RESUMEN
After hematopoietic stem cell transplantation, chimerism assay is a useful approach to monitor the success of the transplant and to select the appropriate treatment strategy, such as donor leukocyte infusion or immunosuppressive drug dosage. Short tandem repeat PCR is the method that has been accepted as the gold standard for chimerism. However, it has not yet been sufficient to detect mixed chimerism in patients with minimal residual disease. Simultaneously, recent years have been marked by developing sensitive, high-throughput, and accurate molecular genetic assays. These novel methods have subsequently been adapted for the analysis of post-transplant chimerism. In this review, we discuss the technical features of both novel and conventional gold standard chimerism assays. We also discuss their advantages and disadvantages.
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Quimerismo , Trasplante de Células Madre Hematopoyéticas , Humanos , Recurrencia Local de Neoplasia/genética , Trasplante Homólogo , Repeticiones de Microsatélite , Quimera por Trasplante/genéticaRESUMEN
BACKGROUND: Mesenchymal stem cells (MSCs) have the ability to self-renew and are multi-potent. They are a primary candidate for cell-based therapy due to their potential anti-cancer effects. The aim of this study was to evaluate the in vitro anti-leukemic effect of Wharton's Jelly-derived MSC (WJ-MSC) on the leukemic cell lines K562 and HL-60. METHODS: In this present study, WJ-MSCs were isolated from human umbilical cord. The cells were incubated according to the standard culture conditions and characterized by flow cytometry. For experiments, WJ-MSC and leukemic cells were incubated in the direct co-culture at a ratio of 1:5 (leukemia cells: WJ-MSC). HUVEC cells were used as a non-cancerous cell line model. The apoptotic effect of WJ-MSCs on the cell lines was analyzed using Annexin V/PI apoptosis assay. RESULTS: After the direct co-culture of WJ-MSCs on leukemic cell lines, we observed anti-leukemic effects by inducing apoptosis. We had two groups of determination apoptosis with and without WJ-MSCs for all cell lines. Increased apoptosis rates were observed in K562 and HL-60 cell lines, whereas the apoptosis rates in HUVEC cells were low. CONCLUSIONS: MSCs are known to inhibit the growth of tumors of both hematopoietic and non-hematopoietic origin in vitro. In our study, WJ-MSC treatment strongly inhibited the viability of HL-60 and K562 and induced apoptosis. Our results also provided new insights into the inhibition of tumor growth by WJ-MSCs in vitro. In the future, WJ-MSCs could be used to inhibit cancer cells in clinical applications.
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Apoptosis , Técnicas de Cocultivo , Células Endoteliales de la Vena Umbilical Humana , Células Madre Mesenquimatosas , Gelatina de Wharton , Humanos , Células Madre Mesenquimatosas/metabolismo , Gelatina de Wharton/citología , Células K562 , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Células HL-60 , Cordón Umbilical/citología , Leucemia/patología , Leucemia/terapia , Proliferación CelularRESUMEN
BACKGROUND: Mesenchymal stem cells (MSCs), are a novel therapeutic option as the most common cell source, play an important role in the immunomodulation. In this study, it was aimed to determine the effect of MSCs on cytokines secreted by the immune system cells. METHODS: Intracellular cytokine levels (Interleukin-4 (IL-4), Interferon-γ (IFN-γ), and Interleukin-17 (IL-17)) detected by flow cytometry before and after co-culture between peripheral blood mononuclear cells (PBMCs) and MCSs. At the same time, supernatant cytokine levels were measured using the ELISA. RESULTS: In our study, MSCs were isolated from cord blood (CB) and Wharton's Jelly (WJ), and their surface markers (CD44 (100%), CD73 (99.6%), CD90 (100%), CD105 (88%)) shown by flow cytometry method. Both CB-MSCs and WJ-MSCs were used in co-culture MSC/PBMC ratios of 1/5 and 1/10, incubation times of 24 h and 72 h. In the present study, when we compared co-cultures of CB-MSC or WJ-MSC with PBMCs, intracellular levels of cytokines IFN-γ, IL-17 (pro-inflamatory) and IL-4 (anti-inflamatory) were increased, and supernatant levels were decreased significantly (p < 0.05). The level of transforming growth factor beta (TGF-ß) (anti-inflamatory) was significantly decreased for both CB-MSC and WJ-MSC in supernatant (p < 0.05). CONCLUSIONS: It was investigated pro-inflammatory and anti-inflammatory effects of CB-MSCs and WJ-MSCs on PBMCs with the obtained results. According to the results, MSCs demonstrated different immunologic effects after the incubation time and ratios. For further studies, it should be known between interaction of MSCs and immune system.
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Leucocitos Mononucleares , Células Madre Mesenquimatosas , Interleucina-17/metabolismo , Interleucina-4/metabolismo , Células Madre Mesenquimatosas/metabolismo , Citocinas/metabolismo , Interferón gamma/metabolismo , Diferenciación Celular , Células Cultivadas , Proliferación CelularRESUMEN
Mesenchymal stem cell (MSC) has recently generated interest in regenerative medicine. For the definition of MSC, three criteria have been proposed - plastic adherent property, specific surface antigens, and multipotent differentiation capacity. MSC exists in almost all tissues such as synovium, fat, liver, dental pulp, cord blood, Wharton's jelly, and also differentiates into osteoblast, chondrocyte, adipocyte, epithelial, and neuron cells originating from three germ layers. The use of different MSCs for regenerative therapies has been studied over the years as a promising option for treatment of tissue damages and various diseases. Here, the most frequently applied and newly developed stem cell-based techniques are designated, and recent MSC applications knowledge for regenerative medicine in the field are explained.
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Células Madre Mesenquimatosas , Gelatina de Wharton , Diferenciación Celular , Sangre Fetal , Medicina Regenerativa , Cordón UmbilicalRESUMEN
Allogeneic stem cell transplantation (SCT) offers a potential cure for some hematological malignancies. For those patients without a family donor, unrelated donor (MUD) registries serve for identifying the best donor. In the present study, we aimed to give a cross-sectional report of our registry's activity and experience as the first established national MUD registry in the country. The study is retrospective and covers the period of 2016 to 2019. A total of 1855 donor searches were performed, and 642 were included in the study. All data were electronically obtained from the institutional database system. All SCTs were either 10/10 or 9/10 HLA matched and originated from an international registry. The most preferred stem cell source was peripheral blood (70.2%). A quarter of transplants were performed using bone marrow, and cord blood was used with a rate of 1.4%. The pandemic-related problems were similar for the other two national registries. During the pandemic, 71 of 432 patients who were searched for donors underwent stem cell transplant(SCT). The low number was related mostly with postponing of SCTs and/also difficulties in continuing of volunteering and in achievement of stem cells from international registry. During the Covid19 pandemic, the SCT activity of centers decreased according to the national, and international guidelines. The study revealed an organized, and multidirectional capacity of the registry and also the adaptation to unpredicted conditions such as pandemic. On the other hand, there is a need for more effective strategies for donor recruitment and retention programme.
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Médula Ósea , COVID-19 , COVID-19/epidemiología , Estudios Transversales , Documentación , Docentes Médicos , Humanos , Sistema de Registros , Estudios Retrospectivos , Donantes de Tejidos , TurquíaRESUMEN
BK virus infections which usually remains asymptomatic in healthy adults may have different clinical manifestations in immunocompromised patient population. BK virus reactivation can cause BK virus nephropathy in 8% of kidney transplant patients and graft loss may be seen if not treated. Clathrin or Caveolar system is known to be required for the transport of many viruses from Polyomaviruses family including BK viruses. In this study, kidney transplant patients with BK virus viremia were divided into two groups according to the BK virus nephropathy found in kidney biopsy (Group I: Viremia+, Nephropathy+ / Group II: Viremia+, Nephropathy-). Kidney biopsies were examined with immunohistochemical staining to determine the distribution and density of the Caveolin-1 and Clathrin molecules. Immunohistochemical staining of the 31 pathologic specimens with anti-caveolin-1 immunoglobulin revealed statistically significant difference between group-I and group-II. The number of the specimens stained with anti-caveolin-1 was less in group I. On the other hand, we did not find any difference between the groups regarding the anti-clathrin immunochemical analysis. According to these findings, caveolin-1 expression differences in kidney transplant patients may be important in disease progression.
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Virus BK , Enfermedades Renales , Infecciones por Polyomavirus , Infecciones Tumorales por Virus , Adulto , Biopsia , Caveolina 1 , Humanos , Inmunosupresores , Riñón , Coloración y Etiquetado , ViremiaRESUMEN
BACKGROUND: RhD typing has remained of primary importance, as being the leading cause of hemolytic disease of the newborn. Among Rh system's 55 blood group antigens, RhD is the most immunogenic. We aimed with this study to determine weak D/partial D variant frequency in blood donors who were admitted to our blood center and have serologically designated blood group weak D. MATERIALS AND METHODS: We screened blood donors who admitted between 2011 and 2017 to our blood center. Sixty-seven serologically weak D phenotyped donors have participated in the study. These donors' samples were studied further by Polymerase Chain Reaction Sequence- Specific Primers (PCR-SSP) for determining D variants. RESULTS: Weak D phenotype was detected in 228(0.12 %) out of 177,554 donors. Sixty-seven of them agreed to take part in the study. The frequency of weak D and partial D was 68.7 % (n = 46), and 22.4 % (n = 15), in order. The most encountered weak D and partial D variant was type 15 and DFR type, respectively. CONCLUSIONS: The prevalence of serologically weak D phenotypes varies by race and ethnicity. Turkey is a country covering a mixture of European and Asian DNA with different ethnic groups. Thus, our research as giving the overall distribution of RHD variants from the largest city of Turkey, which may reflect the general ethnic background of the country, would help to the establishment of a databank for blood banking. This paper is the first molecular study on RHD variants in Turkey. New molecular research would be more reliable and precise.
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Donantes de Sangre/estadística & datos numéricos , Sistema del Grupo Sanguíneo Rh-Hr/metabolismo , Adulto , Humanos , Persona de Mediana Edad , Turquía , Adulto JovenRESUMEN
Istanbul Medical Faculty, Department of Medical Biology, as the first EFI-accredited HLA laboratory in Turkey since 1999 has been organizing both national and international quality control tests for HLA typing under the banner of the "Balkan external proficiency testing (BEPT)" encompassing countries from EFI regions 8. The first round of BEPT in 2004 was organized for low-resolution HLA-A,-B typing by molecular methods and 12 centres participated in the exercise. In 2005, low-resolution HLA-DR typing was added, and in 2007, low-resolution HLA-C and DQ typing were added to the exercise and 28 centres participated. In 2015, high-resolution (four digits or higher typing) HLA-A,-B,-C,-DR and -DQ typing added to the exercise and 40 centres participated. In the last 2 years, 2017 and 2018, the number of participating centres increased to 48 and 52, respectively. When the distribution of low-resolution typing methods applied in the exercises were investigated, it was found that 82% of the centres in the first round of BEPT in 2004 used PCR-SSP, whereas in the last round in 2018, 26% of the centres preferred SSP, while the rest used SSO (50%) or SSP and SSO (24%) methods. Methods for high-resolution typing were SBT (41%), NGS (18%), SSO (18%), SSP (6%), SBT + NGS (6%) and SBT + SSP (11%). In 15th trial for BEPT, nomenclature mistake rate was 12%, and erroneous result rate was 6% for HLA samples. The most common mistakes in the exercises were nomenclature mistakes. The EFI Standards Version 7.0 is stated "HLA type can be reported using a hyphen if homozygosity is not proven by family studies." In order to provide standard results among HLA laboratories and since accurate HLA typing leads to significant increase of graft and patient survival, quality control exercises should be performed in all HLA-based tests periodically.
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Antígenos HLA/genética , Prueba de Histocompatibilidad , Control de Calidad , Peninsula Balcánica , Antígenos HLA-C , Antígenos HLA-DR , HumanosRESUMEN
BACKGROUND: Recently molecular chimerism analysis after allogeneic hematopoietic stem cell transplantation (AHSCT) has become more important. The use of quantitative chimerism methods aims to assess the kinetics of engraftment to determine graft rejection and failure or relapse of the underlying disease after AHSCT. An accurate and sensitive determination of chimerism status is mandatory after AHSCT. This study aimed to compare two chimerism methods: Multiplex Short Tandem Repeat-Polymerase Chain Reaction (STR-PCR) and quantitative Real Time-PCR (qRT-PCR). METHODS: Thirty-nine blood samples at +28 day were used to extract DNA. Most patients had been diagnosed with acute leukemia (74.3 %) and other hematological diseases. For Multiplex STR-PCR method, PCR products were separated on an ABI 3130 Genetic Analyzer (Applied Biosystem, USA) and for qRT-PCR, an ABI 7500 (Applied Biosystem, USA) Plate System Real Time Analyzer was used to determine the quantification of chimerism per-centage. RESULTS: Of the 39 analyzed samples, 82% concordant chimerism results were detected for both STR-PCR and qRT-PCR methods. Ten mixed chimerisms (MC) were found by qRT-PCR whereas of only 3 MC cases were detected by STR-PCR. In the discordant group of 7 by qRT-PCR, we observed Acute Graft versus Host Disease (aGVHD). Three MC cases that were detected by both STR- and qRT-PCR methods died because of relapse. CONCLUSIONS: The quantitative chimerism method along with multiplex STR-PCR method is important for early detection of MC. qRT-PCR methods can be valuable options in the prevention of graft failure and assisting with fast and early treatment strategies for patients undergoing AHSCT.
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Quimerismo , Trasplante de Células Madre Hematopoyéticas/métodos , Leucemia/terapia , Repeticiones de Microsatélite/genética , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Adolescente , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Quimera por Trasplante/genética , Trasplante Homólogo , Adulto JovenRESUMEN
AIM: The aim of the study was define the normal values of tympanic and axillary body temperature in healthy children. METHODS: This observational cross-sectional study was performed in healthy children aged 0 to 17 years who visited the ambulatory general pediatric of Istanbul Medical Faculty. RESULTS: Of 1364 children, 651 (47.7%) were girls and 713 were boys, the mean (SD, range) age was 72.5 (53.6, 1-204) months. The mean (SD) axillary body temperature was 36.04°C (0.46°C; minimum, 35.0°C; maximum, 37.6°C). The 95th and 99th percentiles were 36.8°C and 37.0°C, respectively. The mean (SD) tympanic body temperature was 36.91°C (0.46°C; minimum, 35.15°C; maximum, 37.9°C). The 95th and 99th percentiles were 37.6°C and 37.8°C, respectively. There were statistically significant differences between sexes for only tympanic body temperatures. Both axillary and tympanic body temperatures were statistically higher in 0 to 2 months compared with other age groups. For this age group, the 99th percentile was 37.5°C for axillary and 37.85°C for tympanic temperature. CONCLUSIONS: Axillary and tympanic body temperatures should be considered as fever when they are more than 37.0°C and 37.8°C, respectively. For 0 to 2 months, fever is 37.5°C and 37.85°C in axillary and tympanic temperatures, respectively.
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Temperatura Corporal/fisiología , Fiebre/diagnóstico , Termómetros/estadística & datos numéricos , Adolescente , Axila/fisiología , Niño , Preescolar , Estudios Transversales , Oído Medio/fisiología , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Valores de Referencia , TurquíaRESUMEN
BACKGROUND: The increasing incidence of obesity in children is a significant risk factor for nonalcoholic fatty liver disease and obesity-associated morbidity. Vitamin D has a major role in bone mineral metabolism and has antimicrobial, antioxidant properties. In this study we aimed to investigate the role of vitamin D in children with obesity with hepatosteatosis. METHODS: A total of 101 children with obesity were included in this study. Hepatosteatosis was diagnosed and graded using ultrasonography. Serum levels of 25-hydroxyvitamin D (25-(OH) vitamin D), calcium, phosphate, alkaline phosphatase, and parathormone were tested. Two-sided t test and Pearson χ tests were used for the relation between vitamin D and hepatosteatosis. RESULTS: In our study group, 45.5% were girls (n=46) and the mean age was 11.5 ± 2.8 years (range 3-17 years). Hepatosteatosis was identified in 58 children (57.4%). The diagnosis of grade 1 and grade 2 hepatosteatosis was made in 41 (40.6%) and 17 (16.8%) children, respectively. Median serum 25-(OH) vitamin D levels in children without hepatosteatosis was 16.4 ng/mL (interquartile range 12.4-24.8 ng/mL), whereas children with grade 1 and grade 2 hepatosteatosis had 25-(OH) vitamin D levels of 14.2 ng/mL (interquartile range 9.5-21.2 ng/mL) and 11.5 ng/mL (interquartile range 7.5-16.7 ng/mL), respectively (P=0.005). There was a positive correlation between insulin resistance and the grade of hepatosteatosis (P=0.03). CONCLUSIONS: Serum vitamin D levels in children with obesity with hepatosteatosis are significantly lower than vitamin D levels in children with obesity without hepatosteatosis. In this observational study we only refer to the association of vitamin D deficiency/insufficiency with hepatosteatosis.
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Enfermedad del Hígado Graso no Alcohólico/sangre , Obesidad/sangre , Vitamina D/análogos & derivados , Adolescente , Fosfatasa Alcalina/sangre , Proteínas de Arabidopsis/sangre , Índice de Masa Corporal , Calcio/sangre , Estudios de Casos y Controles , Niño , Preescolar , Femenino , Humanos , Resistencia a la Insulina , Masculino , Enfermedad del Hígado Graso no Alcohólico/complicaciones , Enfermedad del Hígado Graso no Alcohólico/diagnóstico por imagen , Obesidad/complicaciones , Hormona Paratiroidea/sangre , Índice de Severidad de la Enfermedad , Factores Sexuales , Ultrasonografía , Vitamina D/sangre , Circunferencia de la CinturaRESUMEN
The Histocompatibility and Immunogenetics laboratories provide disease association and pharmacogenetic analyses as well as the tests required for transplantation immunology and transfusion medicine. They perform Human Leukocyte Antigen (HLA) genotyping in patients/recipients and potential donor candidates for solid organ and stem cell transplants using various molecular methods, and determine mismatches. In addition, they also perform HLA antibody tests to detect anti-HLA antibodies in patients and flow cross-matches to evaluate donor-recipient compatibility. Evidence-based clinical guidelines have emphasized the importance of laboratory tests in clinical practices for a long time. Understanding the principles of Quality Control and External Quality Assurance is a fundamental requirement for the effective management of Tissue Typing laboratories. When these processes are effectively implemented, errors in routine assays for transplantation are reduced and quality is improved. In this review, the importance of Quality Assurance, Quality control and proficiency testing in Histocompatibility and Immunogenetic testing, the necessity of external proficiency testing (EPT) for accreditation, and existing and potential EPT programmes will be reviewed and evaluated in the light of the literature.
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Mesenchymal stem cells (MSCs) are promising for clinical studies owing to their self-renewal, multipotency, trophic, and immunomodulatory properties. This study aimed to investigate the cytokine levels of human umbilical cord blood (CB) and Wharton's Jelly-(WJ) derived MSCs relevant to immune modulation on different passage levels in vitro. Umbilical CB MSCs were isolated using the ficoll-paque gradient method, and WJ-MSCs were isolated by the explant method. After isolation, the MSCs were characterized using flow cytometry. The supernatant cytokine levels (interferon-gamma (IFN-γ), interleukin 4 (IL-4), interleukin 17 (IL-17)) of MSCs at each passage were evaluated using the ELISA assay. MSCs exhibited different cytokine levels with each passage number. In WJ-MSC culture supernatants, IL-17 levels significantly increased at P4 and P5 compared to the first passage (p < 0.005), while the other passages showed a decrease. IFN-γ levels increased at passage P1 and P4 and decreased at other passages (p < 0.005). IL-4 levels significantly increased only at passage P3 (p < 0.005). In CB-MSC culture supernatants, IL-17 and IL-4 cytokines decreased compared to P0, while IFN-γ cytokine increased from P0 (p < 0.005). The changing ratio of cytokine levelsfor both CB-MSCs and WJ-MSCs were similarly maintained from early to late passages. More research is needed to understand the immunomodulatory functions of MSCs.
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Thrombotic thrombocytopenic purpura (TTP) is one of the rare group disorders classified as thrombotic microangiopathies (TMAs). Approximately 90% of TTP developed immune-mediation by the formation of antibodies against the enzyme ADAMTS-13. The exact cause is unknown. To establish an association between human leukocyte antigen (HLA) and autoimmune basis, as susceptibility or protection against the disease, we contributed a study aiming to evaluate the role of HLA in immune-mediated TTP (iTTP). Considering epidemiological factors such as age, sex, ethnicity, and geographical origins, we contributed the study in our country, Turkey, which consist of a very heterogeneous population. Patients' data collection was retrospectively from electronic database on two University hospitals having big therapeutic apheresis service. Control arm was healthy people registered as stem cell donors matched in terms of age and sex. The frequency of HLA-DRB1 and HLA-DQB1 alleles between acquired TTP and the control group was compared using the chi-square method. Yates correction and logistic regression were performed on these results. A total of 75 iTTP patients and 150 healthy individuals enrolled to the study. HLA-DRB1∗11, HLA-DQB1∗03, HLA-DRB1∗11:01, HLA-DRB1∗14:01, HLA-DRB1∗13:05, HLA-DRB1∗11 + HLA-DQB1∗03 allele pair and HLA-DRB1∗15 + HLA- DQB1∗06 were proved to be susceptibility allele pairs for iTTP. HLA-DRB1∗15, HLA-DRB1∗01:01, HLA-DRB1∗07:01, HLA-DRB1∗13:01, HLA-DRB1∗14:54, HLA-DQB1∗05:01, HLA-DQB1∗02:02 and HLA-DRB1∗07 + HLA-DQB1∗02 allele pair were found to be protective against iTTP. Our findings support an association with iTTP across very heterogenous populations in Turkey.
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Alelos , Cadenas HLA-DRB1 , Púrpura Trombocitopénica Trombótica , Humanos , Turquía/epidemiología , Púrpura Trombocitopénica Trombótica/genética , Púrpura Trombocitopénica Trombótica/inmunología , Femenino , Masculino , Adulto , Persona de Mediana Edad , Cadenas HLA-DRB1/genética , Estudios Retrospectivos , Cadenas beta de HLA-DQ/genética , Estudios de Casos y Controles , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Adulto Joven , Antígenos HLA/genética , AdolescenteRESUMEN
The effect of minor H antigen mismatching on the occurrence of graft-versus-host disease (GvHD) and graft-versus-leukemia (GvL) after HLA-matched hematopoietic stem cell transplantation (HSCT) has mainly been demonstrated in single-center studies. Yet, the International Histocompatibility and Immunogenetics Workshops (IHIW) provide a collaborative platform to execute crucial large studies. In collaboration with 20 laboratories of the IHIW, the roles of 10 autosomal and 10 Y chromosome-encoded minor H antigens were investigated on GvHD and relapse incidence in 639 HLA-identical related donor (IRD) and 210 HLA-matched unrelated donor (MUD) HSCT recipients. Donor and recipient DNA samples were genotyped for the minor H antigens HA-1, HA-2, HA-3, HA-8, HB-1, ACC-1, ACC-2, SP110, PANE1, UGT2B17, and HY. The correlations with the primary outcomes GvHD (acute or chronic GvHD), survival, and relapse were statistically analyzed. The results of these multicenter analyses show that none of the HLA class I-restricted HY antigens were found to be associated with any of the primary outcomes. Interestingly, of the HLA class II-restricted HY antigens analyzed, HLA-DQ5 positive recipients showed a significantly increased GvHD-free survival in female-to-male HSCT compared with male-to-female HSCT (P = .013). Yet, analysis of the overall gender effect, thus independent of the known HY antigens, between the gender groups demonstrated an increased GvHD incidence in the female-to-male transplantations (P < .005) and a decreased GvHD-free survival in the female-to-male transplantations (P < .001). Of all autosomally encoded minor H antigens, only mismatching for the broadly expressed minor H antigen HA-8 increased the GvHD incidence in IRD HSCT (Hazard ratio [HR] = 5.28, P < .005), but not in MUD HSCT. Most striking was the influence of hematopoietic restricted minor H antigens on GvL as mismatching for hematopoietic minor H antigens correlated with lower relapse rates (P = .078), higher relapse-free survival (P = .029), and higher overall survival (P = .032) in recipients with GvHD, but not in those without GvHD. In conclusion, the significant GvHD effect of the broadly expressed minor H antigen HA-8 favors matching for HA-8 in IRD, but not in MUD, patient/donor pairs. The GvHD-GvL association demonstrating a significant lower relapse in hematopoietic minor H antigen mismatched patient/donor pairs underlines their clinical applicability for adoptive immunotherapy, enhancing the GvL effect in a GvHD controllable manner.
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Enfermedad Injerto contra Huésped/inmunología , Efecto Injerto vs Leucemia/inmunología , Trasplante de Células Madre Hematopoyéticas/métodos , Histocompatibilidad/inmunología , Antígenos de Histocompatibilidad Menor/inmunología , Adulto , Femenino , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Prueba de Histocompatibilidad , Humanos , Masculino , Donante no EmparentadoRESUMEN
The aim of this study was to evaluate the incidence of total aflatoxin (AF), ochratoxin A, T-2 toxin, deoxynivalenol (DON), zearalenone (ZEA), and fumonisin (FB) in dairy cattle, beef cattle, and lamb-calf feeds. A total of 180 dairy cattle, beef cattle, and lamb-calf feeds (60 samples each) were randomly collected from farms, feed mills, and villages in Burdur province, between September 2006 and August 2007. All samples were analyzed by the competitive Enzyme Linked Immuno Sorbent Assay (ELISA). The most frequent mycotoxin detected was total AF, which was found in 108 samples (60 %) in concentrations ranging from 3.82 to 116.83 µg kg(-1), followed by DON that was detected in 87 samples (48.3 %), in concentrations ranging from 18.50 to 500 µg kg(-1). Ochratoxin A (OTA), T-2 toxin, ZEA, and FB were found in 84 (46.7 %), 85 (47.2 %), 57 (31.7 %), and 19 (10.6 %) samples, respectively, in concentrations of 1.01 to 15.85 µg kg(-1) for OTA, 3.85 to 52.36 µg kg(-1) for T-2 toxin, 2.10 to 29.30 µg kg(-1) for ZEA, and 2.69 to 4.96 mg kg(-1) for FB. It was concluded that feed samples in Burdur province were contaminated by mycotoxins, and the levels of total aflatoxin in the samples were considered a risk to animal health.
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Alimentación Animal/análisis , Contaminantes Ambientales/análisis , Contaminación de Alimentos/análisis , Micotoxinas/análisis , Animales , Monitoreo del Ambiente , Microbiología de Alimentos , TurquíaRESUMEN
BACKGROUND: We have performed a retrospective analysis of anti-HLA class I MHC and class II MHC antibodies measured using a single antigen bead (SAB) assay and a panel reactive antibody (PRA) assay. MATERIAL AND METHODS: A group of 256 patients with end-stage renal disease (ESRD) was tested for anti-HLA antibodies in the tissue typing laboratory between 2017 and 2020. In the cohort, the serum samples of patients waiting for transplantation were tested. Both the PRA and SAB tests of these patients were analyzed using the Luminex (Immucor) method. The threshold of positivity was accepted as median fluorescence intensities (MFI) ≥1000 for PRA screening and MFI ≥750 for SAB screening. RESULTS: Overall, antibodies to HLA antigens were detected in 202 (78.9%) out of 256 patients in the PRA study. Antibodies against both class I/II antigens were detected only in 15.6% of these patients, whereas antibodies against only against class I HLA in 31.3% and only against class II HLA in 32.0%. By comparison, the SAB study found that 66.8% of patients were positive for HLA antigens. Furthermore, donor-specific antibodies (DSA) were detected in 52.0% of PRA-positive patients and 52.6% of SAB-positive patients. It was shown that 168 patients (83.2%) out of 202 PRA-positive patients were found to be SAB-positive. In addition, 51 patients negative in the SAB assay (94.4%) were also negative in the PRA assay. Statistical analysis established a significant correlation between the PRA and SAB positivity (p > 0.001). It was also shown that MFI ≥3000 PRA positivity for class I HLA antigens (p = 0.049) and MFI ≥5000 PRA positivity for class II antigens (p < 0.001) correlated with the SAB positivity in patients. CONCLUSION: Our results showed the importance of both PRA and SAB assays to define the status of sensitization in patients.
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Anticuerpos , Antígenos HLA , Humanos , Estudios Retrospectivos , Prueba de Histocompatibilidad/métodos , Antígenos de Histocompatibilidad Clase II , IsoanticuerposRESUMEN
Giardia duodenalis (G. duodenalis) is an important zoonotic protozoan agent that causes foodborne and waterborne diarrhea in humans and other mammals. Molecular-based tests are critical in diagnosing giardiasis in humans and animals, identifying species, understanding the zoonotic potential and transmission routes, and evaluating taxonomy. Therefore, this study aimed to investigate the molecular characterization of G. duodenalis in buffaloes in the Van region in Türkiye. Buffaloes are a species that has been poorly studied in this regard. For this purpose, 100 fecal samples were collected from buffaloes in the Van region. The DNA extraction was performed using the GeneMATRIX STOOL DNA Purification Kit from stool samples. The nested PCR test was performed with the appropriate primers from the obtained DNA samples. The obtained bands suitable for sequencing were sent for sequence analysis, and the sequence results were aligned bidirectionally and compared with the database of GenBank by BLAST. As a result of the study, an 11% positivity rate for G. duodenalis was found in buffaloes, and assemblage E and assemblage B were isolated. To our knowledge, assemblage B in buffaloes was reported for the first time in this study. As a result, it was concluded that buffaloes are an important reservoir for waterborne and foodborne giardiasis.
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PURPOSE: The impact of core 1,3-galactosyltransferase-specific molecular chaperon (COSMC) gene expression and methylation profile on clinical progression of IgA nephropathy (IgAN) is unclear. The aim of this study was to determine the clinical significance and the relation of the COSMC gene expression and methylation pattern with the progression of IgAN. METHODS: Thirty-nine biopsy-confirmed IgAN patients, 11 healthy relatives and 20 healthy controls were recruited. The COSMC mRNA levels and methylation profile of COSMC gene promoter were measured using the quantitative real-time PCR. The galactose-deficient IgA1 (Gd-IgA1) levels were measured using ELISA in serum and cell culture supernatant. The effect of IL-4 and AZA on COSMC expression and methylation and the correlation of COSMC gene expression and methylation levels with baseline kidney function tests, histology and long-term outcomes were examined. RESULTS: The mean COSMC mRNA level was significantly lower, and serum Gd-IgA1 level was higher in IgAN patients compared with the control groups (p < 0.001, and p = < 0.001, respectively). The COSMC mRNA levels were correlated with intensity of hematuria (r = - 0.41, p = 0.009), serum creatinine level (r = - 0.37, p = 0.002) and eGFR (r = 0.36, p = 0.002). The COSMC methylation levels were correlated with age (r = 0.25, p = 0.04) and baseline eGFR (r = - 0.326, p = 0.006). Twenty IgAN patients (51.3%) reached to complete (5, 12.8%) or partial remission (15, 38.5%) after a median of 34.5 months (IQR, 13.75-71). In multivariable Cox regression analysis, COSMC mRNA expression (adjusted HR (aHR) 1.871, 95% CI 1.287-2.722, p = 0.001) and Oxford T score (aHR 0.355, 95% CI 0.146-0.859, p = 0.022) predicted the remission. CONCLUSION: COSMC mRNA level is a novel biomarker candidate to predict the remission in IgAN patients.
Asunto(s)
Glomerulonefritis por IGA , Humanos , Inmunoglobulina A/metabolismo , Chaperonas Moleculares/genética , ARN Mensajero/metabolismoRESUMEN
BACKGROUND: Thromboangiitis obliterans (TAO) causes vascular insufficiency due to chronic inflammation and abrupt thrombosis of the medium and small arteries of the extremities. In our study, we aimed to determine biomarkers for the diagnosis of TAO by evaluating 15 male TAO patients with Shinoya diagnostic criteria and 5 healthy controls who did not have TAO-related symptoms in their family histories. METHODS: The Clariom D Affymetrix platform was used to conduct microarray analysis on total RNA extracted from whole blood. A total of 477 genes (FC ≤ 5 or >5) common to the fifteen patient and five control samples were selected using comparative microarray analysis; among them, 79 genes were upregulated and 398 genes were downregulated. RESULTS: According to FC ≤ 10 or >10, in the same TAO patient and control group, 13 genes out of 28 were upregulated, whereas 15 genes were downregulated. The 11 key genes identified according to their mean log2FC values were PLP2, RPL27A, CCL4, FMNL1, EGR1, EIF4A1, RPL9, LAMP2, RNF149, EIF4G2, and DGKZ. The genes were ranked according to their relative expression as follows: FMNL1 > RNF149 > RPL27A > EIF4G2 > EIF4A1 > LAMP2 > EGR1 > PLP2 > DGKZ > RPL9 > CCL4. Using protein-protein interaction network analysis, RPL9, RPL27A, and RPL32 were found to be closely related to EIF4G2 and EIF4A1. The Reactome pathway found pathways linked to 28 genes. These pathways included the immune system, cellular responses to stress, cytokine signaling in the immune system, and signaling by ROBO receptors. CONCLUSIONS: By figuring out the protein expression levels of the genes that have been found to explain how TAO disease works at the molecular level, it will be possible to figure out how well these chosen transcripts can diagnose and predict the disease.