[Indications and contraindications for platelet transfusions in patients with thrombocytopenia]. / Indikacije i kontraindikacije za primjenu trombocitnih transfuzija u bolesnika s trombocitopenijom.

For more than 40 years now, platelet transfusion has provided life-saving supportive therapy to hematological patients with impaired hematopoiesis, bone marrow aplasia induced by chemotherapy, surgical patients and patients with a variety of disorders of platelet count and function. More than 2.9 million platelet components are transfused each year in Europe and 57000 in Croatia. Patients with thrombocytopenia and coagulopathy treated at intensive care units pose special challenges. It is essential to assess the potential risk of thrombogenic side effects over the benefit of stopping and preventing bleeding before platelet transfusion in these patients. Although transfusion practices including indications and contraindications for transfusion, the dose of platelets transfused and ztransfusion trigger' are variable and in some cases the best practices are not fully known, greater harmonization of policies might promote the quality, safety and optimal use of platelet products.

Alloimmune neonatal neutropenia in Croatia during the 1998-2008 period.

PROBLEM: The aim of this study was to estimate the incidence of the disease and to analyze laboratory data of 23 newborns undergoing serologic testing for alloimmune neonatal neutropenia (ANN) during the 1998-2008 period in Croatia. METHOD OF STUDY: Laboratory data on 23 newborns undergoing serologic testing for ANN during the 1998-2008 period and epidemiologic data on the number of live births in Croatia were analyzed. Laboratory testing for ANN included serologic screening of maternal and neonatal sera and granulocytes (neutrophils) by immunofluorescence (IF) method. The monoclonal antibody immobilization of neutrophil antigens (MAINA) was employed to determine anti-HNA antibody specificity. RESULTS: Anti-HNA antibodies were detected in seven (54%) of 13 cases of serologically positive ANN. Only anti-HLA class I antibodies were demonstrated in four (31%) of 13 cases In the 2007-2008 period of prospective data collection, the number of serologically verified ANN cases was one case per 17,323 live births. Results of the prospective study conducted at Maternity Ward, Department of Gynecology and Obstetrics, Sestre milosrdnice University Hospital Center yielded the ANN incidence of one case per 2843 live births. CONCLUSION: Monitoring of neutrophil count in neonatal blood and serologic testing for ANN in case of isolated neutropenia in the newborn contributed considerably to timely detection of ANN. DESCRIPTORS: Neonatal alloimmune neutropenia-incidence, serologic diagnosis, antineutrophil antibodies, anti-HNA, anti-HLA class I, Croatia.

[Hyperhaemolytic siyndrome in patient without haemoglobinopathies]. / Sindrom hiperhemolize u bolesnice bez hemoglobinopatije.

Hyperhemolysis syndrome usually occurs in patients with sickle cell disease and possibly thalassemia who receive multiple transfusions. There are only few clinical reports on patients without hemoglobinopathies as in this report. Our patient was diagnosed with hyperhemolytic reaction and was infused with IVIG and methylprednisolone for several days. Signs of tissue hypoxia developed along with increased cardiac enzymes, hepatocellular and cerebrovascular injury, and finally death. On autopsy, there was no evidence for hemolytic uremic syndrome or thrombotic thrombocytopenic purpura.

A Case of Neonatal Neutropenia Due to Anti-Fc Gamma Receptor IIIb Isoantibodies Treated with Recombinant Human Granulocyte Colony Stimulating Factor.

Alloimmunization to granulocyte-specific antigens can occur during pregnancy. Maternal antibodies of IgG class can cross the placenta to result in alloimmune neonatal neutropenia. Antibodies to human neutrophil antigens anti-HNA-1a, HNA-1b, and HNA-2a have been most commonly reported to cause alloimmune neonatal neutropenia. Isoantibodies to Fc gamma RIIIb (CD16) if mother is a HNA-null phenotype are rarely involved in neonatal neutropenia. We report on a case of severe neutropenia (440 neutrophils/muL) due to anti-Fc gamma RIIIb (CD16) isoimmunization. On day 14 severe omphalitis developed, which was treated for 7 days by an antibiotic (ceftriaxone in a dose of 80 mg/kg/d) according to umbilical swab finding. Omphalitis persisted for 10 days in spite of antibiotic therapy and only resolved upon the introduction of rhG-CSF therapy. Therapy with rh-GCSF proved efficient and led to neutrophil count increase to 1970/muL and cure of omphalitis. However, therapeutic effect on granulocyte count was of transient nature, as granulocyte count fell to 760 n/muL on day 4 of therapy discontinuation. Neutropenia persisted for 2 months. The newborn was discharged from the hospital on day 26 with normal clinical status with clinical and laboratory control examinations at 2-week intervals. No additional infections were observed during the course of neutropenia.

Correlation between synovial fluid and serum IL-1beta levels after ACL surgery-preliminary report.

The possibility of controlling the harmful intra-articular influence of elevated interleukin (IL)-1beta synovial fluid concentration after anterior cruciate ligament (ACL) surgery could be useful. We investigated the correlation between serum and synovial fluid IL-1beta levels following ACL reconstruction. We measured IL-1beta concentration periodically in three synovial fluid and four serum samples in each of 20 patients receiving either autologous conditioned serum (ACS) containing endogenous anti-inflammatory cytokines including IL-1Ra and several growth factors (group A) or placebo (group B). A decrease in IL-1beta synovial fluid concentration appeared to be more pronounced in absolute terms in group A. In eight patients serum IL-1beta was detected on the 6th postoperative day. In four of them whose synovial fluid levels were over 10 pg/ml on the 6th postoperative day, serum IL-1beta was detected on the 10th postoperative day. The results were different in group B. Correlation between serum and synovial fluid IL-1beta appearance persists in patients after ACL surgery and ACS application. This study is an example of ACS influence on the ACL healing process controlling the IL-1beta levels on the basis of the serum IL-1beta detection.

[Neonatal alloimmune thrombocytopenia in Croatia in 1997-2007]. / Neonatalna aloimuna trombocitopenija u Hrvatskoj od 1997. do 2007. godine.

Neonatal alloimmune thrombocytopenia (NATP) is caused by maternal sensitization to paternal alloantigens on fetal platelets during pregnancy. Although the disease is rare, the severity of clinical picture and its sequels associated with central nervous system hemorrhage impose the need of an early diagnosis, and timely and specific treatment of the disease. Based on these and literature data on the prevalence of NATP in Caucasians of 1-2 cases per 1000-5000 live births, it is estimated that 10 to 50 serologically verified cases of NATP and approximately a twofold number of requests for serologic testing for suspected NATP could be expected in Croatia per year. In the present study, results of serology testing and clinical laboratory data of twenty five cases of NATP in Croatia during the 1997-2007 period are evaluated. In 20/25 cases platelet antibody screening was positive. Specific platelet antibodies were confirmed in 14/20 (70%) cases with positive screening (anti-HPA-la in 7/14, anti-HPA-5b in 5/14, and anti GP Ib-IX in 2/14 cases). Only fourteen serologically confirmed cases of NATP per -45,000 live births per year in Croatia indicate the prevalence of the disease to be considerably lower than expected.

Alloimmune neonatal neutropenia due to anti-HNA-2a alloimmunization with severe and prolonged neutropenia but mild clinical course: two case reports.

Alloimmunization to granulocyte-specific antigens can occur during pregnancy. Maternal IgG can cross the placenta and result in neonatal neutropenia. The clinical course of alloimmune neonatal neutropenia is usually self-limiting with only mild infection. However, in severe cases complicated with bacterial sepsis it is a potentially life-threatening disorder. The effect of intravenous (IV) immunoglobulin, prophylactic antibiotic therapy, and recombinant human granulocyte-colony stimulating factor is variable and may prove useful in some cases. Two cases of alloimmune neonatal neutropenia due to anti HNA-2a alloimmunization in two siblings are reported. The first neonate was administered IV gammaglobulins to increase the blood neutrophil count, at a standard dosage (0.4 g/kg body weight) for 5 days without response. The second neonate did not receive specific therapy for blood neutrophil count increase. Neutropenia persisted for 2 and 6 months, respectively. The choice and efficacy of specific therapy for neutrophil count increase in the management of alloimmune neonatal neutropenia have not yet been fully defined and require additional evaluation in the majority of cases.

Plasma cytokines as markers of aseptic prosthesis loosening.

The proinflammatory cytokines IL-1beta, IL-8, and TNF-alpha play a major role in the process of bone resorption during aseptic loosening of large joint prostheses. These cytokines secreted locally during bone resorption in aseptic loosening may enter peripheral circulation. Increased concentration of IL-1gamma, IL-8, and TNF-alpha in peripheral circulation may indicate aseptic loosening. We determined whether bone resorption could be verified by cytokine presence in plasma. We recruited 50 patients with aseptic prosthesis loosening, 50 with stable prostheses, 50 with osteoarthritis, and 50 healthy individuals. Cytokine levels were determined in plasma by ELISA tests. Patients with prosthesis loosening had higher plasma levels (IL-10, 3.7 +/- 5.5 pg/mL; IL-8, 14.7 +/- 9 pg/mL; TNF-alpha, 32.7 +/-+/- 32.4 pg/mL) than patients with stable prostheses (IL-1beta, 1.5 +/- 2 pg/mL; IL-8, 8.1 +/- 4.7 pg/mL; TNF-alpha, 22.9 +/- 18.7 pg/mL), patients with osteoarthritis (IL-1beta, 0.7 +/- 1.1 pg/mL; IL-8, 5.8 +/- 3.8 pg/mL; TNF-alpha, 9.8 +/- 7.7 pg/mL) and healthy individuals (IL-1beta, 0.7 +/- 1.1 pg/mL; IL-8, 4.2 +/- 1.3 pg/mL; TNF-alpha, 3.9 +/- 3.9 pg/mL). Our data suggest elevated plasma levels of proinflammatory cytokines may be useful as markers of bone resorption in the laboratory diagnosis of prosthesis loosening.

Frequency of HPA-15a and HPA-15b (Gov a/b) human platelet alloantigens in the Croatian population.

BACKGROUND: Human platelet antigen (HPA) genotyping is important for epidemiological studies because the prevalence of particular HPA allotypes differs among various populations and plays a major role in the occurrence of HPA alloimmunization. In Caucasians, antibodies to HPA-1a are the most important causes of neonatal alloimmune thrombocytopenia (NATP). Recent studies suggest that anti-HPA 15a/15b (Gov b, Gov a) might be the most likely candidate antibodies following anti-HPA-1a in inducing NATP. METHODS: In the present study, HPA-15 system genotype was determined by PCR-SSP method in 279 unrelated subjects from the Croatian population, yielding an HPA-15a and HPA-15b frequency of 0.53 and 0.47, respectively. RESULTS: Retrograde testing for the presence of anti-HPA-15 antibodies by use of MAIPA in 39 frozen serum samples from serologically negative cases of clinically suspect NATP produced negative results. CONCLUSION: The clinical role of anti-HPA-15 alloantibodies was unable to be confirmed.

Anti-Diego a red blood cell alloantibody as a possible cause of anemia in a 3-week-old infant.

Our aim in this case report was to describe anemia caused by anti-Diego(a) red blood cell (RBC) antibody in a 3-week-old infant derived during pregnancy to low-frequency Diego(a) RBC antigen. Pre- and postnatal maternal serum screening for unexpected RBC antibodies and determination of RBC antibody specificity in the sera of the mother and child and in the elute of the child were performed by use of microcards (Diamed, Basel, Switzerland; BioVue, Ortho Clinical Diagnosis, Raritan, NJ, USA) with commercially prepared test RBCs (Diamed, Ortho Clinical Diagnosis, and Gamma Biologicals, Houston, TX, USA) at 37 degrees C and indirect antiglobulin test (IAT) according to manufacturer instructions. Direct antiglobulin test (DAT) was performed by use of microcards (Diamed, Ortho Clinical Diagnosis) with both polyspecific and monospecific IgG anti-human globulin. Di(a) antigen was determined on maternal, paternal, and infant's red cells by commercial reagent (Gamma Biologicals). Determination of RBC antibody specificities in maternal and child sera and in the child's RBC eluate showed 2+ positive reactions only with two Di(a+) test RBCs. Father and baby were positive and mother was negative for Di(a) antigen. When a newborn has positive DAT and there are no clinical reasons for this, the possibility of positive DAT resulting from alloimmunization to low-frequency RBC antigens should be considered.

[Overview and methods for determination of antiplatelet antibodies and thrombocyte antigens]. / Pregled i primjena metoda za odredivanje antitrombocitnih protutijela i trombocitnih antigena.

The last twenty years have been characterized by great interest in the study of the role of antiplatelet antibodies and platelet antigens in the mechanism of thrombocytopenia. The use of numerous serologic methods for the determination of antiplatelet antibodies has contributed to the better understanding and differential diagnosis of immunologically induced thrombocytopenias. Development of the methods of molecular biology has allowed for a more accurate determination of platelet antigens and assessment of the prevalence of particular antigens in the population. These concepts have improved the possibilities of evaluation of particular antigenic systems in the genesis of auto- and alloimmune thrombocytopenic syndromes. Although majority of tests for determination of antiplatelet antibodies were initially introduced for antibody demonstration in patients with idiopathic thrombocytopenic purpura, these methods are now employed for all diseases associated with platelet impairments, when an immunologic pathomechanism of the disease onset is suspected. The methods are mostly used in serodiagnosis of neonatal thrombocytopenic purpura, posttransfusion purpura and refractoriness to platelet transfusion, and primary and secondary autoimmune thrombocytopenia.