Differential White Cell Counts Research Articles

What's happening in respiratory infection?

Community acquired pneumonia (CAP) continues to be a common cause of general practitioner presentation and of acute admission to the paediatric medical ward. These children often have admission venepuncture for full blood counts (FBC) with differential white cell counts (WCC), C-reactive protein (CRP) or erythrocyte sedimentation rates (ESR) and blood cultures as well as chest X-ray (CXR) in an attempt to differentiate a viral from bacterial cause. Several recent papers have reported data from the USA and Europe that investigate the aetiology and most useful diagnostic tests in CAP and to compare responses with treatment with different antibiotics. Drummond et al. reported a group of 136 children aged two weeks to 16 years. At least one pathogen was identified as a definite or probable cause of the pneumonia in 70 children (51%). Fifty (37%) had a virus implicated (mostly respiratory syncytial virus, followed by influenza A, cytomegalovirus and adenovirus) and 19 a bacterium (7% group A streptococcus, 4% Streptococcus pneumoniae). Admission blood tests and CXR did not assist in differentiating viral from bacterial pneumonia but viral immunofluorescence and paired serology were the most useful diagnostic tests. Wubbel et al. reported on 168 children with radiographic and clinical evidence of pneumonia aged 6 months to 16 years from Dallas, Texas. All blood cultures were negative and infecting organisms were identified by acute and convalescent serology, nasopharyngeal and pharyngeal swabs for culture and polymerase chain reaction (PCR) for Chlamydia pneumoniae and Mycoplasma

Autologous transfusion in surgical patients at Kenyatta National Hospital, Nairobi.

To identify autotransfusion strategies and their basis in elective surgery patients. A cross sectional prospective study. General surgery and orthopaedic wards, Kenyatta National Hospital, Nairobi. Adult patients of both sexes planned for elective surgery. Forevery patient, the following were enquired about and documented: age in years, sex, ethnicity, religion, occupation and educational standard. Blood values of haemoglobin, platelet counts, total and differential white cell counts, urea, electrolytes and liver function tests were assayed. Others were the number of units of blood donated before the operation, the type of surgery performed, time taken from diagnosis to performing the operation and whether the blood was transfused preoperatively, intraoperatively and postoperatively. A total of sixty three cases constituting five per cent of all surgical patients admitted during the period of study were evaluated. Of these 53 (84%) were males and ten (16%) females. The age range was 15 to 65 years with a peak at 45-49 years. There were more Christians (90%) than Muslims (10%). In all, 32 (51.6%) had primary school education, 23 (36.5%) secondary school education, seven (11.3%) no formal education and one (1.6%) had attained college level. Employment pattern showed 50% were civil servants, 30% were self employed and 20% were unemployed. The duration of disease ranged from 1-24 weeks with two peaks at two weeks and six weeks. Orthopaedic cases constituted 78.7% and general surgery 21.3%. Preoperative haemoglobin ranged from 13.5-14.2 g/dl. Transfusions were given intraoperatively to 41 (66.1%) and to 12 (33.9%) postoperatively. Mean duration of hospitalisation was 13 days (range 5 to 21 days). 98.4% deposited only one unit while 1.6% deposited four units of blood. Only one patient required additional transfusion from homologous donors. The strategies and basis for autotransfusion have been demonstrated among a majority of adult patients requiring orthopaedic procedures. Major determinants are shown to be baseline blood count profiles and time to operation.

Therapeutic drug monitoring of azathioprine and 6-mercaptopurine metabolites in Crohn disease.

6-Mercaptopurine (6-MP) and its prodrug azathioprine (AZA) have proven efficacy in the treatment of Crohn disease (CD). The immunosuppressive properties of AZA/6-MP are mediated by the intracellular metabolism of 6-MP into its active metabolites, 6-thioguanine nucleotides (6TGN) and 6methylmercaptopurine (6-MMP). Preliminary studies have suggested that the red blood cell concentration of 6TGN (RBC 6TGN) is a potential guide to therapy. The aims of the study were to evaluate the RBC 6TGN concentrations in adult patients with CD under long-term AZA/6-MP therapy and to correlate it with response to treatment and haematological parameters. Twenty-eight CD patients treated for at least 3 months with AZA/6-MP were prospectively studied. Patients were separated into three main groups: group 1 (n = 19), corresponding to quiescent CD receiving AZA (dose: 2.05 +/- 0.4 mg/kg/day for a mean of 28.6 +/- 25 months) or 6-MP (dose: 1.4 +/- 01 mg/kg/day for a mean of 7.5 +/- 3.5 months) alone; group 2 (n = 6), corresponding to quiescent CD treated by AZA (dose: 2.14 +/- 0.5 mg/kg/day for a mean of 29.5 +/- 22 months) with oral steroids; and group 3 (n = 3), corresponding to active CD on AZA (dose: 1.94 +/- 0.6 mg/kg/day for a mean of 31.3 +/- 35 months) as the only treatment. An assessment was also made by merging groups 1 and 2 forming a larger group of patients (n = 25) defined by clinical remission and groups 2 and 3 forming a larger group of patients (n = 9), non-complete responders with AZA/6-MP alone. Crohn disease index activity (CDAI), blood samples for full blood count and differential white cell count and measurement of RBC 6TGN and 6-MMP concentrations were evaluated at inclusion and at 6 months (n = 17). RBC 6TGN were measured using high performance liquid chromatography (HPLC) on heparinized blood. The baseline characteristics of the three groups of patients were similar. There was no significant difference among the three groups of patients regarding the dose and the duration of immunosuppressive treatment. There was no significant difference between groups according to various parameters tested. Particularly, the median RBC 6TGN concentration at inclusion was similar in the three groups of patients (166 (105-688), 183 (90-261) and 160 (52-194) pmol/8 x 10(8) RBC, respectively). The majority of patients had no detectable level of 6-MMP metabolite, except for 3 patients. There was also no difference between merging groups. Furthermore, there was no significant correlation between RBC 6TGN concentrations and the various biological parameters tested except for the mean erythrocyte volume. At 6 months, all patients of group 1 remained in remission and median RBC 6TGN concentration remained stable. No side effects were observed. There is, contrary to preliminary studies, a broad overlap in RBC 6TGN levels as well as for haematological parameters in patients in remission or not and responders or not to AZA/6-MP therapy. This suggests, beside a variability in the metabolism of these drugs, the existence of complex mechanisms of action. Nevertheless, beside the use of RBC 6TGN determination to confirm compliance to therapy, this dosage could be useful in non-responding patients, allowing, in absence of leukopenia, to increase the dose of AZA/6-MP safely.

Occurrence of Anguillicola crassus in the Water Reservoir Koryčany (Czech Republic) and its Influence on the Health Condition and Haematological Indices of Eels

Palikova M., S. Navratil: Occurrence of Anguillicola crassus in the Water Reservoir Koryaany (Czech Republic) and its Influence on the Health Condition and Haematological Indices of Eels. Acta Vet. Brno 2001, 70: 443-449. The occurrence of the nematode Anguillicola crassus in eels in the Water Reservoir Koryaany was studied from 1994 to 2000. In 1998 and 1999 the study was performed at more frequent intervals in order to evaluate possible seasonal changes of the disease and the influence of the parasite on the health condition of eels in this water reservoir. In all, 143 eels were sampled. Haematological examinations were performed using 46 specimens in 1999. Prevalence of nematodes showed seasonal dynamics and varied from 43 to 100%. Intensity of infection by young and adult nematodes in the swim bladder of eels varied from 1 to 66 specimens, with the mean being 6.6 nematodes per individual. Prevalence of larval stages in the swim bladder amounted from 38.5 to 100%. There were patho-anatomical changes in the swim bladder as well as in the ductus pneumaticus found. Severe damage to the swim bladder was observed especially during autumn months. No significant differences were found between infected and non-infected eels, or in dependence on the numbers of nematodes in the swim bladder with regards to the haematological parameters examined (i.e., hematocrit, red and white blood cell counts). The obtained values are also mentioned in relation to the date of sampling. There was a rise (p < 0.05 and p < 0.01) in the hematocrit value sampled on October 10, 1999 (0.29 L.L-1) in comparison to sampling on June 7, 1999 (0.25 L.L -1 ), September 16, 1999 (0.25 L.L -1 ) and June 30, 1999 (0.27 L.L -1 ), respectively. Red blood cells were significantly (p < 0.05 and p < 0.01) lowest on September 16, 1999 (1.04 T.L -1 ) as compared to sampling on June 7, 1999 (1.28 T.L -1 ), October 6, 1999 (1.24 T.L -1 ) and June 30, 1999 (1.41 T.L -1 ), respectively. White blood cell counts were lower (p < 0.05 and p < 0.01) on October 6, 1999 (27.6 G.L-1) as compared to values obtained on June 7, 1999 (56.2 G.L -1 ), and September 16, 1999 (41.2 G.L -1 ) and June 30, 1999 (39.0 G.L-1), respectively. Differential counts of white blood cells showed no significant changes. Infected as well as non-infected eels were characterised by lymphocytic type of blood (88.02 and 90.34 % of lymphocytes, 10.27 and 7.38 % of granulocytes, 0.52 and 0.94 % of monocytes, 1.17 and 1.34 % of blasts, respectively). Anguillicolosis under conditions of the Koryaany Water Reservoir did not cause mortality and clinical signs of the disease in the population of eels. Prevalence, intensity of infection, hematocrit, erythrocytes, leukocytes, differential leukocyte count

Automated Analysis of Bone Marrow Fluid with the Sysmex XE-2100TM Blood Cell Counter

Background: Until recently, automated analysis of the cellular composition of bone marrow has not been possible, owing to problems in erythroblast identification, contamination by fat particles, and the heterogeneity of cell types. The XE-2100 automated blood cell counter can effectively count erythroblasts and granulocytes using laser light flow cytometry and DNA fluorochromes. In this study we have evaluated the performance of the XE-2100 for analyzing bone marrow cells. Material and Methods: The XE-2100 hematology analyzer performs white cell differential counts by measuring radiofrequency, direct current resistance, forward and side scatter of laser light, and fluorescence. Erythroblasts are identified by means of their specific fluorescent labelling. We have assessed i) the imprecision of the XE-2100 nucleated red blood cell (NRBC) counting using linear regression and the National Committee for Clinical Laboratory Standards (NCCLS) document H20A method of differences between duplicates, and ii) the comparability of NRBC count with microscopy counts using linear regression and analysis of differences. Results: Reproducibility of the XE-2100 measurements was excellent with coefficients of variation below 15%. We compared bone marrow granulocyte and erythroblast proportions by the XE-2100 with microscopy counts using linear regression and analysis of differences on 207 samples: Correlation between XE-2100 and the microscope was excellent for granulocyte percentage but slightly lower for erythroblasts. There was a tendency for the XE-2100 to slightly underestimate erythroblast percentage in most samples with a mean difference (XE-2100 – microscope) of −13.4%.

Secondary myelodysplastic syndrome after fludarabine therapy of a low-grade non-Hodgkin's lymphoma

We diagnosed a probable fludarabine-induced secondary MDS ∼18 months after treatment of a low grade non-Hodgkin's lymphoma. After diagnosis of a B-cell lymphoma composed of relatively small cells, fludarabine was administered between May and October, 1997, to a 64-year-old female patient. In December 1998, a mild bicytopenia was present with a leukocyte count of 3800/μl and platelets of 142 000/μl. The white cell differential count was normal. The hemoglobin level was normal, but MCV was elevated. Bone marrow cytology revealed normal cellularity with dyserythropoesis and dysmegakaryopoesis. PAS staining showed scattered positivity in early erythroid cells. In 12 of 20 mitoses, the karyotype showed complex rearrangements, described as 46,XX,t(4;11)(q23?24;q13),del(5q),del(7)(q22),+mar[8]/45,−3. A diagnosis of treatment-related MDS was made. While there was no evidence of bone marrow infiltration by the lymphoma, CT scans demonstrated paraaortic lymph nodes up to 10 cm in diameter. After one course of CHOP chemotherapy, prolonged bone marrow aplasia and septic complications occurred. Chemotherapy was abandoned and Rituximab was administered. In July and November, 1999, bilateral pneumonia and urinary tract infection, respectively, were treated with antibiotics. NHL was in complete remission, but peripheral blood counts deteriorated markedly, and transfusions of packed red cells had to be started in November, 1999. The suspicion of leukemic transformation could not be confirmed because the patient declined further bone marrow biopsies. In December, 1999, the patient died from pneumonia.

Antenatal endotoxin and glucocorticoid effects on lung morphometry in preterm lambs.

In utero inflammation may accelerate fetal lung maturation but may also play a role in the pathogenesis of chronic lung disease. We examined the impact of endotoxin, a potent proinflammatory stimulus, on structural and functional maturation of preterm sheep lungs. Date bred ewes received 20 mg Escherichia coli endotoxin or saline by ultrasound guided intra-amniotic injection at 119 d gestation. A comparison group of animals received 0.5 mg/kg betamethasone, a known maturational agent, at 118 d gestation. Lambs were delivered by cesarean section at 125 d (term = 150 d) and ventilated for 40 min. Lung function data are reported elsewhere. Total and differential white cell counts were performed on amniotic fluid and fetal lung fluid samples. Morphometric analyses were performed on inflation fixed right upper lobes. Total cell count increased slightly but not significantly in both amniotic fluid and fetal lung fluid. Both endotoxin and betamethasone had similar effects on alveolarization: average alveolar volume increased by approximately 20% and total alveolar number decreased by almost 30%. Both treatments led to thinning of alveolar walls, although this was statistically significant in the betamethasone-treated group only. Although antenatal endotoxin leads to striking improvements in postnatal lung function, this may be at the expense of normal alveolar development.

General versus regional anaesthesia for cataract surgery: effects on neutrophil apoptosis and the postoperative pro-inflammatory state

At clinically relevant concentrations, volatile anaesthetic agents influence neutrophil function. Our hypothesis was that sevoflurane would inhibit neutrophil apoptosis and consequently influence the postoperative pro-inflammatory state. In order to identify selectively the effect of the anaesthetic agent sevoflurane, we studied patients undergoing minimally stimulating (cataract) surgery randomly allocated to receive either sevoflurane (n = 11) or local anaesthesia (n = 12). Venous blood samples were taken immediately prior to anaesthesia and at 1, 8 and 24 h thereafter. The rate of neutrophil apoptosis, plasma concentration of cytokines and differential white cell count were measured. The rates of neutrophil apoptosis and plasma concentrations of IL-1β, TNF-α and IL-8 at each time point were similar in the two groups. IL-6 concentrations increased significantly and to a similar extent compared to preanaesthetic levels at 8 and 24 h. This study demonstrates that sevoflurane does not influence the rate of neutrophil apoptosis, cytokine concentrations and neutrophil count following cataract surgery.

Immunologic and epidemiologic characterization of non-responders/low-responders to hepatitis B vaccine

An important reduction in morbidity and mortality due to hepatitis B was achieved with the discovery of an effective vaccine. However, 2 to 10% of healthy adults do not respond to vaccination with the production of protective levels of antibody anti-HBs (assumed as protective, concentrations of ab. anti-HBs > 10 UI/L). Therefore, the aim of the present work was to study epidemiological (sex, age, obesity, alcoholic and smoking habits, previous diseases) and immunological factors (white cell count, immunoglobulins and sub-classes of IgG, lymphocytic populations and sub-populations) in a group of 20 healthcare workers with a low response (anti-HBs < 50 UI/L) to a genetically-engineered vaccine (Engerix B). The results were compared to the ones found in an identical sample of responders (anti-HBs > 100 UI/L) from the same population. No statistically significant differences regarding the epidemiological data, differential white cell counts and immunoglobulin quantification (IgG, IgA, IgM, IgG1, IgG2, IgG3) were detected. The mean value of IgG4 (mean = 54.53 +/- 59.8 mg/dl) in non/hyporesponders was significantly higher (p = 0.038) when compared to the same result in responders (mean = 33.76 +/- 31.30 mg/dl). A statistically significant difference (p < 0.05) in the quantification of double negative lymphocytic T sub-populations was also found, the mean value being higher in the responders (mean = 6.5 +/- 4.1% versus 4.6 +/- 2.3%). The effectiveness of recombinant hepatitis B vaccines is well known. However, a number of apparently healthy people are unable to achieve protective titres of anti-HBs after vaccination. As our study groups did not have a considerable number of cases, we can not establish definitive conclusions based on differences found in the quantification of IgG4 and double negative lymphocytic T sub-populations. It seems to us that an investment in future research into the eventual causes of nonprotective response and into new strategies of immunization of non/hyporesponders are appropriate. In the meantime, the occupational daily risk of exposure to infected body fluids makes adequate hepatitis B immunization of healthcare workers a priority.

Pretreatment with tumor necrosis factor-alpha attenuates arterial hypotension and mortality induced by endotoxin in pigs.

Tumor necrosis factor (TNF)-alpha administration in large amounts can induce a state of shock similar to that seen during severe sepsis. The objective of this study was to determine whether small doses of TNF-alpha might decrease the disposition for the development of shock induced by a subsequent infusion of endotoxin and to determine whether the mechanism of this protective effect of TNF-alpha pretreatment could be associated with up-regulation of the anti-inflammatory cytokine interleukin (IL)-10. Prospective, randomized, experimental study. Investigative intensive care unit at an academic medical center. A total of 14 female Yorkshire pigs, weighing 20-25 kg. We studied two groups of animals-pigs treated with 500 ng/kg recombinant porcine TNF-alpha (n = 7) and pigs given diluent alone (n = 7). At 24 hrs after treatment, both groups of pigs were subjected to a 24-hr continuous infusion of lipopolysaccharide (LPS) at a rate of 80 ng/kg/min. The mortality rate was determined in both groups. Hemodynamic indices, oxygen transport variables, total and differential white cell counts, and serum concentrations of TNF and IL-10 were determined at frequent intervals before and after TNF-alpha administration and during the LPS infusion. Additionally, peripheral blood mononuclear cells were collected for determination of messenger ribonucleic acid expression of IL-10 by reverse transcription-polymerase chain reaction. The administration of TNF-alpha at the dose used in this study did not have any profound effect. No pig treated with TNF-alpha died in response to the LPS infusion. In contrast, three of seven control pigs died during the LPS infusion. Lipopolysaccharide-induced arterial hypotension and arterial hypoxemia were attenuated in the TNF-alpha-treated group. Both groups had significant increases in serum concentrations of TNF-alpha in response to LPS, with no significant difference in peak serum TNF-alpha between groups. Neither serum concentrations of IL-10 nor expression of IL-10 messenger ribonucleic acid in circulating mononuclear cells differed between groups. The administration of TNF-alpha attenuated the severity of hyperdynamic shock induced by a subsequent infusion of endotoxin. This effect could not be associated with increased expression or elaboration of the anti-inflammatory cytokine IL-10.

Normal hematologic and serum biochemical reference intervals for juvenile wild turkeys.

Blood samples taken from 48 4-mo-old wild turkeys (Meleagris gallopova silvestris) were used to establish reference intervals for hematology and serum chemistry values. The study was conducted during September and October 1996. Packed cell volume, total and differential white cell counts, total protein, albumin, glucose, calcium, uric acid, triglyceride concentrations, as well as aspartate transaminase (AST) and lactate dehydrogenase (LDH) activities were assayed. Reference intervals from wild turkeys are similar to those reported for domestic turkeys.

Les pleurésies conduites diagnostique et thérapeutique

Diagnosis of pleural effusion is difficult in children. The etiologies are numerous; however, infectious agents are more frequent. Thoracocentesis proves to be the first-line diagnostic tool. Light's criteria are the best for distinguishing whether the effusion is a transudate or an exudate. If the patient has an exudative pleural effusion, other tests are indicated to determine the etiology and in some cases the treatment: macroscopic appearance, cytology and differential white cell count (level of glucose, lactate dehydrogenase, adenosine deaminase, pH, bacterial cultures). Others investigations--biopsy of pleura by thoracoscopy or video-assisted thoracoscopy, bronchofibroscopy, CT scan--are sometimes useful. Intrapleural instillation of urokinase appears to be useful and safe. Evaluation is necessary for video-assisted thoracoscopy used early.

Churg-Strauss syndrome.

A syndrome comprising asthma, allergic rhinitis, pulmonary and systemic small-vessel vasculitis, and extravascular granulomas was described in 1951 by Churg & Strauss, mainly from autopsied cases (1). Clinical studies of patients with polyarteritis nodosa, associated asthma, pulmonary in®ltrates, and eosinophilia had been reported previously (2±5). Churg & Strauss described the pathologic lesions and distinguished this entity from polyarteritis nodosa. They described basic anatomic changes consisting of ``widespread vascular lesions of the type seen in polyarteritis nodosa, and of characteristic tissue alteration in the vessel wall and in the extravascular system. This tissue alteration . . . comprised necrosis of eosinophilic exudate, severe `®brinoid' collagen change, and granulomatous proliferation of epithelioid and giant cells.'' Churg & Strauss considered it to be a distinct histopathologic entity which they termed ``allergic granuloma''.

Inflammatory cytokines and their receptors in arterial and mixed venous blood before, during and after infusion of drained untreated blood.

Wound blood for postoperative autologous transfusion is drained through an area of damaged tissue, the surgical wound, and contains inflammatory mediators. The inflammatory cytokines interleukin-1-beta (IL-1beta), interleukin-6 (IL-6), interleukin-8 (IL-8) and tumour necrosis factor-alpha (TNF-alpha) and their modulators interleukin-1-receptor antagonist (IL-1Ra), interleukin 6 soluble receptor (IL-6sR), soluble tumour necrosis factor receptor 1 (sTNF-R1) and interleukin 10 (IL-10), together with white cell count (WCC) and white cell differential count were measured in arterial and mixed venous blood before, during and after infusion of postoperatively drained untreated blood in nine patients operated for thoracic scoliosis. We found a transient increase in IL-6, an increase in TNF-RI, an increase in IL-8 with granulocytosis and a decrease in IL-10 in the systemic circulation. The increase in IL-6 was higher in mixed venous than in arterial blood.

Hormonal and metabolic responses of juvenile alligators to cold shock

Juvenile alligators became completely immobile 5 min after immersion in ice water and remained in rigor for 40 min when removed from the water, but recovered righting responses within 5 min after immersion in tepid water. A blood sample was taken prior to the treatment, at 1 hr post-treatment and at 24 and 48 hr after recovery. Plasma norepinephrine, epinephrine, and dopamine were measured using high-pressure liquid chromatography (HPLC), and corticosterone by radioimmunoassay (RIA). Plasma ions, phosphate, and lipids were measured on an autoanalyzer and blood smears were taken for differential white cell counts. Norepinephrine and epinephrine were close to 4 ng/ml at the initial bleed: at 1 hr post-treatment epinephrine increased to 7 ng/ml and norepinephrine rose to over 40 ng/ml. Mean plasma dopamine was less than 0.7 ng/ml at the initial bleed and post-treatment means were as high as 10 ng/ml, but values were too variable to show statistical significance. Plasma corticosterone rose significantly at 1 hr and returned to levels not significantly different from initial at 24 and 48 hr. Despite the massive increase in catecholamines, plasma glucose did not change throughout the experiment. Plasma triglyceride increased significantly at 24 and 48 hr and plasma cholesterol decreased significantly at 24 and 48 hr. All other plasma components with the exception of calcium and sodium showed changes. Both lymphocytes and heterophils increased at 48 hr and other white cell types showed a decrease. Overall, these results suggest that short-term cold exposure is less stressful to alligators than simple restraint. J. Exp. Zool. 283:566–572, 1999. © 1999 Wiley-Liss, Inc.

Hormonal and metabolic responses of juvenile alligators to cold shock

Juvenile alligators became completely immobile 5 min after immersion in ice wa- ter and remained in rigor for 40 min when removed from the water, but recovered righting re- sponses within 5 min after immersion in tepid water. A blood sample was taken prior to the treatment, at 1 hr post-treatment and at 24 and 48 hr after recovery. Plasma norepinephrine, epinephrine, and dopamine were measured using high-pressure liquid chromatography (HPLC), and corticosterone by radioimmunoassay (RIA). Plasma ions, phosphate, and lipids were measured on an autoanalyzer and blood smears were taken for differential white cell counts. Norepinephrine and epinephrine were close to 4 ng/ml at the initial bleed: at 1 hr post-treatment epinephrine increased to 7 ng/ml and norepinephrine rose to over 40 ng/ml. Mean plasma dopamine was less than 0.7 ng/ml at the initial bleed and post-treatment means were as high as 10 ng/ml, but values were too variable to show statistical significance. Plasma corticosterone rose significantly at 1 hr and returned to levels not significantly different from initial at 24 and 48 hr. Despite the massive increase in catecholamines, plasma glucose did not change throughout the experiment. Plasma triglyceride increased signifi- cantly at 24 and 48 hr and plasma cholesterol decreased significantly at 24 and 48 hr. All other plasma components with the exception of calcium and sodium showed changes. Both lymphocytes and heterophils increased at 48 hr and other white cell types showed a decrease. Overall, these results suggest that short-term cold exposure is less stressful to alligators than simple restraint. J. Exp. Zool. 283:566n572, 1999. © 1999 Wiley-Liss, Inc.

Compound deletion of the rhoGAP C1 and V2 vasopressin receptor genes in a patient with nephrogenic diabetes insipidus

The function of small GTPases is fine-tuned by a complex network of regulatory proteins such as GTPase-activating proteins. The C1 gene at Xq28 encodes a protein assumed to function as a Rho GTPase-activating protein (rhoGAP). Characterization of the molecular defect causing X-linked nephrogenic diabetes insipidus (NDI) in a patient revealed a submicroscopic deletion of a 21.5-kb genomic fragment encompassing the entire arginine-vasopressin V2 receptor gene (AVPR2) and most of the C1 gene locus. In the absence of detailed information about the physiological relevance and specific functions of rhoGAP C1, a thorough clinical and laboratory investigation of the patient was performed. Besides clearly defined NDI symptoms caused by deletion of the AVPR2 gene, no major morphological abnormalities as determined by physical examination, radiography, ultrasound, and computed tomographic scan were detected. Extensive analysis of blood chemical, enzyme, and hormone values over a period of 16 years showed no deviations from normal ranges. On the basis of our observations, the rhoGAP C1 protein is not essential for normal development in the human. Because of a predominant expression pattern of the C1 gene in hematopoietic cells, we focused on immunologic and hematologic laboratory parameters of the affected boy and the mother who was found to be heterozygous. Differential white cell counts, including lymphocyte typing, determination of lymphokines, cytokines, and immunoglobulins, as well as numerous leukocyte function tests, showed no pathological findings. Therefore, we postulate that the loss of rhoGAP C1 function is most likely compensated by other members of the GAP family. Hum Mutat 14:163–174, 1999. © 1999 Wiley-Liss, Inc.

Interrelationship of the kinin system, nitric oxide and eicosanoids in the antigen-induced arthritis in rabbits.

The aim of the present study was to investigate the interrelationship of the kinin system, nitric oxide and eicosanoids in the acute phase of antigen-induced arthritis (AIA) in rabbits. The arthritis was induced in immunized rabbits and the following parameters were evaluated 24 hours later: leukocyte influx (total and differential white cell count), vascular permeability (Evans's blue method), and synovial PMN cell infiltrate. PGE2 and LTB4 (radioimmunoassay) levels were quantified in the synovial fluid. The animals were pre-treated with 20mg/kg/day during 14 days with L-NAME or D-NAME and/or Enalapril (0.12 mg/kg/day-14 days), and/or the B2 antagonist of Bradykinin HOE 140 (0.9 mg/kg). Our results showed that L-NAME was effective in the prevention of AIA with reduction of all Inflammatory parameters analyzed. Enalapril partially reverted the L-NAME anti-inflammatory effects. The simultaneous treatment with HOE 140 abolished this reversion and returned the inflammatory parameters to the levels observed in L-NAME treated animals. Our results suggest that pressoric alterations induced by L-NAME could not account for all its anti-inflammatory action in this model of experimental arthritis. Additionally the contribution of the kinin system in AIA was characterized as well as its interaction with eicosanoids and nitric oxide.

A congenital persistent infection of bovine virus diarrhoea virus in pigs: Clinical, virological and immunological observations

We report on a lifelong ‘carrier’ state of non‐cytopathic bovine virus diarrhoea virus (BVDV) in an otherwise healthy pig. Three out of 13 pigs of a litter congenitally infected with BVDV survived for more than 3 months. One pig was BVDV seropositive at 1 month, the second seroconverted between 6 and 8 months, and the third remained viraemic and BVDV‐immunotolerant until slaughter at 26 months. The latter pig, a boar, excreted virus in oropharyngeal fluid, urine and semen. Ejaculates, however, did not contain spermatozoa. The persistently infected pig had leucopenia from 3 months onwards, though differential white cell counts and thrombocyte counts remained normal. The antibody response of this pig after vaccination against foot‐and‐mouth disease and after infection with porcine parvovirus was not affected. The antibody response after vaccination against hog cholera, however, was delayed. In contrast, superinfection with antigenically related cytopathic BVDV strains did not alter the status of BVDV immunotolerance, nor did it induce clinical signs resembling mucosal disease as observed in cattle persistently infected with BVDV. Although the role of pigs in the epizootiology of bovine virus diarrhoea infections may be limited, the existence of healthy BVDV carrier pigs should be noted wherever control or eradication measures are applied.

Peripheral blood lymphocyte subsets in fleece rot-resistant and -susceptible sheep.

To compare haematological values and lymphocyte phenotypes in the peripheral blood of fleece rot-resistant and -susceptible sheep. Experiments were conducted on 2- and 3-year-old Merino rams, flock 1 (17 rams) and flock 2 (32 rams), respectively. Within each flock, individual rams were classified as fleece rot-resistant or -susceptible, based on established criteria. Total and differential white cell counts, and indirect fluorescent antibody tests specific for B cells and T cells were performed on all sheep. The concentration of various subsets of circulating lymphocytes was then determined in each sheep. There were no significant differences between fleece rot-resistant and -susceptible sheep from either flock in the mean total or differential white cell counts. However, fleece rot-resistant rams in flock 1 did have a significantly higher concentration of circulating SBU-T1+ cells than fleece rot-susceptible rams from the same flock. No such difference was noted in the rams from flock 2. While all rams in flock 1 were free of clinical fleece rot, 24 rams in flock 2 (comprising all 17 fleece rot-susceptible and 7 of 15 fleece rot-resistant animals) had clinical signs of the disease. Fleece rot-free rams in this flock (irrespective of their classification as fleece rot-resistant or -susceptible) had significantly higher concentrations of circulating SBU-T1+ cells compared with fleece rot-affected animals. They also had significantly higher concentrations of circulating B cells, and total lymphocytes. An examination of peripheral blood lymphocyte subsets in fleece rot-resistant and -susceptible sheep revealed a possible association between resistance to fleece rot and the concentration of circulating SBU-T1+ cells.