Physical Characterization and Innate Immunogenicity of Aggregated Intravenous Immunoglobulin (IGIV) in an In Vitro Cell-Based Model.

PURPOSE: To investigate in vitro the innate immune response to accelerated stress-induced aggregates of intravenous immunoglobulin (IGIV) using a well-defined human cell-line model, and to correlate the innate response to physical properties of the aggregates. METHODS: IGIV aggregates were prepared by applying various accelerated stress methods, and particle size, count and structure were characterized. Immune cell activation as tracked by inflammatory cytokines released in response to aggregates was evaluated in vitro using peripheral blood mononuclear cells (PBMC), primary monocytes and immortalized human monocyte-like cell lines. RESULTS: IGIV aggregates produced by mechanical stress induced higher cytokine release by PBMC and primary monocytes than aggregates formed by other stresses. Results with the monocytic cell line THP-1 paralleled trends in PBMC and primary monocytes. Effects were dose-dependent, enhanced by complement opsonization, and partially inhibited by blocking toll-like receptors (TLR2 and TLR4) and to a lesser extent by blocking Fc gamma receptors (FcγRs). CONCLUSIONS: Stress-induced IGIV aggregates stimulate a dose-dependent cytokine response in human monocytes and THP-1 cells, mediated in part by TLRs, FcγRs and complement opsonization. THP-1 cells resemble primary monocytes in many respects with regard to tracking the innate response to IgG aggregates. Accordingly, the measurement of inflammatory cytokines released by THP-1 cells provides a readily accessible assay system to screen for the potential innate immunogenicity of IgG aggregates. The results also highlight the role of aggregate structure in interacting with the different receptors mediating innate immunity.

An international collaborative study to establish the WHO 1st international standard for alpha-1-antitrypsin.

BACKGROUND AND OBJECTIVES: The aim was to establish the 1st International Standard (IS) for alpha-1-antitrypsin (AAT) to standardise potency assignment of therapeutic products, calibrated in moles and mg active AAT in line with product labelling practice. Assigning total protein and antigen values to the IS was also investigated. MATERIALS AND METHODS: The active concentration of four candidate AAT preparations was determined in an international collaborative study by inhibition of trypsin (calibrated by active-site titration). Total protein and antigen content were determined for each candidate using local methods and in-house standards, and a common AAT preparation. The total protein content of the IS was also determined by amino acid analysis. Potency determination of recombinant and transgenic materials against the IS was investigated in a follow-up study. RESULTS: Data analysis for potency determination indicated no statistical difference between any of the candidates, or between the results for recombinant and plasma-derived products. Total protein content of the IS determined by amino acid analysis was consistent with the potency value. The variability in the total protein and antigen results for the other candidates was reduced when the data were recalculated relative to the IS. CONCLUSIONS: Candidate C (05/162) was established by the WHO Expert Committee on Biological Standardization (ECBS) in 2006 as the WHO 1st IS for AAT with a potency of 243 nmoles (12·4 mg) active inhibitor per ampoule. In 2008, ECBS approved the IS for potency determination of recombinant material and assigned a total protein and antigen value of 12·4 mg.

Do children with Adams-Oliver syndrome require endocrine follow-up? New information on the phenotype and management.

Adams-Oliver syndrome (AOS) is a rare genetic condition in which the main diagnostic criteria are terminal transverse limb defects and aplasia cutis congenita. Within the spectra of the clinical phenotype of AOS, anthropometric abnormalities have also been reported. We present growth pattern along with hormonal assays in three patients with AOS, one being treated with growth hormone (GH). In Patient 1 (a boy, age 1.9 years), with delayed psychomotor development, epilepsy, deficits of body mass and height, cryptorchidism, low insulin-like growth factor (IGF-1) levels were found and magnetic resonance imaging (MRI) revealed hypoplasia of midline structures of the central nervous system (CNS). In Patient 2 (a girl, age 3.6 years) no significant abnormalities in development, body mass, height or neuroimaging were found. In Patient 3 (a girl, age 8.2 years), with delayed psychomotor development and short stature, low IGF-1 levels and partial GH deficiency were found; MRI revealed small pituitary and polymicrogyria. The girl started GH treatment, improving height velocity and gross coordination. Based on these observations, it seems that intensity of auxologic and hormonal deficits in children with AOS is associated with CNS lesions. Hence, there are indications for neuroimaging and interdisciplinary follow-up of psychomotor development, growth and puberty in this subset of patients with AOS.

Tolerable risk guidelines.

Safety instrumented systems (SIS) should be designed to reduce the amount of risk in a process to a tolerable level. Expressing the tolerable level of risk is one of the most difficult tasks facing organizations trying to comply with the standards that govern SIS use. Organizations are basing their risk decision-making criteria on a variety of benchmarks including industry standards, local and foreign government regulations, practices of industry partners, and a qualitative assessment of what is fair and reasonable. This paper explores the ways that industry and government have made decisions on what risks are tolerable. The paper begins by reviewing qualitative criteria that are evaluated to determine what amount of risk can reasonably be tolerated. The paper also reviews some methods that have been used in government and industry to include financial aspects into the decision-making process. It then proceeds to review some of the quantitative ways that risk is represented. After discussing risk presentation, the paper reviews and compares tolerable risk guidelines that have been set by government and industry. The paper concludes by comparing the amount of risk reduction that can be justified by sociopolitical tolerability guidelines and that which can be justified by cost-benefit analysis.

[Progressive encephalopathies in children]. / Encefalopatie postepujace u dzieci.

The Authors describe progressive encephalopathies in children hospitalized at the Department of Child Neurology Silesian Medical School from 1980 to 2000. They present ethiology, clinical symptoms and diagnostic protocol based on literature data and their own experience.

Agenesis of corpus callosum: clinical description and etiology.

In 135 children (aged 3 months to 15 years) with structural defects of the central nervous system found on magnetic resonance imaging, agenesis of the corpus callosum was evident in 7. The etiology of agenesis of the corpus callosum has been established in four children: partial trisomy of chromosome 13, partial duplication of the long arm of chromosome 10, Aicardi's syndrome, and intracranial bleeding during the fetal period as a result of injury. Agenesis of the corpus callosum coexisted with a Dandy-Walker malformation in one other patient, which suggests a genetic etiology. In spite of these variable etiologies, dysmorphic features were identified in all seven patients, as was psychomotor retardation. Epileptic seizures had occurred in six patients, and all manifested abnormalities on neurologic examination.

[Cerebral tumors as a cause of refractory epilepsy in children]. / Procesy rozrostowe wewnatrzczaszkowe jako przyczyna padaczki lekoopornej u dzieci.

Cerebral tumours are a cause of seizures in less than 1-2% of children epilepsy. Seizure symptoms usually precede the diagnosis by several years and are often the only symptoms of an ongoing process. The symptomatology of the seizures often correlates with localization of a neoplastic lesion. The authors present six children aged 3 to 18 in whom epileptic seizures not susceptible to treatment were caused by cerebral tumours. The duration of epilepsy until the determining of the aetiology was various--from half a year to 13 years. In each of our six patients focal epilepsy occurred with simple or complex seizures with secondary generalization. It was only one patient in whom the tomography of the head turned out to be sufficient enough to establish the diagnosis of a brain tumour; in the other ones MR was necessary. The final diagnosis in four of the children was supported by histopathologic examination carried out during a neurosurgical procedure, whereas in one of them--by means of biopsy of the brain.

[The evaluation of selected factors in the prognosis of drug-resistant epilepsy in children with brain malformations]. / Ocena wybranych czynników w prognozowaniu padaczki lekoopornej u dzieci z wadami mózgu.

Brain malformations are important and frequent epilepsy reason in children and adolescents. During the last six years in neurological department of Pediatrics Clinic in Katowice were treated 106 children with brain malformations demonstrated in magnetic resonanse imaging. The main clinical symptoms in these patients were following: mental retardation, epilepsy, abnormalities in neurological examinations, dysmorphic features. Epilepsy were observed in above 3/4 of patients (84 children). In most of them there was intractable epilepsy (55 children). The aim of study was evaluation of selected factors in prognosis of epilepsy intractibility. The children with brain malformations and epilepsy were divided into two groups: with intractable epilepsy and with good response for pharmacotherapy. The type of malformation, the pre- i perinatal history, an age at which the first seizures appeared, abnormalities in neurological examination and IQ were compared in both groups. The differences weren't significant statistically apart two data. Normal pre- and perinatal history and early manifestation of seizures (during the first half of life) were confirmed significant statistically more often in group of patients with intractable epilepsy. There was limited value of most of the analysed parameters in prognosis of epilepsy intractibility. Further accumulating of data and increasing of number of the patients group with different types of malformations as well as progress in diagnostics, particularly molecular genetics, may be helpful in correct prognosis.

[Munchausen's syndrome by proxy--a malignant form of child abuse]. / Zespól Munchhausena per proxy--szczególna forma maltretowanego dziecka.

Munchhausen syndrome by proxy is a malignant form of child abuse in which illness in a child is fabricated and/or induced by a parent. It can result in serious illness and even death of the child and it is difficult to detect. The authors give a significant amount of literature examples. They try to find out the difference in ways of hurting by perpetrators. Various types of personalities and emotional disturbances in Munchhausen on proxy syndrome are shown. Child maltreatment and Munchhausen by proxy syndrome need to be part of the differential diagnosis when the clinical picture is atypical or does not appear medically plausible.

Variability of localization and intensity of damage of the white matter of the brain and cerebellum in genetically conditioned diseases.

The investigations were based on 3 cases with Leigh, 5 cases with Krabbe's, 4 cases of Alpers, 2 cases with Sandhoff, 1 case with Alexander's disease and 1 case with metachromatic leukodystrophy. In 1 case included into the study we have diagnosed nonketotic hyperglycinemia II. All the diseases under examination are recognized as genetically conditioned or are supposed to be of genetic origin. Damage of the white matter in a more delineated form in certain regions was found in Leigh disease. The changes demonstrated a variable degree of intensity from demyelination to necrosis. More extensive lesions of white matter in gyri and semivoal centrum were found in diseases with simultaneously damaged gray matter e.g. in Alpers and Sandhoff disease. The most extensive changes of diffuse demyelination were found in Krabbe's and Alexander's disease. In these diseases demyelination was accompanied with specific morphological structures e.g. globoidal cells (Krabbe's disease) and Rosenthal fibers (Alexander's disease). The peculiar type of demyelination was characteristic for nonketotic hyperglycinemia of type II. It was expressed by demyelination with vacuolization.

[Huntington chorea in a boy aged 14 years]. / Plasawica Huntingtona u 14-letniego chlopca.

The authors present a case of Huntington disease in a 14 year old boy. The basis of genetic, pathoetiology, clinical course of disease, significance of molecular analysis of DNA are discussed.

[Lyme disease in children]. / Borelioza u dzieci.

In the recent years the number of diagnosed cases of Lyme Disease has tended to increase. This is due to the possibility of serological examinations. Between 1992-95 at the Neurological Department of 2nd Chair of Pediatrics Silesian Medical Academy in Katowice 7 children with Lyme Disease were hospitalized. The authors analysed the clinical course, important laboratory and serological data and treatment effects.

Deletion screening and carrier detection in Duchenne muscular dystrophy in Polish population via direct analysis of DNA and RNA transcripts.

Analysis of 102 Polish Duchenne/Becker muscular dystrophy (D/BMD) patients was performed by 'multiplex' amplification of 22 fragments of the DMD/BMD gene and deletions were found in 55% of the patients. The data obtained using PCR were compared with results of 25 Southern blotting and hybridization experiments with cDNA probes and with immunostaining using anti-dystrophin antibodies. In order to determine more precise deletion breakpoints, additional experiments were performed on dystrophin transcripts isolated from peripheral blood lymphocytes. These data found direct application in carrier analysis in the respective families by detection or exclusion of aberrant cDNA fragments. Carrier detection was also performed by RFLP-PCR, analysis of polymorphic (CA)n repeats and single stranded conformational polymorphism (SSCP) for selected exons of the DMD gene.

[Silver-Russel Syndrome in aa 13-month old girl]. / Zespól Silvera-Russela u 13-miesiecznej dziewczynki.

The authors present a case of 13-month girl with Silver-Russel syndrome, which is an example of intrauterine growth retardation. The etiology of the disorder remains unknown, although many data suggest the embryopathy of the early gestational period. The main clinical features are: low height, decreased birth weight and normal gestational age. Body asymmetry and craniofacial dysmorphy.

Dihydrofolate reductase synthesis in the presence of immobilized methotrexate. An approach to a continuous cell-free protein synthesis system.

Dihydrofolate reductase was synthesized in a batch system in the presence of the affinity ligand methotrexate, bound to various matrices. Two types of gel were used: commercial methotrexate-agarose with pores inaccessible for translation machinery and methotrexate-POROS with pores easily accessible for translation reaction mixture components. The transcription/translation reaction was not inhibited by either the immobilized methotrexate or the matrix. The enzyme was synthesized with a high yield and could simultaneously be removed from the reaction mixture by the affinity matrix during the synthesis. With methotrexate-POROS present the reaction probably proceeded mainly in the pores of the gel. Kinetic limitations to the reaction in the presence of the gel were not observed. Active dihydrofolate reductase was eluted from methotrexate-POROS. The activity recovered was higher than dihydrofolate reductase activity synthesized in free solution system. The influence of the presence of immobilized methotrexate on dihydrofolate reductase synthesis will be further studied in a novel type of a continuous protein synthesis system.

Study of cell-free protein synthesis in aqueous two-phase systems.

Protein synthesis in an aqueous two-phase system is reported here as a novel type of extractive bioconversion. Translation of BMV RNA was studied using either rabbit reticulocyte lysate or wheat-germ extract in aqueous dextran-PEG systems. Both polymers inhibited protein synthesis and the translation system partitioned into both phases. In the rabbit reticulocyte two-phase system, protein synthesis reached 30% of that in free solution, while in wheat-germ extract it was 60% of that in free solution. Protein was found mainly in the dextran (lower) phase but its partitioning was related to the polymer concentration, and molecular weight, as well as the ionic strength of the system. Protein synthesis was highly influenced by PEG concentration, potassium chloride concentration, and dextran molecular weight.

Ultrastructure of peripheral blood lymphocytes in some degenerative central nervous system disease.

The ultrastructure of peripheral blood lymphocytes of 30 children with degenerative central nervous system diseases was analyzed. The affected children were divided into four groups. Lysosomal storage were characterized by the storage of membrane-bound inclusions in peripheral lymphocytes. Ceroidlipofuscinosis was manifested by the presence of curvilinear bodies. The appearance of abnormal mitochondria was found in mitochondrial encephalopathies. The tubuloreticular structures seen in lymphocytes of some unclassified cases suggested viral factors that acted in prenatal life. All findings confirm the role of peripheral lymphocytes analysis in the diagnosis of degenerative central nervous system diseases.

[Congenital toxoplasmosis in a 6-week-old infant]. / Toksoplazmoza wrodzona u 6-tygodniowego niemowlecia.

A case of congenital toxoplasmosis was observed in an infant aged 6 weeks with fatal outcome. Histological changes produced by Toxoplasma gondii in the brain are described.

Fibrinoidal leucodystrophy (Alexander's disease) at 13 months girl. A case report.

The case of Alexander's fibrinoidal leucodystrophy that was presented below is an exemplar of extremely rare degenerative disease of the CNS. On the grounds of the clinical course of the disease it seems that our case can be reckoned as an early childhood form of Alexander's disease. An interesting difference that pays attention is the marked hydrocephalus. In the most cases of Alexander's disease the volume of ventricular system is normal however most of authors expresses that it sometimes can be slightly and insignificantly enlarged.

[Leigh disease in a 3-year-old girl]. / Choroba Leigh'a u trzyletniej dziewczynki.

A case of Leigh disease in a 3-year-old girl is reported. The child had regression of the psychomotor development, muscular hypotonia, weak tendinous reflexes, opsoclonus, tremor of the whole body, hypertrichosis, autonomic system disturbances. Laboratory investigations demonstrated raised serum lactic acid level. Postmortem histological examination of the brain confirmed the diagnosis of Leigh disease established before death.