Flow cytometric determination of RBC survival in autoimmune hemolytic anemia

BACKGROUND: In cases of warm autoimmune hemolytic anemia (WAIHA), crossmatch incompatible RBCs are most often used for transfusion. The determination of the in vivo survival of transfused and autologous RBCs in WAIHA is helpful in the assessment of the efficacy of transfusion and other therapeutic interventions. CASE REPORT: A 38-year-old man presented with acute WAIHA, thrombocytopenia, and neutropenia. Steroids and IVIG therapy were ineffective, and the patient received RBCS: Because of increasing hemolysis and persisting thrombocytopenia, splenectomy was performed, resulting in partial remission. Further improvement was achieved by immunosuppressive therapy. MATERIALS AND METHODS AND RESULTS: Survival of transfused and autologous RBCs was determined, using a flow cytometric method based on the determination of different blood group antigens of patient and donor RBCS: The survival of autologous and transfused RBCs before splenectomy was determined on two consecutive days. The life span of autologous RBCs remained rather stable at 69 and 64 hours on Days 10 and 11, respectively, whereas the life span of transfused RBCs decreased from 186 hours to 25 hours. After splenectomy, the life span of transfused RBCs almost normalized: 43 days at postsplenectomy Day 3 and 87 days at postsplenectomy Day 69. CONCLUSION: Flow cytometry was successfully used to determine changing hemolytic activity during the clinical course of WAIHA. Additionally, the survival of transfused RBCs could be measured, which may be helpful to judge for the compatibility of allogeneic RBCS: Thus, we were able to show the therapeutic inefficacy of steroids and immunoglobulins, and quick improvement after splenectomy.     

Status of oral contraceptives as a risk factor in cerebrovascular diseases

A critical review of the relevant literature revealed that the incidence of subarachnoid haemorrhage is increased in women taking oral contraceptives and the mortality rate is higher. The data concerning correlation between the use of oral contraceptives and other cerebrovascular disorders are less conclusive. Taking oral contraceptives seems to result in a higher risk of stroke, but there is no definite correlation to the incidence of strokes with a fatal outcome. However, in the case of the coexistence of more than one risk factor, including cigarette smoking, the risk of cerebrovascular disease is considerably increased for women using oral contraceptives. Age also seems to be a significant factor. Women older than 44 should avoid oral contraceptives in general; women between 35 and 44 should use oral contraceptives only if additional risk factors are absent, i.e. if they are non-smokers. Indeed, in the presence of this or other risk factors younger women should also avoid taking oral contraceptives.     

Psychological distress and caregiving experience during the first two years of the COVID-19 pandemic among parents of an offspring with anorexia nervosa

Objective

To compare different aspects of caregiving distress and experience in parents of patients with anorexia nervosa (AN) before and during the COVID-19 pandemic.

Methods

Three cohorts of parents of AN patients (cohort 1-pre-pandemic: N = 78, cohort 2-first pandemic year: N = 51, cohort 3-second pandemic year: N = 119) were recruited from child and adolescent psychiatry wards and cross-sectionally assessed as part of the clinical routine. Quantitative measures of psychological distress, psychopathology, eating disorder (ED)-related burden, expressed emotion and caregiver skills were obtained at the beginning of the child's inpatient or outpatient treatment.

Results

Cohort 2 showed lower levels of anxiety and a tendency of lower emotional overinvolvement and higher caregiving skills compared to the pre-pandemic cohort. In contrast, the levels of general psychological distress, depression, ED-related burden and criticism observed in cohort 3 significantly exceeded pre-pandemic levels. The prevalence of clinically relevant depression was higher in cohort 3 (41.5%) compared to cohorts 1 (24.4%) and 2 (21.6%).

Discussion

The pandemic effects on parents seem to be time-specific. Lower distress in the early phase of the pandemic may be associated with improvements in parent-child-relationships reported in previous studies. However, the pandemic may has negative consequences in the long-term emphasising the need of ongoing parental support.     

Pharmacokinetics of an oral contraceptive containing oestradiol valerate and dienogest

Objective?To evaluate the pharmacokinetics of a combined oral contraceptive (OC) containing oestradiol valerate/dienogest (E₂V/DNG) administered according to a four-phasic dosing regimen with an oestrogen step-down and a progestin step-up over 26 days of active treatment.

Methods?This Phase I, open-label study included healthy women aged 18–50 years. Treatment consisted of the administration of E₂V 3 mg for 2 days, E₂V 2 mg/DNG 2 mg for 5 days, E₂V 2 mg/DNG 3 mg for 17 days, E₂V 1 mg for 2 days, and placebo for 2 days.

Results?Pharmacokinetic data were analysed in 15 women. Stable E₂ concentrations were maintained throughout the study. Minimum mean serum E₂ levels were 33.6–64.7 pg/ml during E₂V administration. The ratio of oestrone:E₂ in serum was approximately 5:1. Minimum mean serum DNG levels were 6.8–15.1 ng/ml during DNG administration. Minimum concentrations of DNG increased only slightly during each phase of the regimen during which DNG was being administered. On day 24 the geometric mean C_max, C_ave and t_½ of DNG were 82.9 ng/ml, 33.7 ng/ml and 12.2 hours, respectively; the median t_max was 1.5 hours. Serum sex hormone-binding globulin concentrations increased by 40% (within the normal range). Cortisol binding-globulin levels remained almost unchanged. Treatment was well tolerated.

Conclusions?Treatment with an OC containing E₂V and DNG was well tolerated and was associated with stable E₂ concentrations over 28 days. The pharmacokinetics of DNG were consistent with previous findings. Minimum serum concentrations of DNG increased only slightly during phases of the regimen during which DNG was administered.     

Collection of WBC-reduced single-donor PLT concentrates with a new blood cell separator: results of a multicenter study

BACKGROUND: A new cell separator (COM.TEC, Fresenius) was recently developed aimed at efficient collection of WBC-reduced single-donor PLT concentrates (SDPs). STUDY DESIGN AND METHODS: Five German centers collected 554 WBC-reduced SDPs with help of the COM.TEC cell separator. Two multicenter cell counting studies were performed at the beginning and at the end of the study to document uniform counting results among the participating centers. RESULTS: A total of 441 (79.6%) PLT collections were included in the study according to the protocol. A total of 342 single-dose and 99 double-dose SDPs were collected. For single-dose SDPs, an average blood volume of 2826 +/- 409 mL was processed in a donation time of 55 +/- 11 minutes. Mean PLT yield of these products was 3.11 x 1011+/- 0.40 x 1011 and the WBC contamination was 0.11 x 106+/- 0.20 x 106. For double-dose SDPs (PLT count, 5.29 +/- 0.93 x 1011), 3943 +/- 639 mL was processed. The average difference between the target and the collected PLT concentration was -2.8 +/- 12.0 percent for single-dose SDPs and -1.8 +/- 9.5 for double-dose SDPs, respectively. The collection efficiency was 53.7 +/- 5.8 percent for single-dose SDPs and 58.2 +/- 6.2 percent for double-dose SDPs. If all results of each sample from the counting study were set to unity (to the mean over all centers), most PLT determinations were very similar to the mean, for example, near or 1 if set to unity. CONCLUSION: The COM.TEC machine makes it possible to obtain WBC-reduced SDPs that comply with current standards.     

Cerebral angiography with the contrast medium vasobrix 32 (ioxitalamate) (author's transl)

On the basis of angiograms carried out 168 patients, the contrast qualities achieved with the contrast medium, Vasobrix 32 were compared with those of other contrast media used so far. In the case of carotid angiography, Vasobrix 32 allowed better visualisation in the arterial phase than achieved with the contrast media used so far. The superiority of Vasobrix 32 over other substances was even more evident in the venous phase. As regards visualisation of the branches of the small arterial vessels such as the striothalamic branches, the results achieved with Vasobrix 32 were better than with the contrast media used so far and, therefore, a general improvement in visualization can be expected from Vasobrix 32 (Ioxitalamate). Complications with neurological sequelae were not observed, subjective complaints occurring to a slightly lesser extent than in the case of contrast media with a high sodium salt content.     

Automated Processing of Human Bone Marrow Grafts for Transplantation

Prior to purging or cryopreservation, we concentrated 21 bone marrow (BM) harvests using a modification of the ‘grancollect-protocol’ of the Fresenius AS 104 cell separator with the Pl-Y set. Within 40–70 min, the initial marrow volume of 1,265 ml (±537 ml) was processed two to three times. A mean of 47% (±21%) of the initial mononuclear cells was recovered in a mean volume of 128 ml (+36 ml). The recovery of clonogenic cells, measured by CFU-GM assays, was 68% (±47%). Red blood cells in the BM concentrates were reduced to 7% (±4%) of the initial number. The procedure was efficient and yielded a BM cell fraction suitable for purging, cryopreservation and transplantation. At this time, 10 of the 21 patients whose BM was processed using this technique have been transplanted. Seven of these 10 patients have been grafted using the BM alone. Three of the 10 patients showed reduced cell viability and colony growth in the thawed BM samples, and therefore obtained BM and peripheral blood-derived stem cells. All transplanted patients showed an evaluable engraftment, achieving 1,000 granulocytes per μl of peripheral blood in a mean of 18 days.     

Significance of left-sided heart disease for the detection of patent foramen ovale by transesophageal contrast echocardiography.

Detection of patent foramen ovale by contrast echocardiography is based on transient inversion (right atrial pressure higher than left atrial pressure) of the interatrial pressure gradient. Therefore, the presence of left-sided heart disease with potential elevation of left atrial pressure might obscure the diagnosis of patent foramen ovale. Accordingly, 150 patients (88 men, 62 women; mean age 51.7 +/- 15.2 years) were evaluated for a patent foramen ovale by transesophageal contrast echocardiography. Additionally, atrial septal motion during normal respiration and during the Valsalva maneuver was analyzed. Patency of the foramen ovale was observed in 20 (27%) of 74 patients without left-sided heart disease and with previous arterial embolism, in none (0%) of 25 patients with left-sided heart disease and embolism, in 7 (39%) of 18 patients without left-sided heart disease and without embolism and in 3 (9%) of 33 patients with left-sided heart disease and without embolism. The detection rate of patent foramen ovale was lower in patients with than without left-sided heart disease (5% vs. 29%, p = 0.0007) but was similar in patients with and without embolism (20% vs. 19.5%, p = NS). Abnormal atrial septal motion was more frequently observed in patients with left-sided heart disease (p = 0.0003) and was inversely correlated to detection of patent foramen ovale (p = 0.0003). Multivariate analysis revealed an independent association between the absence of left-sided heart disease and the detection of patent foramen ovale (p = 0.0003). These data suggest that in patients with left-sided heart disease, patency of the foramen ovale may be missed even by transesophageal contrast echocardiography.