Hyaluronan in mesenchymal stromal cell lineage differentiation from human pluripotent stem cells: application in serum free culture.

BACKGROUND: Hyaluronan (HA) is an extracellular glycosaminoglycan polysaccharide with widespread roles throughout development and in healthy and neoplastic tissues. In pluripotent stem cell culture it can support both stem cell renewal and differentiation. However, responses to HA in culture are influenced by interaction with a range of cognate factors and receptors including components of blood serum supplements, which alter results. These may contribute to variation in cell batch production yield and phenotype as well as heighten the risks of adventitious pathogen transmission in the course of cell processing for therapeutic applications. MAIN: Here we characterise differentiation of a human embryo/pluripotent stem cell derived Mesenchymal Stromal Cell (hESC/PSC-MSC)-like cell population by culture on a planar surface coated with HA in serum-free media qualified for cell production for therapy. Resulting cells met minimum criteria of the International Society for Cellular Therapy for identification as MSC by expression of. CD90, CD73, CD105, and lack of expression for CD34, CD45, CD14 and HLA-II. They were positive for other MSC associated markers (i.e.CD166, CD56, CD44, HLA 1-A) whilst negative for others (e.g. CD271, CD71, CD146). In vitro co-culture assessment of MSC associated functionality confirmed support of growth of hematopoietic progenitors and inhibition of mitogen activated proliferation of lymphocytes from umbilical cord and adult peripheral blood mononuclear cells, respectively. Co-culture with immortalized THP-1 monocyte derived macrophages (Mɸ) concurrently stimulated with lipopolysaccharide as a pro-inflammatory stimulus, resulted in a dose dependent increase in pro-inflammatory IL6 but negligible effect on TNFα. To further investigate these functionalities, a bulk cell RNA sequence comparison with adult human bone marrow derived MSC and hESC substantiated a distinctive genetic signature more proximate to the former. CONCLUSION: Cultivation of human pluripotent stem cells on a planar substrate of HA in serum-free culture media systems is sufficient to yield a distinctive developmental mesenchymal stromal cell lineage with potential to modify the function of haematopoietic lineages in therapeutic applications.

Therapeutic potential of lithium chloride and valproic acid against neuronopathic types of mucopolysaccharidoses through induction of the autophagy process.

Mucopolysaccharidoses (MPS) are a group of inherited disorders, caused by mutations in the genes coding for proteins involved (directly or indirectly) in glycosaminoglycan (GAG) degradation. A lack or drastically decreased residual activity of a GAG-degrading enzyme leads to the storage of these compounds, thus damaging proper functions of different cells, including neurons. The disease leads to serious psycho-motor dysfunctions and death before reaching the adulthood. Until now, induction of the autophagy process was considered as one of the therapeutic strategies for treatment of diseases caused by protein aggregation (Alzheimer's, Parkinson's, and Huntington's diseases). However, this strategy has only been recently suggested as a potential therapy for MPS. In this work, we show that the pharmacological stimulation of autophagy, by using valproic acid and lithium chloride, led to accelerated degradation of accumulated GAGs. Cytotoxicity tests indicated the safety of the use of the investigated compounds. We observed an increased number of lysosomes and enhanced degradation of heparan sulfate (one of GAGs). Induction of the autophagy process was confirmed by measuring abundance of the marker proteins, including LC3-II. Moreover, inhibition of this process resulted in abolition of the valproic acid- and LiCl-mediated reduction in GAG levels. This is the first report on the possibility of using valproic acid and lithium chloride for reducing levels of GAGs in neuronopathic forms of MPS.

X-chromosome inactivation patterns depend on age and tissue but not conception method in humans.

Female somatic X-chromosome inactivation (XCI) balances the X-linked transcriptional dosages between the sexes, randomly silencing the maternal or paternal X chromosome in each cell of 46,XX females. Skewed XCI toward one parental X has been observed in association with ageing and in some female carriers of X-linked diseases. To address the problem of non-random XCI, we quantified the XCI skew in different biological samples of naturally conceived females of different age groups and girls conceived after in vitro fertilization (IVF). Generally, XCI skew differed between saliva, blood, and buccal swabs, while saliva and blood had the most similar XCI patterns in individual females. XCI skew increased with age in saliva, but not in other tissues. We showed no significant differences in the XCI patterns in tissues of naturally conceived and IVF females. The gene expression profile of the placenta and umbilical cord blood was determined depending on the XCI pattern. The increased XCI skewing in the placental tissue was associated with the differential expression of several genes out of 40 considered herein. Notably, skewed XCI patterns (> 80:20) were identified with significantly increased expression levels of four genes: CD44, KDM6A, PHLDA2, and ZRSR2. The differences in gene expression patterns between samples with random and non-random XCI may shed new light on factors contributing to the XCI pattern outcome and indicate new paths in future research on the phenomenon of XCI skewing.

Biological aspects of phage therapy versus antibiotics against Salmonella enterica serovar Typhimurium infection of chickens.

Phage therapy is a promising alternative treatment of bacterial infections in human and animals. Nevertheless, despite the appearance of many bacterial strains resistant to antibiotics, these drugs still remain important therapeutics used in human and veterinary medicine. Although experimental phage therapy of infections caused by Salmonella enterica was described previously by many groups, those studies focused solely on effects caused by bacteriophages. Here, we compared the use of phage therapy (employing a cocktail composed of two previously isolated and characterized bacteriophages, vB_SenM-2 and vB_Sen-TO17) and antibiotics (enrofloxacin and colistin) in chickens infected experimentally with S. enterica serovar Typhimurium. We found that the efficacies of both types of therapies (i.e. the use of antibiotics and phage cocktail) were high and very similar to one another when the treatment was applied shortly (one day) after the infection. Under these conditions, S. Typhimurium was quickly eliminated from the gastrointestinal tract (GIT), to the amount not detectable by the used methods. However, later treatment (2 or 4 days after detection of S. Typhimurium in chicken feces) with the phage cocktail was significantly less effective. Bacteriophages remained in the GIT for up to 2-3 weeks, and then were absent in feces and cloaca swabs. Interestingly, both phages could be found in various organs of chickens though with a relatively low abundance. No development of resistance of S. Typhimurium to phages or antibiotics was detected during the experiment. Importantly, although antibiotics significantly changed the GIT microbiome of chickens in a long-term manner, analogous changes caused by phages were transient, and the microbiome normalized a few weeks after the treatment. In conclusion, phage therapy against S. Typhimurium infection in chickens appeared as effective as antibiotic therapy (with either enrofloxacin or colistin), and less invasive than the use the antibiotics as fewer changes in the microbiome were observed.

Do Body Composition and Values of Selected Nutritional Status Indices Influence the Glycaemic Index Values of Vegetarian Dishes? A Pilot Study in a Group of Older Women.

An ageing population brings with it the need for public policy to respond to the demands and health needs of this group of people. The ageing process has been shown to be associated with changes in body composition. These mainly concern a decrease in muscle mass and an increase in body fat. Body composition and other indicators of nutritional status are important factors differentiating carbohydrate management. Glycaemic index (GI) values may be affected by differences resulting from individual metabolism. The rate of carbohydrate digestion is also influenced by a number of factors, including the degree to which the product is processed, the structure of the starch, and the presence of protein, fat and dietary fibre. Available studies do not provide information on the glycaemic response following the consumption of specific products by older people with varying BMI and body composition. Therefore, the aim of this study was to evaluate the effect of the body mass index (BMI) values of women aged 50-80 years on the glycaemic response after eating vegetarian meals and the influence of selected indices of nutritional status on their GI values. It has been shown that the areas under the glycaemic curves after the consumption of the tested foods, both traditional and modified, are higher in the group of overweight and obese women. Nevertheless, the GI of meals consumed by those with a BMI ≥ 25.0 kg/m2 is lower than that of foods consumed by women with normal values of this index. In the group of women with BMI 18.5-24.9 kg/m2, on the basis of an analysis of the obtained correlations, it was observed that the GI value of modified products depends on the percentage of body fat (FM%) (p = 0.0363) and the percentage of fat free mass (FFM%) (p = 0.0363), and, in the case of traditional products, also on the percentage of total body water (%) (p = 0.0133). In the group of women with a BMI ≥ 25.0 kg/m2, significant correlations were only found between the GI of modified foods and the waist-to-hip ratio (WHR) (p = 0.0363) and the ratio of waist circumference to height (WHtR) (p = 0.0369) indices. The GI values of food set solely with the participation of young, healthy people should not be the basis for the nutrition planning of all groups of people.

Behavioral- and blood-based biomarkers for Huntington's disease: Studies on the R6/1 mouse model with prospects for early diagnosis and monitoring of the disease.

Background: Huntington's disease (HD) is a genetic neurodegenerative disorder characterized by a triad of cognitive, psychiatric and motor symptoms. One of the main mechanisms of the disease, besides the aggregation of mutant proteins, is the chronic inflammation that occurs in patients long before the onset of motor dysfunction. Currently, no effective therapy is available for HD. As the progression of HD is usually slow and its first clinical manifestations are non-specific, a correct diagnosis is difficult. Moreover, any attempts to develop therapies for HD require monitoring of unequivocal markers of the disease which are also hardly available. Objective: Using a mouse model of HD, we aimed to determine a battery of biomarkers in peripheral blood, as well as those related to cognitive or anxiety disturbances, corresponding to the most characteristic breakpoints in HD. Methods: The R6/1 mouse line was used as an animal HD model. Levels of cytokines and hormones in blood were estimated by ELISA. A series of behavioral and movement tests was applied. Results: Significant elevation of levels of specific inflammatory markers (IL-6, TNF-α, IL-1ß, IL-12) was observed in the course of HD. At early stages of the disease, an impairment of anti-inflammatory defense mechanisms was evident, expressed by a significant decrease in IL-10 levels. The disturbances were faster in females than in males which might also translate into a potentially worse response to any administered drugs. Impaired memory and anxiety could be detected. Conclusion: A set of simple behavioral- and blood-based biomarkers for HD has been detected.

Probiotics and Postbiotics as Substitutes of Antibiotics in Farm Animals: A Review.

Since 2006, the use of growth-promoting antibiotics has been banned throughout the European Union. To meet the expectations of livestock farmers, various studies have been carried out with the use of lactic acid bacteria. Scientists are trying to obtain the antimicrobial effect against the most common pathogens in large-scale farms. Supplementing the diet of broilers with probiotics (live, nonpathogenic microorganisms) stabilized the intestinal microbiota, which improved the results of body weight gain (BWG) and feed intake (FI). The positive effect of probiotics based on lactic acid bacteria has been shown to prevent the occurrence of diarrhea during piglet weaning. The antagonistic activity of postbiotics (inanimate bacteria, cell components, or post-fermentation by-products) from post-culture media after lactobacilli cultures has been proven on Staphylococcus aureus-the pathogen most often responsible for causing mastitis among dairy cows. The article aims to present the latest research examining the antagonistic effect of lactic acid bacteria on the most common pathogens in broilers, piglets, pigs, and cow farms.

Dosage Compensation in Females with X-Linked Metabolic Disorders.

Through the use of new genomic and metabolomic technologies, our comprehension of the molecular and biochemical etiologies of genetic disorders is rapidly expanding, and so are insights into their varying phenotypes. Dosage compensation (lyonization) is an epigenetic mechanism that balances the expression of genes on heteromorphic sex chromosomes. Many studies in the literature have suggested a profound influence of this phenomenon on the manifestation of X-linked disorders in females. In this review, we summarize the clinical and genetic findings in female heterozygotic carriers of a pathogenic variant in one of ten selected X-linked genes whose defects result in metabolic disorders.

Effect of eating habits, BMI value, physical activity and smoking cigarettes on blood lipid indices of adolescent boys from Poland.

BACKGROUND: The lifestyle of young boys has impact on the risks of cardiovascular diseases. OBJECTIVE: The aim of the study was to evaluate the effect of atherosclerosis risk factors determined by overweight and obesity and lifestyle, i.e.: eating habits, low physical activity and smoking cigarettes, on blood lipid profile of boys at the age of 16 to 18. MATERIAL AND METHODS: The study covered 369 boys from secondary schools. They were evaluated for the supply of dietary constituents with atherogenic and protective actions, for nutritional status acc. to Cole's criteria, the level of physical activity, and smoking cigarettes. Lipid metabolism was determined based on criteria recommended by the American National Cholesterol Education Program (NCEP). Logistic regression analysis was conducted and risk odds ratio [OR] was determined. RESULTS: Analyses showed the boys to be characterized by overweight (10.8%) and obesity (2.7%), and by inappropriate concentration of total cholesterol (26.5%), LDL (13.3%), HDL (21.7%) and triglycerides (41.7%). High BMI turned out to be a significant risk factor of an elevated total cholesterol concentration: [aOR]=2.27; triglycerides: [aOR]=2.35 and LDL: [aOR]=2.41. Low physical activity was found to negatively affect the concentration of LDL: [aOR]=1.88. The boys smoking cigarettes were shown to have a reduced HDL: [aOR]=1.65. The total content of fat and saturated fatty acids in diet exerted a significantly negative impact on blood lipid profile of the boys. CONCLUSIONS: The lifestyle of the young boys was demonstrated to determine the risk of cardiovascular diseases. Overweight and obesity, abdominal obesity in particular, were found to be a significant risk factor of disorders in their lipid metabolism.

Changes in expressions of genes involved in the regulation of cellular processes in mucopolysaccharidoses as assessed by fibroblast culture-based transcriptomic analyses.

Recent studies indicated that apart from lysosomal storage of glycosaminoglycans (GAGs), secondary and tertiary changes in cellular processes may significantly contribute to development of disorders and symptoms occurring in mucopolysaccharidoses (MPS), a group of lysosomal storage diseases in which neurodegeneration is specific for most types and subtypes. In this report, using transcriptomic data, we demonstrate that regulation of hundreds of genes coding for proteins involved in regulations of various cellular processes is changed in cells derived from patients suffering from all types and subtypes of MPS. Among such genes there are 10 which expression is significantly changed in 9 or more (out of 11) MPS types/subtypes; they include IER3IP1, SAR1A, TMEM38B, PLCB4, SIN3B, ABHD5, SH3BP5, CAPG, PCOLCE2, and MN1. Moreover, there are several genes whose expression is changed over log2 > 4 times in some MPS types relative to control cells. The above analysis indicates that significant changes in expression of genes coding for various regulators of cellular processes may considerably contribute to development of cellular dysfunctions, and further appearance of specific symptoms of MPS, including neurodegeneration.

The New Model of Snail Expression Regulation: The Role of MRTFs in Fast and Slow Endothelial-Mesenchymal Transition.

Endothelial-mesenchymal transition (EndMT) is a crucial phenomenon in regulating the development of diseases, including cancer metastasis and fibrotic disorders. The primary regulators of disease development are zinc-finger transcription factors belonging to the Snail family. In this study, we characterized the myocardin-related transcription factor (MRTF)-dependent mechanisms of a human snail promoter regulation in TGF-ß-stimulated human endothelial cells. Although in silico analysis revealed that the snail promoter's regulatory fragment contains one GCCG and two SP1 motifs that could be occupied by MRTFs, the genetic study confirmed that MRTF binds only to SP1 sites to promote snail expression. The more accurate studies revealed that MRTF-A binds to both SP1 elements, whereas MRTF-B to only one (SP1near). Although we found that each MRTF alone is capable of inducing snail expression, the direct cooperation of these proteins is required to reinforce snail expression and promote the late stages of EndMT within 48 hours. Furthermore, genetic and biochemical analysis revealed that MRTF-B alone could induce the late stage of EndMT. However, it requires a prolonged time. Therefore, we concluded that MRTFs might cause EndMT in a fast- and slow-dependent manner. Based on MRTF-dependent Snail upregulation, we recognized that TGF-ß1, as an MRTF-B regulator, is involved in slow EndMT induction, whereas TGF-ß2, which altered both MRTF-A and MRTF-B expression, promotes a fast EndMT process.

Frequency of occurrence of metabolic syndrome risk factors in children and adolescents from the city of Wroclaw and surroundings

Background: A recently growing number of children and adolescents with overweight or obesity is indicative of the need for diagnosing their complications that may appear in the early childhood. For this reason, diagnostic criteria were developed for components of the metabolic syndrome (MS) also for these groups of the population. Objective: This study was aimed at evaluating the frequency of metabolic syndrome risk factors occurrence in children and adolescents from the city of Wroclaw and surroundings depending on gender, age and physical activity. Material and methods: Investigations on the frequency of occurrence of dietary and non-dietary risk factors of the metabolic syndrome were carried out in the years 2010-2017 among 771 children and adolescents aged 10-18 year, attending to primary schools, gymnasiums and secondary schools in Wroclaw. Results: The lack of any components of the metabolic syndrome was demonstrated in 14.78% of the children aged 10-12 years as well as in 17.38% of both adolescents aged 13-15 and 17-18 years. One risk factor was most frequently diagnosed in children aged 10-12 years (17.89%) and it was arterial hypertension (16.08% of the whole surveyed population). Three MS components were demonstrated in 15 persons (1.95% of the whole surveyed population), including in 7 girls and 8 boys. The persons with three MS risk factors from the age category 13-15 years constituted 0.26% whereas these from the age category 17-18 years constituted 1.69% of the whole surveyed group. Conclusions: It shall be concluded that the incidence of the fully symptomatic MS depended significantly on the age of the surveyed, but not on their gender. Among the three adopted components of MS, the most frequently demonstrated disorders included: arterial hypertension, abdominal obesity and increased concentration of triglycerides in blood serum.

Assessment of dietary habits and lifestyle among people with HIV.

BACKGROUND: The aim of this study was to determine the consumption of specific food groups by people with HIV and to determine the quality of their diets. OBJECTIVES: To assess the relationship between selected eating habits and lifestyles of people infected with HIV. The research was conducted at the HIV/AIDS Preventative and Therapeutic Clinic of the Infectious Disease Prevention and Therapy Center at Wroclaw Health Center (SPZOZ Wroclaw), Poland. MATERIAL AND METHODS: The study was conducted in 2019 among 31 patients of a counselling center in Wroclaw. To determine the frequency of food consumption and eating habits, the KomPAN® questionnaire, prepared by employees of the Polish Academy of Sciences, was used. RESULTS: All study participants were characterized by a small degree of unhealthy features in their diets; 87% of the respondents also demonstrated a small degree of healthy features in their diets, although the responses they gave showed that they assessed their nutritional knowledge and diet highly. Consumption of sweet snacks and adding salt to cooked meals were prevalent. The respondents took part in moderate physical activity and rarely consumed highly processed fast food products, though they ate fish and legumes - an important part of the diet - with similar frequency. CONCLUSIONS: More attention should be given to the nutritional issues of patients treated for HIV, and emphasis should be placed on promoting healthy eating habits among this population. In the scientific literature, few such studies are available that address issues related to the diet of HIV-infected people.

The Role of Dimethyl Sulfoxide (DMSO) in Gene Expression Modulation and Glycosaminoglycan Metabolism in Lysosomal Storage Disorders on an Example of Mucopolysaccharidosis.

Obstacles to effective therapies for mucopolysaccharidoses (MPSs) determine the need for continuous studies in order to enhance therapeutic strategies. Dimethyl sulfoxide (DMSO) is frequently utilised as a solvent in biological studies, and as a vehicle for drug therapy and the in vivo administration of water-insoluble substances. In the light of the uncertainty on the mechanisms of DMSO impact on metabolism of glycosaminoglycans (GAGs) pathologically accumulated in MPSs, in this work, we made an attempt to investigate and resolve the question of the nature of GAG level modulation by DMSO, the isoflavone genistein solvent employed previously by our group in MPS treatment. In this work, we first found the cytotoxic effect of DMSO on human fibroblasts at concentrations above 3%. Also, our results displayed the potential role of DMSO in the regulation of biological processes at the transcriptional level, then demonstrated a moderate impact of the solvent on GAG synthesis. Interestingly, alterations of lysosomal ultrastructure upon DMSO treatment were visible. As there is growing evidence in the literature that DMSO can affect cellular pathways leading to numerous changes, it is important to expand our knowledge concerning this issue.

Autoantibodies to heat shock proteins 60, 70, and 90 in patients with rheumatoid arthritis.

Heat shock proteins (HSP) have been reported to impact immune responses and to be associated with rheumatoid arthritis (RA). Recently, we provided evidence for a role of autoantibodies to Hsp40 in patients with RA. In this study, we aimed at investigating the humoral autoimmune response to Hsp60, Hsp70, and Hsp90 in RA patients (n = 39). In comparison with healthy controls (n = 40), circulating IgG, IgM, and IgA autoantibodies against Hsp60, Hsp70, and Hsp90 were significantly increased in RA patients. Non-parametric statistical analysis, however, revealed no significant association between anti-HSP and disease activity or disease progression. On the other hand, positive correlations between serum levels of anti-Hsp60 IgG and IL-4 (Th2-like cytokine) or between serum levels of anti-Hsp90 IgG and IFN-É£ (Th1-like cytokine) were found to be statistically significant in RA. In addition, a significant inverse correlation was found for serum levels of anti-Hsp70 IgM and TNF-α (Th1-like cytokine) in RA. Our results suggest a pronounced anti-Hsp60, anti-Hsp70, and anti-Hsp90 humoral autoimmune response in RA patients that seems not to be directly linked to RA pathophysiology, however, may have a potential modulatory impact on inflammatory status in this disease. Further investigations are needed to clarify the role of anti-HSP autoantibodies in RA.

Differential effects of various soy isoflavone dietary supplements (nutraceuticals) on bacterial growth and human fibroblast viability.

Flavonoids, polyphenolic compounds present in many food products, affect growth of different bacterial species when tested as purified or synthetic substances. They can also influence gene expression in human cells, like fibroblasts. Here, we asked if soy isoflavone extracts, commonly used in many products sold as anti-menopausal dietary supplements, influence bacterial growth similarly to a synthetic isoflavone, genistein. Four commercially available products were tested in amounts corresponding to genistein concentrations causing inhibition of growth of Vibrio harveyi (a model bacterium sensitive to this isoflavone) and Escherichia coli (a model bacterium resistant to genistein). Differential effects of various extracts on V. harveyi and E. coli growth, from stimulation, to no changes, to inhibition, were observed. Moreover, contrary to genistein, the tested extracts caused a decrease (to different extent) in viability of human dermal fibroblasts. These results indicate that effects of various soy isoflavone extracts on bacterial growth and viability of human cells are different, despite similar declared composition of the commercially available products.

Inhibition of Shiga toxin-converting bacteriophage development by novel antioxidant compounds.

Oxidative stress may be the major cause of induction of Shiga toxin-converting (Stx) prophages from chromosomes of Shiga toxin-producing Escherichia coli (STEC) in human intestine. Thus, we aimed to test a series of novel antioxidant compounds for their activities against prophage induction, thus, preventing pathogenicity of STEC. Forty-six compounds (derivatives of carbazole, indazole, triazole, quinolone, ninhydrine, and indenoindole) were tested. Fifteen of them gave promising results and were further characterized. Eleven compounds had acceptable profiles in cytotoxicity tests with human HEK-293 and HDFa cell lines. Three of them (selected for molecular studies) prevent the prophage induction at the level of expression of specific phage genes. In bacterial cells treated with hydrogen peroxide, expression of genes involved in the oxidative stress response was significantly less efficient in the presence of the tested compounds. Therefore, they apparently reduce the oxidative stress, which prevents induction of Stx prophage in E. coli.

Autophagy stimulation as a promising approach in treatment of neurodegenerative diseases.

Autophagy is a process of degradation of macromolecules in the cytoplasm, particularly proteins of a long half-life, as well as whole organelles, in eukaryotic cells. Lysosomes play crucial roles during this degradation. Autophagy is a phylogenetically old, and evolutionarily conserved phenomenon which occurs in all eukaryotic cells. It can be found in yeast Saccharomyces cerevisiae, insect Drosophila melanogaster, and mammals, including humans. Its high importance for cell physiology has been recognized, and in fact, dysfunctions causing impaired autophagy are associated with many severe disorders, including cancer and metabolic brain diseases. The types and molecular mechanisms of autophagy have been reviewed recently by others, and in this paper they will be summarized only briefly. Regulatory networks controlling the autophagy process are usually described as negative regulations. In contrast, here, we focus on different ways by which autophagy can be stimulated. In fact, activation of this process by different factors or processes can be considered as a therapeutic strategy in metabolic neurodegenerative diseases. These aspects are reviewed and discussed in this article.

How close are we to therapies for Sanfilippo disease?

Sanfilippo disease is one of mucopolysaccharidoses (MPS), a group of lysosomal storage diseases characterized by accumulation of partially degraded glycosaminoglycans (GAGs). It is classified as MPS type III, though it is caused by four different genetic defects, determining subtypes A, B, C and D. In each subtype of MPS III, the primary storage GAG is heparan sulfate (HS), but mutations leading to A, B, C, and D subtypes are located in genes coding for heparan N-sulfatase (the SGSH gene), α-N-acetylglucosaminidase (the NAGLU gene), acetyl-CoA:α-glucosaminide acetyltransferase (the HGSNAT gene), and N-acetylglucosamine-6-sulfatase (the GNS gene), respectively. Neurodegenerative changes in the central nervous system (CNS) are major problems in Sanfilippo disease. They cause severe cognitive disabilities and behavioral disturbances. This is the main reason of a current lack of therapeutic options for MPS III patients, while patients from some other MPS types (I, II, IVA, and VI) can be treated with enzyme replacement therapy or bone marrow or hematopoietic stem cell transplantations. Nevertheless, although no therapy is available for Sanfilippo disease now, recent years did bring important breakthroughs in this aspect, and clinical trials are being conducted with enzyme replacement therapy, gene therapy, and substrate reduction therapy. These recent achievements are summarized and discussed in this review.

Female Fabry disease patients and X-chromosome inactivation.

Fabry disease is an X-linked inherited lysosomal storage disorder caused by mutations in the gene encoding α-galactosidase A (GLA). Once it was thought to affect only hemizygous males. Over the last fifteen years, research has shown that most females carrying mutated allele also develop symptoms, demonstrating a wide range of disease severity, from a virtually asymptomatic to more classical profile, with cardiac, renal, and cerebrovascular manifestations. This variable expression in females is thought to be influenced by the process of X-chromosome inactivation (XCI). The aim of this study was to assess severity of the clinical phenotype, to analyze XCI patterns, and to estimate their effect on disease manifestation in twelve female Fabry disease patients from five unrelated Polish families. Our analyses revealed that patients presented with the broad range of disease expression - from mild to severe, and their clinical involvement did not correlate with XCI profiles. Female carriers of the mutation in the GLA gene with the random XCI may present with the wide range of disease signs and symptoms. Thus, XCI is not a main factor in the phenotype variability of Fabry disease manifestation in heterozygous females.