Monocyte Concentration Research Articles

Characterization of Intra‐Cellular Ionic Concentrations of Monocytes in Contact with Bioactive Glasses and Hydroxyapatite Particles. A TEM Cryo‐X‐ray Analysis of Diffusible Ions

A cells viability can be altered by the presence of bioactive materials particles and their features. This paper compares the influence of hydroxyapatite particles and bioactive glass particles on intracellular ionic concentrations of the monocytes exposed to them. However, microanalysis of intracellular ions like sodium, potassium, and chlorine is a major problem. In order to limit diffusion phenomenon and to preserve the chemical identity of specimens, we used cryomethods (cryo‐preparation, and cryo‐x‐ray microanalysis) to study cells in contact with bioactive materials particles. We measured intracellular ionic concentrations by scanning transmission electron microscopy (STEM) associated with energy dispersive x‐ray spectroscopy (EDXS). Monocytes were exposed to bioactive glass particles and to hydroxyapatite particles during 2 days. The K/Na ratio was used as a sensitive criteria of viability of cells. Bioactive glass particles and, in a less manner, hydroxyapatite particles have an effect on intracellular ionic concentrations. These two types of bioactive particles alter cells. However, bioactive glass particles appear to alter cells more than hydroxyapatite particles.

Caffeine suppresses TNF-α production via activation of the cyclic AMP/protein kinase A pathway

This study investigated the effect of in vitro exposure to caffeine, and its major metabolite paraxanthine, at concentrations relevant to typical caffeine consumption in humans, on lipopolysaccharide (LPS)-stimulated cytokine production in human whole blood. In addition, a role for the cyclic AMP/protein kinase A (PKA) pathway in the immunomodulatory effect of caffeine was investigated. Diluted whole blood (taken following ≥15 h abstinence from caffeine-containing food and beverages) was preincubated with caffeine or paraxanthine (10–100 μM) and stimulated with LPS (1 ∝g/ml) for 24 h. The proinflammatory cytokines tumour necrosis factor (TNF)-α, interleukin (IL)-1β and IL-12, and the antiinflammatory cytokine IL-10 were measured in cell-free supernatants. Whilst caffeine and paraxanthine had little or no effect on IL-10, IL-1β, or IL-12 production, TNF-α production was suppressed in all individuals studied. The effect was statistically significant at 100 μM and consistent across seven experiments performed. Although not statistically significant, a similar effect was observed with paraxanthine. Caffeine (100 μM) also increased intracellular cyclic AMP concentrations in LPS-stimulated monocytes isolated from whole blood. Moreover, the effect of caffeine on TNF-α production was abolished by pretreatment with the protein kinase A inhibitor Rp-8-Br-cAMPS (10−4 and 10−5M). To conclude, this study demonstrates that concentrations of caffeine that are relevant to human consumption consistently suppress production of the proinflammatory cytokine TNF-α in human blood and that this effect is mediated by the cyclic AMP/protein kinase A pathway.

EFFECTS OF PERINATAL EXPOSURE TO LOW DOSES OF PCB 153 AND PCB 126 ON LYMPHOCYTE PROLIFERATION AND HEMATOLOGY IN GOAT KIDS

Pregnant does (10 goats/group) were dosed orally with either PCB 153 or PCB 126 dissolved in corn oil or only corn oil (control group) from day 60 of gestation until delivery. Effects on in vitro mitogen-induced lymphocyte proliferation and blood cell counts in their goat kids exposed to low levels of PCB 153 and PCB 126 during gestation and lactation were assessed. The concentrations of PCB 153 and PCB 126 in adipose tissue in the goat kids 9 mo postpartum were 5800 ng/g (fat weight) and 0.49 ng/g (fat weight), respectively. Kids exposed to PCB 153 had a significantly higher number of white blood cells, neutrophils, and lymphocytes at 2 wk of age compared to controls. In the kids exposed to PCB 126 there was a significantly lower concentration of monocytes at 2, 4, and 8 wk of age. The mean lymphocyte response to phytohemagglutinin (PHA) and to concanavalin A (Con A) was significant lower in the PCB 153 compared to the control group at wk 2, 4, and 8 postnatally. The results of the present study support previous reports on immunotoxic effects of PCB exposure in animals. However, this is the first report to demonstrate immunotoxicity in animals by using low doses of PCB 153. The difference in results between PCB 126 and PCB 153 treatment groups may strengthen the hypothesis that PCBs mediate immunotoxic effects through both AhR-dependent and independent mechanisms.

Effect of intravenous endotoxin on blood cell profiles of broilers housed in cages and floor litter environments

Commercial broilers are constantly exposed to airborne microorganisms and endotoxin (lipopolysaccharide, LPS). It has been shown that microbial contamination of the air was higher in broiler houses using floor litter than in broiler houses using netting-type floors. The current study evaluated the effect of housing conditions on blood leukocyte profiles and tested the hypothesis that, when compared to broilers reared in clean stainless steel cages (Cage group), broilers raised on floor litter (Floor group) should experience a higher environmental challenge and have a desensitized immune system that may exhibit better tolerance/resistance to subsequent intravenous LPS challenge. Hematological parameters were evaluated prior to and following i.v. administration of 1 mg/kg BW Salmonella typhimurium LPS (dissolved at 1 mg/0.25 mL in PBS) or i.v. injection of 0.25 mL/kg BW PBS alone. The results showed that prior to LPS/PBS injection, broilers in the cage group had higher heterophil and monocyte concentrations, a higher B cell percentage within the lymphocyte population, and a higher heterophil to lymphocyte (H:L) ratio in the blood. The i.v. LPS injection resulted in 25% mortality in the cage group and 42% mortality in the floor group within 8 h post-injection. LPS reduced the concentrations of total white blood cells (WBC) and all differential WBC except eosinophils and increased thrombocyte concentrations within 1 h post-injection in both groups. All of these values returned to their respective pre-injection levels within 48 h post-injection in the surviving birds. The two groups exhibited similar overall hematological changes after LPS injection except that the cage group showed a higher H:L ratio at 8 h post-injection and a lower B-cell percentage within the lymphocyte population at 48 h post-injection when compared with the floor group. We concluded that the immune systems of broilers reared on floor litter were desensitized and exhibited less pronounced leukocyte responses to i.v. LPS when compared with those of broilers reared in clean stainless steel cages. However, such desensitization of the immune system did not help broilers survive subsequent i.v. LPS challenge.

Critical role of the complement system in group B streptococcus-induced tumor necrosis factor alpha release.

Group B Streptococcus (GBS) is a major cause of newborn sepsis and meningitis and induces systemic release of tumor necrosis factor alpha (TNF-alpha), believed to play a role in morbidity and mortality. While previous studies have shown that GBS can induce TNF-alpha release from monocytes and macrophages, little is known about the potential modulating effect of plasma or serum on GBS-induced TNF-alpha release, and there are conflicting reports as to the host receptors involved. In a human whole-blood assay system, GBS type III COH-1 potently induced substantial monocyte TNF-alpha release in adult peripheral blood and, due to a higher concentration of monocytes, 10-fold-greater TNF-alpha release in newborn cord blood. Remarkably, GBS-induced TNF-alpha release from human monocytes was enhanced approximately 1000-fold by heat-labile serum components. Experiments employing C2-, C3-, or C7-depleted serum demonstrated that C3 activation via the alternative pathway is crucial for potent GBS-induced TNF-alpha release. Accordingly, whole blood from C3-deficient mice demonstrated significantly reduced GBS-induced TNF-alpha release. Preincubation with human serum enhanced the TNF-alpha-inducing activity of GBS in a C3- and factor B-dependent manner, implying deposition of complement components via the alternative pathway. GBS-induced TNF-alpha release was inhibited by monoclonal antibodies directed against each of the components of CR3 and CR4: the common integrin beta subunit CD18 and the alpha subunits CD11b (of CR3) and CD11c (of CR4). Blood derived from CR3 (CD11b/CD18)-deficient mice demonstrated a markedly diminished TNF-alpha response to GBS. We conclude that the ability of plasma and serum to greatly amplify GBS-induced TNF-alpha release reflects the activity of the alternative complement pathway that deposits fragments on GBS and thereby enhances CR3- and CR4-mediated monocyte activation.

Pulmonary and hematological inflammatory responses to intravenous cellulose micro-particles in broilers

When injected intravenously, cellulose micro-particles become lodged in pulmonary arterioles. The current study investigated the systemic and pulmonary inflammatory responses triggered by cellulose micro-particles at 3, 24, and 48 h postinjection in 6-wk-old broilers. Proportions and concentrations of circulating white blood cells were assessed in saline-injected (control group) and cellulose-injected (particle group) birds. Hematoxylin-eosin (HE)-stained cross-sections of the lungs were used to count the number of granuloma/lymphocyte aggregates, which is indicative of the severity of the inflammatory response to the trapped particles. The cellular components of the aggregates were identified by immunohistochemical staining of frozen cross sections of the lungs. Results showed that cellulose micro-particles trapped in the pulmonary vasculature initiated a dynamic, localized inflammatory response within the surrounding lung parenchyma. Monocytes and basophilic granulocytes closely surrounded the particles. CD4, CD8, TCR1, TCR2, and TCR3 subsets of T cells and B cells were present in the outer rim of the granuloma/lymphocyte aggregates. Circulating total white blood cell (WBC, leukocytes) concentrations were similar in both groups at all times postinjection, whereas at 48 h post-injection the percentages of eosinophils and basophils among circulating WBC were higher in the particle group than in the control group (P < or = 0.05). The circulating monocyte concentration also increased within 24 h postinjection (P < or = 0.05). These observations demonstrate that cellulose micro-particles trapped in the pulmonary vasculature initiated acute focal inflammatory responses in the lungs and that the proportions of WBCs in the blood are modulated within 48 h postinjection.

Blood monocyte concentration is critical for enhancement of collateral artery growth.

Arteriogenesis has been associated with the presence of monocytes/macrophages within the collateral vessel wall. We tested the hypothesis that arteriogenesis is functionally linked to the concentration of circulating blood monocytes. Monocyte concentrations in peripheral blood were manipulated by single injections of the antimetabolite 5-fluorouracil (5-FU), resulting in a marked rebound effect in New Zealand White rabbits. Collateral artery growth was assessed by the use of a model of acute femoral artery ligation. Seven days after ligation, collateral conductance and the number of visible collateral arteries were increased in the rebound group. This increase was accompanied by an increased monocyte accumulation as demonstrated by immunohistology in the thigh 3 days after surgery. In a second animal model (129S2/SvHsd mice), 5-FU treatment caused a remarkable decrease in blood monocyte numbers at day 4, followed by a rebound effect at day 12. Foot blood flow, assessed by laser-Doppler imaging before and at various time points after surgery, increased from day 7 through day 21 in mice from the rebound group. In contrast, ligation during the phase of monocyte depletion resulted in a reduction of blood flow reconstitution. This inhibition could be reversed by an injection of isolated monocytes. In conclusion, we have demonstrated a functional link between the monocyte concentration in the peripheral blood and the enhancement of arteriogenesis.

Immunologic Profile of Highly Exposed Yet HIV Type 1-Seronegative Men

The host immune factors that determine susceptibility to HIV-1 infection are poorly understood. We compared multiple immunologic parameters in three groups of HIV-1-seronegative men: 14 highly exposed (HR10), 7 previously reported possibly to have sustained transient infection (PTI), and a control group of 14 low risk blood bank donors (BB). Virus-specific cellular immune assays were performed for CD4(+) T helper cell responses, CD8(+) cytotoxic T lymphocyte activity, CD8(+) cell chemokine release, and CD8(+) cell-derived antiviral soluble factor activity. General immune parameters evaluated included CCR5 genotype and phenotype, interferon alpha production by PBMCs, leukocyte subset analysis, and detailed T lymphocyte phenotyping. Comparisons revealed no detectable group-specific differences in measures of virus-specific immunity. However, the HR10 group differed from the BB group in several general immune parameters, having higher absolute monocyte counts, higher absolute CD8(+) T cell counts and percentages, lower naive and higher terminal effector CD8(+) cells, and lower levels of CD28(+)CD8(+) cells. These changes were not associated with seropositivity for other chronic viral infections. The PTI men appeared to have normal levels of monocytes and slightly elevated levels of CD8(+) T cells (also with increased effector and decreased naive cells). Although we cannot entirely exclude the contribution of other chronic viral infections, these findings suggest that long-lived systemic cellular antiviral immunity as detected by our assays is not a common mechanism for resistance to infection, and that resistance may be multifactorial. General immune parameters reflected by CD8(+) T cell levels and activation, and monocyte concentrations may affect the risk of infection with HIV-1, and/or serve as markers of exposure.

Physiopathological studies in septic rats and the use of fructose 1,6-bisphosphate as cellular protection.

The aim of this research project was to test the ability of fructose 1,6-bisphosphate (FBP), which has anti-inflammatory effects and maintains cellular energy levels, to inhibit the septic process in an experimental model in rats. Prospective, controlled animal trial. Research laboratory. Fed male Wistar rats. Three experimental groups were formed for the test: control group, untreated septic group, and septic group treated with FBP (500 mg/kg). In the control group, there were no deaths; in the untreated septic group, the mortality rate was 100% within 15 hrs; in the septic group treated with FBP, the mortality rate reached 20% within 15 hrs. The blood cell tests revealed that concentrations of hematocrit, leukocytes, monocytes, and immature cells increased significantly in the untreated septic group compared with both the FBP-treated septic group and the control group. The histologic lesions verified in the heart, lungs, liver, and kidneys of septic animals were smaller and even absent in those treated with FBP. FBP reduced the mortality rate provoked by experimental sepsis and ameliorated hematologic and histologic alterations.

Arteriogenesis dependency on peripheral blood monocyte concentration

Arteriogenesis dependency on peripheral blood monocyte concentration

Evaluation of menstrual and haematological parameters, after 36 months of NorplantR contraception

This ongoing prospective longitudinal study involved 23 women who had complete records, of 37 healthy non-breastfeeding informed volunteers recruited from our family planning clinic since August 1997 to the Norplant training programme. Packed cell volume (PCV), white blood cell (WBC) concentration (total and differential and platelet concentration were analysed at pretreatment and at 12, 24 and 36 months' followup. Statistical analysis was with paired t-tests. The level of significance was set at 5%. Each acceptor received a menstrual calendar to document all events of bleeding. The mean PCV was 40·5 ± 2·4% at pretreatment. This rose to a statistically significant (P < 0·001) mean value of 44·9 ± 4·4% at 36 months. The mean WBC concentration at pretreatment was 5552 ± 1423 per mm3 which declined statistically (P < 0·001) to a mean value of 4400 ± 1281 per mm3 at 12 months without any further significant changes at 24 months (P > 0·6; 4143 ± 1301 per mm3 and at 36 months (P > 0·9; 4070 ± 875 per mm3). At 24 months of study the mean concentration of neutrophils, lymphocytes, monocytes eosinophils and basophils did not change significantly from their respective mean concentrations at 12 months. These insignificant changes were also manifested at 36 months of study in the lymphocytes (P > 0·2), eosinophils (P > 0·5) and basophils (P > 0·2), even though there was a significant decrease in the neutrophils (P < 0·05) with a corresponding significant increase (P < 0·001) in the monocyte concentration. However, there was still a significant decrease (P < 0·001) in the WBC at 36 months of study compared with the mean preinsertion value, which was manifested mainly in the neutrophil concentration (1403 ± 517 per mm3 (P < 0·001). The mean value of platelet count (136260 ± 27664 per mm3) at 24 months showed no significant change compared with the 12 months value (126174 ± 37977 per mm3). The value at 36 months (125391 ± 18858 per mm3), however, still showed a significant decline over the pretreatment mean concentration (208043 ± 27250 per mm3). None of the acceptors had thrombocytopenia at 24 and 36 months. At 2 years and 3 years of studies 60·9% and 47·8% of Norplant users reported irregular bleeding patterns, mainly of reduced episodes. The continuation rate at 36 months was 100%. Women using the Norplant implant are at an advantage, because in spite of the variable aberrations in their menstrual pattern, the packed cell volume increased. The changes ob-served in the white cell and the platelet concentrations did not lead to adverse effects.

Antioxidants Prevent the Lymphocyte DNA Damage Induced by PMA-Stimulated Monocytes

Oxidative stress has been related to several degenerative diseases such as cancer and coronary heart disease. Reactive oxygen species can damage different cellular macromolecules, including DNA, which is directly responsible for mutation and carcinogenesis. In this study, monocytes that were activated by phorbol 12-myristate 13-acetate (PMA) were coincubated with lymphocytes, and the DNA damage was measured by single-cell gel electrophoresis (comet) assay. Stimulation of monocytes with PMA activates the "respiratory burst," which evokes DNA damage in lymphocytes. The extent of the damage is related to the concentration of monocytes and the exposure time. Exogenous addition of superoxide dismutase did not prevent the DNA damage, which suggests that superoxide ions are not directly responsible for the damage. Partial protection was observed when catalase was included (60% protection), which indicates that other reactive species, in addition to H2O2, are responsible for the damage. In this system, the protective activity of natural antioxidants at different concentrations was also investigated. After coincubation of PMA-activated monocytes with lymphocytes in the presence of each antioxidant for one hour at 37°C, the lymphocyte DNA damage was determined. All the compounds protected the lymphocytes to a certain degree, with a maximum effect at different concentrations: 41% protection with 1 μM ascorbic acid, 55% protection with 40 μM α-tocopherol, 50% protection with 3 μM β-carotene, and 56% protection with 5 μM quercetin. On the basis of these results, we maintain that this "ex vivo model," more closely related to physiological conditions, could be used to test the antioxidant activity of different compounds.

Application of the IFN-γ ELISPOT assay to quantify T cell responses against proteins

The ELISPOT assay has been established for the direct ex vivo quantification of peptide-reactive T lymphocytes from peripheral blood mononuclear cells (PBMC). In this report we studied the ELISPOT assay conditions for the detection of T cell responses against protein antigens including Tetanus toxoid (TT), purified-protein-derivative (PPD) and a synthetic 30 mer peptide derived from influenza matrix protein (IMP) containing the HLA-A*0201-restricted 9 mer peptide epitope GILGFVFTL as a model for a viral protein. We found several aspects to be crucial for a sensitive detection of T cell responses against proteins including a pellet preincubation step and the monocyte concentration. Using optimized assay conditions specific CD4+ T cell responses against TT and PPD as well as CD8+ T cell responses against IMP can be quantified directly ex vivo from peripheral blood mononuclear cells.

Effect of hydroxyapatite sintering temperature on intracellular ionic concentrations of monocytes: a TEM-cryo-X-ray microanalysis study.

Hydroxyapatite used as bone replacement can lead to particle release in the implantation site. These particles interact with monocytes, which are the first immune cells to colonize the implant and an inflammatory site. Thanks to cryo-X-ray microanalysis, we can observe cells in a state close to the physiological one and we have access to diffusible ions. We paid particular attention to the potassium-to-sodium ratio, which is one of the best viability criteria. We used this method to study the interaction between three hydroxyapatite particles treated at three different temperatures (not treated, treated at 600 degrees C and 1180 degrees C), and monocytes. In the culture condition, the hydroxyapatite treated at 1180 degrees C underwent the least dissolution. We demonstrate that monocytes were altered by the three hydroxyapatite particles. The hydroxyapatite particules treated at 600 degrees C were found to be more toxic.

Effect of hydroxyapatite sintering temperature on intracellular ionic concentrations of monocytes: A TEM‐cryo‐x‐ray microanalysis study

Hydroxyapatite used as bone replacement can lead to particle release in the implantation site. These particles interact with monocytes, which are the first immune cells to colonize the implant and an inflammatory site. Thanks to cryo-X-ray microanalysis, we can observe cells in a state close to the physiological one and we have access to diffusible ions. We paid particular attention to the potassium-to-sodium ratio, which is one of the best viability criteria. We used this method to study the interaction between three hydroxyapatite particles treated at three different temperatures (not treated, treated at 600 °C and 1180 °C), and monocytes. In the culture condition, the hydroxyapatite treated at 1180 °C underwent the least dissolution. We demonstrate that monocytes were altered by the three hydroxyapatite particles. The hydroxyapatite particules treated at 600 °C were found to be more toxic. © 2001 John Wiley & Sons, Inc. J Biomed Mater Res (Appl Biomater) 58: 238–246, 2001

Changes in menstrual and haematological indices among norplant® acceptors

A longitudinal study of women using Norplant® was conducted to determine the effects of levonorgestrel implants on menstrual and haematological indices among the acceptors. Packed cell volume (PCV), white blood cell (WBC) concentrations (total and differential), and platelet concentration were performed at pre-insertion and at the 12-month follow-up. Each acceptor received a menstrual calendar to chart all bleeding, and spotting events. The mean PCV was 40.5 ± 2.4 at insertion and had a statistically significant (p <0.01) rise to a mean value of 42.2 ± 2.6 at 12 months. The mean WBC concentration had a significant decrease at 12 months (p <0.001) when compared with the pre-insertion value. Similarly, the neutrophil and lymphocyte concentration had a statistically significant decrease (p <0.05 and p <0.01, respectively) at 12 months of use. The reduced values in the concentrations of monocytes, eosinophils and basophils did not reach significant levels. The mean value of the platelet concentrations had a significant decline at 12 months of use (p <0.001). Reduced bleeding patterns were more commonly reported compared to increased bleeding episodes. This pattern was associated with increased PCV, a change that may prevent anaemia in developing countries. The decline in the platelet concentration at 12 months of study was, however, a source of concern that will require follow-up.

In vivo administration of GM-CSF promotes the clearance of apoptotic cells: effects on monocytes and polymorphonuclear leukocytes.

The clearance of apoptotic cells is crucial to avoid chronic inflammation and autoimmunity. Little is known about the factors that regulate it in vivo. We show that granulocyte-macrophage colony-stimulating factor (GM-CSF) administration to carcinoma patients confers to their leukocytes a significantly higher ability to phagocytose apoptotic cells than before (P < 0.005). GM-CSF increased the concentration of monocytes and polymorphonuclear leukocytes in the peripheral blood and activated circulating polymorphonuclear leukocytes. Both effects abated early after treatment, whereas phagocytosis of apoptotic cells was still significantly higher after 18 days compared with basal values (P < 0.005 and P < 0.025 for monocytes and polymorphonuclear leukocytes, respectively). On in vitro phagocytosis of apoptotic cells monocytes, but not polymorphonuclear leukocytes, up-regulated MHC class II membrane expression. These findings are consistent with the possibility that GM-CSF endows both scavenger and antigen-presenting leukocytes with the ability to internalize apoptotic tumor cells.

Intracellular hyperinsulinism: a metabolic characteristic of obesity with and without type 2 diabetes: Intracellular insulin in obesity and Type 2 diabetes

There is evidence that intracellular insulin may carry out some insulin mediated actions, including glucose transport. As intracellular insulin has never been quantitatively assessed in human cells, we evaluated its concentrations in monocytes from normal subjects ( n=7) and obese patients without ( n=9) and with Type 2 diabetes mellitus ( n=10). After the incubation of cells with labeled insulin for 60 min at 37°C, intracellular intact insulin concentrations were measured by HPLC and expressed as pmol .10 −6. Insulin concentrations were higher (ANOVA P<0.01) within cells from obese (115.4±26.4 pmol · 10 −6/2 · 10 5 cells) and obese diabetic patients (93.2±36.3 pmol·10 −6/2 · 10 5 cells) compared with normal cells (28.5±13.1 pmol · 10 −6/2 · 10 5 cells). Moreover, after insulin was removed from the incubation medium the decrease of intracellular insulin was significantly lower ( P<0.01) in cells from both obese and obese diabetic patients than in normal subjects. Intracellular undissociated insulin-insulin receptor complexes on average, increased 2-fold ( P<0.01) in cells from insulin resistant patients compared with normal cells. Finally, in downregulated cells from obese and obese diabetic patients, the recycling of the internalized insulin receptor was completely disrupted. In conclusion, monocytes from obese patients with and without Type 2 diabetes mellitus, present increased intracellular insulin concentrations and these conditions are associated with a significant impairment of insulin receptor processing. Increased intracellular insulin concentration in cells from these patients may be necessary in order to overcome insulin resistance.

Distribution of life cycle stages of Sarcoptes scabiei var wombati and effects of severe mange on common wombats in Victoria.

Seven female and three male common wombats (Vombatus ursinus) collected from forested areas of Victoria (Australia) over a 10 mo period, 10 April 1997 to 22 February 1998 had at least 30% of their skin affected by severe hyperkeratotic sarcoptic mange. Mangy wombats were grazing during the day, could be readily approached, were in poor body condition, and lacked subcutaneous fat. The anterolateral surface of the body was most heavily parasitised with Sarcoptes scabiei var wombati followed by the posterolateral surface, the dorsal region between the ears, the ears, ventral abdomen, medial aspect of the legs, axillary and inguinal areas, and the dorsal midline. Larvae were the most prevalent life-cycle stage followed by eggs, nymphs, females, and males. Mite numbers and the severity of clinical signs, namely thickness of scale crust and the degree of alopecia, were correlated and were symmetrical on each side of the body. Fissuring of crust and skin only occurred when scale crust was present. Bacterial infections occurred in three of 10 wombats within lymph nodes or the pleural cavity. Lymphoid depletion did not occur in lymph nodes or spleens and prescapular lymph nodes contained a greater amount of nuclear debris in germinal centres than non-mangy wombats. Seven wombats had fatty change in their livers. Gonads of mature wombats were not active or had minimal activity. Significant histopathological changes were not seen in the gastrointestinal tract, kidney, brain, myocardium, spleen, thyroid, reproductive tract, and gonads. Hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, and concentrations of hemoglobin, lymphocytes, calcium, glucose, creatinine, total solids, total protein, albumin determined both colormetrically and electrophoretically, and globulins were significantly lower and concentrations of neutrophils, monocytes, phosphorus, urea, glutamate dehydrogenase, aspartate aminotransferase and creatine kinase were significantly higher in mangy versus captive wombats. Concentrations of erythrocytes, mean corpuscular hemoglobin, leucocytes, band neutrophils, eosinophils, nucleated erythrocytes, sodium, potassium, chloride, total bilirubin, alkaline phosphatase, and gamma glutamyltransferase for mangy wombats were not significantly different from that reported for captive wombats. Hematological and pathological changes in mangy wombats were consistent with anemia, inflammation, and changes seen with starvation.

SERUM CORTISOL ELEVATIONS AND LEUKOCYTE SUBSET CONCENTRATIONS AFTER ACUTE HEAVY RESISTANCE EXERCISE IN WOMEN

116 Cortisol has been shown to cause neutrophilia, lymphocytopenia, and monocytopenia. However, few studies are available regarding the role of serum cortisol elevation in the immune system of women after resistance exercise. To examine the high (> 1000 nmol L−1) and low (< 500 nmol L−1) concentrations of serum cortisol for the leukocyte subset concentrations after acute heavy resistance exercise, eight high (23.5 ± 3.2 yrs) and eight low (22.1 ± 3.4 yrs) responders were studied from a population of 46 healthy, but non-strength trained women. The resistance test consisted of six sets of 10 RM squats with two minutes rest between sets utilizing a computerized Plyometric Power System. Blood samples were obtained at pre-exercise and immediately post-exercise. Serum cortisol was measured using an 125I solid-phase radioimmunoassay. Leukocyte subset proportions were measured by immunofluorescent staining with subset specific monoclonal antibodies and analyzed using flow cytometry. Results from a two-way ANOVA analysis indicated that the squat exercise significantly (p ≤ 0.05) increased lymphocyte, monocyte, and granulocyte concentrations of both groups. However, only granulocyte concentration of high cortisol group (pre: 4.88; post: 6.95: × 106 ml−1) was significantly greater than one of low cortisol group (pre: 3.69; post: 5.23: × 106 ml−1). Thus, the high cortisol levels may have played a role in the increased granulocyte concentration which could be related to muscle tissue damage and inflammation after acute heavy resistance exercise. Supported by DOD US Army Grant DAMD 17-95-C-5069 to WJK