Frequencies of fetal nucleated red blood cells in maternal blood during different stages of gestation

High molecular weight forms of tyrosinase have been found to be expressed during spontaneous remelanization of the amelanotic B-16 melanoma cells in culture as well as in melanotic tumors formed from amelanotic melanoma cells grown in C57BL/6J mice. Overnight extraction of the crude melanosomal fractions from such tumors and cultured melanoma cells reveal the presence of an additional DOPA-MBTH positive band well below the stacking gel. This band has been found to be alpha-PEP7 (antibody specific for tyrosinase) positive and alpha-PEP1 (antibody specific for TRP-1) negative on Western blot analysis. Heat treatment at 60 degrees C for 60 min results in the loss of this band and considerable loss of activity of the melanosomal extract. Trypsin treatment of these melanosomal extracts resulted in a minor change in the mobility of the high molecular weight band. SDS-PAGE under reduced conditions followed by Western blotting revealed that the high molecular weight band was lost and not detected by alpha-PEP7 or alpha-PEP1. These findings indicate that high molecular weight, heat sensitive and trypsin resistant forms of tyrosinase are transiently expressed in B-16 melanoma cells and tumors that are initiating remelanization following phenotypic drift towards the amelanotic state.     

Frequency of nucleated red blood cells in maternal blood during the different gestational ages

PURPOSE: To study the course and visual outcome of Vogt-Koyanagi-Harada (VKH) syndrome in children. METHODS: The medical records of 97 consecutive patients with VKH syndrome were reviewed. Patients had been seen at King Khaled Eye Specialist Hospital in Riyadh, Saudi Arabia. The records were divided into two groups: those in whom the onset of the disease occurred at the age of 14 years or younger (pediatric group) and those in whom the disease manifested after the age of 14 years (adult group). RESULTS: Eight (61%) of the 13 children required cataract surgery compared to 14(17%) of 84 adult patients. Despite medical and surgical therapy, eight (61%) of the pediatric group had a final visual acuity of 20/200 or worse compared with 22 (26%) of the 84 adult patients (p=0.0103). CONCLUSION: The course of early-onset VKH (before the age of 14 years) appears to be aggressive. The ocular complications are more severe in children than in adults, leading to rapid deterioration in vision. Despite therapy, the final visual outcome of VKH in children is poor.     

Charge flow separation: quantification of nucleated red blood cells in maternal blood during pregnancy

We set out to ascertain the numbers of fetal cells that enter the maternal blood stream during pregnancy. Samples of 15-16 ml of whole blood were collected from 225 women--mostly 10-18 weeks pregnant--and then processed by charge flow separation, a novel method based on free flow electrophoresis in a buffer counterflow gradient. After their recovery in four different separation instruments, nucleated red blood cells (NRBC) were enumerated histologically. In some cases fetal NRBC were identified and enumerated by fluorescence in situ hybridization with probes for the X and Y chromosomes and fetal haemoglobin mRNA. Recoveries were consistent among the four separation instruments: the median numbers of NRBC obtained were 4190, 1590, 2805 and 3860. Our data show that approximately 30 per cent of those cells were fetal. Thus, recent reports on the separation of fetal NRBC by other methods, give underestimates of their frequency in maternal blood.     

Current red blood cell transfusion practices

The appropriate use of blood transfusions remains variable among health-care institutions and patient populations. Transfusion practices are discussed in this article in relation to medical practice guidelines and utilization review. Specific transfusion practices in the settings of intensive care, orthopedic surgery, and open heart surgery are reviewed. A new, promising approach to improving transfusion outcomes is the use of transfusion algorithms. Transfusion algorithms may prove especially useful if they incorporate point-of-care testing that is both physiologic and patient-specific for transfusion decisions. Transfusion algorithms are discussed and data presented for cardiac surgical adults.     

Normal variation in cutaneous blood content and red blood cell velocity in humans

This study was designed to determine normal variations in cutaneous blood content, red blood cell content and skin thickness in healthy human volunteers. The blood content (BC) of human skin and the velocity of red blood cells (RBV) has been comprehensively mapped over 30 anatomical sites in 28 volunteers between the age of 7 and 77 years using reflectance spectrophotometry (RFS) and laser Doppler (LD) techniques respectively. Total skin thickness, which may have a relationship with blood content, has been mapped at the same anatomical sites in six of the volunteers using two-dimensional array, 12 MHz ultrasound equipment. The major determinant of BC and RBV was found to be anatomical site. Regardless of age, gender or prior sun exposure, anatomical sites above the waist produced higher readings than below. BC and RBV were found to be dependent on age and gender at most sites with higher readings being observed in males and younger volunteers. The largest difference was seen in males under 55 years whose RFS readings above the waist were significantly higher than in older males or females of any age. Heavy prior sun exposure and skin thickness could not be demonstrated to impact on either RFS or LD readings independently of the effects of anatomical site, gender or age. In general, RFS and LD readings paralleled one another and correlated linearly at most anatomical sites. While RFS readings tended to decrease in the cranio-caudal direction, LD readings were relatively uniform across the body except for the head and groin where they were higher, and the abdomen where they were lower. The site-to-site variations in RFS readings observed in this study correspond well to capillary density variations noted in previous studies.     

Bone-marrow imaging in pure red blood cell aplasia

Bone-marrow scintigraphy with indium chloride in 111 was performed on a patient with pure red blood cell aplasia before and after successful treatment with immunosuppressive drugs. The return of erythroid precursors to the bone marrow was accompanied by a substantial increase in the marrow uptake of 111In. The distribution of 111In in the posttreatment scan was indistinguishable from that of 52Fe. These results indicate that indium chloride in 111 is a useful agent for the delineation of erythroid cellularity within bone marrow.     

Effect of bone xenograft transplantation on total T lymphocyte and its subset counts in mouse blood and spleen

Fresh calf cancellous bone xenograft (FX), antigen free cancellous bone carrier (BC) and reconstituted bone xenograft (RBX) were inplanted in the thigh muscle pouches of Balb/c mice. Histological examinations were done at 7, 14 and 28 days postoperation, and indirect immunofluorescence was used to determine the positive counts of Thy1, L3T4, Lyt2, and Tac T lymphocyte of the blood and spleen at the same time, with the normal mouse lymphocytes as control. There was no significant difference between the BC implanted group and the normal (control) group, while the FX group had a significant increase rates of Thy1, L3T4 and Lyt2, especially of Tac lymphocytes. The RBX implanted group had no increase of Tac but increase of Thy1 and Lyt2. Th-Ts ratio decreased, and it was probably caused by bovine bone morphogenetic protein (bBMP) in the RBX. Histologically, intense immune rejection was noted in FX implanted group but not in the other two groups. Heterotopic ossification was noted in the RBX implanted group. FX increased intense immunologic rejection in the host, bat RBX did not, that might be due to the decrease of Th/Ts caused by bBMP. Tac lymphocyte count may indicate the immunologic rejection of bone xenograft transplantation.     

Delayed hypersensitivity in untreated patients with Hodgkin''s disease--with special reference to small lymphocyte counts in the blood

A hundred and thirty patients have been treated with Amitriptylin in the "Clinique de Psychiatrie" in Marseille during the last eight years. The review of the hundred cases for which the information are the most detailed show that Amitriptylin remains one of the most powerful, manageable and well-tolerated anti-depressant drugs available at the present time. Its therapeutic indications are numerous, but the best ones are proceeding from depressed states with high anxious participation in subjects of less that 60 years of age.     

Intrauterine transfusions influence fetal leukocyte counts and subsets

Histamine is an important mediator in allergic reactions, gastric acid secretions, and neurotransmission in the central nervous system. Basophils and mast cells are the main sources of histamine, which is formed from L-histidine by histidine decarboxylase (HDC). However, the regulatory mechanism of HDC in these cells remains unclear. We examined the regulation of HDC activity and gene expression using a unique human mast cell line, HMC-1, after stimulation with phorbol 12-myristate 13-acetate (PMA) or ionomycin. HDC activity was increased from 52.1+/-0.4 (mean+/-standard deviation) to 154+/-6.9, or 105.6+/-6.2 pmol/min/mg protein (n = 3), 4 hours after stimulation with PMA (10 ng/mL) or ionomycin (10[-6] M). Although actinomycin D had no effect on this increase, cycloheximide completely inhibited the increase caused by these stimuli. The population of HMC-1 cells containing HDC protein was increased after stimulation with either PMA or ionomycin as evaluated by immunocytochemical analysis with anti-HDC antibody as a marker. HMC-1 constitutively expressed HDC mRNA, and its level was not increased with these stimuli. These results suggest that the increase of HDC activity in HMC-1 induced by PMA or ionomycin is regulated at the translational level.     

Rabbit red blood cell hexokinase. Purification and properties

Rabbit red blood cell hexokinase (EC 2.7.1.1.) has been purified 300,000-fold by a combination of ion exchange chromatography, affinity chromatography, and preparative polyacrylamide gel electrophoresis. The hexokinase activity has been isolated in 35% yield as a protein that is homogeneous by polyacrylamide and sodium dodecyl sulfate gel electrophoresis. The highest specific activity obtained was 145 units/mg of proteins. The native protein has a molecular weight of 110,000 by gel filtration on Ultrogel AcA 44 and 112,000 by sedimentation velocity on sucrose density gradients. Sodium dodecyl sulfate-polyacrylamide gels gave a molecular weight of 110,000 indicating that hexokinase is a monomer. The enzyme had a pI of 6.20 to 6.30 pH units by isoelectric focusing. The enzyme was specific for Mg . ATP and Mg . ITP as the nucleotide substrates. Several hexokinase with different affinities.     

Imipramine distribution among red blood cells, plasma and brain tissue

Hypertension complicates the treatment of anaemia of chronic renal failure with recombinant human erythropoietin (EPO) in some patients. We conducted a prospective study measuring changes in cardiac index (CI) and systemic vascular resistance index (SVRI) in 29 patients from before commencement of EPO to attainment of target haemoglobin concentration. We used the operator-independent technique of trans-thoracic bioimpedance. The group of patients who developed EPO-induced hypertension (EpHT) were separately analysed and compared with the group who had no change in blood pressure (NC). Our results showed there was a significant rise in SVRI after treatment in EpHT group patients but in the NC group there was a small fall. CI increased significantly in the NC group after treatment but no change was recorded in the EpHT group. These findings clearly demonstrate how the cardiovascular changes differ in patients who develop EPO-induced hypertension.     

Frequency of fetal cells in sorted subpopulations of nucleated erythroid and CD34+ hematopoietic progenitor cells from maternal peripheral blood

Fetal cells that circulate in maternal peripheral blood (PB) during pregnancy offer a potential source of nucleated fetal material for noninvasive prenatal diagnosis. Fluorescence-activated cell sorting was used to target two populations of fetal cells: nucleated erythroid cells (NECs; CD71/glycophorin-A+ CD45(lo-int) CD34-) and hematopoietic progenitor cells (CD34+ cells; CD34++ CD71/glycophorin-A- CD45(int)). Fetal cells were detected by fluorescence in situ hybridization (FISH) using directly conjugated chromosome X and Y probes in 65% (13 of 20) of the maternal PBs (fetal karyotype 46,XY). The frequency of fetal cells isolated from the NEC and CD34+ fractions was, respectively, 0 to 14 and 0 to 7 cells per 2 x 10(7) previously frozen maternal cells (approximately 20 mL of blood). In nonfrozen samples, the yield and recovery of fetal cells was moderately improved. Culturing the CD34+ sorted fractions in serum-free media with cytokines improved the quality of the FISH preparations and resulted in a slight expansion in detectable fetal cells. The frequency of fetal cells isolated from cultured CD34+ fractions was 0 to 35 and 0 to 93 cells per 2 x 10(7) previously frozen and nonfrozen maternal PB cells, respectively. These results document the isolation, characterization, and enumeration of fetal cells from the maternal periphery that appear to be present in most, but not all, samples analyzed.     

Electrophoretic difference between fetal and adult acetylcholinesterase of human red cell membranes

The membrane bound enzyme acetylcholinesterase (AChE) of human erythrocytes shows a lower level of activity in the newborn than in the adult. To evaluate if the decreased activity is correlated with changes in other properties of the enzyme, an electrophoretic method, recently devised to detect AChE activity, was used. Our results revealed an electrophoretic difference between adult and fetal enzymes. Sialic acid removal through neuraminidase treatment reduced the electrophoretic mobility of both forms rendering them identical in migration without, however, altering the activity.     

Low dose flunarizine protects the fetal brain from ischemic injury in sheep

This retrospective long-term study analyzes the clinical function, failures, and radiographic status of 131 Richards Series 2 prostheses in total hip arthroplasties (THAs) performed during 1981 and 1982. After a minimum follow-up period of 11 years, the results were evaluated. Sixty two (54%) patients with 71 THAs were available for follow-up; 44 (38%) patients with 49 THAs had died. Three patients with three THAs were lost to follow-up, and seven patients with eight THAs were unable to participate in the examination due to causes not related to their present hip status. Ten patients underwent revision THA during the observation period. The median hip functional index (HFI) was 8.6 preoperatively and 15.8 at follow-up. Hip functional index correlated to radiographic signs of loosening. With revision as endpoint, the Kaplan-Meier survivorship for all 131 hips was 93% after 10 years and 91% after 12 years. The radiographic examination at follow-up revealed 5 (8%) definite loose cups and 12 (19%) definite loose stems. Including the number of aseptic loosened cups and stems confirmed at revision, aseptic loosening was found in 6% of all cups and 13% of all stems. This radiographic evaluation confirmed that lack of containment of the cup and varus position of the stem were factors influencing the long-term stability of the prostheses, whereas etiology, type of arthritis, and ectopic ossification did not. A fixation and stability score that was modified for cemented prostheses was developed and was of value in detecting the prostheses at risk. The long-term results of the Richards Series 2 THA are acceptable and comparable with other long-term studies on conventional prosthetic designs. Ten percent to 20% revision THAs due to aseptic loosening can be expected. In addition, patients younger than age 60 at surgery and especially men were found to belong to a high-risk group with regard to revision THA. These patients must be evaluated carefully so that revision THA can be performed before the bone bed is destroyed.     

The ontogeny of key endoplasmic reticulum proteins in human embryonic and fetal red blood cells

Recently, using immunohistochemical methods, we surprisingly found that endoplasmic reticulum glucose-6-phosphatase is present in human embryonic and fetal red blood cells (RBCs) but not in adult RBCs. The fact that an endoplasmic reticulum enzyme, whose major site of expression in adults is the liver, is present in human embryonic and fetal RBCs, particularly nucleated cells, indicated that it would be sensible to determine whether these cells also contain other endoplasmic reticulum enzyme systems normally found in adult liver. Therefore, we have studied the expression of other endoplasmic reticulum proteins and found that human embryonic and fetal RBC precursors contain other protein components of the glucose-6-phosphatase system, ie, the phosphate and glucose transport proteins as well as other enzymes (eg, uridine diphosphate-glucuronosyltransferases, cytochrome P450 isozymes, nicotinamide adenine dinucleotide phosphate cytochrome P450 oxidoreductase, and prostaglandin H synthase). In addition, we also found the predominantly cytosolic markers 15-hydroxyprostaglandin dehydrogenase, prostaglandins PGE2 and 13,14-dihydro-15-keto-PGE2. The expression of key enzymes that control glucose production, detoxification of endobiotics and xenobiotics, and the regulation of prostaglandin levels in embryonic and early fetal RBCs means that these cells may have an important role in protecting the developing conceptus before it establishes an efficient circulation and before all tissues fully express their normal complement of these enzymes.     

The mechanism of chlorpromazine-induced red blood cell swelling

1. Chlorpromazine (CPZ)-induced red blood cell (RBC) swelling was investigated by determination of electrolyte, mean cell volume (MCV) and water content changes in CPZ-treated cells. 2. CPZ-induced RBC swelling is dose-, hematocrit- and pH-dependent, and is associated with a net increase in RBC monovalent cation and Cl- contents. 3. The partition coefficient (Kp) for the CPZ-RBC membrane interaction is pH dependent (Kp = 2500 at pH 7.8; Kp = 840 at pH 6.8). 4. Despite the pH dependence of Kp values an equal number of CPZ molecules is required to induce a 12% increase in MCV at pH 7.8 and 6.8.