Human umbilical cord stroma-derived mesenchymal stem cells (hUCS-MSCs) are considered as a remark... more Human umbilical cord stroma-derived mesenchymal stem cells (hUCS-MSCs) are considered as a remarkable and promising stem cell source to be potentially used in cellular therapies. While no graft rejection has been reported in the recipient organism even in xeno-transplantation studies, attenuate tumor cell growth and gene transfers have been experimentally shown. In this study, we have demonstrated a reliable, reproducible and efficient cryopreservation method of hUCS-MSCs resulting in one of the highest cell survival rates reported so far. Conventional, computer-controlled multistep slow freezing (MSSF), and vitrification methods were comparatively tested using cell permeable [dimethylsulfoxide (DMSO), ethylene glycol] and impermeable [trehalose, sucrose, hydroxyethyl starch (HES), human serum albumin] cryoprotectant agents (CPAs). After determining the ice nucleation point for each solution, latent heat evolution was suppressed during freezing, followed by a cooling process to -40°C at 1°C/min or 0.3°C/min. The efficiency of the cryopreservation techniques used was determined by cell viability and proliferation assays, the expression of cell surface markers, cytoskeletal proteins and chromosome alignments. The cell survival rate was found to be highest (87±5%) by MSSF with sucrose (0.1 M) +DMSO (10%) at 1°C/min freezing rate. In this group, no significant difference was noted before and after the cryopreservation in cell morphology, cytokeratin, vimentin, and -smooth muscle actin profiles and the expressions of CD105, CD90, CD73, CD29 and HLA-DR. Second highest cell survival ratio (85±6%) was obtained in DMSO (10%) alone at 1°C/min freezing rate. Interestingly, poor (18±15%) cell survival rates were obtained after vitrification. Cumulatively, results indicated that MSSF favors the other freezing protocols with an addition of sucrose or DMSO alone depending on the freezing rate used.
Yetiskin insan vucudunda ve embriyonda varligi gosterilen kok hucrelerin in vitro ve in vivodeney... more Yetiskin insan vucudunda ve embriyonda varligi gosterilen kok hucrelerin in vitro ve in vivodeneylerinde bircogunun cesitli dokulari olusturmak uzere farklilasabildigi gosterilmistir. Bunlardan bir grubu da, gobek kordonu icinde yer alan stroma hucreleridir (IGKS-MKH). Bu hucreler ozellikle mezenkimal seri hucreleri olan adiposit, kondrosit, osteosit ve myosite olan farklilasmalar dikkati cekecek kadar umit vaat edici bicimde ortaya konmustur. Bunun yaninda bu hucreler alicida immun yanit olusturmamisve doku reddi ile karsilasilmamistir ve bu hucrelerin immun yetmezligi olan SCID farelere verildiginde teratom olusturmadigi gozlenmistir. Bu hucre kaynagi yuksek miktarda elde edilebilme ve multipotent ozellige sahip olmasiyla onem kazanmistir. Fakat yaptigimiz calismalarda in vitro ortama alindiklarinda replikatif omurlerinin sinirli (P10-13) oldugu gozlenmistir. Yaptigimiz calismalar bu hucrelerin telomeraz aktivitesinin dokudan cikarildiklarinda ve kulture edildiklerinde azaldigi yonundedir. Bu nedenle, yaptigimizcalismada IGKS-MKH’lerine retroviral yontemle Telomerazin katalitik alt birimi olan TERT (Telomerase Reverse Transcriptase) geni kalici olarak aktarilarak, hucreler olumsuz hale getirilip, sahip olduklari kok hucre ozelliklerini koruyup korumadiklari test edilmistir. Sonuclarimiz, TERT geni ile enfekte edilen IGKS-MKH’lerinde TERT geni ifadesinin arttigi ve telomeraz aktivitesinin yukseldigini gostermektedir. T-IGKS-MKH’leri 30 pasaj sonra bile MKH ozelliklerini ve tipik hucre morfolojisini korurken, yuksek proliferasyon yetenegi gostermektedir. Ote yandan son donemde ozellikle allojeneik transplantasyonlarda kullanilmak uzere bir dizi MKH serilerinin gelistirildigi izlenmektedir. Insanda immun yanit olusturmadigi kanitlanan IGKS-MKH’nin immortal serilerinin yapimi onemlidir. Bu calismada ozgun degere sahip bu degerli hucreleretelomeraz aktivitesi kazandirip olumsuzlestirerek bu kaynagin potansiyel ozelliklerini kaybetmeden korunmasi saglanmistir.
While septic shock is a syndrome of circulatory in-sufficiency which can be caused by a variety o... more While septic shock is a syndrome of circulatory in-sufficiency which can be caused by a variety of illnesses it may result in prostration, hypotension, pallor, diaphoresis, cool skin and oliguria. The terms bacteremia and septicemia refer to the presence of bacteriae and their toxins in the blood. Bacteremia is used when bacteriae are recovered from blood cultures of a patient who does not appear to be seriously ill and who may be afebrile. In con trast, septicemia implies that blood cultures are positive and that the patient appears seriously or critically ill. In some patients bacteremia or septi cemia may be related to focal infection (e.g. pneu monia, osteomyelitis, endocarditis, meningitis) the presence of which can be suspected or confirmed rapidly by history, physical examination and roentgenographic or other laboratory studies. In such cases, bacteremia or septicemia may be suspected with a high degree of likelihood (5, 13, 17).
Mesenchymal stem cells (MSCs) have been well identified in cultures obtained from various human t... more Mesenchymal stem cells (MSCs) have been well identified in cultures obtained from various human tissues. However, they give no clue as to their native identity, frequency, or anatomical location. Based on in vivo and in vitro experimental studies, the most promising candidate for the MSC niche is the vicinity of blood vessels. Capillaries to large-caliber arteries and veins house multipotent progenitor cells that share many morphological, phenotypical, and developmental features with freshly isolated or cultured MSCs. In this mini-review, results from our and other laboratories are summarized and suggest that MSCs originate from tissue sites where pericytes reside, although we do not rule out the possibility that only a small portion of pericytes give rise to MSCs. Understanding the MSC niches can definitely help us to take many parameters into account when designing isolation, expansion, and differentiation protocols for using these cells in future therapeutic applications.
Although many countries banned of its usage, carbofuran (CF) is still one of the most commonly us... more Although many countries banned of its usage, carbofuran (CF) is still one of the most commonly used carbamate derivative insecticides against insects and nematodes in agriculture and household, threatening the human and animal health by contaminating air, water, and food. Our goal was to evaluate the potential toxic effects of CF on mammalian oocytes besides mitotic cells. Caspase-dependent apoptotic pathway was assessed by immunofluorescence and western blot techniques. Alterations in the meiotic spindle formation after CF exposure throughout the in vitro maturation of mice oocyte-cumulus complexes (COCs) were analyzed by using a 3D confocal laser microscope. Maturation efficiency and kinetics were assessed by direct observation of the COCs. Results indicated that the number of TUNEL-positive cells increased in CF-exposed groups, particularly higher doses (>250 mM) in a dose-dependent fashion. The ratio of anticleaved caspase-3 labeled cells in those groups positively correlated with TUNEL-positivity. Western blot analysis confirmed a significant increase in active caspase-3 activity. CF caused a dose-dependent accumulation of oocytes at prometaphase-I (PM-I) of meiosis. Partial loss of spindle microtubules (MTs) was noted, which consequently gave rise to a diamond shape spindle. Aberrant pericentrin foci were noted particularly in PM-I and metaphase-I (M-I) stages. Conclusively, CF (1) induces programmed cell death in a dose-dependent manner, and (2) alters spindle morphology most likely through a mechanism that interacts with MT assembly and/or disorientation of pericentriolar proteins. Overall, data suggest that CF could give rise to aneuploidy or cell death in higher doses, therefore reduce fertilization and implantation rates.
Centrosomes, major regulatory sites for the microtubule (MT) nucleation, are regulated in a dynam... more Centrosomes, major regulatory sites for the microtubule (MT) nucleation, are regulated in a dynamic manner throughout the process of meiotic maturation. Recently, centrosome orientation in mouse oocytes has been demonstrated in metaphase I through metaphase II. However, centrosomal protein expression in concordance with MT polymerization in earlier stages of oocyte maturation from germinal vesicle stage (GV) to prometaphase I still remains unclear. The present study aims to assess the centrosome±microtubule remodelling during the onset of meiosis based on strict criteria of nuclear maturation. Six consecutive stages were determined for scoring the oocytes as unrimmed nucleolus (UR), partially rimmed nucleolus (PR), fully rimmed nucleolus (FR), nuclear lamina dissolution (NLD), disappearance of nucleolus (DON), and chromatin condensation (CC). A centrosomal protein, pericentrin, was found tightly localized adjacent to nuclear lamina in UR, lacking any MT nucleation activity. In concordance with the competency to resume meiosis, an increase in the amount and nucleation capacity of pericentrin is noted. In FR, cytoplasmic MT almost disappeared while de-novo microtubule polymerization was found in small aggregates of pericentrin localized around the nucleus. Towards the end of DON and CC, a sudden burst of pericentrin was noted with an extreme MT nucleation activity in an organized fashion that is essential for the rapid formation of ®rst meiotic spindle. The results show that centrosomes display precisely controlled spatio-temporal changes during the onset of meiotic maturation. Accumulation of centrosomal proteins to a single locus followed by a sequestration to several spots might be evidence of a mechanism by which the proper distribution of centrosomal material during nuclear breakdown and subsequently formation of spindle are regulated in concordance with the nuclear maturation.
As the collection and isolation of human bone marrow-derived mesenchymal stem cells (MSCs) requir... more As the collection and isolation of human bone marrow-derived mesenchymal stem cells (MSCs) require invasive and often undesirable procurement procedures, investigators have begun to seek alternative sources of human MSC including the umbilical cord stroma. Here we describe the noninvasive isolation, culture, and basic characterization of human umbilical cord stroma-derived mesenchymal stem cells (hUCS-MSCs). Although some technical and observational variations exist between laboratories, there has been a relatively common consensus regarding the immunological and functional characteristics of hUCS-MSCs. Successful in vitro and in vivo differentiation to several lineages makes these cells an invaluable stem cell source, deserving further testing as a cellular therapy or other applications in regenerative medicine. Therefore, the isolation and culture of hUCS-MSCs still need better clarification to ultimately build an optimal standard procedure among laboratories, tissue banks, and clinics.
The advances and success of umbilical cord-derived mesenchymal stromal cells (UC-MSCs) in experim... more The advances and success of umbilical cord-derived mesenchymal stromal cells (UC-MSCs) in experimental disease animal models have fueled the development of targeted therapies in humans. The therapeutic potential of allogeneic transplantation of UC-MSCs has been under examination since 2009.The purpose of this systematic analysis was to review the published results, limitations and obstacles for UC-MSC transplantation. An extensive search strategy was applied to the published literature, 93 peer-reviewed full-text articles and abstracts were found published by early August 2017 that investigated the safety, efficacy and feasibility of UC-MSCs in 2001 patients with 53 distinct pathologies including many systemic/local, acute/chronic conditions. Few data were extracted from the abstracts and/or Chinese-written articles (n = 7, 8%). Importantly, no long-term adverse effects, tumor formation or cell rejection were reported. All studies noted certain degrees of therapeutic benefit as evidenced by clinical symptoms and/or laboratory findings. Thirty-seven percent (n = 34) of studies were found published as a single case (n = 10; 11%) or 2-10 case reports (n = 24; 26%) with no control group. Due to the nature of many stem cell-based studies, the majority of patients also received conventional therapy regimens, which obscured the pure efficacy of the cells transplanted. Randomized, blind, phase 1/2 trials with control groups (placebocontrolled) showed more plausible results. Given that most UC-MSC trials are early phase, the internationally recognized cell isolation and preparation standards should be extended to future phase 2/3 trials to reach more convincing conclusions regarding the safety and efficacy of UC-MSC therapies.
The printing, publishing and sales rights of this book belong to Akademisyen Kitabevi A.Ş. All or... more The printing, publishing and sales rights of this book belong to Akademisyen Kitabevi A.Ş. All or parts of the book may not be reproduced, printed or distributed by mechanical, electronic, photocopying, magnetic paper and/or other methods without the permission of the said institution. Tables, figures and graphics cannot be used for commercial purposes without permission. This book is sold by the Turkish Ministry of Culture banner.
w w w . a k a d e m i s y e n . c o m Bu üründe yer alan bilgiler sadece lisanslı tıbbi çalışanla... more w w w . a k a d e m i s y e n . c o m Bu üründe yer alan bilgiler sadece lisanslı tıbbi çalışanlar için kaynak olarak sunulmuştur. Herhangi bir konuda profesyonel tıbbi danışmanlık veya tıbbi tanı amacıyla kullanılmamalıdır. Akademisyen Kitabevi ve alıcı arasında herhangi bir şekilde doktor-hasta, terapist-hasta ve/veya başka bir sağlık sunum hizmeti ilişkisi oluşturmaz. Bu ürün profesyonel tıbbi kararların eşleniği veya yedeği değildir. Akademisyen Kitabevi ve bağlı şirketleri, yazarları, katılımcıları, partnerleri ve sponsorları ürün bilgilerine dayalı olarak yapılan bütün uygulamalardan doğan, insanlarda ve cihazlarda yaralanma ve/veya hasarlardan sorumlu değildir. İlaçların veya başka kimyasalların reçete edildiği durumlarda, tavsiye edilen dozunu, ilacın uygulanacak süresi, yöntemi ve kontraendikasyonlarını belirlemek için, okuyucuya üretici tarafından her ilaca dair sunulan güncel ürün bilgisini kontrol etmesi tavsiye edilmektedir. Dozun ve hasta için en uygun tedavinin belirlenmesi, tedavi eden hekimin hastaya dair bilgi ve tecrübelerine dayanak oluşturması, hekimin kendi sorumluluğundadır. Akademisyen Kitabevi, üçüncü bir taraf tarafından yapılan ürüne dair değişiklikler, tekrar paketlemeler ve özelleştirmelerden sorumlu değildir.
Amaç: Yaşayan hücrelere gen salımı yapmak üzere pek çok viral vektör geliştirilmiştir. Sendai vir... more Amaç: Yaşayan hücrelere gen salımı yapmak üzere pek çok viral vektör geliştirilmiştir. Sendai viral (SeV) vektörleri geçici gen ifadesi, geniş konak özgüllüğü, düşük patojenite ve yüksek immünojenite gibi özellikleri sayesinde gen aktarımı için önemli vektörlerdir. SeV vektörleri gen tedavisi, aşı teknolojileri ve rejeneratif amaçlı moleküler tıpta sıklıkla kullanılır. Yöntem: Bu çalışmada, farklı melanoma hücre dizilerinde SeV vektörlerinin gen aktarım verimlilikleri floresan mikroskop ve konfokal lazer taramalı mikroskop görüntüleme teknikleri ile değerlendirilmiştir. A375, MDA-MB-435, G361 ve WM115 hücreleri yeşil flüoresan proteini (GFP) ifade eden SeV vektörleri tarafından farklı virüs derişimlerinde (enfeksiyon çarpanı (MOI): 1, 3 ve 9) transdükte edilmiştir. GFP ifadesi virüs inkübasyonundan 24 ve 48 saat sonrasında kontrol edilmiştir. Konfokal lazer taramalı mikroskop görüntüleme ile gen salım verimliliği hesaplanmıştır. Bulgular: Floresan mikroskop görüntüleme ile düşük virüs derişimlerinde dahi (enfeksiyon çarpanı: 1), A375, MDA -MB-435, G361 ve WM115 hücrelerinin SeV tarafından verimli şekilde transdükte edildiği gösterilmiştir. Viral transdüksiyonu takiben, GFP kontrol gen aktivitesi 24 saat içerisinde gözlemlenmeye başlanmış ve 48 saatte artış göstermiştir. Transdüksiyondan 24 saat sonrasında
The aim of the present study is to develop microemulsion and liposome carrier systems for oral ad... more The aim of the present study is to develop microemulsion and liposome carrier systems for oral administration of transforming growth factor alpha (TGF-) and to investigate the effects of these carrier systems on the gastrointestinal efficiency in rats. Microemulsion (w/o) and liposomes (MLV) were developed and characterised. The carrier systems were administered intragastrically by gastric cannula to male Wistar rats. The highest reduction in the basal acid secretion was observed in the microemulsion containing TGF-and aprotinin group (TAME).The gastric mucus secretion in microemulsion containing TGF-(TME) and TAME treatment groups increased significantly compared to the other groups. TGF-levels in both stomach and duodenum were significantly increased in the TAME group. As a result, it was determined through confocal laser scanning microscope (CLSM) studies that exogenous-applied TGF-attached to endogenous EGF receptors. The microemulsion formulation was found to be a more suitable carrier system for oral administration of TGF-.
Introduction: Human umbilical cord stroma is a rich source of primitive multipotent stromal cells... more Introduction: Human umbilical cord stroma is a rich source of primitive multipotent stromal cells (hUC-MSCs). However, the methods for hUC-MSC isolation and propagation remain controversial and vary among laboratories. Our group previously demonstrated that two cell types emerge upon enzymatic isolation of hUC-MSCs, which subsequently undergo a transition towards a fibroblastoid phenotype in later passages. The aim of this study was to further analyse cultured hUC-MSCs by evaluating the cytoskeletal and cell adhesion proteins and by comparing the remodelling of those proteins in umbilical cord sections to determine the cell alterations due to enzymatic and explant methods. Methods: Tissue sections and cultured cells isolated by enzymatic or explant methods were analysed morphologically and by labelling cytokeratin, vimentin, alpha-smooth muscle actin, E-cadherin and Ncadherin profiles. Results: The present observations confirmed that wide, flat cells (type-1) share myofibroblastic features, appear exclusively in enzymatically isolated early cultures; gradually diminish or are replaced by fibroblastoid cells (type-2) in later passages. In contrast, the explant method does not result in the existence of type-1 cells in vitro. Among the tested CK subtypes, CK18 expression is upregulated, whereas CK19 expression is downregulated upon culturing after both protocols. Vimentin and a-SMA, as the major intermediate filaments of hUC-MSCs were found unaltered throughout the culturing period regardless of the cell isolation technique used. Discussion: The data presented confirm and further elucidate the previously observed phenotypic change in hUC-MSCs as illustrated by alterations in structural proteins during enzymatic isolation and subsequent culturing of cells compared with in situ equivalents.
The present study reports the occurrence of three Indo-Pacific gastropod species at new localitie... more The present study reports the occurrence of three Indo-Pacific gastropod species at new localities in the Mediterranean Sea. Chromodoris quadricolor was recorded on the 29 May 2004 in the eastern Medi-terranean, which is the third report of the species from the Mediterranean Sea. Finella pupoides and Syrnola fasciata are collected from the several sites in the Aegean Sea between 1997 and 2002. The morphological, ecological and distributional characteristics of these species are described.
Stem cell biology and regenerative medicine is a relatively young field. However, in recent years... more Stem cell biology and regenerative medicine is a relatively young field. However, in recent years there has been a tremendous interest in stem cells possibly due to their therapeutic potential in disease states. As a classical definition, a stem cell is an undifferentiated cell that can produce daughter cells that can either remain a stem cell in a process called self-renewal, or commit to a specific cell type via the initiation of a differentiation pathway leading to the production of mature progeny cells. Despite this acknowledged definition, the classification of stem cells has been a perplexing notion that may often raise misconception even among stem cell biologists. Therefore, the aim of this brief review is to give a conceptual approach to classifying the stem cells beginning from the early morula stage totipotent embryonic stem cells to the unipotent tissue-resident adult stem cells, also called tissue-specific stem cells.
Objective: Several studies have demonstrated that human dental pulp from deciduous teeth and huma... more Objective: Several studies have demonstrated that human dental pulp from deciduous teeth and human umbilical cord stroma contains tissue-specific stem cells having varying potentials to differentiate into certain lineages. Both are multipotent, ethically available in large numbers, immunocompatible and developmentally primitive. Also, ease of isolation and high expansion potential in vitro have made them very attractive as a model system for many researchers. However, there are many controversies still existing concerning their functional roles. A comparison of their patterns to multilineage differentiation might give a clear idea for their further usage in tissue engineering.
Human umbilical cord stroma-derived mesenchymal stem cells (hUCS-MSCs) are considered as a remark... more Human umbilical cord stroma-derived mesenchymal stem cells (hUCS-MSCs) are considered as a remarkable and promising stem cell source to be potentially used in cellular therapies. While no graft rejection has been reported in the recipient organism even in xeno-transplantation studies, attenuate tumor cell growth and gene transfers have been experimentally shown. In this study, we have demonstrated a reliable, reproducible and efficient cryopreservation method of hUCS-MSCs resulting in one of the highest cell survival rates reported so far. Conventional, computer-controlled multistep slow freezing (MSSF), and vitrification methods were comparatively tested using cell permeable [dimethylsulfoxide (DMSO), ethylene glycol] and impermeable [trehalose, sucrose, hydroxyethyl starch (HES), human serum albumin] cryoprotectant agents (CPAs). After determining the ice nucleation point for each solution, latent heat evolution was suppressed during freezing, followed by a cooling process to -40°C at 1°C/min or 0.3°C/min. The efficiency of the cryopreservation techniques used was determined by cell viability and proliferation assays, the expression of cell surface markers, cytoskeletal proteins and chromosome alignments. The cell survival rate was found to be highest (87±5%) by MSSF with sucrose (0.1 M) +DMSO (10%) at 1°C/min freezing rate. In this group, no significant difference was noted before and after the cryopreservation in cell morphology, cytokeratin, vimentin, and -smooth muscle actin profiles and the expressions of CD105, CD90, CD73, CD29 and HLA-DR. Second highest cell survival ratio (85±6%) was obtained in DMSO (10%) alone at 1°C/min freezing rate. Interestingly, poor (18±15%) cell survival rates were obtained after vitrification. Cumulatively, results indicated that MSSF favors the other freezing protocols with an addition of sucrose or DMSO alone depending on the freezing rate used.
Yetiskin insan vucudunda ve embriyonda varligi gosterilen kok hucrelerin in vitro ve in vivodeney... more Yetiskin insan vucudunda ve embriyonda varligi gosterilen kok hucrelerin in vitro ve in vivodeneylerinde bircogunun cesitli dokulari olusturmak uzere farklilasabildigi gosterilmistir. Bunlardan bir grubu da, gobek kordonu icinde yer alan stroma hucreleridir (IGKS-MKH). Bu hucreler ozellikle mezenkimal seri hucreleri olan adiposit, kondrosit, osteosit ve myosite olan farklilasmalar dikkati cekecek kadar umit vaat edici bicimde ortaya konmustur. Bunun yaninda bu hucreler alicida immun yanit olusturmamisve doku reddi ile karsilasilmamistir ve bu hucrelerin immun yetmezligi olan SCID farelere verildiginde teratom olusturmadigi gozlenmistir. Bu hucre kaynagi yuksek miktarda elde edilebilme ve multipotent ozellige sahip olmasiyla onem kazanmistir. Fakat yaptigimiz calismalarda in vitro ortama alindiklarinda replikatif omurlerinin sinirli (P10-13) oldugu gozlenmistir. Yaptigimiz calismalar bu hucrelerin telomeraz aktivitesinin dokudan cikarildiklarinda ve kulture edildiklerinde azaldigi yonundedir. Bu nedenle, yaptigimizcalismada IGKS-MKH’lerine retroviral yontemle Telomerazin katalitik alt birimi olan TERT (Telomerase Reverse Transcriptase) geni kalici olarak aktarilarak, hucreler olumsuz hale getirilip, sahip olduklari kok hucre ozelliklerini koruyup korumadiklari test edilmistir. Sonuclarimiz, TERT geni ile enfekte edilen IGKS-MKH’lerinde TERT geni ifadesinin arttigi ve telomeraz aktivitesinin yukseldigini gostermektedir. T-IGKS-MKH’leri 30 pasaj sonra bile MKH ozelliklerini ve tipik hucre morfolojisini korurken, yuksek proliferasyon yetenegi gostermektedir. Ote yandan son donemde ozellikle allojeneik transplantasyonlarda kullanilmak uzere bir dizi MKH serilerinin gelistirildigi izlenmektedir. Insanda immun yanit olusturmadigi kanitlanan IGKS-MKH’nin immortal serilerinin yapimi onemlidir. Bu calismada ozgun degere sahip bu degerli hucreleretelomeraz aktivitesi kazandirip olumsuzlestirerek bu kaynagin potansiyel ozelliklerini kaybetmeden korunmasi saglanmistir.
While septic shock is a syndrome of circulatory in-sufficiency which can be caused by a variety o... more While septic shock is a syndrome of circulatory in-sufficiency which can be caused by a variety of illnesses it may result in prostration, hypotension, pallor, diaphoresis, cool skin and oliguria. The terms bacteremia and septicemia refer to the presence of bacteriae and their toxins in the blood. Bacteremia is used when bacteriae are recovered from blood cultures of a patient who does not appear to be seriously ill and who may be afebrile. In con trast, septicemia implies that blood cultures are positive and that the patient appears seriously or critically ill. In some patients bacteremia or septi cemia may be related to focal infection (e.g. pneu monia, osteomyelitis, endocarditis, meningitis) the presence of which can be suspected or confirmed rapidly by history, physical examination and roentgenographic or other laboratory studies. In such cases, bacteremia or septicemia may be suspected with a high degree of likelihood (5, 13, 17).
Mesenchymal stem cells (MSCs) have been well identified in cultures obtained from various human t... more Mesenchymal stem cells (MSCs) have been well identified in cultures obtained from various human tissues. However, they give no clue as to their native identity, frequency, or anatomical location. Based on in vivo and in vitro experimental studies, the most promising candidate for the MSC niche is the vicinity of blood vessels. Capillaries to large-caliber arteries and veins house multipotent progenitor cells that share many morphological, phenotypical, and developmental features with freshly isolated or cultured MSCs. In this mini-review, results from our and other laboratories are summarized and suggest that MSCs originate from tissue sites where pericytes reside, although we do not rule out the possibility that only a small portion of pericytes give rise to MSCs. Understanding the MSC niches can definitely help us to take many parameters into account when designing isolation, expansion, and differentiation protocols for using these cells in future therapeutic applications.
Although many countries banned of its usage, carbofuran (CF) is still one of the most commonly us... more Although many countries banned of its usage, carbofuran (CF) is still one of the most commonly used carbamate derivative insecticides against insects and nematodes in agriculture and household, threatening the human and animal health by contaminating air, water, and food. Our goal was to evaluate the potential toxic effects of CF on mammalian oocytes besides mitotic cells. Caspase-dependent apoptotic pathway was assessed by immunofluorescence and western blot techniques. Alterations in the meiotic spindle formation after CF exposure throughout the in vitro maturation of mice oocyte-cumulus complexes (COCs) were analyzed by using a 3D confocal laser microscope. Maturation efficiency and kinetics were assessed by direct observation of the COCs. Results indicated that the number of TUNEL-positive cells increased in CF-exposed groups, particularly higher doses (>250 mM) in a dose-dependent fashion. The ratio of anticleaved caspase-3 labeled cells in those groups positively correlated with TUNEL-positivity. Western blot analysis confirmed a significant increase in active caspase-3 activity. CF caused a dose-dependent accumulation of oocytes at prometaphase-I (PM-I) of meiosis. Partial loss of spindle microtubules (MTs) was noted, which consequently gave rise to a diamond shape spindle. Aberrant pericentrin foci were noted particularly in PM-I and metaphase-I (M-I) stages. Conclusively, CF (1) induces programmed cell death in a dose-dependent manner, and (2) alters spindle morphology most likely through a mechanism that interacts with MT assembly and/or disorientation of pericentriolar proteins. Overall, data suggest that CF could give rise to aneuploidy or cell death in higher doses, therefore reduce fertilization and implantation rates.
Centrosomes, major regulatory sites for the microtubule (MT) nucleation, are regulated in a dynam... more Centrosomes, major regulatory sites for the microtubule (MT) nucleation, are regulated in a dynamic manner throughout the process of meiotic maturation. Recently, centrosome orientation in mouse oocytes has been demonstrated in metaphase I through metaphase II. However, centrosomal protein expression in concordance with MT polymerization in earlier stages of oocyte maturation from germinal vesicle stage (GV) to prometaphase I still remains unclear. The present study aims to assess the centrosome±microtubule remodelling during the onset of meiosis based on strict criteria of nuclear maturation. Six consecutive stages were determined for scoring the oocytes as unrimmed nucleolus (UR), partially rimmed nucleolus (PR), fully rimmed nucleolus (FR), nuclear lamina dissolution (NLD), disappearance of nucleolus (DON), and chromatin condensation (CC). A centrosomal protein, pericentrin, was found tightly localized adjacent to nuclear lamina in UR, lacking any MT nucleation activity. In concordance with the competency to resume meiosis, an increase in the amount and nucleation capacity of pericentrin is noted. In FR, cytoplasmic MT almost disappeared while de-novo microtubule polymerization was found in small aggregates of pericentrin localized around the nucleus. Towards the end of DON and CC, a sudden burst of pericentrin was noted with an extreme MT nucleation activity in an organized fashion that is essential for the rapid formation of ®rst meiotic spindle. The results show that centrosomes display precisely controlled spatio-temporal changes during the onset of meiotic maturation. Accumulation of centrosomal proteins to a single locus followed by a sequestration to several spots might be evidence of a mechanism by which the proper distribution of centrosomal material during nuclear breakdown and subsequently formation of spindle are regulated in concordance with the nuclear maturation.
As the collection and isolation of human bone marrow-derived mesenchymal stem cells (MSCs) requir... more As the collection and isolation of human bone marrow-derived mesenchymal stem cells (MSCs) require invasive and often undesirable procurement procedures, investigators have begun to seek alternative sources of human MSC including the umbilical cord stroma. Here we describe the noninvasive isolation, culture, and basic characterization of human umbilical cord stroma-derived mesenchymal stem cells (hUCS-MSCs). Although some technical and observational variations exist between laboratories, there has been a relatively common consensus regarding the immunological and functional characteristics of hUCS-MSCs. Successful in vitro and in vivo differentiation to several lineages makes these cells an invaluable stem cell source, deserving further testing as a cellular therapy or other applications in regenerative medicine. Therefore, the isolation and culture of hUCS-MSCs still need better clarification to ultimately build an optimal standard procedure among laboratories, tissue banks, and clinics.
The advances and success of umbilical cord-derived mesenchymal stromal cells (UC-MSCs) in experim... more The advances and success of umbilical cord-derived mesenchymal stromal cells (UC-MSCs) in experimental disease animal models have fueled the development of targeted therapies in humans. The therapeutic potential of allogeneic transplantation of UC-MSCs has been under examination since 2009.The purpose of this systematic analysis was to review the published results, limitations and obstacles for UC-MSC transplantation. An extensive search strategy was applied to the published literature, 93 peer-reviewed full-text articles and abstracts were found published by early August 2017 that investigated the safety, efficacy and feasibility of UC-MSCs in 2001 patients with 53 distinct pathologies including many systemic/local, acute/chronic conditions. Few data were extracted from the abstracts and/or Chinese-written articles (n = 7, 8%). Importantly, no long-term adverse effects, tumor formation or cell rejection were reported. All studies noted certain degrees of therapeutic benefit as evidenced by clinical symptoms and/or laboratory findings. Thirty-seven percent (n = 34) of studies were found published as a single case (n = 10; 11%) or 2-10 case reports (n = 24; 26%) with no control group. Due to the nature of many stem cell-based studies, the majority of patients also received conventional therapy regimens, which obscured the pure efficacy of the cells transplanted. Randomized, blind, phase 1/2 trials with control groups (placebocontrolled) showed more plausible results. Given that most UC-MSC trials are early phase, the internationally recognized cell isolation and preparation standards should be extended to future phase 2/3 trials to reach more convincing conclusions regarding the safety and efficacy of UC-MSC therapies.
The printing, publishing and sales rights of this book belong to Akademisyen Kitabevi A.Ş. All or... more The printing, publishing and sales rights of this book belong to Akademisyen Kitabevi A.Ş. All or parts of the book may not be reproduced, printed or distributed by mechanical, electronic, photocopying, magnetic paper and/or other methods without the permission of the said institution. Tables, figures and graphics cannot be used for commercial purposes without permission. This book is sold by the Turkish Ministry of Culture banner.
w w w . a k a d e m i s y e n . c o m Bu üründe yer alan bilgiler sadece lisanslı tıbbi çalışanla... more w w w . a k a d e m i s y e n . c o m Bu üründe yer alan bilgiler sadece lisanslı tıbbi çalışanlar için kaynak olarak sunulmuştur. Herhangi bir konuda profesyonel tıbbi danışmanlık veya tıbbi tanı amacıyla kullanılmamalıdır. Akademisyen Kitabevi ve alıcı arasında herhangi bir şekilde doktor-hasta, terapist-hasta ve/veya başka bir sağlık sunum hizmeti ilişkisi oluşturmaz. Bu ürün profesyonel tıbbi kararların eşleniği veya yedeği değildir. Akademisyen Kitabevi ve bağlı şirketleri, yazarları, katılımcıları, partnerleri ve sponsorları ürün bilgilerine dayalı olarak yapılan bütün uygulamalardan doğan, insanlarda ve cihazlarda yaralanma ve/veya hasarlardan sorumlu değildir. İlaçların veya başka kimyasalların reçete edildiği durumlarda, tavsiye edilen dozunu, ilacın uygulanacak süresi, yöntemi ve kontraendikasyonlarını belirlemek için, okuyucuya üretici tarafından her ilaca dair sunulan güncel ürün bilgisini kontrol etmesi tavsiye edilmektedir. Dozun ve hasta için en uygun tedavinin belirlenmesi, tedavi eden hekimin hastaya dair bilgi ve tecrübelerine dayanak oluşturması, hekimin kendi sorumluluğundadır. Akademisyen Kitabevi, üçüncü bir taraf tarafından yapılan ürüne dair değişiklikler, tekrar paketlemeler ve özelleştirmelerden sorumlu değildir.
Amaç: Yaşayan hücrelere gen salımı yapmak üzere pek çok viral vektör geliştirilmiştir. Sendai vir... more Amaç: Yaşayan hücrelere gen salımı yapmak üzere pek çok viral vektör geliştirilmiştir. Sendai viral (SeV) vektörleri geçici gen ifadesi, geniş konak özgüllüğü, düşük patojenite ve yüksek immünojenite gibi özellikleri sayesinde gen aktarımı için önemli vektörlerdir. SeV vektörleri gen tedavisi, aşı teknolojileri ve rejeneratif amaçlı moleküler tıpta sıklıkla kullanılır. Yöntem: Bu çalışmada, farklı melanoma hücre dizilerinde SeV vektörlerinin gen aktarım verimlilikleri floresan mikroskop ve konfokal lazer taramalı mikroskop görüntüleme teknikleri ile değerlendirilmiştir. A375, MDA-MB-435, G361 ve WM115 hücreleri yeşil flüoresan proteini (GFP) ifade eden SeV vektörleri tarafından farklı virüs derişimlerinde (enfeksiyon çarpanı (MOI): 1, 3 ve 9) transdükte edilmiştir. GFP ifadesi virüs inkübasyonundan 24 ve 48 saat sonrasında kontrol edilmiştir. Konfokal lazer taramalı mikroskop görüntüleme ile gen salım verimliliği hesaplanmıştır. Bulgular: Floresan mikroskop görüntüleme ile düşük virüs derişimlerinde dahi (enfeksiyon çarpanı: 1), A375, MDA -MB-435, G361 ve WM115 hücrelerinin SeV tarafından verimli şekilde transdükte edildiği gösterilmiştir. Viral transdüksiyonu takiben, GFP kontrol gen aktivitesi 24 saat içerisinde gözlemlenmeye başlanmış ve 48 saatte artış göstermiştir. Transdüksiyondan 24 saat sonrasında
The aim of the present study is to develop microemulsion and liposome carrier systems for oral ad... more The aim of the present study is to develop microemulsion and liposome carrier systems for oral administration of transforming growth factor alpha (TGF-) and to investigate the effects of these carrier systems on the gastrointestinal efficiency in rats. Microemulsion (w/o) and liposomes (MLV) were developed and characterised. The carrier systems were administered intragastrically by gastric cannula to male Wistar rats. The highest reduction in the basal acid secretion was observed in the microemulsion containing TGF-and aprotinin group (TAME).The gastric mucus secretion in microemulsion containing TGF-(TME) and TAME treatment groups increased significantly compared to the other groups. TGF-levels in both stomach and duodenum were significantly increased in the TAME group. As a result, it was determined through confocal laser scanning microscope (CLSM) studies that exogenous-applied TGF-attached to endogenous EGF receptors. The microemulsion formulation was found to be a more suitable carrier system for oral administration of TGF-.
Introduction: Human umbilical cord stroma is a rich source of primitive multipotent stromal cells... more Introduction: Human umbilical cord stroma is a rich source of primitive multipotent stromal cells (hUC-MSCs). However, the methods for hUC-MSC isolation and propagation remain controversial and vary among laboratories. Our group previously demonstrated that two cell types emerge upon enzymatic isolation of hUC-MSCs, which subsequently undergo a transition towards a fibroblastoid phenotype in later passages. The aim of this study was to further analyse cultured hUC-MSCs by evaluating the cytoskeletal and cell adhesion proteins and by comparing the remodelling of those proteins in umbilical cord sections to determine the cell alterations due to enzymatic and explant methods. Methods: Tissue sections and cultured cells isolated by enzymatic or explant methods were analysed morphologically and by labelling cytokeratin, vimentin, alpha-smooth muscle actin, E-cadherin and Ncadherin profiles. Results: The present observations confirmed that wide, flat cells (type-1) share myofibroblastic features, appear exclusively in enzymatically isolated early cultures; gradually diminish or are replaced by fibroblastoid cells (type-2) in later passages. In contrast, the explant method does not result in the existence of type-1 cells in vitro. Among the tested CK subtypes, CK18 expression is upregulated, whereas CK19 expression is downregulated upon culturing after both protocols. Vimentin and a-SMA, as the major intermediate filaments of hUC-MSCs were found unaltered throughout the culturing period regardless of the cell isolation technique used. Discussion: The data presented confirm and further elucidate the previously observed phenotypic change in hUC-MSCs as illustrated by alterations in structural proteins during enzymatic isolation and subsequent culturing of cells compared with in situ equivalents.
The present study reports the occurrence of three Indo-Pacific gastropod species at new localitie... more The present study reports the occurrence of three Indo-Pacific gastropod species at new localities in the Mediterranean Sea. Chromodoris quadricolor was recorded on the 29 May 2004 in the eastern Medi-terranean, which is the third report of the species from the Mediterranean Sea. Finella pupoides and Syrnola fasciata are collected from the several sites in the Aegean Sea between 1997 and 2002. The morphological, ecological and distributional characteristics of these species are described.
Stem cell biology and regenerative medicine is a relatively young field. However, in recent years... more Stem cell biology and regenerative medicine is a relatively young field. However, in recent years there has been a tremendous interest in stem cells possibly due to their therapeutic potential in disease states. As a classical definition, a stem cell is an undifferentiated cell that can produce daughter cells that can either remain a stem cell in a process called self-renewal, or commit to a specific cell type via the initiation of a differentiation pathway leading to the production of mature progeny cells. Despite this acknowledged definition, the classification of stem cells has been a perplexing notion that may often raise misconception even among stem cell biologists. Therefore, the aim of this brief review is to give a conceptual approach to classifying the stem cells beginning from the early morula stage totipotent embryonic stem cells to the unipotent tissue-resident adult stem cells, also called tissue-specific stem cells.
Objective: Several studies have demonstrated that human dental pulp from deciduous teeth and huma... more Objective: Several studies have demonstrated that human dental pulp from deciduous teeth and human umbilical cord stroma contains tissue-specific stem cells having varying potentials to differentiate into certain lineages. Both are multipotent, ethically available in large numbers, immunocompatible and developmentally primitive. Also, ease of isolation and high expansion potential in vitro have made them very attractive as a model system for many researchers. However, there are many controversies still existing concerning their functional roles. A comparison of their patterns to multilineage differentiation might give a clear idea for their further usage in tissue engineering.
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