Search

This review describes the very specific role of Sigma1 receptor in different types of muscle cells. Sigma1 receptor is a transmembrane protein residing in such structures like MAM. It has chaperoning activity supporting function of many proteins, particularly ion channels, including Ca2+ channels. This latter function is of particular meaning for muscle cells, due to their calcium-based/regulated metabolism. Here we discuss new reports pointing to participation of Sigma1 receptor in muscle specific processes like contraction, EC-coupling, calcium currents and in diseases like left ventricular hypertrophy, transverse aortic stenosis and hypertension-induced heart dysfunction.

Simpson-Golabi-Behmel syndrome (SGBS) is a rare genetic condition and is inherited in an X-linked recessive manner. The disease is caused by a change in the nucleotide sequence of an X-linked gene encoding glypican 3, a protein belonging to the heparan-sulfate membrane proteoglycan family. SGBS case studies are almost entirely restricted to the pediatric population. Scarce literature describing SGBS course in adults may be due to both the high mortality of SGBS patients in childhood and low rate of SGBS diagnosis in adults. We present a case of a 39-year-old man with an initial diagnosis of acromegaly. Genetic tests revealed a hitherto unreported deletion in the GPC3 gene. SGBS manifestations in our patient included tall stature, dysmorphic features, and central nervous system (CNS) anatomical pathology. MRI of the head visualized abnormalities of median line structures, a feature consistent with SGBS: an unclosed craniopharyngeal canal, a sellar-suprasellar cyst, dysmorphic pituitary gland, and a cyst of the septum pellucidum. Moreover, cardiomyopathy complicated by life-threatening paroxysmal ventricular tachycardia was diagnosed. Although various cardiac anomalies are often found in SGBS, their pathogenesis is unclear and may be multifactorial. We believe that the presented case contributes to a better understanding of SGBS and may help clinicians in introducing prophylaxis and treatment for its comorbidities.

Barker’s thrifty phenotype hypothesis suggests a relationship between low birth weight and obesity, type 2 diabetes mellitus and atherosclerosis in later life. Aim: The aim of the study was to assess the nutritional status and the incidence of selected atherosclerotic risk factors among people in their third decade of life in relation to their birth weight and term of delivery. Material and methods: A total of 119 subjects aged between 19 and 29 years were classified into the following groups: born prematurely (group I – 38 subjects), born at term, but with low birth weight (group II – 39 subjects), and born at term with normal birth weight (group C – 42 subjects) as well as into groups with birth weight

The occurrence of overweight and obesity in children and adolescents can be associated with increased incidence of hypertension in this population. Aim of the study: Assessment of the relationship between 24-hour blood pressure parameters in hospitalised teenagers with primary hypertension and their nutrition status at the stage of diagnosis. Material and methods: The study group consisted of 112 patients aged 11–17 years (42 girls and 70 boys) with diagnosed primary hypertension. Nutrition indicators were taken into account. The study group was divided into group I – persons with normal nutrition status (40 patients) and group II – persons with overweight/obesity (72 patients). The groups were compared using the t-test with a separate variance estimation or with the Mann–Whitney U test. The relationship between blood pressure parameters and nutrition status indicators was assessed based on Spearman’s rank correlation. Results: The study groups were significantly different in terms of the nutrition status indicators, but no significant differences were observed in the evaluated blood pressure parameters. The nutrition status indicators in group I were not significantly correlated with the blood pressure parameters. Significant correlations were found in group II. Positive correlation with the nutrition indicators were observed for nocturnal systolic blood pressure load and average nighttime value of this load. Body mass index exhibited a negative correlation with nocturnal systolic load decline. Conclusions: 1) 24-hour blood pressure parameters in hospitalised adolescents with primary hypertension that correlated with the nutrition status include nighttime systolic blood pressure parameters (nocturnal load, mean measure, nocturnal fall). 2) The relationship of these parameters with the nutrition status was observed only in patients with overweight/obesity.

Cancer cells exhibit specific metabolism allowing them to survive and proliferate in various oxygen conditions and nutrients' availability. Hepatocytes are highly active metabolically and thus very sensitive to hypoxia. The purpose of the study was to investigate the effect of oxygen on the expression of phase II detoxification enzymes in hepatocellular carcinoma cells (HepG2) cultured in minimal and rich media (with nonessential amino acids and GSH). The cells were cultured at 1% hypoxia, 10% tissue normoxia, and 21% atmospheric normoxia. The total cell count was determined by trypan blue exclusion dye and the expression on mRNA level by RT-PCR. The result indicated that the expression of glutathione-dependent enzymes (GSTA, M, P, and GPX2) was sensitive to oxygen and medium type. At 1% hypoxia the enzyme expression (with the exception of GSTA) was higher in minimal compared to rich medium, whereas at 10% normoxia it was higher in the rich medium. The expression was oxygen-dependent in both types of medium. Among phenol sulfotransferase SULT1A1 was not sensitive to studied factors, whereas the expression of SULT1A3 was depended on oxygen only in minimal medium. It can be concluded that in HepG2 cells, the detoxification by conjugation with glutathione and, to a lower extent with sulfate, may be affected by hypoxia and/or limited nutrients' availability. Besides, because the data obtained at 10% oxygen significantly differ from those at 21%, the comparative studies on hypoxia should be performed in relation to 10% but not 21% oxygen.

Cu/Zn superoxide dismutase (SOD1) gene mutations are the most frequently reported genetic causes of amyotrophic lateral sclerosis (ALS). The objective of the study was to describe a clinical phenotype and haplotype background of Polish and Japanese ALS patients harbouring the K3E SOD1 mutation. The K3E mutation was identified by direct sequencing, high resolution melting analysis or high-throughput microarray-based resequencing system. Microsatellite polymorphic markers flanking SOD1 were genotyped in members of six kindreds and two SALS patients. Results demonstrated that the K3E mutation was responsible for classic ALS. The median age of onset was 54 years. The clinical phenotype did not substantially differ between SALS and FALS cases of either ethnic origin, with some intrafamiliar variabilities. There was a limb onset in 92% of patients. In patients with bulbar syndrome, dysphagia predominated over dysarthria. Respiratory insufficiency was found in 61.1% of patients (19-84 months after the first symptoms onset). Median survival was 101 months with age of death ranging from 45 to 77 years. K3E was the most frequent SOD1 mutation among Polish FALS patients. It originated independently, on different haplotype background in the Polish and Japanese populations. In conclusion, recurrent K3E mutation results in a relatively slowly progressing limb onset ALS with classic phenotype.

Summary Diffusion tensor imaging is unique in its ability to non-invasively visualize white matter fiber tracts in the human brain in vivo. White matter fiber bundles of the human brain form a spatial defined by the anatomical and functional architecture. Determination of axonal pathways provides an invaluable means to study the connectivity of human brain and its functional network. Comparison of fiber tract properties across subjects requires comparison at corresponding anatomical locations. In this paper, we present application of white matter tractography method based on incoherent motion of water molecules in fiber tissue, mainly in central nervous system. This motion is itself dependent on the micro-structural environment that restricts the movement of the water molecules. In white matter fibers there is a pronounced directional dependence on diffusion. With white matter fiber tracking or tractography, projections among brain regions can be detected in the three-dimentional diffusion tensor dataset according to the directionality of the fibers. The authors indicate diagnostic possibilities of MR tractography in otolaryngology for imaging the nervous tracts of sense of hearing, smell and taste with particular consideration of otoneurological evaluation of patients with cochlear implants and bone anchored hearing aid (BAHA). White matter tracts can be evaluated independently by using diffusion tensor tractography, which appears to be a promising technique for determining changes in white matter in degenerative disease. The authors also indicate that method as a beneficial in the surgical planning for patients with intrinsic brain tumors.

The work is a continuation of studies on tau expression and alternative splicing in the central nervous system of transgenic mice harboring human SOD1 with G93A amyotrophic lateral sclerosis (ALS)-associated mutation. Since age is an important risk factor for ALS, we expanded the studies into younger animals (age 5 and 25 days). We also included cerebellum, a structure not studied in the context of neurodegeneration in ALS. We found decreased total tau-mRNA expression in hippocampus but not in cortex and spinal cord of young transgenics, and a lack of exon 10 in 5-day-old mice. In cerebellum, the total tau-mRNA expression was increased in transgenic animals during the whole period of life, however at the symptomatic stage of ALS (age 120 days) the level of protein was decreased. It can be concluded that the SOD1 G93A mutation causes early alterations of tau expression in cns, which are not exclusively restricted to the upper and lower motor neuron.

Glutathione S-transferase pi (GSTP1) is a crucial enzyme in detoxification of electrophilic compounds and organic peroxides. Together with Se-dependent glutathione peroxidase (Se-GSHPx) it protects cells against oxidative stress which may be a primary factor implicated in motor neuron disease (MND) pathogenesis. We investigated GSTP1 polymorphisms and their relationship with GST and Se-GSTPx activities in a cohort of Polish patients with MND. Results were correlated with clinical phenotypes. The frequency of genetic variants for GSTP1 exon 5 (I105V) and exon 6 (A114V) was studied in 104 patients and 100 healthy controls using real-time polymerase chain reaction. GST transferase activity was determined in serum with 1-chloro-2,4-dinitrobenzene, its peroxidase activity with cumene hydroperoxide, and Se-GSHPx activity with hydrogen peroxide. There were no differences in the prevalence of GSTP1 polymorphism I105V and A114V between MND and controls, however the occurrence of CT variant in codon 114 was associated with a higher risk for MND. GSTP1 polymorphisms were less frequent in classic ALS than in progressive bulbar palsy. In classic ALS C* (heterozygous I /V and A /V) all studied activities were significantly lower than in classic ALS A* (homozygous I /I and A/A). GST peroxidase activity and Se-GSHPx activity were lower in classic ALS C* than in control C*, but in classic ALS A* Se-GSHPx activity was significantly higher than in control A*. It can be concluded that the presence of GSTP1 A114V but not I105V variant increases the risk of MND, and combined GSTP1 polymorphisms in codon 105 and 114 may result in lower protection of MND patients against the toxicity of electrophilic compounds, organic and inorganic hydroperoxides.

Background: Sporadic amyotrophic lateral sclerosis (SALS) is a fatal motor neuron degenerative disease of unclear pathogenesis. Disturbances of intracellular transport are possible causes of the disease. Objective: We evaluated the expression of motor proteins involved in the anterograde (kinesins KIF1B, KIF5C) and retrograde (KIFC3, dynactin subunits DCTN1 and DCTN3) intracellular transport in peripheral blood mononuclear cells (PBMCs). Materials and Methods: PBMCs were obtained from 74 SALS patients with different clinical phenotypes, 65 blood donors (healthy control I), and 29 cases with other neurological diseases (disease control II) divided into subgroups IIA (atypical parkinsonism) and IIB (ALS-mimicking disorders). mRNA expression was studied by real-time qPCR, and protein level by Western blotting. Results: In SALS, KIF5C and KIFC3 expression was significantly lower and DCTN1 higher than in control I, and dependent of age. KIF1B expression was significantly higher in SALS than in subgroup IIB, whereas DCTN1 and DCTN3 were higher in SALS than in subgroup IIA. All changes in the studied proteins were statistically significant in classic ALS but not in progressive muscular atrophy. Conclusion: In SALS, and especially in classic ALS, the changes in motor protein expression may alter bidirectional intracellular transport in PBMCs. More studies are needed to find out whether the levels of KIF5C and DCTN1 may be useful in ALS diagnosis, and whether KIF1B expression may discriminate ALS from ALS-mimicking disorders.

The expression of glutathione S-transferase pi (GST pi), an enzyme responsible for inactivation of a large variety of toxic compounds was studied in spinal cord, motor and sensory brain cortex obtained from patients who died in the course of amyotrophic lateral sclerosis (ALS). The studies were performed on formalin-fixed, paraffin-embedded (FFPE) and freshly frozen tissues. The method of RNA isolation from FFPE was modified. A significant decrease of GST pi-mRNA expression was found in cervical spinal cord and motor brain cortex of ALS subjects comparing to analogue control tissues (P

Survival Motor Neuron 1 (SMN1) is a causative gene for autosomal recessive infantile and juvenile proximal spinal muscular atrophy. SMN1 duplications have recently been found to increase susceptibility to amyotrophic lateral sclerosis. The role of centromeric SMN copy (SMN2) has been postulated in progressive muscular atrophy (PMA). The aim of this study was to analyse the SMN1 and SMN2 copy number variations in patients with PMA. SMN1 and SMN2 genotype was studied in 87 patients with PMA, diagnosed at the Department of Neurology, Medical University of Warsaw, between 1992 and 2012 and in 600 healthy controls. Results demonstrated that three copies of SMN1 were found in 8.1% of PMA patients and in 24% of PMA patients with disease duration above 48 months compared to 4.6% of the general population. Patients with three SMN1 copies had a limb onset, lower median age of onset and longer disease duration compared to patients with two SMN1 copies. There were no significant differences in the SMN2 copy numbers. In conclusion, the increased copy number of SMN1 may be a susceptibility factor to PMA and influence the clinical phenotype.

To evaluate foveal function, retinal circulation and foveal thickness before and after intravitreal ranibizumab injections in eyes with wet type of age-related macular degeneration (AMD).The study group consisted of 21 eyes (20 patients) with choroidal neovascularisation (CNV) due to AMD. Inclusion criteria were based on fluorescein angiography (FA) and distance best corrected visual acuity (DBCVA)--log MAR scale. In each eye, 3 consecutive injections of ranibizumab every 4 weeks were administered and then individual course for re-injections according to DBCVA and optical coherence tomography (OCT) up to 12 months was applied. At baseline, 3, 6 and 12 months follow-up, the following tests were performed: DBCVA, multifocal electroretinogram (mfERG) and OCT. Additionally, FA was carried out before the treatment, 3 and 12 months from the baseline.At baseline, FA revealed mainly minimally occult choroidal neovascularisation--57% (12/21) of eyes. At 3 months choroidal neovascularisation diameter was stable; no leakage from active choroidal neovascularisation was seen in 76% (16/21) of eyes. After 12 months follow-up, increase in choroidal neovascularisation diameter was seen in 43% (9/21) of eyes and no leakage in 57% (12/21) of cases. The mean DBCVA significantly improved only after 3 months (p0.02). Significant decrease of mean foveal thickness was observed in each follow-ups (p0.01). The mfERG data from the macular region remained stable or improved slightly in some cases.In our series of patients with the wet type of AMD after intravitreal injections of ranibizumab in 12 months follow-up, the reduction of foveal thickness was noted while DBCVA and the bioelectrical function from the macular region measured by the mfERG remained stable.

Dysfunction of fast axonal transport, vital for motor neurons, may lead to neurodegeneration. Anterograde transport is mediated by N-kinesins (KIFs), while retrograde transport by dynein 1 and, to a minor extent, by C-kinesins. In our earlier studies we observed changes in expression of N- and C-kinesins (KIF5A, 5C, C2) in G93ASOD1-linked mouse model of motor neuron degeneration. In the present work we analyze the profile of expression of the same kinesins in mice with a dynein 1 heavy chain mutation (Dync1h1, called Cra1), presenting similar clinical symptoms, and in Cra1/SOD1 mice with milder disease progression than SOD1 transgenics. We found significantly higher levels of mRNA for KIF5A and KIF5C but not the KIFC2 in the frontal cortex of symptomatic Cra1/+ mice (aged 365 days) compared to the wild-type controls. No changes in kinesin expression were found in the spinal cord of any age group and only mild changes in the hippocampus. The expression of kinesins in the cerebellum of the presymptomatic and symptomatic mice (aged 140 and 365 days, respectively) was much lower than in age-matched controls. In Cra1/SOD1 mice the changes in KIFs expression were similar or more severe than in the Cra1/+ groups, and they also appeared in the spinal cord. Thus, in mice with the Dync1h1 mutation, which impairs dynein 1-dependent retrograde transport, expression of kinesin mRNA is affected in various structures of the CNS and the changes are similar or milder than in mice with double Dync1h1/hSOD1G93A mutations.

Heroin is an illicit narcotic abused by millions of people worldwide. In our earlier studies we have shown that heroin intoxication changes the antioxidant status in human brain. In the present work we continued our studies by estimating the effect of heroin abuse on reduced glutathione (GSH) and enzymes related to this cofactor, such as glutathione S-transferase detoxifying electrophilics (GST) and organic peroxides (as Se-independent glutathione peroxidase—GSHPx), and Se-dependent glutathione peroxidase (Se-GSHPx) specific mainly for hydrogen peroxide. Studies were conducted on human brains obtained from autopsy of 9 heroin abusers and 8 controls. The level of GSH and the activity of glutathione-related enzymes were determined spectrophotometrically. The expression of GST pi on mRNA and protein level was studied by RT-PCR and Western blotting, respectively. The results indicated significant increase of GST and GSHPx activities, unchanged Se-GSHPx activity, and decreased level of GSH in frontal, temporal, parietal and occipital cortex, brain stem, hippocampus, and white matter of heroin abusers. GST pi expression was increased on both mRNA and protein levels, however the increase was lower in brain stem than in other regions. Heroin affects all regions of human brain, and especially brain stem. Its intoxication leads to an increase of organic rather then inorganic peroxides in various brain regions. Glutathione S-transferase plays an important role during heroin intoxication, however its protective effect is lower in brain stem than in brain cortex or hippocampus.

Diffusion tensor imaging is unique in its ability to non-invasively visualize white matter fiber tracts in the human brain in vivo. White matter fiber bundles of the human brain form a spatial defined by the anatomical and functional architecture. Determination of axonal pathways provides an invaluable means to study the connectivity of human brain and its functional network. Comparison of fiber tract properties across subjects requires comparison at corresponding anatomical locations. In this paper, we present application of white matter tractography method based on incoherent motion of water molecules in fiber tissue, mainly in central nervous system. This motion is itself dependent on the micro-structural environment that restricts the movement of the water molecules. In white matter fibers there is a pronounced directional dependence on diffusion. With white matter fiber tracking or tractography, projections among brain regions can be detected in the three-dimentional diffusion tensor dataset according to the directionality of the fibers. The authors indicate diagnostic possibilities of MR tractography in otolaryngology for imaging the nervous tracts of sense of hearing, smell and taste with particular consideration of otoneurological evaluation of patients with cochlear implants and bone anchored hearing aid (BAHA). White matter tracts can be evaluated independently by using diffusion tensor tractography, which appears to be a promising technique for determining changes in white matter in degenerative disease. The authors also indicate that method as a beneficial in the surgical planning for patients with intrinsic brain tumors.

Glutathione S-transferase (GST, EC 2.5.1.18) is one of the most important enzymes protecting cells against toxic, electrophilic compounds. It occurs in isoenzymes, expression of which changes in various pathological conditions including neoplasia. The aim of the present study was to analyze the expression pattern of GST pi, mi4 and alpha in human tongue and floor of mouth squamous cell carcinomas.The studies were conducted on tissue samples obtained from surgery. RT-PCR was used to determine the GSTs-mRNA expression, and Western blotting to study GST pi protein expression.Indicated statistically significant differences in GST pi mRNA expression in tumor and control tissues. GST pi expression was raised in tumors on both mRNA and protein levels, whereas mRNA-GSTmi4 and alfa expression was diverse.Increased level of GST pi expression indicates its role in carcinogenesis.

Tau is a protein involved in regulation of microtubule stability, axonal differentiation and transport. Alteration of retrograde transport may lead to motor neuron degeneration. Thus alternative mRNA splicing and expression of tau isoforms were studied in a transgenic mouse model harboring the human SOD1 G93A mutation. The studies were performed on cortex, hippocampus and spinal cord of 64- and 120-day-old animals (presymptomatic and symptomatic stage) and wild type controls. Exon 10 was found in all studied tissues. The 2N isoform containing exons 2 and 3 (+2+3) and the 1N (+2-3) predominated over the 0N (-2-3) in brain regions of the studied mice. The 2N expression was significantly lower in cortex and hippocampus of symptomatic animals compared to analogue control tissues. The decrease in 2N expression resulted in lower levels of total tau mRNA and tau protein. No changes in tau expression were observed in spinal cord of studied animals.

Glutathione S-transferase (GST) is a part of the most important enzymatic systems protecting organisms from toxic, electrophilic compounds. Expression of numerous GST isoenzymes is changed in various pathological conditions, including neoplasia. The aim of present study was to analyze the expression pattern of GST pi, mu (mu4 and mu5) and alpha at the mRNA and protein levels in human primary gliomas. The studies were conducted on tissue samples obtained from surgery.There were no changes in GST pi mRNA expression, determined by RT-PCR, between gliomas and tumor adjacent tissues. However, using Western blotting method, an increase in GST pi protein in tumors was observed, which might be caused by the lower rate in protein degradation. Decrease in the expression of GST mu at mRNA level (mu4), as well as at protein level was shown in gliomas compared to control tissues. mRNA for GST mu5 isoform was demonstrated only in two tumor cases, but not in normal tissues, and for GST alpha, in one sample of control tissue. At the protein level, GST pi expression was increased in gliomas, whereas GST mu was decreased.Changes in GST isoenzymes expression can play an important role in the susceptibility of central nervous system to carcinogenesis.

Dynactin is a complex motor protein involved in the retrograde axonal transport disturbances of which may lead to amyotrophic lateral sclerosis (ALS). Mice with hSOD1G93A mutation develop ALS-like symptoms and are used as a model for the disease studies. Similar symptoms demonstrate Cra1 mice, with Dync1h1 mutation. Dynactin heavy (DCTN1) and light (DCTN3) subunits were studied in the CNS of humans with sporadic ALS (SALS), mice with hSOD1G93A (SOD1/+), Dync1h1 (Cra1/+), and double (Cra1/SOD1) mutation at presymptomatic and symptomatic stages. In SALS subjects, in contrast to control cases, expression of DCTN1-mRNA but not DCTN3-mRNA in the motor cortex was higher than in the sensory cortex. However, the mean levels of DCTN1-mRNA and protein were lower in both SALS cortexes and in the spinal cord than in control structures. DCTN3 was unchanged in brain cortexes but decreased in the spinal cord on both mRNA and protein levels. In all SALS tissues immunohistochemical analyses revealed degeneration and loss of neuronal cells, and poor expression of dynactin subunits. In SOD1/+ mice both subunits expression was significantly lower in the frontal cortex, spinal cord and hippocampus than in wild-type controls, especially at presymptomatic stage. Fewer changes occurred in Cra1/SOD1 and Cra1/+ mice.It can be concluded that in sporadic and SOD1-related ALS the impairment of axonal retrograde transport may be due to dynactin subunits deficiency and subsequent disturbances of the whole dynein/dynactin complex structure and function. The Dync1h1 mutation itself has slight negative effect on dynactin expression and it alleviates the changes caused by SOD1G93A mutation.

Glutathione S-transferase pi (GST pi) is an enzyme involved in cell protection against toxic electrophiles and products of oxidative stress. GST pi expression was studied in transgenic mice hybrids (B6-C3H) with symptoms of neurodegeneration harboring SOD1G93A (SOD1/+), Dync1h1 (Cra1/+) and double (Cra1/SOD1) mutations, at presymptomatic and symptomatic stages (age 70, 140, 365 days) using RT-PCR and Western blotting. The main changes in GST pi expression were observed in mice with the SODG93A mutation. In SOD1/+ and Cra1/SOD1 transgenics, with the exception of cerebellum, the changes in GST pi-mRNA accompanied those in GST pi protein. In brain cortex of both groups the expression was unchanged at the presymptomatic (age 70 days) but was lower at the symptomatic stage (age 140 days) and at both stages in hippocampus and spinal cord of SOD1/+ but not of Cra1/SOD1 mice compared to age-matched wild-type controls. In cerebellum of the presymptomatic and the symptomatic SOD1/+ mice and presymptomatic Cra1/SOD1 mice, the GST pi-mRNA was drastically elevated but the protein level remained unchanged. In Cra1/+ transgenics there were no changes in GST pi expression in any CNS region both on the mRNA and on the protein level. It can be concluded that the SOD1G93A but not the Dync1h1 mutation significantly decreases detoxification efficiency of GST pi in CNS, however the Dync1h1 mutation reduces the effects caused by the SOD1G93A mutation. Despite similarities in neurological symptoms, the differences in GST pi expression between SOD1/+ and Cra1/+ transgenics indicate a distinct pathogenic entity of these two conditions.