Incorporation of osteoblasts (MG63) into 3D nanofibre matrices by simultaneous electrospinning and spraying in bone tissue engineering.

Nanofibre scaffolds are suitable tools for bone tissue engineering. Mimicking the extracellular matrix, they allow for cell growth and differentiation. However, in large 3D scaffolds, uniform cell colonisation presents an unsolved problem. Our aim was to design and analyse a method of colonising nanofibre scaffolds, combining electrospinning of fibres and electrospraying of cells, to determine its impact on cell survival, growth, and gene expression. The osteoblast-like cell line MG63 was suspended in medium and electrosprayed into growing scaffolds of poly-(l-lactic acid) (PLLA) or PLLA/Col-I blend nanofibres. Fluorescein diacetate (FDA) staining was used to determine survival and growth over a 22 d culture period. Expression of osteocalcin (OC) and type I collagen (Col-I) genes was determined by real time PCR. Fluorescence microscopy was used to analyse Col-I and OC deposition, as well as cell densities. While spraying distance and cell density in the spraying solution influenced survival and cell density, the combination of electrospinning and electrospraying did not negatively influence the maintenance of the osteoblast phenotype. Furthermore, VEGF induction in response to hypoxia was not suppressed, but modulated by polymer composition. Therefore, simultaneous electrospinning and electrospraying is a suitable tool in producing nanofibre based 3D cell seeded scaffolds.

The plasminogen activator inhibitor-1 gene polymorphism in determining the risk of pediatric ischemic stroke--case control and family-based study.

Pediatric ischemic stroke, though relatively rare, remains an important medical problem since 20-40% of patients have recurrent strokes and 50-85% of them suffer from long-term neurological deficits. Approximately 20-50% of the affected children have prothrombotic disorders, therefore upon looking for possible genetic causes of the disease we focused on the plasminogen activator inhibitor (PAI-1)--the major inhibitor of fibrinolysis. The aim of the present study was to investigate a possible association between the -675_-674insG PAI-1 gene polymorphism and pediatric ischemic stroke. The study population consisted of 343 individuals: 70 children with ischemic stroke, 140 their biological parents and 133 control children. The PAI-1 gene polymorphism was genotyped using the restriction fragment length polymorphism and was visualized by AgNO3 staining. The transmission/disequilibrium test showed exactly the same transmission of alleles from parents to the affected children (37:37). The case-control model also did not reveal any statistical significance in alleles and genotypes distribution between patients and control children. The obtained results suggest that the 4 G/5 G polymorphism of the PAI-I gene is not a risk factor of ischemic stroke in Polish children.

Organic nanofibers containing insect pheromone disruptants: a novel technical approach to controlled release dispensers with potential for process mechanization.

Beginning fifty years ago, the search for suitable dispensers containing insect pheromones grew with the availability of these synthetic biotechnical tools. Many economic entomologists and application engineers dearly wish they had the "smart, intelligent and ideal dispenser". More or less suitable approximations are available commercially, but none so far meets all demands. Under economic strictures, novel inexpensive systems would be advantageous with release characteristics tailored to the specific life histories of pest insects, the plants considered and the numerous requirements of growers alike. Simultaneously, their field distribution should be mechanizable and be accomplished by one (or very few) application runs. The dispensers should be biodegradable, biocompatible, sustainably applicable, and they should be based on renewable resources. This report presents first results of a novel organic, electrospun nanofiber dispenser with dimensions in the upper nanometer range. Its load of pheromone can be adjusted to be sufficient for 7 weeks of constant disruptive action in vineyards and can be directed against the European Grape Vine Moth Lobesia botrana (Lepidoptera: Tortricidae) which here serves as a readily available model. Mating disruption in L. botrana and the related Eupoecilia ambiguella is a well studied and developed engineering process. Equally, nanofiber production by electrospinning (for a comprehensive review see Greiner and Wendorff, 2007A, B) is well known and already has numerous applications in filtration technology, air conditioning, and medical wound dressing. Our goal was to bring together and successfully mate these (partly incompatible) technologies via technical tricks of a proprietary nature. Even though the lifetime and effectiveness of currently available nanofibers still must be doubled, the rather complicated system of their production and analysis is known well enough to identify the parameters that need future adjustment. Another challenge is the mechanical distribution of the fibers in the vineyards by suitable machinery. Also, in this respect, certain technical leads are available for future development.

Organic electrospun nanofibers as vehicles toward intelligent pheromone dispensers: characterization by laboratory investigations.

Organic nanofibers have a history of technical application in various independent fields, including medical technology, filtration technology, and applications of pharmaceuticals via inhalation into the lungs. Very recently, in a joint effort with polymer chemists, agricultural applications have been added to this list of priorities. The aim is finding novel approaches to insect control. Pheromones, dispensed in a quantifiable way, are being used here in disrupting the mating communication between male and female pest insects, e.g. the European grapevine moth Lobesia botrana (Lepidoptera: Tortricidae), where current dispenser technology does not fully meet the high expectations of growers and environmentalists with respect to longevity of constant release, self decomposition, mechanical distribution, renewability as well as sustainability of resources. The methodology of electrospinning is exhaustively covered by Greiner and Wendorff (2007), with technical details reported by Hellmann et al. (2009), Hein et al. (2011), and Hummel et al. (2010). Wind tunnel studies were run within a tunnel with adjustable laminar flow and 0.5 m/sec air velocity. Mass losses of the electrospun fiber bundles were determined with a sensitive analytical balance 2-3 times per week and recorded as time vs. mass change. CLSA experiments were performed with a self developed glass apparatus (Lindner, 2010) based on various suggestions of previous authors. Microgram quantities of volatile pheromone (E,Z)-7,9-Dodecadienylacetate were absorbed on a filter of rigorously purified charcoal and desorbed by repeated micro extraction with a suitable solvent mixture. Aliquots of the solution were subjected to temperature programmed capillary GLC. Retention times were used for identification, whereas the area covered by the pheromone peak originating from a FID detector signal was integrated and compared with a carefully calibrated standard peak. Since these signals were usually in the low nanogram range, several replications were averaged for statistical improvement. - Thermogravimetric analysis between ambient temperature and 500 degrees C provided a series of degradation curves where the diagram contained information on the evaporation of pheromone alone, polymer fiber alone and pheromone included in the fiber.- Microscopic investigations resulted in pictures of nanofibers from which the overall morphology and the fiber dimensions could be quantified. Organic nanofibers loaded with the grapevine moth pheromone have been well characterized by 5 different lab methods, followed by field bioassays reported elsewhere in these communications volumes (HUMMEL et al., 2011). This comprehensive analytical approach to fiber characterization is new and will be further refined. The federal agency JKI Berlin subjected the pheromone loaded organic fibers to various independent toxicological and ecotoxicological tests and found no adverse side effects.

Lack of obvious influence of PLLA nanofibers on the gene expression of BMP-2 and VEGF during growth and differentiation of human mesenchymal stem cells.

Growth factors like bone morphogenetic protein 2 (BMP-2) and vascular endothelial growth factor (VEGF) play an important role in bone remodeling and fracture repair. Therefore, with respect to tissue engineering, an artificial graft should have no negative impact on the expression of these factors. In this context, the aim of this study was to analyze the impact of poly(L-lactic acid) (PLLA) nanofibers on VEGF and BMP-2 gene expression during the time course of human mesenchymal stem cell (hMSC) differentiation towards osteoblasts. PLLA matrices were seeded with hMSCs and cultivated over a period of 22 days under growth and osteoinductive conditions, and analyzed during the course of culture, with respect to gene expression of VEGF and BMP-2. Furthermore, BMP-2-enwoven PLLA nanofibers were used in order to elucidate whether initial down-regulation of growth factor expression could be compensated. Although there was a great interpatient variability with respect to the expression of VEGF and BMP-2, PLLA nanofibers tend to result in a down-regulation in BMP-2 expression during the early phase of cultivation. This effect was diminished in the case of VEGF gene expression. The initial down-regulation was overcome when BMP-2 was directly incorporated into the PLLA nanofibers by electrospinning. Furthermore, the incorporation of BMP-2 into the PLLA nanofibers resulted in an increase in VEGF gene expression. Summarized, the results indicate that the PLLA nanofibers have little effect on growth factor production. An enhancement in gene expression of BMP-2 and VEGF can be achieved by an incorporation of BMP-2 into the PLLA nanofibers.

One-step production of polymeric microtubes by co-electrospinning.

Herein we demonstrate the ability to fabricate polymeric microtubes with an inner diameter of approximately 3 microm through co-electrospinning of core and shell polymeric solutions. The mechanism by which the core/shell structure is transformed into hollow fibers (microtubes) is primarily based on the evaporation of the core solution through the shell and is described here in detail. Additionally, we present the filling of these microtubes, thus demonstrating their possible use in microfluidics. We also report the incorporation of a protein (green fluorescent protein) within such fibers, which is of interest for sensorics.

Fibrinogen alpha and beta gene polymorphisms in pediatric stroke--case-control and family based study.

BACKGROUND/PURPOSE: Data on the role of the -455G > A polymorphism of the gene encoding ß fibrinogen subunit (FGB) and the Thr312Ala polymorphism of the gene for the α fibrinogen subunit (FGA) in childhood ischemic stroke are insufficient. Therefore the aim of the study was to evaluate a possible association between these two polymorphisms and arterial ischemic stroke. METHODS: The study group consisted of 85 children after ischemic stroke, 146 of their parents and 159 controls. Both polymorphisms were genotyped using the restriction fragment length polymorphism method. Two study designs were used: a case-control model and a family-based transmission-disequilibrium test. Statistica 7.1 and EpiInfo 6 softwares were used in all analyses. RESULTS: In the TDT test, a tendency to a higher transmission of the 312Ala allele of the FGA gene and the -455A allele of the FGB gene was observed, however, it was statistically non-significant. The frequencies of alleles and genotypes of both FGA and FGB genes polymorphisms did not differentiate children from both groups also in the case-control model. Additive or synergistic effects between FGA and FGB genes polymorphisms were not observed. CONCLUSION: An analysis of the results obtained in this study and a critical review of previously published data indicate that examined gene polymorphisms are not related to ischemic stroke in children.

Krabbe disease: an ultrastructural study of globoid cells and reactive astrocytes at the brain and optic nerves.

We report here a detailed ultrastructural study of a brain biopsy along with post-mortem brain and optic nerve specimens from a case of Krabbe disease, a relatively rare leukodystrophy caused by a mutation in the gene for galactocerebrosidase (GALC) mapped to the 14q31 region of chromosome 14. GALC is responsible for lysosomal hydrolysis of several galactolipids including galactosylceramide, a major sphingolipids of the white matter of the central nervous system, galactosylsphingosine (psychosine) and galactosyldigluceride. The main neuropathological features such as accumulation of globoid cells, loss of myelin and marked gliosis were observed in the white matter. The monocytic origin of globoid cells was confirmed by CD-68 and ferritin-positivity and periodic acid Schiff (PAS) positivity. Ultrastructural study of the globoid cells showed the accumulation of tubular crystalloids, which are highly specific for this disease. The differences with Gaucher's disease and the pathomechanism of neuropathological damage are discussed.

[Electrospun poly-l-lactide nanofibres as scaffolds for tissue engineering]. / Elektrogesponnene Poly-L-Laktid-Nanofasern als resorbierbare Matrix für Tissue-Engineering.

Tissue engineering is a promising tool for treating structural and functional defects in bone and cartilage. To provide optimal conditions for three-dimensional cell growth the use of a scaffold is necessary. The aim of the study was to test the potential application of an electrospun poly (l-lactide)-nanostructured scaffold as a matrix for tissue engineering. Matrices were seeded with human osteosarcoma MG-63 cells and cultivated for 14 days. Cells showed a clear preference for growth along the nanofibres, and demonstrated no signs of degeneration or apoptosis. The fine structure of electrospun nanofibres makes them an ideal scaffold for tissue engineering, in particular for cartilage repair. They can be "doped" with growth factors, medications, etc., and are both biocompatible and biodegradable.

[Case of subacute myelo-optico-neuropathy]. / Przypadek neuropatii rdzeniowo-ocznej.

A 52-year-old female patient is reported in whom after taking of about 20 g Clioquinol blindness and paralysis of extremities with urinary incontinence developed. Complete cure ensued.

[Alternating hemiplegia as a migraine equivalent in a small child]. / Porazenie polowicze naprzemienne jako ekwiwalent migreny malego dziecka.

The authors present the diagnostic criteria, clinical course and additional examinations in alternating hemiplegia (ah). According to International Headache Classification ah belongs to the syndromes which precede or accompany migraine. We give a description of the case which is the first published in Polish literature and discuss the etiology supporting the connection of ah with migraine.

[Tiagabine in drug-resistant epilepsy in children: preliminary study]. / Tiagabina w lekoopornej padaczce u dzieci--doswiadczenia wstepne.

UNLABELLED: The aim of the study was the presentation of our preliminary experiences and the data concerning the efficacy and safety of tiagabine in patients below the 12-th year of life. 4 cases of drug--resistant epilepsy--with partial complex attacks in 3 and simple in 1 (frontal epilepsy from supplementary motor area) with multiple seizures in the day were the subject of the study. Tiagabine add-one therapy to CBZ or to VPA was used. Initial period--the dose titration lasted 4 weeks. The stable dosis period amounted to 5 months. The effective dosis of tiagabine was 1 mg/kg/day. In 1 case the seizures completely disappeared and in 3 the frequency decreased to 2-4 times in comparison to frequency per day before the treatment. There were not any side-effects reported which would be the cause of the drug discontinuation. IN CONCLUSION: Tiagabine appeared to be an effective and safe drug in resistant epilepsy in children below 12 years of age.

[Allergy effect on migraine course in older children and adolescents]. / Wplyw alergii na przebieg migreny u dzieci starszych i mlodziezy.

UNLABELLED: The interaction between allergy and migraine has been discussed since many years. The aim of the present study was the evaluation of: 1. allergy prevalence in the studied children with migraine, 2. allergy effect on the clinical course of migraine. MATERIAL AND METHOD: The studied group comprised 30 children and adolescents aged 11-17 years with the diagnosis of migraine with or without aura by IHS criteria. In each case the migraine index was determined and symptoms of allergy were sought. Skin prick test and tests for immunoglobulin E in serum were done. The analysis of results showed the presence of allergy in 12 cases (40%), and on this basis two groups were isolated: with and without allergy. The allergic children were given antiallergic treatment for 6 months (pharmacological or diet restriction). RESULTS: In the allergy group the migraine index decreased significantly from 2.45 to 0.33. Headache intensity decreased as well. CONCLUSIONS: 1. In 40% of cases migraine was associated with allergy. 2. Treatment results suggest that allergy and antiallergic treatment may influence the course of migraine attacks.

[Open multicenter study of the effectiveness and safety of gabitril in epileptic patients with partial seizures]. / Otwarte wieloosrodkowe badanie skutecznosci i bezpieczenstwa stosowania Gabitrilu u chorych z padaczka z napadami czesciowymi.

The paper presents the results obtained by 53 investigators implementing the first Polish multicentre study of the effectiveness and safety of tiagabine (Gabitril). The study included 81 patients with refractory epilepsy with partial seizures. The duration of the study was 16 weeks. For the initial 6 weeks, Gabitril was gradually introduced till a dose of 30 mg/day was achieved. Within the subsequent 10 weeks the treatment effectiveness was observed and monitored, with the provision that the dose could be increased. The final analysis included 62 patients, while in 12 subjects the treatment was discontinued in less than 16 weeks. The results indicate a very beneficial effect of Gabitril on the frequency of seizures in patients with drug-resistant epilepsy. Almost 1 of the analyzed patients were seizure free. The most beneficial effects with respect to seizure number and intensity reduction were noted in subjects with partial complex and partial seizures with secondary generalization. The dynamic character of the decrease in seizure frequency was best observed between the first and third month of therapy. In 2/3 of patients the recommended dose was achieved and maintained. Less than 15% of subjects were excluded from the study, mainly due to lack of therapeutic effects. The number and character of adverse effects observed in the course of the present study did not differ from these noted in long-term Gabitril trials. The drug was demonstrated to exert no effect on vital functions and laboratory parameters. The results confirm the high effectiveness of Gabitril in treatment of patients with partial seizures and a good tolerance of this agent.

[Chorea gravidarum--case report]. / Stan plasawiczy u ciezarnej--opis przypadku.

A case of a 19 years old pregnant girl with chorea gravidarum is described, the diagnosis assuming rheumatic background to be the direct cause of the disease. Administered were: relanium, penicillin, solu-medrol and encorton with a positive therapeutic outcome.

[Herpes simplex encephalitis in children]. / Opryszczkowe zapalenie mózgu u dzieci.

Clinical course of Herpes Simplex Encephalitis were shown in this paper. 24 children in the age of 4 to 15 were treated at Polish Mother's Memorial Hospital in years 1992-1994. Value of EEG, CT scan, NMR were underlined in early diagnostic and casual treatment of diseases. All children from this analysis group survived, only 8 demonstrated late complications: epileptic attacks (3), spastic tetraparesis (1), encephalopathy and mental disturbances (1). Positive results of treatment were connected with early diagnosis and early antiviral treatment which is very important in decreasing death rate and preventing permanent injury.

[Occurrence of epilepsy, its types and course during the first decade of life in progeny of mothers with epilepsy--prospective study]. / Wystepowanie padaczki, jej rodzaje i przebieg w pierwszej dekadzie zycia u potomstwa matek chorych na padaczke--badania prospektywne.

On the basis of the prospective study, concerning 100 children of mothers with epilepsy, observed in the first decade, it was established that epilepsy appeared more often in this period than it was reported in the retrospective studies. The frequency of epilepsy amounted 7%. The pregnant-perinatal negative factors in mothers whose children suffered from epilepsy, weren't essentially larger than in other mothers with epilepsy. The occurrence of epilepsy in mothers till 10 year's of age increases the risk of early appearance of epilepsy in offspring essentially (p < 0.05). The epilepsy with absence seizures in mothers is related to the increased number of children with epilepsy in the first decade significantly more often than the epilepsy only with generalized tonic-clonic seizures (p < 0.012). Among the children with epilepsy, there were cases with the same type as in mothers epilepsy (absence), and with other generalized idiopathic epileptic syndromes (West syndrome, Dose syndrome, epilepsy with tonic-clinic seizures). The course of epilepsy in offspring of mothers with epilepsy was typical for the relevant epileptic syndroms appearing in childhood.

[Neurological status and psychomotor development of infants with dilated pericerebral space]. / Stan neurologiczny i rozwój psychoruchowy niemowlat z poszerzona przestrzenia okolomózgowa.

Neurologic condition and psychomotoric development were analysed in infants up to 14 months of age with dilated pericerebral space detected by ultrasound scan performed between 2nd and 5th month of life. The good prognosis was stressed in children with mild degree of dilatation of the subarachnoid space.

The unusually large striae distensae all over the body.

Striae distensae or stretch marks are cutaneous changes which may appear everywhere on the body, except for the face, and they usually occur in stretching areas. We want to present unusually large striae that developed nearly all the body of a 14-year-old patient during steroid therapy. Such huge striae are very rare, especially when the doses of steroids are not high. Widely spread striae are not a serious disease or disability, but only a cosmetic problem. However, they may lead to a persistent complex and feeling of inferiority or even cause serious depression states, especially in teenagers, as in this case.