The levels of TNF-alpha in cerebrospinal fluid and serum do not correlate with the counts of the white blood cells in acute phase of ischaemic stroke

Stroke-induced inflammatory reaction, which leads to invasion of leukocytes into the evolving brain infarct, seems to play a key role in the deterioration of brain ischaemic impairment. We have studied CSF and serum levels of tumour necrosis factor-alpha (TNF-alpha), the potent proinflammatory cytokine, and peripheral white blood cells (WBC) counts in patients within the first 24 hours of ischaemic stroke. TNF-alpha levels in CSF and serum as well as WBC counts were increased. There was no correlation between TNF-alpha levels either in CSF and serum or in WBC counts. The results of our study suggest that increased CSF TNF-alpha levels may represent acute intracerebral inflammation in stroke, whereas elevated levels of TNF-alpha in serum may reflect the peripheral proinflammatory state as well as stroke-induced systemic inflammatory reaction. Increased CSF and serum TNF-alpha levels do not correlate with the elevation of WBC counts, suggesting that TNF-alpha overexpression observed in early phase of stroke is not dependent on increased total number of peripheral leukocytes

Adhesion molecules of immunoglobulin gene superfamily in stroke

Stroke-induced inflammatory reaction leads to the accumulation of leukocytes in the brain ischaemic region, where they exert a detrimental effect - promotion and extension of cerebral damage. Intracerebral infiltration of peripheral blood leukocytes requires prior endothelial-leukocyte interactions that are mediated by such cell surface proteins as adhesion molecules. Among adhesion molecules, it is the immunoglobulin gene superfamily (IgSF) that is responsible for strong attachment and transendothelial migration of leukocytes. The principal members of IgSF are: intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1). In this review the following issues were described and discussed: an increased expression of ICAM-1 and VCAM-1 in ischaemic brain as well as a detection of their soluble(s) forms in sera of stroke victims. The presented data suggest the involvement of both ICAM-1 and VCAM-1 in the sequence and timing of the infiltration of leukocytes into the brain ischaemic zone after stroke. They have also revealed changes in serum concentrations of sICAM-1 and sVCAM-1 that are characteristic for stroke. Recently, increase in sPECAM-1 levels in serum and cerebrospinal fluid (CSF) has been shown within 24 h of the onset of stroke, having indirectly suggested involvement of the molecule in the inflammatory events during the early phase of stroke

Acute ischaemic stroke increases the erytrocyte sedimentation rate, which correlates with early brain damage

The acute phase response follows tissue injury and contributes to its exacerbation with pro-inflammatory and pro-thrombotic mechanisms. Acute phase proteins promote erythrocyte aggregation and falling, with the result that the erythrocyte sedimentation rate (ESR) is a measure of the acute phase response. As the acute phase response accompanies ischaemic brain damage, we studied ESR values in patients within the first 24 hours of ischaemic stroke and evaluated whether these values may be related to the volume of anatomically relevant single hemispheric brain computed tomography (CT) areas observed at the same period, indicating early stroke-related cerebral changes. We observed an increase in ESR in stroke patients and a positive correlation between the ESR values and the volume of early brain CT hypodense areas. The results suggest that elevation in ESR values is observed soon after a stroke and may reflect the relationship between the degree of acute phase response in the early phase of ischaemic stroke and the extent of local brain damage

Dwie mutacje w jednym genie dystrofiny

Background and purpose Duchenne/Becker muscular dystrophies (DMD/BMD) lead to progressive irreversible muscle deterioration caused by recessive mutations in the dystrophin encoding gene (Xp21.1). Approximately 60% of mutations are deletions, 10% are duplications and the remaining 30% are point mutations. The aim of the study is to present the rare occurrence of two pathogenic mutations (deletions or duplications) in one allele of the dystrophin gene. Material and methods DNA of patients from 1364 DMD/BMD families was tested. Two techniques – PCR-multiplex and multiplex ligation-dependent probe amplification – were used to search for mutations in the dystrophin gene. Results Deletion was detected in 648 families and duplication was found in 74 families (analysis in progress). In two families, presence of two mutations in one gene was documented – in the first family two deletions were found (exons 45–49 and 60–61), and in the second family two duplications were detected (exons 2–7 and 50–59). One of the deletions disrupted the reading frame, and the other deletion retained the reading frame. Both duplications also retained the reading frame of the gene but in both families the disease took a severe course (DMD). In the family with two duplications prenatal diagnosis was also carried out, and carriership of both mutations was discovered in the female fetus. Conclusions In the analyzed group of DMD/BMD families, the frequency of combined occurrence of two mutations in one gene was 2 per 722 (0.3%). The phenomenon of detected non-contiguous deletions and duplications is presented together with 31 similar cases published so far.Wstęp i cel pracy Dystrofia mięśniowa Duchenne'a/Beckera (DMD/BMD) jest związana z postępującym i nieodwracalnym zanikiem mięśni wywołanym recesywnymi mutacjami w genie dystrofiny (Xp21.1). Szacuje się, że 60% mutacji stanowią delecje, a 10% – duplikacje; pozostałe 30% mutacji ma charakter punktowy. Celem pracy jest przedstawienie rzadkich przypadków współwystąpienia w jednym allelu genu dystrofiny dwóch chorobotwórczych mutacji – delecji lub duplikacji. Materiał i metody Badano DNA pacjentów z 1364 rodzin skierowanych z podejrzeniem DMD lub BMD. Mutacji poszukiwano, używając dwóch technik: PCR-multiplex i MLPA (multiplex ligation-dependent probe amplification). Wyniki W 648 rodzinach wykryto delecję, a w 74 rodzinach – duplikację (badania w toku). W dwóch rodzinach udokumentowano łączne wystąpienie w jednym genie dystrofiny dwóch mutacji – w pierwszej rodzinie w jednym allelu wykryto dwie delecje (eksony 45–49 i 60–61), a w drugiej rodzinie dwie duplikacje (eksony 2–7 i 50–59). Jedna z delecji naruszała fazę odczytu, druga zaś ją zachowywała; obie duplikacje zachowywały fazę odczytu, jednak w obu rodzinach choroba przybierała ostrą postać (DMD). W rodzinie, w której wykryto dwie duplikacje, wykonano diagnostykę prenatalną, stwierdzając u płodu płci żeńskiej nosicielstwo obu mutacji. Wnioski W analizowanej grupie rodzin z delecją lub duplikacją częstość łącznego wystąpienia dwóch mutacji wyniosła 2 na 722 (0,3%). Zjawisko wykrytych nieciągłych delecji i duplikacji przedstawiamy w zestawieniu z opisanymi dotychczas 31 podobnymi przypadkami

On recombination in strong laser fields: effect of a slow drift

The dynamics of the recombination in ultrastrong atomic fields is studied for one-dimensional models by numerical simulations. A nonmonotonic behavior of the bound state final population as a function of the laser field amplitude is examined. An important role of a slow drift of an electron wave packet is observed.Comment: 4 pages, 6 figure

Postać dziedziczna choroby prionowej w Polsce

Background and purpose The aim of the study was to perform molecular analysis in a group of patients affected with prion disease. Diagnosis was based on results of clinical and/or histopathological examination of the brain. This is the largest investigation of this type performed so far in Poland. Material and methods Analysed material contained 36 cases of prion disease, including 35 cases of Creutzfeldt-Jakob disease and one case of Gerstmann-Sträussler-Scheinker disease, as well as two familial cases initially suspected of Huntington disease and Alzheimer disease. The control group consisted of 87 subjects. The most frequent known mutations in the PRNP gene were looked for, namely those in codons 102, 117, 178, 200, 217 and OPRI; the polymorphism Met/Val in codon 129 was also analysed. The methods applied were PCR-RFLP and DNA sequencing. Results The following mutations were found: E200K in 5 families, P102L in one family (previously identified), D178N in one family and 6OPRI in one family. Overall, mutations were detected in 17 persons (including 8 preclinical ones) from 8 pedigrees. Highly significant difference of codon 129 Met/Val heterozygosity frequencies was found between the affected subjects and the controls. Frequency of the familial form of prion disease in the material analysed was 14%. Conclusions Screening for mutations in the PRNP gene should be performed in all diagnosed cases of prion disease and cases of familial occurrence of early onset dementia of unknown aetiology. Families with identified mutations should be offered genetic counselling and informed of risks of blood and organs’ donation.Wstęp i cel pracy Celem pracy była analiza molekularna w grupie osób dotkniętych chorobą prionową, rozpoznaną na podstawie objawów klinicznych i/lub wyniku badania neuropatologicznego mózgu. Było to największe tego typu badanie przeprowadzone dotychczas w Polsce. Materiał i metody W skład analizowanego materiału weszło 36 przypadków choroby prionowej, w tym 35 przypadków choroby Creutzfeldta-Jakoba i jeden przypadek choroby Gerstmanna-Sträusslera-Scheinkera, a także dwa przypadki rodzinne podejrzane o chorobę Huntingtona i chorobę Alzheimera oraz grupa kontrolna (87 osób). Poszukiwano najczęstszych mutacji w genie PRNP: wkodonach 102, 117, 178, 200, 217 i OPRI. Kodon 129 analizowano również pod kątem zygotyczności (walina/metionina). Stosowano metodę PCR-RFLP i sekwencjonowanie. Wyniki Wykryto mutacje: E200K – pięć rodzin, P102L – jedna rodzina (wcześniej zidentyfikowana), D178N – jedna rodzina, 6OPRI – jedna rodzina. Łącznie stwierdzono mutację w 8 rodowodach u 17 osób, w tym u 8 osób w fazie przedobjawowej. Zaobserwowano także bardzo istotną różnicę w częstości występowania heterozygotyczności Met/Val pomiędzy grupą badaną i grupą kontrolną. Częstość dziedzicznej postaci choroby prionowej w analizowanym materiale wynosi 14%. Wnioski Mutacji w genie PRNP należy poszukiwać we wszystkich przypadkach choroby prionowej oraz w przypadkach rodzinnie występującego otępienia o wczesnym początku i niewyjaśnionej etiologii. Przebieg kliniczny i zmiany neuropatologiczne w niektórych przypadkach dziedzicznych chorób prionowych mogą się różnić od spotykanych najczęściej w sporadycznej postaci choroby. Rodziny ze stwierdzoną mutacją winny być objęte poradnictwem genetycznym i poinformowane o zagrożeniu związanym z dawstwem krwi i narządów do przeszczepienia

MLPA based detection of mutations in the dystrophin gene of 180 Polish families with Duchenne/Becker muscular dystrophy

Duchenne/Becker muscular dystrophy (DMD/BMD) is a recessive, X-linked disorder caused by a mutation in the dystrophin gene. Deletions account for approximately 60–65% of mutations, duplications for 5–10%. The remaining cases are mainly point mutations. According to Monaco theory clinical form of the disease depends on maintaining or disrupting the reading frame. The purpose of the study was to determine frequency and location of deletions and duplications in the dystrophin gene, to determine the compliance between maintaining/disrupting the reading frame and clinical form of the disease and to check the effectiveness of MLPA (multiplex ligation-dependent probe amplification) in the detection of these mutations in hemizygous patients and heterozygous female carriers. The material is composed of combined results of molecular diagnosis carried out in years 2009–2012 in 180 unrelated patients referred with the diagnosis of DMD/BMD tested by use of MLPA. We identified 110 deletions, 22 duplication (in one patient two different duplications were detected) and 2 point mutations. Deletions involved mainly exons 45–54 and 3–21, whereas most duplications involved exons 3–18. The compliance with Monaco theory was 95% for deletions and 76% for duplications. Most of mutations in the dystrophin gene were localized in the hot spots – different for deletions and duplications. MLPA enabled their quick identification, exact localization and determination whether or not they maintained or disrupted the reading frame. MLPA was also effective in detection of deletions and duplications in female carriers

Ex vivo measurement of calpain activation in human peripheral blood lymphocytes by detection of immunoreactive products of calpastatin degradation.

Limited proteolysis of multiple intracellular proteins by endogenous Ca-dependent cysteine proteases--calpains--is an important regulatory mechanism for cell proliferation, apoptosis etc. Its importance for cellular functions is stressed by existence of endogenous calpain inhibitors--calpastatins. The calpain-calpastatin system within living cells is in a fragile balance, which depends on both partners. The interdependence of calpain--a protease--and calpastatin--an endogenous inhibitor and at the same time a substrate for this enzyme makes any assessment of actual activity of this enzyme in the cells very difficult. In this work we made an attempt to estimate and compare the activity of calpain in human peripheral blood lymphocytes by assessing the levels of limited proteolysis of calpastatin in these cells by western blot, while at the same time the levels of calpain protein inside these cells was measured by flow cytometry. Our results indicate that it is possible to compare (semi-quantitatively) the activities of calpain in peripheral blood CD4+ and CD19+ lymphocytes from various donors that way. Preliminary results showed that calpain activity is increased in the CD4+ T cells isolated from peripheral blood of rheumatoid arthritis patients as compared to control lymphocytes. Extremely high intrinsic activity of calpain was detected in chronic lymphocytic leukemia (CD19+) cells. All this confirms the detection of immunoreactive products of calpastatin as a good maker of endogenous calpain activity

Photodetachment in superintense fields: the problem of stabilization and a role of rescattering wavepacket's fragments

Photodetachment in ultrastrong laser field in two spatial dimensions is investigated numerically. The problem of an adiabatic stabilization is discussed, in particular it is shown that a quick drift in the direction of the electric field and a magnetic drift cannot be avoided simultaneously. A qualitative behavior of the packet for a short-range binding potential is contrasted with that for a soft-core potential, in particular dynamical effects due to a rescattering of the fragments separated from the main packet are demonstrated.Comment: 14 pages, 7 figure