Effects of bradykinin and some of its fragments on smooth muscles and chemotaxis

Bradykinin (BK) and two of its C-terminal fragments, namely H-Phe-Ser-Pro-Phe-Arg-OH and H-Ser-Pro-Phe-Arg-OH were found to be potent inhibitors of the chemotaxis of rat polymorphonuclear neutrophils (PMN). The activity of the three peptides was significantly enhanced by SQ 14225, a rather specific inhibitor of kininase II. A shorter C-terminal sequence of BK (Phe-Arg-OH) was inactive. The whole peptide (BK) exerted potent actions on blood pressure and on isolated organs, e.g. the rabbit mesenteric vein or the guinea pig ileum, but none of its fragments showed similar effects. The present findings suggest that rat PMN possess membrane receptors which cannot be considered identical to B1- and B2-receptors, previously characterized on isolated smooth muscles.     

Retention in Glass Bead Columns as a Measure of Leucocyte Adhesiveness

Erythrocyte sedimentation significantly influenced test results when leucocyte retention in glass bead columns was used as a measure of leucocyte adhesiveness. In blood with elevated ESR, this influence was especially evident. Erythrocyte sedimentation highly increased leucocyte retention in vertical columns with a downward flow, whereas in slightly tilted columns with an upward flow, the retention was reduced. The error in measurements caused by erythrocyte sedimentation could be corrected by means of the haematocrit values in samples and eluates. Correction was better for results obtained with slightly elevated than with vertical columns. The granulocyte retention rates increased with increasing sample size, whereas lymphocyte retention remained constant.     

Granulocyte Release of Vitamin B12-Binders in vivo and in vitro in Leukaemia and Non-Neoplastic Leucocytosis

The unsaturated vitamin B₁₂-binding capacities of the ‘large molecular size vitamin B₁₂-binding protein’ (LBP) and the ‘small molecular size vitamin B₁₂-binding protein’ (SBP) were determined by a Sephadex G 150 gel filtration method in 9 patients with chronic myelocytic leukaemia (CML), 5 patients with blast cell leukaemia and 12 patients with non-neoplastic leucocytosis. EDTA plasma and serum separated after 20 min and after 120 min were examined. In the 20 min EDTA plasma samples, the mean LBP value was 8,009 pg/ml in CML, 2,468 in blast leukaemia, 175 in non-neoplastic leucocytosis, and 57 in normal controls. The in vitro release of LBP into serum was much smaller in the leukaemias than in non-neoplastic leucocytosis. No correlation was found between the LBP values and the white blood cell counts or lysozyme values, but lysozyme was correlated to white cell count in CML. It is suggested that the plasma LBP levels reflect the fraction of LBP decay taking place at sites, e.g. the spleen, from which the released LBP can enter the circulation.     

Subcellular localization and properties of adenosine triphosphatase in human polymorphonuclear leucocytes

Abstract. Magnesium-dependent adenosine triphosphatase (Mg²⁺-ATPase) activities were studied in human neutrophilic polymorphonuclear leucocytes. Kinetic studies on whole leucocyte homogenates produced curvilinear kinetics suggesting the presence of at least two forms of Mg²⁺-ATPase.
Neutrophils were homogenized in isotonic sucrose and, after low-speed centrifugation, the supernatant was subjected to analytical subcellular fractionation. Gradient fractions were assayed for Mg²⁺-ATPase and for principal organelle marker enzymes. Mg²⁺-ATPase was distributed between the plasma membrane, mitochondrial and cytosol fractions. Kinetic and inhibitor studies on Mg²⁺-ATPase from each localization indicated the presence of three forms of the enzyme. The plasma membrane and mitochondrial activities had a K _m value of 0.2 mmol/1 for ATP, whilst the K _m for the cytosolic enzyme was 1 -8 mmol/I. Inhibitor studies showed further differences between the three enzymes.
Neutrophils were isolated from control subjects, patients with chronic granulocytic leukaemia and patients in the third trimester of pregnancy. The specific activities (mUnits/mg protein) of Mg²⁺-ATPase, in contrast to those of alkaline phosphatase, were similar in all three patient groups. This result, together with the fractionation experiments and inhibitor studies, strongly suggest that the ATPase is not attributable to neutrophil alkaline phosphatase.     

Effect of Mimosa pudica root powder on oestrous cycle and ovulation in cycling female albino rat, Rattus norvegicus

Mimosa pudica root powder (150 mg/kg body weight) when administered intragastrically, altered the oestrous cycle pattern in female Rattus norvegicus. Nucleated and cornified cells were absent in all rats. The smear was characterized by leucocytes only, as in dioestrus, which persisted for 2 weeks. There was a significant reduction in the number of normal ova in rats treated with the root powder compared with the control rats, and a significant increase in the number of degenerated ova.     

解脲支原体感染与精液白细胞及精子密度关系探讨

目的探讨解脲支原体,精液白细胞与精子密度的关系。方法对275例男性不育患者精液样本进行分析,解脲支原体采用培养法进行检测,白细胞采用联苯胺染色方法进行检测,精子密度检测采用计算机辅助系统进行分析。结果UU阳性率为31．64％,白细胞阳性率为22．55％。UU阳性组精子密度显著低于UU阴性组（P〈0．05）,白细胞异常组精子密度显著低于白细胞正常组（P〈0．05）,UU阳性组白细胞异常例数与UU阴性组相比,有显著性差异（P〈0．05）,UU阳性自细胞异常组精子密度显著低于其对应的白细胞正常组及UU阴性组（P〈0．001）,而UU阴性白细胞异常组精子密度与其对应的正常组无显著性差异（P〉0．05）。结论UU感染、白细胞、精子密度关系密切,UU感染可通过引起白细胞异常,进而使ROS水平增高,使精子密度下降。     

Cerebrospinal fluid leucocyte counts in healthy neonates

OBJECTIVES: To determine the cerebrospinal fluid (CSF) white blood cell (WBC) count of normal term neonates, and compare the CSF WBC profile of normal and symptomatic infants without infection of the central nervous system (CNS). METHOD: Neonates were included if (a) they were at risk of congenital Toxoplasma infection and had undergone a lumbar puncture to assess CNS involvement, and (b) serial specific serum IgG and IgM determinations had ruled out congenital infection. According to neonatal chart reviews, 30 consecutive patients without CNS infection were classified as normal (absolutely asymptomatic) or symptomatic (any kind of symptoms). RESULTS: CSF WBC count was higher in 11 symptomatic (7/mm(3), 0-30/mm(3)) than in 19 normal (1/mm(3), 0-5/mm(3)) neonates (p<0.01). CONCLUSION: Normal neonatal CSF contains up to 5 WBCs/mm(3). Mild pleocytosis can be found in symptomatic infants without CNS infection.     

Flow cytometry technique for analysing Leishmania promastigote phagocytosis by human polymorphonuclear leucocytes and monocytes

In this study, we developed a flow cytometry technique for studying Leishmania (L.) mexicana phagocytosis by human polymorphonuclear leucocytes (PMNs) and monocytes. Leishmania promastigotes are elongated in shape and flagellated. This influences the light scatter when phagocytosis is measured by flow cytometry. Accordingly, we developed an oxidative burst method for measuring the phagocytic process. As this is an indirect marker of phagocytosis, we used confocal, light and electron microscopy to verify that promastigotes were, indeed, internalized by the phagocytes. For both PMNs and monocytes, the optimal conditions for achieving high sensitivity in flow cytometry detection were 5% pooled human serum and 15 min. incubation time. Incubations at 35, 37 and 39°C were also equally efficient for both PMNs and monocytes. Optimal parasite ratios were 10 parasites per PMN and 20 parasites per monocyte. Under these conditions, Leishmania were readily phagocytosed by human PMNs and monocytes and the effects of other influences, such as treatment, would be readily detectable. This indicated that these cells may play a role in the immune response against Leishmania.     

Effect of antioxidant supplementation on leucocyte expression of reactive oxygen species in athletes

Objective. Previous studies have shown that leucocytes, in particular granulocytes, have an enormous capacity for reactive oxygen species (ROS) production, and that this can be influenced by the physical activity of the individual. Theoretically, endurance‐trained athletes could profit by increasing their intake of antioxidants, thus neutralizing increased ROS production. The aim of the present investigation was to study the effect of dietary antioxidant supplementation on leucocyte ROS expression and total plasma antioxidant status (TAS) in endurance‐trained athletes over the course of 4 weeks. Methods. Eighteen athletes were recruited for the study. A randomized, double‐blinded, placebo‐controlled crossover study of 4 weeks of antioxidant supplementation (BiO‐Antioxidant 2.1 (400?mg vitamin C and 180?mg vitamin E d^?1) and BiO‐Quinon Q10 (200?mg?d^?1)) was performed. Flow cytometry was applied to examine the leucocyte expression of ROS using the ROS‐sensitive probes dihydroethidium and dihydrorhodamine 123. The Randox Total Antioxidant Status kit was used to measure the plasma TAS of the athletes. Results. After 4 weeks of antioxidant supplementation, we observed no significant differences in ROS levels in granulocytes and monocytes either basally or after in vitro stimulation with phorbol myristate acetate. Plasma TAS did not change significantly during the intervention period. Conclusions. We observed no influence of 4 weeks of dietary antioxidant supplementation on oxidative stress status. Based on these findings, there is no rationale advising athletes to ingest antioxidant supplements in addition to their regular diet if that includes daily recommended doses of vitamins.