Genetic Basis of Hearing Loss

Etiology of hearing impairment (HI) is complex and comprises genetic and environmental factors. Currently, the background of genetic hearing impairment is an area of intensive research and we are witnessing fast progress in this field. The story has begun in 1997 when the DFNB1 locus was discovered with GJB2 and GJB6 genes causative for almost 50% of cases of recessive, profound, prelingual hearing loss. Nowadays, we have much more possibilities for dissecting the reason of HI, but proper assessment of clinical symptoms is essential for selecting the most optimal diagnostic pathway. In the first stage, the detailed characteristic of hearing loss including its level established by pure tone audiometry (PTA) or auditory brainstem responses (ABR), age of onset, and other helpful features as progressive or no progressive type should be provided. Subsequently, the presence or absence of accompanying symptoms should be established and followed by a detailed analysis of pedigree. In addition, modern assistive algorithms such as AudioGene, Face2Gene, and POSSUM are also discussed. Taking into account the variety of causative genes and pathogenic variants underling hearing loss, searching for causative genes, after exclusion of the DFNB1 variants, should be performed with multigenic panels based on next-generation sequencing technology

Nonhematopoietic stem cells of fetal origin--how much of today's enthusiasm will pass the time test?

Stem cells originating at fetal age are for many reasons superior as a material for the regenerative medicine purposes, when compared to their adult counterparts. While hematopoietic cells, isolated from fetal liver or cord blood, have been well known for a long time and have passed practical tests as clinical transplantation material, the non-hematopoietic cells are newly recognized, and the knowledge of their phenotype and differentiation potential is rather insufficient. We, and the others, have identified a subpopulation of cord blood cells phenotypically different from hematopoietic cells (CD34-, CD45-, CD29+, CD44+, CD51+, CD105+, SH-2, SH-3), in vitro plastic adherent, and capable of multilineage differentiation. The other candidates for multipotential stem cells are cells extracted from umbilical cord or placental tissue. The preliminary observations suggest, that these cells, phenotypically similar to the nonhematopoietic cord blood cells, are capable of extensive replication in vitro and of multilineage differentiation into a variety of tissues including cardiac muscle, bone and cartilage, adipocytes, and nerve cells. The other possible medical applications include "rejuvenation" of selected tissues and systems in senile patients, and therapeutical cloning - for both purposes, cells at the fetal stage of genetic regulation may be more useful than cells collected from adult donors. There is still, however, a high level of uncertainty concerning future medical applications of fetal stem cells. Their numbers and characteristics may differ from the preliminary observations, and their behavior in vivo may not fulfill the expectations originating from the in vitro studies. Finally, the autologous applications of stem cells collected at the stage of birth may need the involvement of technical and financial resources for the storage of frozen cell samples throughout the period of life of their potential user. Such procedure seems possible from technical point of view, but may be inadequately substantiated by the eventual advantages

Maternal predictors and quality of umbilical cord blood units

The aim of the study was to determine the relationship between the maternal age at delivery and selected properties of the cord blood stem cells. The study included 50 pregnant women aged between 18 and 38 years in which spontaneous labors or elective cesarean sections were performed. Umbilical cord blood was collected immediately after the women were delivered of newborns. The samples were analyzed in the Polish Stem Cells Bank in Warsaw. The highest mean WBC level (p < 0.05) was observed in the umbilical blood collected from patients aged 35 years and more. Similarly, the highest mean cell viability was observed in the umbilical cord blood collected from patients aged 35 and more. There were no statistically significant correlations between the CD34+ cells count and mean cell viability in the umbilical cord blood and the maternal age. With the significance level at p < 0.001, the females after spontaneous labor revealed a visibly higher WBC level than patients after a cesarean section. The higher mean WBC concentration (24.95 thousand/μl) was observed in the umbilical cord blood of patients aged 35 and more after spontaneous labors. In the same group, the umbilical cord blood was also characterized by the highest mean cell viability (98.72%). The number of nucleated cells in the umbilical cord blood collected in the perinatal period increases together with the maternal age. In the course of physiological spontaneous labors, the collected umbilical cord blood has more nucleated cells as compared with elective caesarian sections

Wpływ pH i pCO2 krwi pępowinowej uzyskiwanej okołoporodowo na wybrane parametry komórek macierzystych

Objective: The aim of the study was to demonstrate a correlation between pH and pCO2 levels in umbilical cord blood CD34+ cells and their vitality was analyzed. Material and methods: The study included 50 pregnant women after vaginal delivery at term or elective cesarean section. Umbilical cord blood was collected immediately after birth. The probes were analyzed at the Polish Stem Cell Bank in Warsaw. Results: The number of CD34+ cells ranged from 0.1-0.2 in white blood cells count over 12 thousand/ml and pH of >7.3. If pH ranged between 7.35-7.40, the number of CD34+ was 0.3-0.4. The highest number of CD34+ cells was noted for pH of 7.30-7.35 and amounted to 0.4-0.5. Analysis of stem cell vitality showed that the highest level, over 98%, was obtained when pH was 40.0 mmHg. For pCO2Cel pracy: Celem badania było wykazanie korelacji pomiędzy poziomem pH i pCO2 krwi pępowinowej pobieranej od dawcow, a jakością pozyskanego materiału. Dodatkowo postanowiono znaleźć zależność wpływu pH i pCO2 krwi pępowinowej na ilość komorek CD34+, ich żywotność oraz wpływ sposobu zakończenia ciąży na właściwości krwi pępowinowej. Materiał i metody: Do badania zakwalifikowano 50 ciężarnych kobiet, u ktorych wystąpił porod siłami natury lub elektywne cięcie cesarskie w terminie porodu. Krew pępowinowa została pobrana bezpośrednio po urodzeniu noworodka. Probki poddano analizie w Polskim Banku Komorek Macierzystych w Warszawie. Wyniki: Przy liczbie leukocytow powyżej 12 tys./μl i jednoczesnym pH >7,3 ilość CD34+ mieściła się w przedziale 0,1-0,2. W przypadku gdy pH wynosi 7,35 -7,40 zakres wartości CD34+ wyniosł 0,3-0,4. Największą ilość CD34+ uzyskano dla pH 7,30-7,35 i wynosiła ona 0,4-0,5. Badając żywotność komorek macierzystych, jej największy poziom >98% uzyskano gdy pH wyniosło poniżej 7,3 oraz gdy pH było ≥7,4. Badania wykazały, że żywotność komorek macierzystych obniżała się do 97-98% przy pH 7,3-7,4. Zaobserwowano niskie wartości CD34+ (0,01-0,09) przy wartościach pCO2 >40,0 mmHg. Przy pCO

Surgical Treatment of Wounds Using Stem Cells in Epidermolysis Bullosa (EB)

Epidermolysis bullosa (EB) is a group of hereditary skin diseases, or genodermatoses, characterized by the formation of severe, chronic blisters with painful and life-threatening complications. Despite the previous and ongoing progress in the field, there are still no effective causative treatments for EB. The treatment is limited to relieving symptoms, which—depending on disease severity—may involve skin (blisters, poorly healing wounds caused by the slightest mechanical stimuli, contractures, scarring, pseudosyndactyly) and internal organ abnormalities (esophageal, pyloric, or duodenal atresia; renal failure; and hematopoietic abnormalities). The last decade saw a series of important discoveries that paved the way for new treatment methods, including gene therapy, bone marrow transplantation, cell therapy (allogenic fibroblasts, mesenchymal stem cells [MSCs], and clinical use of induced pluripotent stem cells. Tissue engineering experts are attempting to develop skin-like structures that can facilitate the process of healing to promote skin reconstruction in injuries that are currently incurable. However, this is incredibly challenging, due to the complex structure and the many functions of the skin. Below, we characterize EB and present its potential treatment methods. Despite the cure for EB being still out of reach, recent data from animal models and initial clinical trials in humans have raised patients’, clinicians’, and researchers’ expectations. Consequently, modifying the course of the disease and improving the quality of life have become possible. Moreover, the conclusions drawn based on EB treatment may considerably improve the treatment of other genetic diseases

Lokale Soziale Ökonomie: Lern- und Studienmaterial ; ein Europäisches Curriculum für Praktiker, Unterstützer und Multiplikatoren in Sozialen Unternehmen

Obwohl in einigen europäischen Ländern durchaus eine ganze Reihe von Lehr- und Lernmaterialien für Praktiker und Unterstützer in der Sozialen Ökonomie verfügbar sind, besteht ein wachsender Bedarf an spezialisiertem Lern- und Studienmaterial. Das vorliegende Curriculum zeichnet sich in erster Linie dadurch aus, dass es sich methodisch von traditionellen Formen durch eine explizit partizipative Arbeitsweise unterscheidet und sich inhaltlich vor allem auf jene Aspekte oder Spezifika konzentriert, welche für soziale Unternehmen und die weitere Lokale Soziale Ökonomie typisch sind

Novel Missense Mutation A789V in IQSEC2 underlies X-Linked intellectual disability in the MRX78 family

Disease gene discovery in neurodevelopmental disorders, including X-linked intellectual disability (XLID) has recently been accelerated by next-generation DNA sequencing approaches. To date, more than 100 human X chromosome genes involved in neuronal signaling pathways and networks implicated in cognitive function have been identified. Despite these advances, the mutations underlying disease in a large number of XLID families remained unresolved. We report the resolution of MRX78, a large family with six affected males and seven affected females, showing X-linked inheritance. Although a previous linkage study had mapped the locus to the short arm of chromosome X (Xp11.4-p11.23), this region contained too many candidate genes to be analyzed using conventional approaches. However, our X-chromosome exome resequencing, bioinformatics analysis and inheritance testing revealed a missense mutation (c.C2366T, p.A789V) in IQSEC2, encoding a neuronal GDP-GTP exchange factor for Arf family GTPases (ArfGEF) previously implicated in XLID. Molecular modeling of IQSEC2 revealed that the A789V substitution results in the insertion of a larger side-chain into a hydrophobic pocket in the catalytic Sec7 domain of IQSEC2. The A789V change is predicted to result in numerous clashes with adjacent amino acids and disruption of local folding of the Sec7 domain. Consistent with this finding, functional assays revealed that recombinant IQSEC2A789V was not able to catalyze GDP-GTP exchange on Arf6 as efficiently as wild-type IQSEC2. Taken together, these results strongly suggest that the A789V mutation in IQSEC2 is the underlying cause of XLID in the MRX78 family

Determination of potentially characteristic pro-inflammatory and pro-angiogenic proteins as a new area of development of SPRi matrix biosensors

Niniejsza dysertacja doktorska składa się z cyklu 6 artykułów naukowych: Lukasz Oldak, Anna Sankiewicz, Beata Zelazowska-Rutkowska, Bogdan Cylwik, Zenon Lukaszewski, Marcin Skoczylas, Ewa Gorodkiewicz, Two SPRi biosensors for the determination of cathepsin S in blood plasma. Talanta 225 (2021) 121900; Lukasz Oldak , Beata Zelazowska-Rutkowska, Anna Lesniewska, Piotr Mrozek, Marcin Skoczylas, Zenon Lukaszewski and Ewa Gorodkiewicz, Two Biosensors for the Determination of VEGF-R2 in Plasma by Array SPRi. Molecules 28 (2023) 155; Lukasz Oldak, Anna Lesniewska, Beata Zelazowska-Rutkowska, Eryk Latoch, Zenon Lukaszewski, Maryna Krawczuk-Rybak and Ewa Gorodkiewicz, An Array SPRi Biosensor for Simultaneous VEGF-A and FGF-2 Determination in Biological. Applied Sciences 12 (2022) 12699; Lukasz Oldak, Patrycja Milewska, Sylwia Chludzinska-Kasperuk, Kamil Grubczak, Joanna Reszec and Ewa Gorodkiewicz, Cathepsin B, D and S as Potential Biomarkers of Glioma Malignancy. Journal of Clinical Medicine 11 (2022) 6763; Lukasz Oldak, Sylwia Chludzinska-Kasperuk, Patrycja Milewska, Kamil Grubczak, Joanna Reszec and Ewa Gorodkiewicz, Laminin-5, Fibronectin, and Type IV Collagen as Potential Biomarkers of Brain Glioma Malignancy. Biomedicines 10 (2022) 2290; Lukasz Oldak , Sylwia Chludzinska-Kasperuk, Patrycja Milewska, Kamil Grubczak, Joanna Reszec and Ewa Gorodkiewicz. Biomolecules 12 (2022) 1477.Przedmiotem rozprawy doktorskiej jest konstrukcja trzech nowych bioczujników SPRi, czułych na czynniki proangiogenne i prozapalne oraz opracowanie trzech testów analitycznych opartych na bioczujnikach SPRi, służących do diagnostyki glejaka mózgu. Dla nowo opracowanych metod przeprowadzono procedurę walidacyjną i weryfikacyjną, które doprowadziły do wyznaczenia parametrów analitycznych i stwierdzenia, że nowe metody mogą być przydatne do celów diagnostycznych. Opracowane testy analityczne z przeznaczeniem do diagnostyki glejaka mózgu, ze względu na niemożność rozróżnienia grupy kontrolnej od łagodnych stopni choroby, umożliwiają diagnostykę tej choroby w bardziej zaawansowanych stopniach złośliwości. Mimo to, stanowią jednak wartościową propozycję metod wspierających ubogą jak dotąd diagnostykę laboratoryjną glejaka mózgu, co potwierdza analiza ROC.The subject of the doctoral dissertation is the construction of three new SPRi biosensors, sensitive to pro-angiogenic and pro-inflammatory factors, and the development of three analytical tests based on SPRi biosensors for the diagnosis of brain glioma. A validation and verification procedure was carried out for the newly developed methods, which led to the determination of analytical parameters and the conclusion that the new methods may be useful for diagnostic purposes. Analytical tests developed for the diagnosis of brain glioma, due to the inability to distinguish the control group from mild stages of the disease, enable the diagnosis of this disease in more advanced stages of malignancy. Despite this, they constitute a valuable proposition of methods supporting the so far poor laboratory diagnosis of brain glioma, which is confirmed by the ROC analysis.Uniwersytet w Białymstoku. Wydział Praw