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  • Hellberg, Carina (författare)
  • β2 integrin-induced signal transduction in granulocytic cells
  • 1997
  • Doktorsavhandling (övrigt vetenskapligt)abstract
    • <p>Neutrophil granulocytcs are highly mobile, phagocytic white blood cells whose motile functions are critically dependent on adhesion receptors belonging to the B2 integrin subfamily. These integrins are vital for neutrophil functions due to their ability to generate intracellular signals. The B2 integrin~rnediated phagocytosis of complement-opsonised particles is believed to be regulated by PKC. Particle engulfment has previously been found to be accompanied by accumulation of DG, a lipid known to activate PKC. In the present experiments, this DO originated mainly from PLD-mediated hydrolysis of phosphatidylcholinc. The subsequent activation of PKC was demonstrated by the ability of this kinase to phosphorylate the PKC~ specific substrate MARCKS. Moreover, PKC regulated the activity ofPLD in a positive way and thereby maintained its own activity. PKC has also been shown to phosphorylate the B2 subunit of the integrin. Although this phosphorylation does not regulate the avidity of the integrins, it may be needed for the induction of intracellular signals. However, antibody-induced ligation of B2 integrins did not alter the phosphorylation status of these integrins, indicating that this phosphorylation is not involved in the generation of second messengers. In contrast, pretreatment of cells with phorbol esters inhibited the tyrosine kinase activation and the subsequent rise in cytosolic free Ca2+ in a way that correlated with its ability to induce a phosphorylation of the B2 subunit of the integrin. Therefore, a PKC-mediated phosphorylation of this subunit may represent a mechanism for other receptors to regulate the signalling capacity of B2 integrins.</p><p>The B2 integrin-induced ca2+ signal was shown to be due to both a release of ca2+ from intracellular stores and an influx across the plasma membrane, Moreover, integrin ligation induced a tyrosine phosphoryialion of PLCy2 accompanied by a small formation of Ins(1,4,5)P3, indicating that PLCy2 is responsible for the intracellular release of ca2+. Further evidence was provided by the findings that an activation tyrosine kinase(s) precedes, and is a prerequisite for, the rise in cytosolic free ca2+ concentration. The Src family tyrosine kinase Fgr is activated by B2 integrin-dependent neutrophil adhesion, and this occurs through mechanisms that are yet unidentified. The present results show that ligation of B2 integrins elicited a rapid and transient association of Fgr with B2 integrins. The tyrosine kinase inhibitor genistein did not affect this association, but it did prevent the subsequent dissociation. The interaction between these two proteins could represent an initial step in the B2 integrin-induced activation of Fgr, whereas the dissociation of Fgr from B2 integrins appears to be dependent on the activation of a tyrosinekinase, probably Fgr itself.</p>
  • Helmrot, Ebba (författare)
  • Systematic analysis of a radiological diagnostic system : A method for application in the effective use of x-rays in intraoral radiology
  • 1996
  • Doktorsavhandling (övrigt vetenskapligt)abstract
    • <p>The effective use of an imaging system in diagnostic radiology implies an optimisation process. This demands knowledge and relevant descriptions of the various components, the X -ray units, the objects and receptors. Thus, in this work photon energy spectra have been measured, phantomsresembling human tissue constructed and receptor characteristics investigated. Essential image quality parameters (contrast, signal-to-noise ratio) and a radiation risk indicator for the patient (energyimparted) have been defined and their variation with photon energy spectrum determined for different objects and details. Results are presented as basic infonnation to be used by radiologists in collaboration with physicists in optimising examinations according to the patient and the particular diagnostic task.</p> <p>For intraoral radiology with Ultra-speed fihn, it is shown that to achieve equalised radiographic contrasts of an ivory wedge, kV-settings have to be decreased by 5-8 kV when single pulse X-ray generators are replaced by high-frequency constant potential ones. When imaging the ivory wedge in a PMMA phantom with Ektaspeed and Ultra-speed films at equal contrast, 7-9 kV lower kV-setting must be used with the Ektaspeed film. Ektaspeed film then gives a 35-40% decrease in the energy imparted tothe patient which can be compared to 45-55% decrease observed if the loss of contrast is not compensated for by lowering the kV-setting. Comparison of the contrasts of both films shows that Ultra-speed has higher contrast than Ektaspeed, but the latter has a wider dynamic range and higher values of base and fog optical densities, which contribute to its lower contrast at low optical densities. The radiographic contrast of details in the object is the product of object and film contrast. Objectcontrast depends on photon energy and is the same for both films. The energy imparted to the patient is calculated using conversion factors derived in this work.</p> <p>To simulate the large variety of anatomical structures encountered in intraoral radiology, a multimaterial compound hard tissue phantom was constructed. The constituent elements and their fractions by weight were carefully determined so as to allow computational methods to be used tocomplement experimental data. Different types of imaging system imply different optimal photon energy spectra. Strategies of optimising photon energy spectra with respect to image quality parameters and patient dose are described for both fihn and a digital system (Digora) using an imaging plate. In the digital system, the characteristics of the receptor affect image acquisition similarly to film but digitalized image formation and display may limit image quality (contrast resolution).</p>
  • Henriksson, Chris, 1938- (författare)
  • Living with fibromyalgia : A study of the consequences for daily activities
  • 1995
  • Doktorsavhandling (övrigt vetenskapligt)abstract
    • <p>Fibromyalgia (FM) is a chronic generalized muscle pain syndrome. This thesis is a study of 116 patients with FM, twenty living in the USA and the rest in Sweden, all meeting the classification criteria proposed by the American College of Rheumatology, and 39 patients with chronic regio!Jal muscle pain. The purpose was to explore the consequences of the fibromyalgia syndrome for the activities of daily life. The consequences of the syndrome have, been described from the patients' perspective. Data have been collected through different types of questionnaires, 250 semi-structured diaries, and qualitative analysis of 40 semi-structured interviews. A five-year follow-up study was performed to collect data on changes over time in symptoms and psychosocial consequences.</p><p>The results show that the fibromyalgia syndrome influences daily life profoundly. Both pain and muscular fatigue are continuous, though there are changes in severity, both during the day and over longer periods. Most activities are difficult to perform, at least over a prolonged period. The time structure of the day is disrupted and most patients have to adjust habits and roles in order to manage their life situation. There is acontradiction between the level of disability experienced by the patients and their healthy and non-disabled appearance. This leads to misunderstandings and influences the selfimage. The results in this thesis indicate the importance of early intervention, where the patient is given information about the condition and support to adjust to the limitations, thus preventing unnecessary disabilities and handicaps from evolving.</p><p>The comparison between patients with chronic regional muscle pain and fibromyalgia indicates that there is a quantitative, rather than a qualitative, difference between the two conditions.</p><p></p>
  • Henriksson (Lysholm), Marketta (författare)
  • Rehabilitation and Evaluation after Anterior Cruciate Ligament Injury : Function, Stability and Postural Control
  • 1999
  • Doktorsavhandling (övrigt vetenskapligt)abstract
    • <p>Traditionally, the success of treatment after anterior cruciate ligament (ACL) injury has been judged from the recovery of muscle strength and knee range of motion (ROM), but also from the static knee stability after ACL reconstruction. However, since the sensory role of the ACL and surrounding tissues for the functional stability of the knee has been documented, the measurement of proprioceptive function related to knee stability is likely to become important in this evaluation.</p><p>The main purpose of this thesis was to critically examine some generally accepted principles in the treatment after ACL injury and in the way the outcome of this treatment is judged. In addition, we introduce a dynamic posturography system in order to functionally assess the peripheral contribution to postural control in patients with a chronic ACL insufficiency and in patients after ACL reconstruction.</p><p>The hypothesis tested was that dynamic measurement of anterior-posterior (AlP) laxity would be of greater value than the currently used measurement of static AlP laxity for assessing the result of ACL reconstruction and that these measurements could also be used for selection of rehabilitation exercises. We found that open kinetic chain exercises provoked larger sagittal plane translation than closed kinetic chain exercises and therefore should be avoided in early ACL rehabilitation. The present study furthermore shows that dynamic laxity measurements may be clinically useful to identify ACL injured patients, who are unable to limit the anterior translation of tibia during dynamic exercise.</p><p>A prospective randomized comparison in patients after ACL reconstruction did not reveal any differences with respect to ROM, static total AlP laxity and subjective knee score and activity level between patients postoperatively treated with an early ROM training and patients treated with five weeks immobilization in plaster. On the contrary, at the 2-year follow-up, the strength deficit in the hamstring and quadriceps muscles tended to be larger in the early ROM trained than in the plaster group (p&lt;0.05 for hamstrings and p=0.07 for quadriceps), possibly related to the greater need for involvement of the physical therapist in the plaster group.</p><p>It was found that still several years after injury, non-operated ACL injured patients had a significantly greater body sway while standing on the injured limb but also on the uninjured limb on a stable surface. Furthermore, the measurements during perturbations in the sagittal plane showed a longer reaction time and a greater sway amplitude in the single-limb stance among patients than among c:ontrols, with no difference between the injured and the uninjured leg. On the other hand, following ACL reconstruction most measurements of postural control were not different in patients than in controls (36 months postoperatively). The normalization of postural control as well as in subjective knee score and activity level occurred in spite of the total sagittal laxity being significantly higher in the ACL reconstructed knee than in the uninjured knee and in the control group. However, the patients still had significantly longer reaction times for the postural responses to perturbations forwards and backwards, with no difference between the patients' ACL reconstructed and healthy leg. The impaired postural control in ACL deficient patients both while standing on the injured limb and on the uninjured limb, as well as the lack of correlation between the measured laxity of the reconstructed knee and any of the variables in perturbations provide evidence that rehabilitation with proprioceptive and agility training is an important part of the treatment to condition the dynamic restraints and thus improve the functional stability in the ACL reconstructed knee.</p>
  • Hensing, Gunnel (författare)
  • Sickness absence and psychiatric disorder : epidemiological findings and methodological considerations
  • 1997
  • Doktorsavhandling (övrigt vetenskapligt)abstract
    • <p>Psychiatric disorder is an important health problem among people of working age, with consequences for work capacity and sickness absence. Increased knowledge of sickness absence and psychiatric disorder is important for early detection of psychiatric disorders and appropriate rehabilitation measures.</p><p>The occurrence of psychiatric sick-leave (sick-leave spells &gt;7 days) during the years 1985, 1986 and 1987 was analysed in the county of Östergotland (400 000 inhabitants). Women working in male-dominated occupational groups, such as industrial work and as craftsmen, had the highest incidence. Men working in extremely female-dominated occupational groups, social workers and secretaries, had the highest incidence among all men. Occupational groups with an equal sex distribution had the lowest psychiatric sick-leave. More women were sick-listed, but men had more sick-leave days. There were small sex differences in the pattern of sickleave diagnosis.</p><p>The role of psychiatric disorder for sick-leave in all diagnoses (not only psychiatric) was analysed in a stratified sample of 292 women selected from a population-based study of the female general population ofGoteborg (425 000 inhabitants). Women with psychiatric disorders according to a research interview had higher general sickness absence over ten years (1981-1990) both regarding spells and length, also when stratified for age, functional capacity, physical health, marital status, motherhood and socio-economic group. Single women with children did not have any increased sick-leave unless they had a psychiatric disorder. Women with psychiatric disorders also had an increased number of sick-leave days in diseases of the locomotor system, the digestive system and in mental illness. Women with alcohol problems had high sick-leave due to· diseases of the locomotor system. Eighty-nine per cent of the women with psychiatric disorders were sick-listed at least once 1989-1990 but only 16% were sick-listed with mental illness. Unrecognised psychiatric disorder is suggested as an explanation.</p><p>In the methodological studies, critical evaluations of population at risk, sick-leave measures and estimation of person-time were made. Five sick-leave measures; frequency, length, duration, cumulative incidence and incidence rate, were defined and tested in a pilot study. Results from the pilot study showed that women with 1-7 sick-leave days over a year had a better self-reported health than women without any sick-leave and those with more than 8 days. Methods for assessing recurrency, duration and intensity within an epidemiological framework need to be developed.</p><p>In conclusion, psychiatric disorder is an important factor in sickness absence, especially in the number of sick-leave days. The relation between psychiatric morbidity, environmental factors and individual coping behaviour for sickness absence is not clear, and further research is needed. Epidemiological and clinical research from a gender perspective is needed, with a focus on the health care and the social insurance systems. Prevention and rehabilitation programs should focus on the interplay between somatic illness, mental health and the use of health services and the sickness insurance.</p>
  • Herbertsson, Helena, 1967- (författare)
  • Studies on a novel cytosolic/nuclear binding protein for the eicosanoid 12(S)-hydroxyeicosatetraenoic acid
  • 1998
  • Doktorsavhandling (övrigt vetenskapligt)abstract
    • <p>Hydroxyeicosatetraenoic acids (HETEs) are lipoxygenase metabolites of arachidonic acid. HETEs were long considered to have few or no important biological function - a view that proved to be incorrect. These compounds have now been identified in many different types of cells, and they have been attributed several important actions, the molecular mechanisms of which are largely unclear. This thesis describes the discovery of a putative receptor for 12(S)-hydroxy-5,8,10,14-eicosatetraenoic acid (12(S)-HETE).</p><p>Specific high-affinity binding sites for 12(S)-HETE were detected in the cytosol (52%) and the nuclei (18%) of cells of the Lewis lung carcinoma (LLC) line. Similar l2(S)-HETE binding sites were detected in the cytosol of several other kinds of cells and in human platelets. Gel permeation chromatography and density gradient centrifugation indicated an apparent molecular weight of about 650 kDa and a sedimentation coefficient of20.5 S for the binding sites in the cytosol ofLLC cells. Treatment with ATP caused the 650 kDa component to dissociate into subunits. The actuall2(S)-HETE binding subunit was subsequently shown to have a molecular weight of about 50 kDa.</p><p>The subcellular distribution of l2(S)-HETE binding sites in LLC cells was found to resemble the distribution of some nuclear receptors of the steroid hormone receptor superfamily. The untransformed glucocorticoid receptor has been reported to be located in cytosol in association with heat shock proteins 70 and 90, and Western blot analyses and immunoprecipitation revealed the same two proteins in the cytosolic 650 kDa l2(S)-HETE binding complex.</p><p>A possible relationship to the steroid hormone receptor superfamily was also suggested by the observation that the 50 kDa l2(S)-HETE binding protein interacted with steroid receptor coactivator-l (SRC-l), which is known to be recruited to the site of transcriptional activity by several nuclear receptors. The interaction with SRC-1 occurred only when l2(S)-HETE was bound to its 50 kDa binding protein.</p><p>In summary, the research presented in this thesis led to the discovery of a novel type of eicosanoid receptor. This binding site resembles the nuclear receptors for steroids and related compounds by virtue of its subcellular distribution, the inclusion of heat shock proteins in its binding complex, and its strictly ligand-dependent interaction with SRC-l.</p>
  • Hermanson, Ola (författare)
  • Molecular mechanisms of nociception in the rat brain : anatomical connections and trans-synaptic regulation of gene expression of neurons in the pontine parabrachial nucleus
  • 1997
  • Doktorsavhandling (övrigt vetenskapligt)abstract
    • <p>The pontine parabrachial nucleus (PB) is a major recipient of fibers fromnociceptive (pain-responsive) spinal and trigeminal dorsal horn neurons. With the use of combinations of molecular and morphological techniques, the anatomical connections of PB neurons and their expression of neuropeptide genes and transcription factors were studied, both in naive rats and in rats that had been subjected to nociceptive stimulation.</p><p>A large number of neurons in PB expressed preproenkephalin mRNA(ppENK), which implies that these neurons synthesize the opioid enkephalin. One region of PB, the internal lateral subnucleus, contained many ppENKexpressing neurons that projected to the thalamic intralaminar nuclei, suggesting a role for these neurons in attentional mechanisms. Another region, the Kölliker-Fuse subnucleus (K-F), contained many ppENK-expressing neurons with descending projections to regions in the ventral medulla and spinal cord involved in pain control and autonomic modulation. The ppENKexpression in K-F increased after nociceptive stimulation of the neck or tail, suggesting that supraspinal opioidergic neurons with descending projections are activated by pain stimuli.</p><p>Stimulation of the K-F region has been shown to elicit analgesia inanimals and humans. This analgesia has previously been ascribed to theactivation of noradrenergic neurons located in and close to K-F (the A7 cell group). Double-labeling procedures demonstrated that the ppENK-expressing neurons constituted a separate, much larger population, interspersed with the noradrenergic neurons. Thus, analgesia elicited by stimulation of K-F must involve the activation of opioidergic neurons.</p><p>In search for the intracellular proteins that mediate the noxious-inducedup-regulation of ppENK, the expression of FOS in PB was studied after various types of nociceptive stimulation. FOS expression is low in PB of the naive animal, but FOS is rapidly produced after trans-synaptic activation, and it can bind to the DNA site that regulates ppENK transcription. However, noxiousevoked FOS was predominantly expressed in ppENK-negative neurons in PB. Another protein, CREB, can also regulate ppENK transcription. Therefore, the expression of active, phosphorylated, CREB was investigated in PB after nociceptive stimulation and was found to be present in the majority of theppENK-expressing neurons in K-F and the internal lateral subnucleus. These findings suggest that phosphorylation of CREB can mediate noxious-induced up-regulation of ppENK transcription in PB.</p><p>Following nociceptive stimulation of the neck, tail, and hindlimb, most ofthe FOS was found in the dorsal and the superior lateral subnuclei of PB. The results suggest that neurons in these subnuclei are activated trans-synaptically by a direct spinal nociceptive input. The expression of FOS in PB after nociceptive stimulation of the face was found to be different from that seen after nociceptive stimulation of other parts of the body. Most of the FOSexpressing neurons were now detected in K-F, the external lateral, and the external medial subnuclei. This result is coherent with the termination pattern of fibers from the trigeminal dorsal horn, and suggests that neurons that are activated by nociceptive stimulation of the face have other functions than those activated by stimulation of other regions.</p><p>Since FOS can bind to the ppCCK gene and regulate cholecystokininproduction, the parabrachial expression of preprocholecystokinin mRNA(ppCCK), which encodes the neuropeptide cholecystokinin, was investigated. Many ppCCK-expressing neurons were present in the superior lateral subnucleus, and these neurons displayed FOS after noxious stimulation. In addition, many ppCCK-expressing neurons in PBsl displayed the second messenger Ca2+/calmodulin-dependent kinase Il (CaMKII) and, after noxious stimulation, phosphorylated CREB. These findings point to a putative intracellular route for noxious-induced calcium-mediated regulation of ppCCK transcription through FOS induction via CaMKII and CREB. The ppCCKexpressing neurons in the superior lateral subnucleus were shown to project to the ventromedial hypothalamic nucleus, suggesting that they influence blood glucose homeostasis as a response to tissue damage.</p><p>In search for a neuropeptide that is expressed by noxious-activated(FOS-1abeled) neurons in the dorsal lateral subnucleus, the preprodynorphin mRNA (ppDYN) expression was investigated. ppDYN encodes the opioid dynorphin, and the transcription of ppDYN can similar to other neuropeptide genes be regulated by the FOS protein. The dorsal lateral subnucleus was rich in ppDYN-expressing neurons, and many of the ppDYN-expressing neurons contained FOS, which implies that they are activated by nociceptive stimulation. The ppDYN-expressing neurons were found to project to the hypothalamic median preoptic nucleus, suggesting an involvel)lent of these neurons in cardiovascular regulation.</p><p>In summary, the present study demonstrates that both opioidergic andnon-opioidergic neurons in a brainstem region, PB, are trans-synapticallyactivated by nociceptive stimulation in the awake rat. It is shown thatnociresponsive neurons in PB are organized in distinct cell groups that display different anatomical connections, intracellular signaling mechanisms, and gene expressions, suggesting different functions for these cell groups. It is suggested that nociresponsive parabrachial neurons influence the tuning and excitability of autonomic reflex circuits as part of an integrated response to tissue damage.</p>
  • Hermansson, Ann-Charlotte, 1945- (författare)
  • War-wounded refugees : A prospective study of well-being and social integration
  • 1996
  • Doktorsavhandling (övrigt vetenskapligt)abstract
    • <p>This thesis is a prospective study of a culturally heterogenous group of war-wounded refugees, who arrived in Sweden in the late 1980s. The general aim was to describe the situation of warwounded refugees from medical, psychological and social perspectives after arrival and aftertwo years in Sweden and, if possible, to identify factors of significance for their well-being and social integration. The study group comprised 61 war-wounded patients admitted to somatic care. The patients were investigated during hospitalization, shortly after arrival in Sweden, andfollowed up after two years. The data collection covered medical, psychological and social conditions and was performed by personnel on the ward using interviews and rating scales. All data collection was performed with the help of interpreters.</p> <p>From clinical experience and a pilot study, it was expected that war-wounded refugees would be specially vulnerable. In addition, the literature on refugee migration and war trauma suggested that war-wounded refugees could be a high risk group for decreased well-being and adjustment difficulties in countries of resettlement. The type of injuries and medical complications observed in the study group were representative of small unit operations of war with poor access to early medical care. Characteristics of the refugees' background were:obstructed or interrupted schooling and working life; long periods of duty as guerillas, or other political activities; a high frequency of death and separation of family members; a high frequency of imprisonment and torture; unfavourable conditions prior to the flight. Thus, the group was heavily burdened with respect to risk factors for decreased well-being after resettlement. However, the quantitative data analysis showed that these factors had not had the influence expected on well-being following arrival or after two years in the host country.Instead, the analysis indicated that life circumstances and events related to the present situation, "here and now", were more important for their well-being and social integration. With respect to labour market attachment, the war-wounded group was found to follow wellthe expected labour market integration process for refugees. On group level, well-being had not improved after two years compared with well-being following arrival. The qualitative findings suggest that in this early stage of migration, the refugees are mourning their losses but are not observably occupied by working through the pre-migration war traumas.</p>
  • Hernefalk, Leif (författare)
  • Internal fixation of femoral neck fractures
  • 1996
  • Doktorsavhandling (övrigt vetenskapligt)abstract
    • <p>The incidence of femoral neck fractures in elderly people is rising requiring large health care resources. Internal fixation of the fracture is the treatment of choice in Scandinavia, but in 20-40% of the cases a late complication such as a segmental collapse of the femoral head after initial bone necrosis, or a non-union of the fracture because of non-healing, will occur. Because of the high disability associated with these local complications, revision surgery is needed, and in such a case the cost more than triples. Reduction of these local complications would avoid suffering and costs. The present study focuses on the analysis of technical principles for device design and internal fixation of femoral neck fractures, with the aim to reduce the number of local complications due to non-healing. The analysis is based on animal experiments, evaluation of patients who were treated with different internal fixation techniques, and finally biomechanical in-vitro tests of osteotomized femur specimens, which were stabilized wilh different internal fixation devices.</p><p>The results indicate that late complications are associated with early pronounced femoral shortening; monitoring of femoral length had, therefore, a high accuracy (92%) in predicting the late outcome. A shortening of more than 5 mm at one month predicted a greater lhan 6-fold increase of the incidence oflate complications. Use of a device that enabled a rigid osteosynlhesis reduced the incidence of non-unions, thereby reducing the overall local complication rate by one-third, and the need for revision surgery was halved. The rate of late segmental collapse, which occurred in 14-19%, remained independent of choice of device.</p><p>The healing of the fracture due to a stable fixation may accelerate the course to a segmental collapse in cases with initial osteonecrosis of lhe femoral head, due to vascular impairment at injury. Otherwise a stable fracture fixation, preventing a high postoperative deformation at the fracture site, was found to be a prerequisite for optimal fracture healing. Such a stable fixation can be optimally achieved with a device incorporating two or more threaded pins or screws, preferably cannulated, which are implanted With an a traumatic technique with high precision using guide systems to prevent unnecessary damage to bone. The optimal device for internal fixation of a femoral neck fracture should also have a good grip in osteoporotic cancellous ancl subchondral bone; the good grip of suitable threads may be enhanced by bone cement.</p>
  • Hildén, Jan-Olof (författare)
  • Anti-D antibodies : In vitro tests to predict the severity of hemolytic disease of the newborn
  • 1997
  • Doktorsavhandling (övrigt vetenskapligt)abstract
    • <p>The ability of different in vitro tests to predict the severity of hemolytic disease of the newborn (HDN) has been studied. When properly performed the traditional indirect antiglobulin titers (IAT) on maternal serum can be used to identify a low !iter group ~32 in which the fetus will not suffer from severe hemolytic disease and a high titer group ~1000 where all newboms will be affected. However, in the titer interval64-512, the prediction by IAT titers is poor and additional tests are needed. Widely used are determination of the anti-D concentration in maternal serum and measurement of the bilirubin concentration inamniotic fluid, the latter requiring amniocentesis (AMC). The negative predictive value using bilirubin measurements was 0.53 and using anti-D concentration determination 0.73. This shows that no further information is gained by adding amnion fluid examination if anti-D concentration determination is carried out. Moreover, as invasive procedures such as AMC may be dangerous to the fetus, it is preferable to use measurements of anti-D concentration, requiring only a venous blood sample from the mother.</p> <p>AutoAnalyzers have been used for 25 years and are still the main instrument for determining anti-D concentration. As this test is important in the prediction of HDN, there is an obvious need for a modern and reliable method of anti-D quantitation. A new method using flow cytometry was developed and found to give exactly the same classification as the AutoAnalyzer in 89% of cases. In the !I% that differed, the flow cytometer found more relevant cases than the AutoAnalyzer.</p> <p>The IgG subclass of the maternal anti-D can be of importance for the outcome in the newborn. To perform IgG subclassing of anti-D, a method using the gel technique was elaborated. Among severely affected cases, two or more subclasses were detected in 77%, compared to 20% in moderately or unaffected cases. When used as a complementary test in borderline cases having anti-D concentrations 0.6-1.4 jlg/mL_, no cases at risk of HDN would be missed and cases where additional invasive procedures must be performed would bereduced by about 60% (in the present series from 22to 9).</p> <p>HLA typing revealed the HLA DQB 1 a!lel *020 I to be four times more common in women with severe immunizations than in those with low levels of anti-D. Typing for this allel may be a means of identifying women with a high risk of developing severe immunizations.</p>
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