin levels were higher (P<0.01) both at fasting and during the course of the OGTT in obese women from both ethnic groups compared to the lean women. CONCLUSIONS: Glucose oxidation is reduced in obese subjects of both ethnic groups; inter- and intra-ethnic differences were observed in visceral fat mass and FFA production and it is possible that such differences may play a role in the differing prevalences of obesity-related disorders that have been reported in these two populations.

Ethnic differences in lipid metabolism in two groups of obese South African women.

There is a higher prevalence of ischemic heart disease (IHD) in South African white than black women. The objective of this study was to determine biochemical explanations for this prevalence. The study group contained 15 obese black women (OBW) and 14 obese white women (OWW), all premenopausal, who were examined after an overnight fast. Anthropometric measurements and blood concentrations of glucose, non-esterified fatty acids (NEFAs), catecholamines, plasminogen activator inhibitor-1, C-peptide, proinsulin, lipograms, cortisol, growth hormone, and post-heparin lipoprotein lipase activity were measured during an oral glucose tolerance test (OGTT). Body composition was measured using bioelectrical impedance analysis, and subcutaneous and visceral fat mass were assessed with CT-scans. Visceral fat area was higher in OWW (139.7 +/- 10.7 cm(2)) than in OBW (72.3 +/- 3.9 cm(2)) (P < 0.01), as were fasting and 3 h triglyceride concentrations (P < 0.05 for all). OWW also had higher NEFA levels than OBW at 3 and 4 h compared with OBW (P < 0.05 for both). Fasting cortisol (266 +/- 24 vs. 197 +/- 19 nmol/l; P < 0.05) was higher in OWW than in OBW. These data demonstrate that OWW have higher visceral fat mass than OBW, which may lead to a more atherogenic fasting and postprandial lipid profile. The higher cortisol levels of the OWW may promote visceral fat deposition.

Variations among unbred heifers in the activities of polymorphonuclear leucocytes from the mammary gland and blood.

The phenotypic characteristics are described for the activity of polymorphonuclear leucocytes NMN) obtained by either lavage of the cavity system of juvenile mammary glands stimulated with a synthetic muramyl dipeptide analogue or isolation from the peripheral blood. Attention was paid to the variability of characteristics and its sources, and to correlations among them. The following characteristics were investigated in 27 clinically healthy, unbred Bohemian Red Pied x Holstein heifers: migration activity in situ, number of phagocytosing PMN, phagocytotic index, bactericidal activity of PMN and unstimulated and zymosan-stimulated luminol-dependent chemiluminescence. Considerable individual variation was found in the characteristics. Significant differences between blood PMN and PMN from lavages after influx induction were found for bactericidal activity (P < 0.05) and chemiluminescence (P < 0.01). A significant correlation between blood PMN and mammary gland PMN was found only for the number of phagocytosing cells (r = 0.329; P < 0.01). Highly significant positive correlations (P < 0.01) were demonstrated between the number of phagocytosing PMN [a], phagocytotic index [b], and bactericidal activity [c] in both blood PMN (r(ab) = 0.602; r(ac) = 0.565; r(bc) = 0.529) and mammary gland PMN (r(ab) = 0.730, r(ac) = 0.618, r(bc) = 0.589). No significant correlation was demonstrated for non-stimulated (NS), zymosan-stimulated (ZS), or opsonized zymosan-stimulated (OZS) chemiluminescence with any of the other characteristics of phagocytotic activity, in either blood PMN or mammary gland PMN (P > 0.05). The animal was a highly significant source of variability for all the phagocytotic activity characteristics (P < 0.01). Udder quarter was a non-significant source of variability for all the characteristics of phagocytotic activity except for NS chemiluminescence (P < 0.05) and ZS or OZS chemiluminescence (P < 0.01). However, udder quarter was a non-significant source of variability of chemiluminescence indices ZS/NS and OZS/NS (P > 0.05). It has been demonstrated that in situ migration activity, the number of phagocytosing PMN, phagocytotic index, bactericidal activity of PMN and chemiluminescence indices of PMN collected from juvenile mammary glands of unbred heifers after influx induction can be regarded as candidate early markers of resistance to mammary infections.

Immunosuppression in bitches with pyometra.

The function of the immune system was studied in 34 bitches affected by pyometra. The mean age of the animals was 8.1 +/- 2.8 years. Samples of peripheral blood were examined using a battery of immunological tests. Data obtained from four sets of animals, grouped according to the severity of their disease, were compared. Leucocytosis (neutrophilia and partial monocytosis) and lymphopenia were detected in 62 per cent and 35 per cent of the bitches, respectively. The increase in the number of leucocytes was higher in the severely affected bitches. Inhibition of mitogen-driven lymphocyte proliferation was the characteristic feature of the immunological profile in bitches affected by pyometra, and this corresponded with the impairment of the general state of health. Sera from bitches showing functional inhibition of lymphocytes suppressed the activity of lymphocytes isolated from normal dogs. Sera from bitches affected by pyometra also had higher levels of immunoglobulins, lysozyme and circulating immune complexes. It can be concluded that pyometra is associated with an inflammatory response, but also with a marked suppression of immune system activity.