Respiratory healthcare by design: Computational approaches bringing respiratory precision and personalised medicine closer to bedside.

Precision medicine represents a potentially powerful means to alleviate the growing burden of chronic respiratory diseases. To realise its potential, however, we need a systems level understanding of how biological events (signalling pathways, cell-cell interactions, tissue mechanics) integrate across multiple spatial and temporal scales to give rise to pathology. This can be achieved most practically in silico: a paradigm that offers tight control over model parameters and rapid means of testing and generating mechanistic hypotheses. Patient-specific computational models that can enable identification of pathological mechanisms unique to patients' (omics, physiological, and anatomical) profiles and, therefore, personalised drug targets represent a major milestone in precision medicine. Current patient-based models in literature, especially medical devices, cardiac modelling, and respiratory medicine, rely mostly on (partial/ordinary) differential equations and have reached relatively advanced level of maturity. In respiratory medicine, patient-specific simulations mainly include subject scan-based lung mechanics models that can predict pulmonary function, but they treat the (sub)cellular processes as "black-boxes". A recent advance in simulating human airways at a cellular level to make clinical predictions raises the possibility of linking omics and cell level data/models with lung mechanics to understand respiratory pathology at a systems level. This is significant as this approach can be extended to understanding pathologies in other organs as well. Here, I will discuss ways in which computational models have already made contributions to personalised healthcare and how the paradigm can expedite clinical uptake of precision medicine strategies. I will mainly focus on an agent-based, asthmatic virtual patient that predicted the impact of multiple drug pharmacodynamics at the patient level, its potential to develop efficacious precision medicine strategies in respiratory medicine, and the regulatory and ethical challenges accompanying the mainstream application of such models.

Microbial symbionts affect Pisum sativum proteome and metabolome under Didymella pinodes infection.

UNLABELLED: The long cultivation of field pea led to an enormous diversity which, however, seems to hold just little resistance against the ascochyta blight disease complex. The potential of below ground microbial symbiosis to prime the immune system of Pisum for an upcoming pathogen attack has hitherto received little attention. This study investigates the effect of beneficial microbes on the leaf proteome and metabolome as well as phenotype characteristics of plants in various symbiont interactions (mycorrhiza, rhizobia, co-inoculation, non-symbiotic) after infestation by Didymella pinodes. In healthy plants, mycorrhiza and rhizobia induced changes in RNA metabolism and protein synthesis. Furthermore, metal handling and ROS dampening was affected in all mycorrhiza treatments. The co-inoculation caused the synthesis of stress related proteins with concomitant adjustment of proteins involved in lipid biosynthesis. The plant's disease infection response included hormonal adjustment, ROS scavenging as well as synthesis of proteins related to secondary metabolism. The regulation of the TCA, amino acid and secondary metabolism including the pisatin pathway, was most pronounced in rhizobia associated plants which had the lowest infection rate and the slowest disease progression. BIOLOGICAL SIGNIFICANCE: A most comprehensive study of the Pisum sativum proteome and metabolome infection response to Didymella pinodes is provided. Several distinct patterns of microbial symbioses on the plant metabolism are presented for the first time. Upon D. pinodes infection, rhizobial symbiosis revealed induced systemic resistance e.g. by an enhanced level of proteins involved in pisatin biosynthesis.

Coarse and fine root plants affect pore size distributions differently.

AIMS: Small scale root-pore interactions require validation of their impact on effective hydraulic processes at the field scale. Our objective was to develop an interpretative framework linking root effects on macroscopic pore parameters with knowledge at the rhizosphere scale. METHODS: A field experiment with twelve species from different families was conducted. Parameters of Kosugi's pore size distribution (PSD) model were determined inversely from tension infiltrometer data. Measured root traits were related to pore variables by regression analysis. A pore evolution model was used to analyze if observed pore dynamics followed a diffusion like process. RESULTS: Roots essentially conditioned soil properties at the field scale. Rooting densities higher than 0.5 % of pore space stabilized soil structure against pore loss. Coarse root systems increased macroporosity by 30 %. Species with dense fine root systems induced heterogenization of the pore space and higher micropore volume. We suggested particle re-orientation and aggregate coalescence as main underlying processes. The diffusion type pore evolution model could only partially capture the observed PSD dynamics. CONCLUSIONS: Root systems differing in axes morphology induced distinctive pore dynamics. Scaling between these effective hydraulic impacts and processes at the root-pore interface is essential for plant based management of soil structure.

Root induced changes of effective 1D hydraulic properties in a soil column.

AIMS: Roots are essential drivers of soil structure and pore formation. This study aimed at quantifying root induced changes of the pore size distribution (PSD). The focus was on the extent of clogging vs. formation of pores during active root growth. METHODS: Parameters of Kosugi's lognormal PSD model were determined by inverse estimation in a column experiment with two cover crops (mustard, rye) and an unplanted control. Pore dynamics were described using a convection-dispersion like pore evolution model. RESULTS: Rooted treatments showed a wider range of pore radii with increasing volumes of large macropores >500 µm and micropores <2.5 µm, while fine macropores, mesopores and larger micropores decreased. The non-rooted control showed narrowing of the PSD and reduced porosity over all radius classes. The pore evolution model accurately described root induced changes, while structure degradation in the non-rooted control was not captured properly. Our study demonstrated significant short term root effects with heterogenization of the pore system as dominant process of root induced structure formation. CONCLUSIONS: Pore clogging is suggested as a partial cause for reduced pore volume. The important change in micro- and large macropores however indicates that multiple mechanic and biochemical processes are involved in root-pore interactions.

Field quantification of wetting-drying cycles to predict temporal changes of soil pore size distribution.

Wetting-drying (WD) cycles substantially influence structure related soil properties and processes. Most studies on WD effects are based on controlled cycles under laboratory conditions. Our objective was the quantification of WD cycles from field water content measurements and the analysis of their relation to the temporal drift in the soil pore size distribution. Parameters of the Kosugi hydraulic property model (rm,Kosugi, σKosugi) were derived by inverse optimization from tension infiltrometer measurements. Spectral analysis was used to calculate WD cycle intensity, number and duration from water content time series. WD cycle intensity was the best predictor (r2 = 0.53-0.57) for the temporal drift in median pore radius (rm,Kosugi) and pore radius standard deviation (σKosugi). At lower soil moisture conditions the effect of cycle intensity was reduced. A bivariate regression model was derived with WD intensity and a meteorological indicator for drying periods (ET0, climatic water balance deficit) as predictor variables. This model showed that WD enhanced macroporosity (higher rm,Kosugi) while decreasing pore heterogeneity (lower σKosugi). A drying period with high cumulative values of ET0 or a strong climatic water balance deficit on the contrary reduced rm,Kosugi while slightly increasing σKosugi due to higher frequency at small pore radius classes. The two parameter regression model was applied to predict the time course of soil pore size distribution parameters. The observed system dynamics was captured substantially better by the calculated values compared to a static representation with averaged hydraulic parameters. The study showed that spectral analysis is an adequate approach for the quantification of field WD pattern and that WD intensity is a key factor for the temporal dynamics of the soil pore size distribution.

Environmental and management influences on temporal variability of near saturated soil hydraulic properties.

Structural porosity is a decisive property for soil productivity and soil environmental functions. Hydraulic properties in the structural range vary over time in response to management and environmental influences. Although this is widely recognized, there are few field studies that determine dominant driving forces underlying hydraulic property dynamics. During a three year field experiment we measured temporal variability of soil hydraulic properties by tension infiltrometry. Soil properties were characterized by hydraulic conductivity, effective macroporosity and Kosugi's lognormal pore size distribution model. Management related influences comprised three soil cover treatment (mustard and rye vs. fallow) and an initial mechanical soil disturbance with a rotary harrow. Environmental driving forces were derived from meteorological and soil moisture data. Soil hydraulic parameters varied over time by around one order of magnitude. The coefficient of variation of soil hydraulic conductivity K(h) decreased from 69.5% at saturation to 42.1% in the more unsaturated range (- 10 cm pressure head). A slight increase in the Kosugi parameter showing pore heterogeneity was observed under the rye cover crop, reflecting an enhanced structural porosity. The other hydraulic parameters were not significantly influenced by the soil cover treatments. Seedbed preparation with a rotary harrow resulted in a fourfold increase in macroporosity and hydraulic conductivity next to saturation, and homogenized the pore radius distribution. Re-consolidation after mechanical loosening lasted over 18 months until the soil returned to its initial state. The post-tillage trend of soil settlement could be approximated by an exponential decay function. Among environmental factors, wetting-drying cycles were identified as dominant driving force explaining short term hydraulic property changes within the season (r2 = 0.43 to 0.59). Our results suggested that beside considering average management induced changes in soil properties (e.g. cover crop introduction), a dynamic approach to hydrological modeling is required to capture over-seasonal (tillage driven) and short term (environmental driven) variability in hydraulic parameters.

Prophylactic dexamethasone for postoperative nausea and vomiting in pediatric strabismus surgery: a dose ranging and safety evaluation study.

In this double-blind, randomized, placebo-controlled study, we evaluated the efficacy and safety of different doses of prophylactic IV dexamethasone for postoperative nausea and vomiting (PONV) in 168 children (aged 2-15 yr) scheduled for strabismus surgery. Patients received IV dexamethasone 0.25 mg/kg (D 0.25), 0.5 mg/kg (D 0.5), 1.0 mg/kg (D 1), or saline (S) immediately after induction of general anesthesia. Patients were discharged 24 h after surgery. Nausea and vomiting were assessed at 0-2, 2-6, and 6-24 h after surgery. Blood glucose was measured preoperatively and at 4 h after study drug administration. Wound healing and infection were assessed after 1 wk. More patients in group S had vomiting at 0-2, 2-6, and 6-24 h (P = 0.001, P = 0.003, and P = 0.04, respectively) and required larger doses of rescue antiemetics compared with the dexamethasone groups. Fewer patients in the dexamethasone groups (6, 3, and 6 in D 0.25, D 0.5, and D 1, respectively) had severe PONV compared with group S (P = 0.001). No significant increase in postoperative blood glucose levels was observed and wound healing was satisfactory in all four groups. The results suggest that dexamethasone 0.25 mg/kg is more effective than saline and equally effective compared with larger doses for preventing PONV for pediatric strabismus surgery.

Efficacy of low-dose dexamethasone for preventing postoperative nausea and vomiting following strabismus repair in children.

We studied the efficacy of a range of doses of dexamethasone for prevention of postoperative nausea and vomiting following strabismus repair in children in a hospital-based, prospective, double-blinded, randomized, placebo-controlled trial. Two hundred and ten children were randomized to receive either dexamethasone in one of four dosages: 50 microg/kg (Group 1), 100 microg/kg (Group 2), 200 microg/kg (Group 3) and 250 microg/kg (Group 4) or normal saline (Group 5) prior to corrective surgery for strabismus. Anaesthesia was standardized and included nitrous oxide, pethidine, intubation and the use of muscle relaxant and reversal with neostigmine. Postoperative nausea and vomiting were evaluated in epochs of 0-2 hours, 2-6 hours and 6-24 hours after surgery. Parent satisfaction was assessed 24 hours after surgery and the operated eye was examined for wound infection and delayed healing one week later Dexamethasone was effective in preventing nausea and vomiting after strabismus repair: 57.1% children in Group 1, 42.9% in Group 2, 52.4% in Group 3, and 59.5% in Group 4 were free from postoperative nausea and vomiting compared with 7.1% in placebo group. The lowest dose of 50 microg/kg was as efficacious as the higher dosages of dexamethasone during the 24 hours studied. Of the children who developed postoperative nausea and vomiting those who received dexamethasone had significantly fewer episodes than those in the placebo group. We conclude that dexamethasone 50 microg/kg is effective for the prevention of postoperative nausea and vomiting following strabismus repair in children.

Effect of epidural morphine on minimum alveolar concentration of isoflurane in humans.

BACKGROUND AND AIMS: The anaesthetic potency of volatile anaesthetic agents is measured by the minimum alveolar concentration (MAC) required to suppress response in 50% of subjects. We studied the effect of epidural morphine on MAC of isoflurane in humans. SETTINGS AND DESIGN: A prospective single-blind study designed to study the effect of epidural morphine on MAC of isoflurane. SUBJECTS AND METHODS: Forty-eight patients were randomly divided into two groups - Group I patients received 3 mg morphine in 10 ml saline, and Group II patients received 10 ml saline epidurally. Anaesthesia was induced with isoflurane in oxygen and nitrous oxide. Later nitrous oxide was discontinued and MAC of isoflurane determined using modified Dixon's method of sequential sampling. RESULTS: Epidural morphine resulted in a significant reduction in MAC of isoflurane, 0.98 vs. 1.14 in control group (p<0.05). CONCLUSIONS: Epidural administration of 3 mg morphine in 10 ml saline decreased the MAC of isoflurane.

Intrathecal midazolam added to bupivacaine improves the duration and quality of spinal anaesthesia.

BACKGROUND: The antinociceptive action of intrathecal midazolam is well documented. In this prospective study, we investigated the addition of midazolam to intrathecal bupivacaine on the duration and quality of spinal blockade. METHODS: Forty ASA I or II adult patients undergoing lower abdominal surgery were selected for the study. The patients were randomly allocated to receive 3 ml of 0.5% hyperbaric bupivacaine intrathecally either alone or with 1 mg of midazolam using a combined spinal epidural technique. The duration and quality of sensory and motor block, perioperative analgesia, haemodynamic changes, and sedation levels were assessed. RESULTS: The duration of sensory block (i.e. time to regression to the S2 segment) was significantly longer in the midazolam group than the control group (218 min vs. 165 min; P < 0.001). The duration of motor block was also prolonged in the midazolam group as compared with the control group (P < 0.01). In 90% of the patients in the midazolam group, the quality of block was adequate during the intra-operative period as compared with only 65% of the patients in the control group (P < 0.05). The duration of effective analgesia was longer in the midazolam group than in the control group (199 vs. 103 min; P < 0.001). Blood pressure, heart rate, oxygen saturation and sedation scores were comparable in both groups. No neurological deficit or other significant adverse effects were recorded. CONCLUSION: The addition of intrathecal midazolam to bupivacaine significantly improves the duration and quality of spinal anaesthesia and provides prolonged perioperative analgesia without significant side-effects.

Stress ulcer prophylaxis in patients on ventilator.

The objective of the study was to assess the efficacy of the H2-receptor antagonists and sucralfate for the prophylaxis of stress ulcer in patients on a ventilator in an intensive care unit in the general intensive care unit of our institute. A randomized, clinical controlled trial was conducted. Fifty-two critically ill patients, who required mechanical ventilation for more than 24 hours, were randomly divided into 3 groups. Group I received ranitidine 50 mg (intravenous) 8 hourly, group II received tablet sucralfate 1 g 8 hourly through a Ryle's tube, whereas group III was not given any drug. The incidence of upper gastrointestinal bleed, change in gastric pH and growth of gram-negative organisms in the gastric juice and bronchoalveolar lavage (BAL) culture were noted and analysed. The treatment groups were similar with respect to the baseline characteristics. The incidence of upper gastrointestinal bleeding was similar in the ranitidine (12.5%) and sucralfate groups (14.35%) but was high in the control group (57.14%). The mean gastric pH was significantly low in the control group (mean pH 2.07) compared to the ranitidine (mean pH 5.25) and sucralfate groups(mean pH 3.54)(p < 0.05). The incidence of positive culture for gram-negative organisms was significantly high in the ranitidine group (75%) in comparison with the sucralfate group (33.33%) (p < 0.002). However, the incidence of positive growth in the BAL culture was similar in all three groups. We conclude that both ranitidine and sucralfate are equally effective in decreasing the incidence of upper gastrointestinal haemorrhage and other stress- related lesions. Though ranitidine was more effective in increasing the gastric pH, the incidence of gastric colonization was higher in the ranitidine group compared to the sucralfate group.

Remifentanil and propofol sedation for retrobulbar nerve block.

We studied remifentanil and propofol for analgesia and sedation during the placement of an ophthalmic block. Eighty ASA I or II patients undergoing elective cataract surgery under a retrobulbar block in a rural camp setting were included in the study. Patients were randomly divided into four groups and received different drug combinations as follows: Group I--remifentanil 1 microg/kg, Group II--remifentanil 0.5 microg/kg and propofol 0.5 mg/kg, Group III--remifentanil 1 microg/kg with propofol 0.5 mg/kg and Group IV--saline 0.1 ml/kg. Patients were observed for degree of movement, sedation, pain, recall and respiratory depression. No patient in the study groups reported pain or displayed movement whereas most of the patients in the control group had significant pain during the placement of the block. Also, seven (35%) patients in the control group showed significant movement which may have led to failure of block in two patients and retrobulbar haemorrhage in one patient. Incidence of significant respiratory depression was maximum in Group III patients (60%), followed by Group I (20%) and least in Group II (5%). All patients in the study groups remained cooperative and obeyed commands except four patients in group III (OAA/S-4). Postoperatively, other than the control group, recall was maximum in Group I (55%) and least in Group II (5%). Hence, a combination of remifentanil 0.5 microg/kg with propofol 0.5 mg/kg as a bolus was considered to provide excellent relief of pain and anxiety with least adverse effects for the placement of ophthalmic blocks.

Effect of addition of clonidine to local anaesthetic mixture for peribulbar block.

Clonidine added to local anaesthetics prolongs the duration of anaesthesia and analgesia of peripheral, neuraxial and retrobulbar blocks. The present randomized blinded controlled study was conducted to evaluate the effect of the addition of clonidine to local anaesthetic mixture on the quality, onset time, duration of peribulbar block, perioperative analgesia and patients' comfort. The study comprised two groups of 12 patients each. Group A (control) patients received 7 ml of a mixture of 2% lignocaine and hyaluronidase with 1 ml normal saline, while group B (clonidine group) patients had clonidine 1 microg/kg added to the above mixture. Onset and duration of lid akinesia, globe anaesthesia and akinesia, time to first analgesic medication and total analgesic requirement were assessed. Patients were monitored for heart rate, blood pressure, sedation and respiratory depression. Addition of clonidine to local anaesthetic mixture resulted in a significant increase in duration of lid akinesia (85.4+/-25.6 vs 173.3+/-35.3 min, P<0.001), globe anaesthesia (63.2+/-6.9 vs 78.8+/-17.5 min, P=0.012) and globe akinesia (161.3+/-24.3 vs 201.2+/-45.7 min, P=0.016). The onset time and quality of block were similar in both the groups. No significant haemodynamic, respiratory or sedative effects were recorded. The perioperative pain scores and the analgesic requirements were significantly (P<0.01) lower in group B patients. We found that addition of clonidine 1 microg/kg to local anaesthetic mixture significantly increases the duration of anaesthesia and analgesia after peribulbar block.

Radiofrequency ablation: a new therapeutic modality for the management of hepatocellular cancer.

We report two patients of hepatocellular cancer who recently underwent radiofrequency ablation at our center. Both underwent successful ablation of the tumour requiring 1-2 sessions of upto 15 minutes. There were no post procedure complications. One of the patients had developed another lesion after 10 months of follow up and underwent another session of RFA, while the second patient is doing well after one year of the procedure.

A dose response study of clonidine with local anesthetic mixture for peribulbar block: a comparison of three doses.

UNLABELLED: Clonidine prolongs anesthesia and analgesia of local anesthetics in various neural blocks as well as the duration of retrobulbar block. We assessed the dose-response relationship of clonidine added to lidocaine in peribulbar block. Sixty patients undergoing cataract surgery were given peribulbar block with 7 mL of 2% lidocaine and hyaluronidase with either saline (Control) or clonidine in 0.5-microg/kg (0.5 Clon), 1.0-microg/kg (1.0 Clon), or 1.5-microg/kg (1.5 Clon) doses. The onset and duration of lid and globe akinesia, globe anesthesia and analgesia, postoperative analgesic requirement, and adverse effects (hypotension, bradycardia, hypoxia, sedation, and dizziness) were recorded. The success rate and onset of block were comparable in all groups. The duration of lid and globe akinesia, globe anesthesia and analgesia was significantly (P < 0.01) prolonged in patients receiving 1.0 and 1.5 microg/kg clonidine as compared with the Control group. Perioperative pain scores and analgesic requirement were significantly less in these groups. 0.5 microg/kg clonidine did not increase the duration of anesthesia and analgesia significantly. Hypotension and dizziness were observed more in patients receiving 1.5 microg/kg clonidine as compared with other groups. We conclude that 1.0 microg/kg clonidine with a mixture of lidocaine (2%) significantly prolonged the duration of anesthesia and analgesia after peribulbar block with limited side effects. IMPLICATIONS: We studied the effect of the addition of 0.5, 1.0 and 1.5 microg/kg clonidine to a lidocaine-hyaluronidase mixture on the onset and duration of peribulbar block and perioperative analgesia. A dose of 1.0 microg/kg produced a significant increase in duration of anesthesia and analgesia with minimal side effects.

The interleukin-10 promoter genotype determines clinical immune function in hemodialysis patients.

BACKGROUND: Immune dysfunction and the impaired hepatitis B vaccination response are complications of chronic renal failure that are tightly associated with inflammation induced by uremia and blood-membrane contacts. Proinflammatory cytokines, such as interleukin (IL)-6, are counter-regulated by IL-10 with a large interindividual variability. Part of the variability of cytokine production is genetically determined since polymorphisms in the cytokine gene promoters lead to high or low production. The aim of this study was to detect the genetic influence of the IL-10 promoter on immune function of chronic hemodialysis patients. METHODS: The IL-10 genotype (polymorphic bases at positions -1082 and -819) was determined in 272 chronic hemodialysis patients using highly specific PCR and related to the patients' response to a triple vaccination against hepatitis B. Secretion of IL-10 and IL-6 by peripheral blood leucocytes in vitro was determined by ELISA. RESULTS: The prevalence of the IL-10 genotypes in dialysis patients with well-preserved immune function (vaccination responders) was similar to the general population. In contrast, prevalence of the -1082G* allele (associated with high production of IL-10) was low in the nonresponders. The relative risk of vaccination nonresponse in patients homozygous for the -1082A* allele was 1.394 (95% CI, 1.091 to 1.781, P < 0.05) compared to those homozygous for -1082G*. There was no relationship between the IL-10 genotype and the type of renal disease. CONCLUSIONS: The IL-10 genotype determines IL-10 production in dialysis patients, which down-regulates uremia- and dialysis-induced chronic inflammation and helps to preserve immune defense functions.

Propofol as a sole agent for paediatric day care diagnostic ophthalmic procedures: comparison with halothane anaesthesia.

BACKGROUND: Our aim was to study the feasibility of total intravenous anaesthesia with propofol in spontaneously breathing children undergoing ophthalmic procedures. METHODS: Fifty-five children (aged 6 months to 5 years) were randomly allocated to receive either propofol bolus (until loss of eyelash reflex) followed by infusion [group P (n=29)] or halothane 3-4% for induction, followed by 1-2% in 70% nitrous oxide and oxygen via face mask [group H (n=28)]. Dose for induction and maintenance, intraoperative adverse events, time to recovery (on an Observer's Assessment of Alertness/Sedation Scale, 5 at each level) and duration of procedure were recorded. All children in both groups, were anaesthetized successfully. RESULTS: 4.0 +/- 0.7 mg x kg(-1) and 5.1 +/- 1.0 mg x kg(-1) of propofol were required for loss of eyelash reflex and tolerance of the ophthalmic speculum, respectively. An infusion rate of 8.3 +/- 1.7 mg x kg(-1) x h(-1) was needed for maintenance of anaesthesia; 3.4 +/- 0.5%, 3.6 +/- 0.4% and 1.4 +/- 0.4% halothane was needed for induction, tolerance of the eye speculum and maintenance of anaesthesia, respectively. Induction and recovery were significantly faster with halothane compared with propofol [induction - 38.3 +/- 6.6 s (group H)/60.9 +/- 15.2 s (group P) (P < 0.001); recovery 12.8 +/- 4.6 min (group H)/27.0 +/- 23.3 min (group P) (P < 0.001)]. Apnoea, coughing and breath-holding were seen only in group H. Group P had significantly higher incidence of involuntary movements (minor degree) (n=6) (P < 0.01). CONCLUSIONS: Propofol is a feasible option for paediatric diagnostic ophthalmic procedures with the advantage over halothane of providing complete access to the eye.

Strong depletion of CD14(+)CD16(+) monocytes during haemodialysis treatment.

BACKGROUND: The immune defect in haemodialysis (HD) patients is associated with a monocytic dysfunction, including an increased production of proinflammatory cytokines. Monocytes fall into subpopulations comprising CD14(++)CD16(-) and CD14(+)CD16(+) cells. Circulating numbers of the latter can rapidly increase during infectious episodes and inflammation. METHODS: We determined the amount of CD14(+)CD16(+) monocytes in HD patients and characterized their fate during HD treatment. In 34 HD patients and 17 healthy controls, the distinct cell populations were determined by differential blood counts and flow cytometry. Cells from 14 HD patients were analysed at the start, 10, 30 and 120 min thereafter, and at the end of HD treatment. RESULTS: Before HD, patients show a monocytosis with a strongly increased CD14(+)CD16(+) subpopulation. Early during HD treatment, circulating leukocyte numbers decrease, with monocytes being most profoundly influenced. Interestingly, among them, sequestration is most pronounced in the CD14(+) CD16(+) subpopulation. After 30 min, approximately 83+/-9% of CD14(+)CD16(+) cells are removed from circulation. This sequestration does not differ between patients treated with polyamide or haemophan membranes. The sequestration is a short-lived temporary effect and cell numbers are replenished within 120 min of treatment for the entire monocyte population. Beyond that time point, cellular activation by the dialyser membrane becomes visible. Reappearence kinetics of CD14(+)CD16(+) monocytes is slower; however, initial numbers are reached by the end of treatment. CONCLUSION: Haemodiaysis leads to temporary removal of monocytes from the bloodstream followed by the reappearance of activated cells. This might contribute to the state of chronic microinflammation, which is reflected by high levels of CD14(+)CD16(+) monocytes.

Postpartum acquired haemophilia: clinical recognition and management.

Postpartum acquired haemophilia is a rare but serious complication of an otherwise normal pregnancy. Patients usually present with postpartum haemorrhage (PPH) or uncontrolled bleeding following surgical interventions, which fail to respond to conservative treatment. A high index of clinical suspicion along with early laboratory diagnosis and prompt institution of appropriate therapy is essential for the management of acute bleeding episodes. Our patient, a 32-year-old female, presented with severe PPH and shock. She had undergone dilation and curettage three times, with subsequent total abdominal hysterectomy and internal iliac artery ligation, before she was diagnosed with acquired haemophilia (factor VIII autoantibodies) and an inhibitor level of 8 Bethesda units (BU). The patient underwent an abdominal laparotomy for removal of the abdominal packing used in the previous operation, and blood and blood clots, and was given FEIBA(R) therapy. The patient responded to these measure and the factor VIII inhibitor level decreased to 2 BU at the time of discharge 10 weeks later.

Selective sequestration of cytokine-producing monocytes during hemodialysis treatment.

Hemodialysis treatment leads to leukocyte activation and cytokine production. Studying this effect has been complicated because cell activation by blood membrane contact also induces adherence factors on leukocytes, leading to margination of cells to the endothelium of the lung. Using single-cell cytokine determination, we studied the relation between cytokine production and cell sequestration during dialysis therapy. Blood was sampled in 11 chronic hemodialysis patients using hemophane dialyzers before hemodialysis and at 20 and 120 minutes of treatment. Lipopolysaccharide (LPS)-induced cytokine production in monocytes was studied by intracellular staining for interleukin-6 (IL-6) and IL-10 and flow cytometry. Results obtained in dialysis patients were compared with samples from an ex vivo dialysis system. Monocyte maturation stage was evaluated by detection of several surface markers through flow cytometry. Within 20 minutes of hemodialysis, the numbers of circulating monocytes decreased to one third of initial values. Before dialysis, 56.7% +/- 15.7% of circulating monocytes responded to LPS by the production of IL-6. This fraction decreased to 21.1% +/- 17.3% (P < 0.001 versus before hemodialysis) at 20 minutes and 32.3% +/- 13.8% (P < 0.001 versus before hemodialysis) at 120 minutes of treatment. A similar decrease occurred for IL-10. Cytokine-positive cells did not decrease during ex vivo dialysis. Surface marker studies showed that mature monocytes expressing HLA-DR or CD86 were predominantly removed. We provide the first evidence for a subtype-specific sequestration of monocytes caused by dialysis treatment. Fully differentiated cells capable of cytokine production and antigen presentation are removed and relatively immature cells remain in circulation.