Whole blood viscosity of preterm infants - differences to term neonates.

Determination of shear stresses at given shear rates with the LS 300 allows approximation of flow curves by mathematical models and to calculate viscosities of non-Newtonian fluids across the entire physiological shear range. Particular in preterm neonates, which in comparison to term neonates and adults have a substantially lower mean arterial blood pressure (MAP), rheological blood properties play a decisive role to maintain the perfusion. Whole blood viscosity was measured in umbilical cord blood taken from 34 preterm neonates using the LS 300 viscometer. In addition aggregation index, plasma viscosity, hematocrit and red blood cell (RBC) deformability was determined. The highest quality of approximation of the flow curve of whole blood was achieved by the method of Ostwald. Shear stresses of whole blood of preterm neonates were significantly lower compared to term neonates at similar shear rates (velocity range 6.16 s-1-50 s-1). With hematocrit calculated to 0.40 the related exponent (n) of the viscosity of preterm neonates (by Ostwald) showed a significant lower exponent (n) (0.71 ± 0.07) than term neonates (0.76 ± 0.06). The highest quality of approximation of the flow curve of plasma was achieved with the model of Ostwald in preterm neonates as well as in term neonates. The viscosity of plasma determined by Newton was lower for preterm neonates (0.89 ± 0.19) than for term neonates (1.04 ± 0.16). Concurrent to term neonates the whole blood of preterm neonates showed a very low aggregation index (term neonates 2.97 ± 2.10; preterm neonates 2.37 ± 1.32) and preterm neonates showed higher RBC deformability than term neonates. Because of the physiologically lower MAP and the particular viscous properties of neonatal blood special attention should be given when treating neonates with conventional blood products derived from adult donors.

Determination of whole blood and plasma viscosity in term neonates by flow curve analysis with the LS300 viscometer1.

Determination of shear stresses at given shear rates allow approximation of flow curves by mathematical models and to calculate viscosities of non-Newtonian fluids. In term neonates, the mean arterial blood pressure (MAP) is markedly below that of adults, therefore rheological properties of blood play an important role in maintaining perfusion. Whole blood viscosity was measured in umbilical cord blood taken from 62 term neonates using the LS 300 viscometer. Individual parameters that influence the viscosity of whole blood were measured: red blood cell (RBC) aggregation, plasma viscosity, hematocrit, and RBC deformability. The flow curve of whole blood of neonates was approximated by the method of Ostwald with the highest quality whereas in adults the best approximation was found by the method of Casson. With hematocrits of 0.40, the viscosity of whole blood in newborns approximated by Ostwald (9.84 ± 5.12 mPa·s) was significantly lower than that of adults (15.34 ± 3.01 mPa·s). The aggregation index of the blood of newborns was markedly lower (2.98 ± 2.12) than in adults (14.63 ± 3.50) whereas RBC deformability was higher in neonates. The viscosity of plasma determined by Ostwald revealed a lower exponent (n) in neonates (0.94 ± 022) compared to adults (1.01 ± 0.12) and the viscosity determined by Newton was lower in neonates (1.04 ± 0.16 mPa·s) than in adults (1.19 ± 0.07 mPa·s). The flow curve of neonatal blood which is best approximated by the model of Ostwald emphasizes its important viscous properties necessary for conditions with physiologically low blood pressure.

Managing neonatal severe sepsis in Germany: a preliminary survey of current practices.

BACKGROUND: In 2002 and 2007, the American College of Critical Care Medicine (ACCCM) provided clinical guidelines for hemodynamic support of pediatric and neonatal patients in septic shock. In 2008 and 2013, the Surviving Sepsis Campaign (SSC) Guidelines Committee offered up-to-date clinical guidelines for the management of severe sepsis and septic shock in adults and in pediatric patients. The aim of this study was to assess the standard of care of neonates with severe sepsis and septic shock in German neonatal intensive care units (NICUs) with regard to variability in management and guideline conformity. METHODS: 199 pediatric clinics in Germany were asked to describe their management of septic neonates in a telephone survey. The questionnaire that was used for the preliminary survey was designed based on the ACCCM and SSC clinical guidelines. RESULTS: A total of 90 (45%) surveys were completed and analyzed. Among all hospitals, the guidelines most commonly included in current practice patterns were obtaining cultures before administering antibiotics (100%), determining capillary refill time (99%), and using crystalloids for initial fluid therapy (97%). The guidelines least commonly included in current practice were determination of ammoniac to rule out inborn errors of metabolism (51%) and the use of dopamine as the first choice of hemodynamic support (48%). CONCLUSIONS: The management of sepsis, severe sepsis, and septic shock in neonates is not always guideline consistent, but quite a number of ACCCM and SSC guidelines were included in the current practice pattern.

A retrospective analysis of systemic propranolol for the treatment of complicated infantile haemangiomas.

AIM: A small percentage of complicated infantile haemangiomas need early, safe and effective treatment, and the option of off-label systemic propranolol treatment has been in existence since 2008. The aim of this study was to provide greater insight into the drug's efficacy and side effects. METHODS: We report our retrospective experiences of 207 paediatric patients treated with systemic propranolol for complicated infantile haemangiomas, which were photographed and analysed with a specific haemangioma score. RESULTS: Systemic treatment was successful in 99.5% of the patients. The haemangioma score before treatment was 8.3 ± 3.3 and the score at the end of the treatment was 1.5 ± 1.4. The reduction in the haemangiomas was significant and did not show any differences when distributed according to different localisations or to the patients' ages. Relevant side effects that may have made it necessary to discontinue the treatment were not observed. However, there was a statistically significant reduction in heart rate during the first six in-hospital drug applications. CONCLUSION: Our study demonstrates that systemic propranolol treatment is a highly effective treatment that is nearly always safe. These findings are a milestone for particularly complicated haemangiomas and provide highly valuable information on this drug treatment.

Flow behavior of fetal, neonatal and adult RBCs in narrow (3-6 µm) capillaries--Calculation and experimental application.

BACKGROUND AND OBJECTIVES: In capillaries with diameters less than those of resting RBCs, the cells have to deform to pass through such narrow vessels. Since RBCs of fetuses, neonates and adults differ in their geometrical properties the flow behavior of RBCs with different sizes in uniform tubes with diameters of 3 to 6 µm were studied by means of a micropipette system and a mathematical model. Assumptions in this model include an RBC flow velocity of 1 mm/s, axisymmetric cell shape and a gap between the RBC and the vessel wall that allows sufficient lubrication. The flow resistance depends on the surface area and volume of RBCs, the plasma viscosity and the vessel diameter. METHODS: Surface area and volume of different RBC populations (20 fetuses, 20 preterm neonates, 10 term neonates and 10 adults) were determined by means of a micropipette system and plasma viscosity was measured using a capillary tube viscometer. The flow behavior of RBCs with different volumes (61, 83 and 127 fl) was studied by direct microscopic observation using a micropipette system. The micropipettes had diameters of 3.5, 4.1, 5.1, and 6.0 µm. The flow velocity of the RBC in the pipettes was 1 mm/s and the calculated and measured cell lengths were compared. RESULTS: Volume and surface area of RBCs were 140 ± 29 fl and 172 ± 20 µm2, respectively, in the fetuses, decreased with increasing maturity (term neonates: 110 ± 20 fl and 149 ± 18 µm2) and reached the lowest values in adults (93 ± 14 fl and 136 ± 12 µm2). Plasma viscosities increased with increasing maturity due to rising plasma protein concentrations. The flow model leads to the following conclusions: During the passage of 3- to 6-µm vessels, the large fetal and neonatal RBCs are more elongated than the smaller adult RBCs. The critical vessel diameter, i.e., when the rear of the RBC becomes convex during passage of a narrow capillary, was 4.1 µm for fetal RBCs, 3.6 µm for neonatal RBCs and 3.3 µm adult cells. Suspended in the same medium, fetal and neonatal RBCs require 27% (term neonates) to 100% (fetuses) higher driving pressures than adult RBCs to achieve the necessary elongation for passing through a 4.5-µm capillary. However, the different RBCs require similar driving pressures if the cells are suspended in the corresponding autologous plasma. Cell lengths of the RBCs with different geometry determined during the flow experiments agreed with the predicted values. CONCLUSION: We conclude that the large size of fetal and neonatal RBCs may cause impaired flow in narrow vessels with diameters below the critical values of 3.6 to 4.1 µm. In vessels with diameters above the critical diameter (Dcr), the disadvantage of the large size of neonatal and fetal RBCs appears to be completely compensated for by the lower plasma viscosity.

Treatment of neonatal abstinence syndrome in preterm and term infants.

OBJECTIVE: Neonatal abstinence syndrome (NAS) is treated with a variety of drug preparations. With the optional treatment of NAS with chloral hydrate, phenobarbital or morphine the cumulative drug consumption of the mentioned drugs, the length of hospital stay and treatment duration was evaluated in preterm and term neonates. METHODS: Retrospective, uncontrolled study which evaluates different therapies of neonatal abstinence syndrome (NAS) in preterm and term neonates. RESULTS: During the past 16 years data were obtained from medical records of 51 neonates with NAS; 9 preterm and 35 term neonates were evaluated and 7 were excluded because of incomplete data sets. 31 (72.1%) received a pharmacological treatment (6 preterm and 25 term neonates). Treatment started at 4.3 [3.3-5.3] d. Mean duration of treatment was 11.7 [6.6-16.7] d. In our study chloral hydrate (ch) and phenobarbital (pb) were first line medication escalated by the morphine (mp) solution. Mean cumulative dosage of ch was 643.5 [260.3-1 026.7] mg, of pb 53.2 [19.7-86.8] mg and of mp 4.22 [0-8.99] mg. CONCLUSION: Our study group showed similar treatment duration and length of hospital stay compared to other studies. The cumulative dose of mp was lower compared to most studies. This benefit resulted at the expense of a further medication with pb and ch. However, 6 of 9 preterm neonates needed significantly less pharmacological therapy compared to term neonates indicating less susceptibility of immature brain to abstinence of maternalo-pioids.

The NO-donor FK409 improves mechanical properties of activated neonatal PMN.

BACKGROUND: Activated polymorphonuclear neutrophils (PMN) play an important role in the microcirculation. Nitric oxide (NO) reduces the sequestration of PMN in the narrow vessels of various organs and, therefore, may reduce organ injury during inflammation. OBJECTIVES: Since PMN of term neonates show various functional differences compared to PMN in adults (decreased chemotaxis, decreased intracellular killing, decreased adhesion), we studied the influence of the semi-synthetical NO-donor FK-409 (4-Ethyl-2-hydroxyimino-5-nitro-3-hexenamide) on the deformability of IL-8 activated PMN in term neonates and adults. METHODS: A cell transit analyzer (CTA) was used to study transit times of individual PMN through 8 µm filter pores, neutrophil elastase concentrations were determined by enzyme-immunoessay and activation of PMN was classified by mircroscopic evaluation. RESULTS: The transit times of PMN activated by IL-8 in adults were 9.3 ± 2.9 s, in term neonates 10.7 ± 3.3 s. FK-409 improved the transit time of activated PMN in adults (5.4 ± 1.6 s) and in term neonates (5.6 ± 1.1 s). Despite of the functional differences of PMN in term neonates and adults, the improvement of the transit times by FK-409 was not different between the two groups. The NO donor decreased the neutrophil elastase concentrations and the morphological signs of activation in neonates and adults. CONCLUSIONS: We conclude that the NO-donor FK-409 improves the microcirculation by increasing the deformability of IL-8 activated PMN. NO may reduce in neonates tissue damage by reduced PMN sequestration due to decreased PMN rigidity.

Shear stress and force required for tether formation of neonatal and adult erythrocytes.

Red blood cells (RBC) of neonates have a shorter survival time and they are more susceptible to mechanical alterations than RBC of adults. Irreversible alteration of the membrane of RBC of preterm neonates, term neonates and adults due to tether formation was studied by means of a micropipette technique. Shear stress and forces were applied with this technique in an axisymmetric configuration and were calculated with an approximation method. The applied shear stress and forces that induced tether formation were lowest for RBC of preterm neonates (1.5 Pa, 1.8 pN), in between for RBC of neonates (1.7 Pa, 2.1 pN) and highest for RBC in adults (1.9 Pa, 2.6 pN). Decreased mechanical stability of the membrane of neonatal RBC may in part cause the shorter life span, the greater tendency to endocytosis and the higher amount of irregularly shaped cells of neonatal RBC compared to RBC in adults.

Respiratory distress syndrome due to a novel homozygous ABCA3 mutation in a term neonate.

The authors report, for the first time in the literature, a case of respiratory distress syndrome in a term baby due to homozygosity for a p.Trp308Arg/W308R substitution in the ATP-binding cassette transporter 3. The sequence was confirmed by genetic analysis of the baby and both parents. Management and long-term outcome of a patient carrying this novel genetic defect have not been reported in the literature before. Currently, lung transplant appears to be the only long-term survival option available, for which, our patient is being evaluated.

Lipid A decreases human erythrocytes deformability by increasing intracellular Ca(2+): effects of verapamil, staurosporine and the rho-kinase inhibitor Y-27632.

There are several reports demonstrating an involvement of bacterial toxins in the rigidity of red blood cells (RBC). The present study investigates the influence of E. coli F-583-Rd lipid A on RBC deformability under mechanical shear stress. Verapamil (Ca(2+) channel inhibitor), staurosporine (protein kinase inhibitor) and Y-27632 (rho-kinase inhibitor) were used to modify the effect of lipid A on RBC deformability. We also determined if E. coli F-583-Rd Lipid A could induce an increase of intracellular Ca(2+) concentration. For the deformation measurements RBC (10 adult donors) were incubated with E. coli F-583-Rd lipid A (100 µg/ml) and also co-incubated with either verapamil (10(-7) mol/l), staurosporine (10(-7) mol/l) or Y-27632 (10(-7) mol/l). The deformation of the RBC under different shear stresses (0.6-60 Pa) was measured by a shear stress diffractometer (Rheodyne SSD). Intracellular Ca(2+) was determinded by flow cytometry in RBC incubated with Lipid A and labeled with fluorescent Fluo-4/AM which binds intracellular Ca(2+) with high affinity resulting in enhanced green fluorescence intensity. At increasing shear stresses Lipid A induced a significantly lower elongation. Co-incubation of the erythrocytes with verapamil or staurosporine inhibited lipid A induced decrease in elongation while Y-27632 had no effect. Verapamil, Staurosporine and Y-27632 did not influence the elongation response of the cells under control conditions. Lipid A induced a marked increase in fluorescence Fluo-4/AM indicating increased intracellular Ca(2+). These results suggest that E. coli F-583-Rd lipid A is able to influence red blood cell rigidity by a rapid and significant increase of intracellular Ca(2+) concentration. Verapamil and staurosporine abolished the decrease in deformability of Lipid A incubated RBC.

Effects of phosphodiesterase (III/IV)-inhibitors and cytokines on mechanical properties of neutrophilic granulocytes in neonates and adults.

Sequestration of activated PMN and enrichment in tissues play a key role in tissue damage during septicaemia and after ischemia/reperfusion. Since polymorphonuclear neutrophilic granulocytes (PMN) of term neonates show various functional differences compared to PMN in adults (decreased chemotaxis, decreased intracellular killing, decreased adhesion) we studied the influence of interleukin 8 (IL-8), tumor necrosis factor-alpha (TNF-alpha) and N-formyl-methionyl-leucyl-phenylalanine (fMLP) on the reduction of deformability of PMN in neonates and adults. The following phosphodiesterase (PDE)-inhibitors were applied to ameliorate the reduction in deformability when the PMN were stimulated with fMLP or IL-8: Enoximone, Milrinone (PDE-III-inhibitors), Pentoxifylline (PTX) and Piclamilast (PDE-IV-inhibitors). The micropipette technique and the cell transit analyzer (CTA) were used and compared. Aspiration times into micropipettes with an internal diameter of 5 microm, transit times through 8 microm filter pores and neutrophil elastase concentrations were determined. Despite of the functional differences of PMN in neonates compared to adults the significant decrease of deformability of PMN activated with cytokines compared to passive PMN was not different in both groups. The neutrophil elastase concentrations reflect the activation of the PMN: highest concentrations during activation, decreased concentrations due to PDE-inhibitors, and PMN in a passive state. The neutrophil elastase concentrations were not different from PMN of neonates and adults. These PDE-inhibitors significantly increased the deformability of activated PMN but significant differences between the deformability of PMN in neonates and adults were not found. Despite the functional differences of PMN in neonates PDE-III/IV-inhibitors lead to similar improvement of mechanical properties of activated PMN in neonates and adults. These drugs may ameliorate impaired microcirculation also in neonates during inflammation.

Gamma-hydroxybutyric acid sedation in neonates and children undergoing MR imaging.

BACKGROUND: Insufficient sedation in pediatric magnetic resonance imaging (MRI) results in prolonged examination time. To describe the efficacy and side effects of sedation with Phenobarbital short-time infusion followed by continuous gamma-hydroxybutyric acid (GHB) infusion in neonates and children for MRI examinations in a retrospective study. PATIENTS: With Institutional Review Board approval 94 children (Group I: 1-4 weeks; Group II: >1 to 6 months; Group III: >6 months) were sedated with phenobarbital 10 mg/kg (maximum 200 mg) intravenously 30 min prior to examination. Than intravenous sedation was maintained with GHB 10 mg/kg/h after a priming dose of 30 mg/kg in 20 min. RESULTS: In group 1 all neonates (n=8) were well sedated without side effect. One of 21 infants in group 2 showed restlessness and the MRI failed. Two of 65 patients of group 3 were not sufficiently sedated and one of them vomited. CONCLUSIONS: Non-invasive diagnostic procedures in neonates and children may be managed by phenobarbital and GHB sedation with side effects or failure of 3%.

Deformability of passive and activated neutrophils in children with Gram-negative septicemia.

Animal experiments suggest that obstruction of small vessels by polymorphonuclear neutrophils (PMNs) may contribute to the disturbed microcirculation in septic shock. The purpose of this investigation was to study deformability and volume of passive and activated PMNs and immature neutrophils in 15 children with Gram-negative septicemia and in 20 healthy children. Membrane cytoplasm tongues of neutrophils were aspirated by means of a micropipette system into 2.5-microm (diameter) pipettes for 60 s. Tongue growth was used as an indicator of deformability and the cell volume was calculated from the cell circumference before aspiration. Septicemic children showed higher percentages of immature neutrophils (38+/-13%) and activated PMN (12+/-5%) than healthy children (3+/-2% and 3+/-2%). In septicemic children, cellular volume of passive neutrophils decreased progressively with increasing maturity from myeloblasts (493+/-105 fL) to mature PMNs (346+/-29 fL) and the final tongue length in the micropipette increased from 2.7+/-1.1 to 8.5+/-1.8 microm during maturation. The final tongue length of activated PMNs was decreased by 60% compared with that of passive PMNs. The increased number of rigid activated and immature neutrophils may contribute to impaired microcirculation in septicemic patients.

Effect of activators and the phosphodiesterase inhibitors pentoxifylline and enoximone on the deformability of neutrophils in neonates and adults.

BACKGROUND: Stimulated polymorphonuclear leukocytes (PMN) are extremely rigid compared to resting PMN. They may obstruct narrow vessels and contribute to ischaemic organ injury. Deformability is a prerequisite for both active and passive movement in the microcirculation. AIM: The investigation was designed to study whether stimulators and inhibitors of stimulation show different effects on deformability of neonatal and adult PMN. METHODS: Deformability of PMN was assessed by complete aspiration of a PMN into a micropipette with an internal diameter of 5 microm. Blood samples from 20 neonates and 20 adults were studied before and after stimulation with N-formyl-methionyl-leucyl-phenylalanine (fMLP), interleukin-8 (IL-8) or tumour necrosis factor-alpha (TNF-alpha). Moreover, effects of the phosphodiesterase inhibitors Pentoxifylline (PTX) and Enoximone on the deformability of stimulated PMN were investigated. RESULTS: fMLP, IL-8 and TNF-alpha significantly delayed aspiration times of PMN in relation to the concentrations of the stimulators. The addition of PTX or Enoximone to stimulated PMN increased the deformability up to 60% depending on the concentration of the inhibitors. No significant differences in the aspiration times were found between neonatal and adult PMN at any of the experimental conditions after activation with the three stimulators and treatment with the two inhibitors. CONCLUSION: Neonatal and adult PMN show similar reduction of passive deformability when stimulated with either fMLP, IL-8 or TNF-alpha compared to resting PMN and a similar improvement of deformability in response to PTX or Enoximone.

Formation and relaxation of erythrocyte membrane tethers in micropipettes.

The red blood cell (RBC) membrane forms tethers in response to shear forces acting on tiny membrane points. Tether formation depends on viscous and elastic membrane properties and has been used as indicator of membrane fragility. A micropipette technique was used to study time dependent tether formation and tether relaxation of individual RBC. Point attached RBC were aspirated at a negative pressure of -5 mm H2O into a micropipette with an internal diameter of 7.8 microm. If tether formation occurred and the tether reached a length of approximately 16 microm, the pipette was carefully pulled back. The RBC left the orifice of the micropipette and the tether relaxed and pulled the main body of the RBC back to the attachment point. The relaxation of the tethers was exponential. The time constant for tether relaxation was 0.144 s which is similar to the time constant for recovery of entire RBC from extensional elastic deformation. Repeated tether formation and relaxation of the same RBC led to an earlier begin of tether formation and changed the behavior of tether growth, although the relaxation time did not change. We conclude that repeated tether formation decreases the resistance of the RBC membrane to form tethers upon given shear forces. Weakening of the membrane due to repeated plastic deformation may play a role in the membrane loss of circulating RBC during aging.

Impaired deformability of erythrocytes and neutrophils in children with newly diagnosed insulin-dependent diabetes mellitus.

AIMS/HYPOTHESIS: Abnormal rheological properties of erythrocytes, leucocytes and plasma may have a role in the development of diabetic microangiopathy. We hypothesized that changed haemorrheological variables may already be found in children with onset diabetes. METHODS: Erythrocyte deformation (rheoscope), neutrophil deformation (micropipette), erythrocyte aggregation, blood and plasma viscosity were measured in 15 children with insulin-dependent diabetes mellitus before initiation of insulin treatment and 4 to 6 weeks later, 15 diabetic children treated with insulin for 5 to 8 years, 15 healthy children and 15 healthy adults. RESULTS: At a low shear stress of 0.6 Pa, erythrocyte deformation was decreased in the diabetic children before (-28%), after 4 to 6 weeks (-22%) and after 5 to 8 years (-17%) of insulin treatment compared with healthy children. More active neutrophils were counted in the untreated diabetic children (9 +/- 6%) than in healthy children (3 +/- 2%). Deformability of passive neutrophils was greatly decreased in the children with onset diabetes and moderately reduced in the diabetic children who were treated with insulin. Neutrophil deformation (r = -0.52) and erythrocyte deformation at 0.6 Pa (r = -0.62) were inversely related to haemoglobin A1c. Haematocrit and blood viscosity were increased in the untreated children and in the children treated with insulin for 5 to 8 years. Plasma viscosity and erythrocyte aggregation were similar in the three groups of children. CONCLUSION/INTERPRETATION: Decreased erythrocyte deformation at low shear force, increased count of active neutrophils and impaired deformability of passive neutrophils may increase the risk for acute cerebro-vascular complications in children with uncontrolled insulin-dependent diabetes mellitus.

Deformability and geometry of neonatal erythrocytes with irregular shapes.

More stomatocytes, spherocytes, and erythrocytes with spicules and protrusions are found in blood from newborn infants than in blood from adults. The objective of this study was to measure volume, surface area, membrane rigidity, and viscosity of red blood cells (RBC) with irregular shapes in preterm and full-term neonates. RBC from 13 full-term and 12 preterm (25-36 wk of gestation) infants were studied by means of a micropipette system. In full-term neonates and in preterm infants, 19 and 26% of RBC, respectively, showed shapes different from those of discocytes. Keratocytes, spherocytes, akanthocytes (in full-term infants), and elliptocytes (in preterm infants) had smaller volumes than did discocytes (p < 0.05). Spherocytes showed decreased and elliptocytes had increased excess surface area and swelling capacity compared with discocytes, indicating both volume and membrane loss for spherocytes and only volume loss for elliptocytes. Immature RBC and knizocytes had similar large volumes and surface areas, suggesting that knizocytes in neonates are relatively young RBC. Increased membrane rigidity was found only in stomatocytes. Membrane viscosity was decreased in immature RBC and knizocytes and increased in stomatocytes, keratocytes, akanthocytes, and elliptocytes when compared with discocytes. There were inverse relationships between RBC volume and membrane rigidity (r=-0.42) and between RBC volume and membrane viscosity (r=-0.72). This suggests that volume loss of neonatal RBC is associated with impaired membrane deformability. Moreover, we conclude that the geometry and membrane deformability of some of the irregularly shaped neonatal RBC resemble properties of RBC in hemolytic anemias (e.g. spherocytosis).

Passive deformability of mature, immature, and active neutrophils in healthy and septicemic neonates.

Obstruction of narrow vessels by rigid neutrophils may contribute to ischemic organ injury. In septicemia, a substantial portion of the neutrophils may become activated and the number of circulating immature neutrophils may rise sharply. Volume and deformability of mature (PMN) and immature neutrophils in healthy preterm and full-term infants and in septicemic neonates were studied by means of a micropipette system. Membrane cytoplasm tongues were aspirated into 2.5-microm (diameter) pipettes over a period of 60 s. Volume and tongue growth of mature resting PMN were similar in healthy preterm and full-term neonates and adults. Compared with mature PMN (about 360 fl), the volumes of band cells (415 fl), metamyelocytes (470 fl), and less mature cells (myeloblasts, promyelocytes, and myelocytes; 490 fl) were significantly increased (p < 0.005). Final tongue lengths of band cells, metamyelocytes, and less mature cells were decreased by about 50, 60, and 70%, respectively, when compared with passive mature cells. In septic neonates, the percentage of immature neutrophils was increased, but the deformability and volume of the cell subpopulations were not affected by septicemia. Active PMN were characterized by pseudopod formation. More active PMN were found in group B streptococcal (14% of total PMN), gram-negative (12%), and Staphylococcus epidermidis septicemia (8%) than in healthy neonates and adults (4%). The main bodies of active PMN were less deformable than passive PMN, and the pseudopods showed very little membrane deformation. The increased number of rigid active and immature neutrophils may contribute to impaired microcirculation and the high risk for organ injury in septic patients.

Cellular and membrane deformability of red blood cells in preterm infants with and without growth retardation.

Membrane and whole red cell deformability are essential for effective blood flow, oxygen uptake and release. Whole cell deformability (rheoscope with shear stresses of 0.6-30 Pa) and membrane shear elastic modulus (flow channel) of red blood cells (RBC) were studied in appropriate and small for gestational age preterm infants (28-32 wk of gestation), in healthy full-term neonates and in adults. No significant differences were found between the two groups of preterm infants. The preterm infants showed significantly increased RBC deformation in the rheoscope when compared to the full-term neonates who, in turn, showed more RBC deformation than adults. Correspondingly, the membrane shear elastic modulus (i.e., the resistance to elastic membrane deformation) was lower in the preterm infants than in the full-term neonates and lower in full-term infants than in adults. Thus, both whole cell and membrane deformability appear to be increased in preterm infants. Improved RBC deformability may aid in the maintenance of adequate blood circulation in preterm infants in spite of low vascular pressures.

Effects of group-A streptolysin O on neonatal and adult red blood cell deformability and haemolysis.

Neonates are more susceptible than adults to many Gram-positive and Gram-negative bacterial infections. Whereas group B streptococcus causes life-threatening infections in neonates, group A beta-haemolytic streptococcus infections rarely occur in neonates. To test the hypothesis that group A streptococcus may have different effects on neonatal and adult red blood cells (RBCs), haemolysis and deformability (rheoscope) of RBCs from adults, full-term and pre-term neonates were studied during 60 min incubation with 1 haemolytic unit (HU) mL-1 group A streptolysin O (SLO). SLO incubation of adult RBCs resulted in almost linearly increasing time-dependent haemolysis reaching 82%, whereas haemolysis of neonatal RBCs was below 60% after 1 h. After 60 min SLO incubation, RBC deformation was significantly (P < 0.05) more reduced in adults than in full-term and preterm neonates. An inverse overall relationship (r = 0.68) between SLO-induced haemolysis and RBC deformation was found after 60 min of SLO incubation. We conclude that SLO causes less haemolysis and less impairment of RBC deformation in neonates than in adults. The decreased RBC deformation of unhaemolysed RBC indicates that, before lysis, mechanical RBC membrane properties are altered by SLO.