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PURPOSE: This study examined SSC proliferation on an epididymosome-enriched decellularized testicular matrix (DTM) hydrogel and spermatogenesis induction in azoospermic mice. METHODS: Epididymosomes were extracted and characterized using SEM and western blotting. After cryopreservation, thawed SSCs were cultured in a hydrogel-based three-dimensional (3D) culture containing 10 ng/mL GDNF or 20 µg/mL epididymosomes. SSCs were assessed using the MTT assay, flow cytometry, and qRT-PCR after two weeks of culture. The isolated SSCs were microinjected into the efferent ducts of busulfan-treated mice. DiI-labeled SSCs were followed, and cell homing was assessed after two weeks. After 8 weeks, the testes were evaluated using morphometric studies and immunohistochemistry. RESULTS: The expression of PLZF, TGF-ß, and miR-10b did not increase statistically significantly in the 3D + GDNF and 3D + epididymosome groups compared to the 3D group. Among the groups, the GDNF-treated group exhibited the highest expression of miR-21 (*P < 0.05). Caspase-3 expression was lower in the epididymosome-treated group than in the other groups (***P < 0.001). Compared to the 3D and negative control groups, the 3D + epididymosomes and 3D + GDNF groups showed an increase in spermatogenic cells. Immunohistochemical results confirmed the growth and differentiation of spermatogonial cells into spermatids in the treatment groups. CONCLUSION: The DTM hydrogel containing 20 µg/mL epididymosomes or 10 ng/mL GDNF is a novel and safe culture system that can support SSC proliferation in vitro to obtain adequate SSCs for transplantation success. It could be a novel therapeutic agent that could recover deregulated SSCs in azoospermic patients.
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Azoospermia , Criopreservación , Epidídimo , Espermatogénesis , Testículo , Animales , Masculino , Ratones , Azoospermia/patología , Testículo/crecimiento & desarrollo , Criopreservación/métodos , Epidídimo/metabolismo , Espermatogonias/citología , Andamios del Tejido/química , Células Madre Germinales Adultas/metabolismo , Proliferación CelularRESUMEN
Bulls with varying freezability exhibit substantial variation in semen characteristics after cryopreservation. Sperm freezability is positively correlated with membrane cholesterol content, membrane integrity, mitochondrial activity and antioxidant content. The purpose of this study was to determine the optimal concentration of hyaluronic acid (HA) in bull sperm with different cryotolerances. Simmental bulls (n = 10) semen samples were taken and categorized based on their progressive motility (PM) after freeze-thawing: Group I, consisting of bulls (n = 5) with progressive sperm motility ≥45%, was considered good freezability ejaculates (GF), and Group II, including bulls (n = 5) with progressive sperm motility ≤30%, was considered poor freezability ejaculates (PF) bulls. Semen samples were diluted with a Tris-egg-yolk-glycerol (TEYG) extender containing various concentrations of HA: without HA (control), 1 mM HA, 2 mM HA and 4 mM HA. After the freeze-thaw process, sperm kinematics, plasma membrane and acrosome integrity, mitochondrial activity and apoptotic status were evaluated. The addition of 1 mM HA to the diluent of bulls with GF increased PM and linearity (LIN) compared to the control group (p < 0.05). Normal morphology was improved after thawing in the samples treated with 1 and 2 mM HA in the GF and PF bulls respectively. The membrane and acrosome integrity of GF bulls treated with 1 mM HA was significantly (p < 0.05) greater than that of the control groups. Adding 1 mM HA to the extender of bulls with GF and PF improved the proportion of viable cells compared with the highest concentration (4 mM) of HA. The mitochondrial activity of PF bulls treated with 1 and 2 mM HA was significantly (p < 0.05) greater than that of the controls and 4 mM HA. Finally, it can be concluded that adding low doses of HA (1 mM) to the TEYG extender of GF and PF bulls ameliorated the post-thaw semen quality.
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Preservación de Semen , Semen , Masculino , Animales , Bovinos , Congelación , Ácido Hialurónico/farmacología , Análisis de Semen/veterinaria , Motilidad Espermática , Preservación de Semen/veterinaria , Espermatozoides , Criopreservación/veterinaria , Glicerol/farmacología , Apoptosis , Crioprotectores/farmacologíaRESUMEN
Pericytes are multifunctional cells wrapped around capillary endothelia, essential for vascular health, development, and blood flow regulation, although their role in human placental chorionic villi has not been fully explored. The second half of normal pregnancy is characterized by a progressive decline in placental and fetal oxygen levels which, by term, comprises a substantial degree of hypoxia. We hypothesized this hypoxia would stimulate pericyte regulation of chorionic villous capillary function. This study's objective was to investigate the role of hypoxia on normal term placental pericytes (PLVP) and their signaling to endothelial cells. First, we confirmed fetoplacental hypoxia at term by a new analysis of umbilical arterial blood oxygen tension of 3,010 healthy singleton neonates sampled at caesarean section and before labor. We then measured the release of cytokines, chemokines, and small extracellular vesicles (PLVPsv), from PLVP cultured at 20%, 8% and 1% O2. As O2 levels decreased, secreted cytokines and chemokines [interleukin-6 (IL-6), interleukin-1α (IL-1α) and vascular endothelial growth factor (VEGF)], and small extracellular vesicle markers, (Alix, Syntenin and CD9) increased significantly in the culture supernatants. When primary human umbilical vein endothelial cells (HUVEC) were cultured with PLVPsv, polygon formation, number, and tube formation length was significantly increased compared to cells not treated with PLVPsv, indicating PLVPsv stimulated angiogenesis. We conclude that adding PLVPsv stimulates angiogenesis and vessel stabilization on neighboring endothelial cells in response to hypoxia in term pregnancy compared to no addition of PLVPsv. Our finding that PLVP can release angiogenic molecules via extracellular vesicles in response to hypoxia may apply to other organ systems.
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Vesículas Extracelulares , Placenta , Recién Nacido , Femenino , Embarazo , Humanos , Placenta/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Pericitos/metabolismo , Cesárea , Hipoxia/metabolismo , Oxígeno/metabolismo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Citocinas/metabolismo , Vesículas Extracelulares/metabolismoRESUMEN
Preeclampsia (PE) is a hypertensive complication of pregnancy that affects 2-8% of women worldwide and is one of the leading causes of maternal deaths and premature birth. PE can occur early in pregnancy (<34 weeks gestation) or late in pregnancy (>34 weeks gestation). Whilst the placenta is clearly implicated in early onset PE (EOPE), late onset PE (LOPE) is less clear with some believing the disease is entirely maternal whilst others believe that there is an interplay between maternal systems and the placenta. In both types of PE, the syncytiotrophoblast (STB), the layer of the placenta in direct contact with maternal blood, is stressed. In EOPE, the STB is oxidatively stressed in early pregnancy (leading to PE later in gestation- the two-stage model) whilst in LOPE the STB is stressed because of villous overcrowding and senescence later in pregnancy. It is this stress that perturbs maternal systems leading to the clinical manifestations of PE. Whilst some of the molecular species driving this stress have been identified, none completely explain the multisystem nature of PE. Syncytiotrophoblast membrane vesicles (STB-EVs) are a potential contributor to this multisystem disorder. STB-EVs are released into the maternal circulation in increasing amounts with advancing gestational age, and this release is further exacerbated with stress. There are good in vitro evidence that STB-EVs are taken up by macrophages and liver cells with additional evidence supporting endothelial cell uptake. STB-EV targeting remains in the early stages of discovery. In this review, we highlight the role of STB-EVs in PE. In relation to current research, we discuss different protocols for ex vivo isolation of STB-EVs, as well as specific issues involving tissue preparation, isolation (some of which may be unique to STB-EVs), and methods for their analysis. We suggest potential solutions for these challenges.
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Vesículas Extracelulares , Preeclampsia , Embarazo , Femenino , Humanos , Trofoblastos , Placenta , Edad GestacionalRESUMEN
INTRODUCTION: Kidney involvement, ranging from mild hematuria and proteinuria to acute kidney injury (AKI) in patients with coronavirus disease-2019 (COVID-19), is a recent finding with various incidence rates reported among hospitalized patients with COVID-19. Given the various AKI rates and their associated risk factors, lack of AKI recovery in the majority of patients hospitalized with COVID-19, and limited data regarding AKI in patients with COVID-19 in Iran, we aim to investigate the potential risk factors for AKI development and its incidence in patients hospitalized with COVID-19. METHODS: In this retrospective cohort study, we enrolled adult patients referred to the Sina Hospital, Iran, from February 20 to May 14, 2020, with either a positive PCR test or a highly susceptible chest computed tomography features consistent with COVID-19 diagnosis. AKI was defined according to the kidney disease improving global outcomes criteria, and patients were stratified based on their AKI staging. We evaluated the risk indicators associated with AKI during hospitalization besides in-hospital outcomes and recovery rate at the time of discharge. RESULTS: We evaluated 516 patients with a mean age of 57.6 ± 16.1 years and a male-to-female ratio of 1.69 who were admitted with the COVID-19 diagnosis. AKI development was observed among 194 (37.6%) patients, comprising 61.9% patients in stage 1, 18.0% in stage 2, and 20.1% in stage 3. Out of all patients, AKI occurred in 58 (11.2%) patients during the hospital course, and 136 (26.3%) patients arrived with AKI upon admission. AKI development was positively associated with all of the in-hospital outcomes, including intensive care unit admissions, need for invasive ventilation, acute respiratory distress syndrome (ARDS), acute cardiac injury, acute liver injury, multiorgan damage, and mortality. Patients with stage 3 AKI showed a significantly higher mortality rate, ARDS, and need for invasive ventilation than other stages. After multivariable analysis, male sex (odds ratio [OR]: 11.27), chronic kidney disease (CKD) (OR: 6.89), history of hypertension (OR: 1.69), disease severity (OR: 2.27), and high urea levels (OR: 1.04) on admission were independent risk indicators of AKI development. Among 117 (28.1%) patients who experienced AKI and survived, only 33 (28.2%) patients made a recovery from the AKI, and 84 (71.8%) patients did not exhibit full recovery at the time of discharge. DISCUSSION/CONCLUSION: We found that male sex, history of CKD, hypertension, disease severity, and high serum urea were independent risk factors associated with AKI in patients with COVID-19. Also, higher stages of AKI were associated with increased risk of mortality and in-hospital complications. Our results indicate a necessity for more precise care and monitoring for AKI during hospitalization in patients with COVID-19, and lack of AKI recovery at the time of discharge is a common complication in such patients.
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Lesión Renal Aguda/etiología , Lesión Renal Aguda/terapia , COVID-19/complicaciones , Lesión Renal Aguda/epidemiología , Adulto , Anciano , COVID-19/epidemiología , Prueba de COVID-19 , Estudios de Cohortes , Femenino , Mortalidad Hospitalaria , Hospitalización , Humanos , Incidencia , Irán/epidemiología , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Estudios Retrospectivos , Factores de Riesgo , Factores Sexuales , Tomografía Computarizada por Rayos X , Resultado del TratamientoRESUMEN
BACKGROUND: With COVID-19 pandemic, concerns about kidney transplant recipients are rising. However, the incidence, clinical course, outcome, and predictive factors of disease severity are obscured. METHODS: We describe clinical and laboratory manifestations, radiologic findings, clinical course, and finally outcome of kidney transplant recipients with COVID-19 pneumonia. RESULTS: Of 2493 kidney transplant recipients under follow-up in our clinic, 19 cases (4 cases diagnosed based on radiologic findings) were admitted. The mean age of patients was 47.6 ± 12.4 years, and the mean time from transplantation was 115.6 ± 70.3 months. Lymphopenia and eosinopenia were 84.2% and 78.9%, respectively. Nine patients did not survive the hospital course. History of acute rejection during the past 12 months, diabetes, higher N/L ratio, lower platelet count, elevated N/L x CRP, higher levels of LDH, positive D-dimer, higher troponin, and prolonged PT were associated with mortality. Among patients with positive COVID-19 test, history of acute rejection, low platelet count, and positive D-dimer were associated with poor outcome. Treatment with cyclosporine was associated with better clinical outcome. CONCLUSIONS: Low rate of admission in transplant recipients specially in the very first years of transplantation might be due to protective effects of immunosuppressive agents against cytokine storm or modification of immunity function. We suggest evaluation of T-cell number, function, and cytokine profile as a guide to manage COVID-19 mainly in patients with higher risk of mortality.
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COVID-19/epidemiología , Síndrome de Liberación de Citoquinas/epidemiología , Rechazo de Injerto/inmunología , Inmunosupresores/uso terapéutico , Trasplante de Riñón/efectos adversos , Adulto , Anciano , Antivirales/uso terapéutico , COVID-19/complicaciones , COVID-19/diagnóstico , COVID-19/inmunología , Síndrome de Liberación de Citoquinas/diagnóstico , Síndrome de Liberación de Citoquinas/inmunología , Síndrome de Liberación de Citoquinas/virología , Citocinas/sangre , Citocinas/inmunología , Femenino , Estudios de Seguimiento , Rechazo de Injerto/epidemiología , Rechazo de Injerto/prevención & control , Humanos , Inmunidad/efectos de los fármacos , Huésped Inmunocomprometido , Irán/epidemiología , Pulmón/diagnóstico por imagen , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Pandemias , SARS-CoV-2/inmunología , SARS-CoV-2/aislamiento & purificación , Linfocitos T/inmunología , Tomografía Computarizada por Rayos X , Receptores de Trasplantes/estadística & datos numéricos , Tratamiento Farmacológico de COVID-19RESUMEN
Therapeutic plasma exchange (TPE) is a process in which plasma containing antibodies, immune complexes, inflammatory moderators, paraproteins and other toxins which are believed to be the cause of disease is removed from a patient. TPE is the first-line treatment (category I, level 1A) in all forms of Acute inflammatory demyelinating polyradiculoneuropathy disease (axonal, demyelinating and miller-fisher variant) as well as in acute myasthenic crisis, chronic inflammatory demyelinating polyradiculoneuropathy and Paraproteinemic neuropathies (category I, level 1B). Moreover, TPE in kidney diseases, for instance: desensitization in renal transplantation(ABO compatible) (living donor)and desensitization in deceased donor, desensitization in renal transplantation(ABO incompatible) (living donor), thrombotic microangiopathy complement Mediated (Factor H autoantibodies), Focal segmental glomerulosclerosis(recurrent in transplanted kidney), ANCA-associated rapidly progressive glomerulonephritis(Dialysis dependence, DAH), Anti-Glomerular basement membrane disease Goodpasture's syndrome)(DAH,Dialysis-independence,) has been utilized as an initial treatment. (category I) TPE has been used as the key therapeutic modality to reduce anti-A or anti-B antibody titers in the liver peri-transplant period with the goal of preventing rejection and facilitating graft survival. Also, plasma exchange is the first-line therapy in Wilson's disease (category I, level1C).
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Sistema del Grupo Sanguíneo ABO , Enfermedades Gastrointestinales/terapia , Enfermedades Renales/terapia , Hepatopatías/terapia , Enfermedades del Sistema Nervioso/terapia , Intercambio Plasmático , Eliminación de Componentes Sanguíneos , Humanos , Trasplante de Riñón , Trasplante de Hígado , Donadores VivosRESUMEN
Many cellular processes in spermatozoa, including apoptosis and motility, are regulated by miRNA. Different miRNAs and molecular pathways are involved in asthenozoospermia (AS) conditions, which are thought to be one of the causes of infertility with reduced sperm motility. Thirty-two semen samples from four Holstein bulls with normozoospermia (NS), total motility ≥ 70%, and progressive motility ≥ 60%, and 32 semen samples from four bulls with AS, total motility ≤ 40%, and progressive motility ≤ 32% were used to investigate the function of apoptosis-related miRNAs in the AS group. Samples were then aspirated into a 0.5 mL straw after dilution with a Tris-egg yolk extender and frozen at -196°C. After freezing, semen samples were thawed for 2 weeks at 37°C and sperm kinematic parameters, plasma membrane integrity, acrosome integrity, DNA fragmentation, apoptosis status, and expression of apoptosis-related miRNAs (miR-2114, miR-296-3p, miR-455-3p, and miR345-3p) were evaluated. Our results showed that the functional and flow cytometric parameters of the NS group were significantly better than those of the AS group. In the NS group, miR-455-3pp and miR-2412 were upregulated, while miR-345-3p was downregulated compared with the AS group. In the AS group, miR-296-39, miR-2412, and miR-345-3p levels were strongly correlated with membrane integrity, DNA fragmentation, and apoptosis status. The findings demonstrated that the selected miRNAs based on bioinformatic analysis in AS and NS samples had a substantial association with functional and flow cytometry indicators and may be involved in regulating apoptosis and motility in AS samples.
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Background: Preeclampsia (PE) is a hypertensive disorder of pregnancy, affecting 2%-8% of pregnancies worldwide, and is the leading cause of adverse maternal and fetal outcomes. The disease is characterized by oxidative and cellular stress and widespread endothelial dysfunction. While the precise mechanisms are not entirely understood, the pathogenesis of PE is closely linked to placental dysfunction and, to some extent, syncytiotrophoblast extracellular vesicle release (STB-EVs). These vesicles can be divided into the less well-studied medium/large EVs (220-1,000â nm) released in response to stress and small EVs (<220â nm) released as a component of intercellular communication. The previously described production of m/lSTB-EVs in response to cellular stress combined with the overwhelming occurrence of cellular and oxidative stress in PE prompted us to evaluate the microRNAome of PE m/lSTB-EVs. We hypothesized that the microRNAome profile of m/lSTB-EVs is different in PE compared to normal pregnancy (NP), which might permit the identification of potential circulating biomarkers not previously described in PE. Methods/study design: We performed small RNA sequencing on medium/large STB-EVs isolated from PE and NP placentae using dual-lobe ex vivo perfusion. The sequencing data was bioinformatically analyzed to identify differentially regulated microRNAs. Identified microRNAs were validated with quantitative PCR analysis. We completed our analysis by performing an in-silico prediction of STB-EV mechanistic pathways. Results: We identified significant differences between PE and NP in the STB-EVs micro ribonucleic acid (microRNA) profiles. We verified the differential expression of hsa-miR-193b-5p, hsa-miR-324-5p, hsa-miR-652-3p, hsa-miR-3196, hsa-miR-9-5p, hsa-miR-421, and hsa-miR-210-3p in the medium/large STB-EVs. We also confirmed the differential abundance of hsa-miR-9-5p in maternal serum extracellular vesicles (S EVs). In addition, we integrated the results of these microRNAs into the previously published messenger RNA (mRNA) data to better understand the relationship between these biomolecules. Conclusions: We identified a differentially regulated micro-RNA, hsa-miR-9-5p, that may have biomarker potential and uncovered mechanistic pathways that may be important in the pathophysiology of PE.
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BACKGROUND: Preeclampsia is a major cause of maternal and perinatal morbidity and mortality worldwide. Identifying women with high risk of developing preeclampsia in early pregnancy remains challenging. Extracellular vesicles released from the placenta offer an attractive biomarker but have been elusive to quantify. METHODS: Here, we tested ExoCounter, a novel device that immunophenotypes size-selected small extracellular vesicles <160 nm, for its ability to perform qualitative and quantitative placental small extracellular vesicles (psEV) analysis. To investigate disease-specific and gestational age-specific changes, we analyzed psEV counts in maternal plasma samples taken at each of the 3 trimesters from women who had (1) normal pregnancy (n=3); (2) women who developed early-onset preeclampsia (EOPE; n=3); and (3) women who developed late-onset preeclampsia (n=4) using 3 antibody pairs, CD10-placental alkaline phosphatase (PLAP), CD10-CD63, and CD63-PLAP. We further validated the findings in first-trimester serum samples among normal pregnancy (n=9), women who developed EOPE (n=7), and women who developed late-onset preeclampsia (n=8). RESULTS: We confirmed that CD63 was the major tetraspanin molecule coexpressed with PLAP-a known placental extracellular vesicles marker on psEV. Higher psEV counts for all 3 antibody pairs were detected in the plasma of women who developed EOPE than the other 2 groups in the first trimester, which persisted through the second and third trimesters. Significantly higher CD10-PLAP (P<0.01) and CD63-PLAP (P<0.01) psEV counts were validated in the serum of the first trimester of women who developed EOPE compared with normal pregnancy. CONCLUSIONS: Application of the ExoCounter assay developed here could identify patients at risk of developing EOPE in the first trimester, thereby providing a window of opportunity for early intervention.
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Vesículas Extracelulares , Preeclampsia , Embarazo , Humanos , Femenino , Placenta , Primer Trimestre del Embarazo , BiomarcadoresRESUMEN
Background: Preeclampsia (PE) is a pregnancy-specific hypertensive disorder affecting 2%-8% of pregnancies worldwide. Biomarker(s) for the disorder exists, but while these have excellent negative predictive value, their positive predictive value is poor. Extracellular vesicles released by the placenta into the maternal circulation, syncytiotrophoblast membrane extracellular vesicles (STB-EVs), have been identified as being involved in PE with the potential to act as liquid biopsies. Objective: The objective of this study was to identify the difference in the transcriptome of placenta and STB-EVs between preeclampsia and normal pregnancy (NP) and mechanistic pathways. Methods/study design: We performed RNA-sequencing on placental tissue, medium/large and small STB-EVs from PE (n = 6) and NP (n = 6), followed by bioinformatic analysis to identify targets that could be used in the future for EV-based diagnostic tests for preeclampsia. Some of the identified biomarkers were validated with real-time polymerase chain reactions. Results: Our analysis identified a difference in the transcriptomic STB-EV cargo between PE and NP. We then identified and verified the differential expression of FLNB, COL17A1, SLC45A4, LEP, HTRA4, PAPP-A2, EBI3, HSD17B1, FSTL3, INHBA, SIGLEC6, and CGB3. Our analysis also identified interesting mechanistic processes via an in silico prediction of STB-EV-based mechanistic pathways. Conclusions: In this study, using comprehensive profiling of differentially expressed/carried genes of three linked sample subtypes in PE, we identified potential biomarkers and mechanistic gene pathways that may be important in the pathophysiology of PE and could be further explored in future studies.
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Introduction and importance: As immunocompromised individuals, kidney transplant recipients (KTR) were more prone to severe and prolonged COVID-19 infection. Case presentation: A 52-year-old man with a history of two kidney transplants for polycystic kidney disease (PKD) was hospitalized due to COVID-19 illness after receiving the first dose of COVID-19 vaccination with Sputnik V. After being admitted to the hospital, the patient was given Remdesivir and oxygen therapy. Clinical discussion: We reported a COVID-19 infection after the first dose of Sputnik V vaccination in an immunocompromised patient who did not follow the protocols after vaccination. Regardless of vaccination, he had been infected, but the vaccination saved him from severe infection despite his comorbidity. Conclusion: To summarise, infection with COVID-19 should be considered after vaccinations, particularly the first dose, in immunocompromised patients such as KTR, and protocols for these patients should be strictly followed. It is worth mentioning that even a full dose of vaccination does not provide full protection from infection to anyone, including KTR.
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AIMS: This study aimed to explore the effect of epididymosomes on the proliferative efficiency of spermatogonial stem cells (SSCs) in vitro and the resumption of spermatogenesis in the azoospermic mice. MAIN METHODS: The epididymosomes were extracted from the epididymis and characterized. SSCs were cultured in 2D (two-dimensional) and hydrogel-based 3D culture in the presence of 20 µg/mL epididymosome or 10 ng/mL GDNF. After two weeks of culture, the proliferation and purity of the separated SSCs were evaluated using the MTT test and flow cytometry, respectively. qRT-PCR was used to analyze PLZF, caspase-3, TGF-ß, miR-10b, and miR-21 expression levels. Then, SSCs grown in the 3D culture system were labeled by DiI and transplanted into azoospermic mice via the efferent duct. After 2 weeks, tracing of DiI and cell homing were evaluated. Subsequently, histomorphometric studies and immunohistochemistry analysis were performed in testes after eight weeks of transplantation. KEY FINDINGS: The expression of PLZF, TGF-ß, miR-10b, and miR-21 increased significantly (*p < 0.05) in the 3D + GDNF and 3D + epididymosomes groups than in the 2D group. Transplanted SSCs migrated into the seminiferous tubules of recipient mice and the number of spermatogenic cells and protein expression of PLZF, SCP3 and ACRBP in the 3D + GDNF and 3D + epididymosomes groups were considerably higher (∗ ∗ ∗ p < 0.001) compared to the azoospermic group. SIGNIFICANCE: This finding indicates that culturing SSCs on decellularized testicular matrix (DTM) hydrogel with 10 ng/mL GDNF or 20 µg/mL epididymosomes could lead to an increase in SSCs proliferation which provides a sufficient number of SSCs for successful transplantation in azoospermic mice.
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Azoospermia , MicroARNs , Animales , Masculino , Ratones , Acrosoma/metabolismo , Azoospermia/terapia , Azoospermia/metabolismo , Proteínas Portadoras/metabolismo , Factor Neurotrófico Derivado de la Línea Celular Glial/genética , Factor Neurotrófico Derivado de la Línea Celular Glial/metabolismo , Hidrogeles/metabolismo , MicroARNs/metabolismo , Espermatogénesis , Espermatogonias/metabolismo , Células Madre , Testículo/metabolismo , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
Introduction: This research was conducted to assess the effect of myo-inositol (MYO) in the freezing extender on the semen quality and oxidative stress parameters of frozen-thawed bull sperm. Materials and Methods: Semen samples were obtained from four bulls (n = 24, six ejaculates per bull), twice a week, and diluted into four equal aliquots in freezing extenders containing different concentrations of MYO (0, 2, 3, and 4 mg/mL). After a freezing/thawing process, velocity parameters, plasma membrane integrity, apoptosis status, malondialdehyde level, and oxidative stress parameters were assessed. Results: Supplementation of freezing extender with 3 mg/mL MYO resulted in higher rapid motility (62.22% ± 2.63%), progressive motility (77.45% ± 2.65%), viability (78% ± 0.91%), plasma membrane integrity (86 ± 0.85), catalase (20.03 ± 0.39 U/mL) activity, and lower significance of lipid peroxidation (3.60 ± 0.15 nmol/dL) than those of the control group (p < 0.05). A significantly lower percentage of normal morphology and intact acrosomes were observed for frozen-thawed semen in the extender supplemented with 4 mg/mL MYO than those of the control group (p < 0.05). Freezing of the sperm in the extender containing 3 mg/mL of MYO leads to a higher percentage of live cells (38.3 ± 2.76). Beat-cross-frequency, amplitude of lateral head displacement, linearity, total antioxidant capacity, total peroxidase activity, early apoptotic status, and superoxide dismutase activities were not affected by MYO levels in the extenders (p > 0.05). Conclusion: The findings of this study suggest that the supplementation of the freezing extender with 3 mg/mL MYO resulted in a higher quality of frozen-thawed bull sperm.
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Análisis de Semen , Preservación de Semen , Animales , Antioxidantes , Bovinos , Criopreservación/métodos , Criopreservación/veterinaria , Crioprotectores/farmacología , Congelación , Inositol/farmacología , Masculino , Estrés Oxidativo , Preservación de Semen/métodos , Preservación de Semen/veterinaria , Motilidad Espermática , EspermatozoidesRESUMEN
OBJECTIVES: COVID-19 has been spreading rapidly throughout the world, with nearly every country thus far documenting this infection. In this study, our aim was to evaluate the risk factors for increased mortality in deceased donor kidney transplant recipients with COVID-19 at a single center in Iran. MATERIALS AND METHODS: This was a retrospective study in a single center. During the 17-month ongoing COVID19 pandemic in Iran, there were 153 deceased donor kidney recipients at our center with suspected COVID19 symptoms. Of these patients, 138 had positive COVID-19 tests, and thus a therapeutic regimen was commenced for these patients. Data were analyzed with SPSS version 16 software. RESULTS: The patients were predominantly male (83, 60.1%) with a median age of 47.09 ± 13.75 years and a median time since transplant of 51 months (IQR, 1-276 months). Among these patients, 84 (60.8%) had hypertension and 43 (31.2%) had diabetes mellitus. We observed a significant relationship between disease severity and mortality (P < .001). After risk adjustments for age, presence of diabetes mellitus and hypertension and blood group type were factors that showed a significantly higher risk of death. CONCLUSIONS: Deceased donor kidney transplant recipients with confirmed COVID-19 experienced less fever as an initial symptom. However, recipients with COVID-19 and an underlying disease had a higher rate of mortality, severity of infection, and progression of disease. Appropriate management of renal complications and vaccinations in deceased donor kidney transplant recipients may help lead to more favorable outcomes.
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COVID-19 , Trasplante de Riñón , Adulto , Humanos , Irán/epidemiología , Trasplante de Riñón/efectos adversos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Receptores de Trasplantes , Resultado del TratamientoRESUMEN
Choriocarcinoma is a rare kind of cancer, which may be either gestational or non-gestational. Choriocarcinoma is responsible for about a quarter of all documented neoplastic aneurysms. It is a descriptive case report of choriocarcinoma transmission from a donor, following kidney donation. A 45-year-old woman got a kidney from a 25-year-old woman who was taken to the hospital due to a non-traumatic cerebral hemorrhage. She delivered a healthy baby 48 days before her brain death. The transplant was successfully done. Five weeks' post-transplantation, the recipient had pain and erythema in the surgical area. Regarding the high level of beta-human chorionic gonadotropin in her blood, diagnostic tests were performed. Following the confirmation of the cancer, a five-phase chemotherapy plan with various pharmaceutical regimens was initiated. Liver function test values rose after the final round of chemotherapy, and the patient developed hepatic encephalopathy. Considering the thrombocytopenia, dialysis, or hemoperfusion, which are normally performed to reduce liver enzymes, were not initiated. Finally, she died due to the hepatic failure and disseminated intravascular coagulation. Although the nephrologists disagree on the optimal course of treatment, it seems that nephrectomy would be helpful in such instances. Physicians should be aware of the possibility of transplant-related choriocarcinoma in female donors of reproductive age who die because of intracerebral brain hemorrhage for unclear reasons. Every donor must undergo a thorough examination. It is critical to get documents, clarify history, and interview relatives.
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Artificial insemination is a valuable and essential tool in genetic improvement programs, and its success requires proper semen collection, freezing, and thawing procedures. Nowadays, despite applying of advanced protocols for semen cryopreservation, post-thawing sperm quantitative and qualitative parameters are not satisfactorily comparable to fresh sperm. The present study was designed to evaluate the effects of the supplementation of an alcoholic extract of Caulerpa sertolarioides alga into the tris-egg yolk-based Simmental bull sperm freezing media. The pooled semen samples were divided into five groups, of which four were supplemented with 500, 1000, 1500, and 2000 ppm alga extract and one allocated as a control. Total motility, progressive motility, plasma membrane integrity, DNA integrity, apoptosis, superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities, and total antioxidant capacity (TAC) of sperm were measured. The frozen sperm from each group were used for IVF on the slaughterhouse-derived oocytes. Fertilization, cleavage, and blastocyst rates were assessed for all groups. Total motility, progressive motility, and velocity curvilinear (VCL) parameters were higher (p ≤ 0.05) in group 1000 ppm than the control group. Velocity in a straight path (VSL) was higher (p ≤ 0.05) in all treatment groups except in 500 ppm compared to the control group. Average path velocity (VAP) was higher (p ≤ 0.05) in 1000 and 1500 ppm groups than in the control group. Straightness (STR) showed a higher value (p ≤ 0.05) in 1000 and 2000 ppm than the control group. Groups 500 and 1000 ppm showed more viable sperm than the control group (p ≤ 0.05). DNA damage was lower (p ≤ 0.05) in group 1000 ppm than in the control group. HOST was higher (p ≤ 0.05) in all groups than in the control group. SOD, GPx, and TAC were higher (p ≤ 0.05) in 1000 ppm than the control and all other groups. Apoptosis was not significantly different among the treatment and control groups. In conclusion, supplementation of alcoholic extract of Caulerpa sertularioides into the Simmental bull freezing extender ameliorated the sperm parameters after the freeze-thawing process. Moreover, the results of this study indicated that the best dose to achieve the antioxidant properties of the alga extract in Simmental bull sperm freezing media was 1000 ppm. It was also evident that 1000 ppm alga extract supplementation into the bull sperm improved fertilization, cleavage, and blastocyst rates.
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Caulerpa , Preservación de Semen , Animales , Antioxidantes/metabolismo , Antioxidantes/farmacología , Caulerpa/metabolismo , Criopreservación/métodos , Criopreservación/veterinaria , Crioprotectores/farmacología , Desarrollo Embrionario , Masculino , Extractos Vegetales/farmacología , Análisis de Semen/métodos , Análisis de Semen/veterinaria , Preservación de Semen/métodos , Preservación de Semen/veterinaria , Motilidad Espermática , Espermatozoides , Superóxido Dismutasa/metabolismoRESUMEN
BACKGROUND: Every year, a large number of people undergo kidney transplants because of various reasons leading to renal failure. These patients usually have low immunoglobulin levels due to the use of immunosuppressive drugs. In recent years, the COVID-19 pandemic has been a major global health risk. Patients who are immunocompromised or who have diabetes are especially at risk. METHODS: In this study, we enrolled 156 patients who had undergone kidney transplant and had received 2 doses of Sinopharm/BIBP-CorV. The serum antibody levels against COVID-19 spike glycoprotein (immunoglobulin [Ig] G and IgM) were measured using a sandwich enzyme-linked immunosorbent assay kit to evaluate whether different immunosuppressive drugs could affect the body's response to the said vaccine. RESULTS: We found that only patients receiving Rapamune had increased IgM secondary to COVID-19 vaccine. None of the immunosuppressive drugs in this study have shown a positive correlation with increased IgG levels. The only factor that showed a significant effect on both IgM and IgG was a positive history of COVID-19, which was correlated with increased levels of serum IgG/M. CONCLUSIONS: Only patients treated with Rapamune showed an acute immune reaction to the vaccine in the form of positive serum IgM levels, and no rise of serum IgM antibody was observed in COVID-19-naive patients. Patients who had a previous history of COVID-19 infection showed an elevated serum IgM and IgG level, suggesting that vaccines in general and Sinopharm/BIBP-CorV in particular are not enough to ensure immunity against COVID-19 in transplant recipients. We recommend further studies using different types of vaccines and immunosuppressive drugs.
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Vacunas contra la COVID-19 , COVID-19 , Trasplante de Riñón , Humanos , Anticuerpos Antivirales , COVID-19/prevención & control , Vacunas contra la COVID-19/efectos adversos , Inmunoglobulina G , Inmunoglobulina M , Inmunosupresores/efectos adversos , Trasplante de Riñón/efectos adversos , Pandemias , SARS-CoV-2 , SirolimusRESUMEN
BACKGROUND: Asthenozoospermia is a usual male infertility factor, characterized by decreased semen quality. It has been revealed that antioxidants improve sperm function, enhance endogenous antioxidant activities, and protect spermatozoa against oxidative damage during cryopreservation. This aimed to evaluate the effects of vitamin D on sperm kinematics and apoptosis in the semen of bulls with normozoospermia and asthenozoospermia after the freeze-thaw process. For this purpose, 32 semen samples of four Holstein bulls (normozoospermic, progressive motility > 70 %) and 32 semen samples of four bull (asthenozoospermic progressive motility < 40 %) were collected and pooled separately (normozoospermic and asthenozoospermic). Samples were then diluted into four equal aliquots of extender containing different vitamin D concentrations (0, 5, 10, and 50 ng/mL) and aspirated into a 0.5 mL straw. RESULTS: The percentages of sperm progressive motility and viability were significantly higher (P < 0.05) in 50 ng/mL of vitamin D in normozoospermic group. Sperm kinematics parameters including curvilinear velocity (VCL), straight-line velocity (VSL), and average path velocity (VAP) were significantly higher in the high dose (50 ng/mL) vitamin D-treated group compared to the low dose vitamin D-treated group (5ng/mL) in normozoospermic bull semen samples. The supplementation of the semen extender with different concentrations of vitamin D could not increase the rate of acrosome integrity in normozoospermic bulls compared to the control group (P < 0.05). In the asthenozoospermic group, 10 ng/mL vitamin D-treated group could increase the rate of plasma membrane integrity compared to 5 ng/mL vitamin D-treated group (P < 0.05). The percentages of early-apoptosis (P = 0.049) and late-apoptosis (P = 0.005) were significantly higher in the asthenozoospermic than the normozoospermic group. CONCLUSIONS: The present study revealed that a high dose (50 ng/mL) of vitamin D protected normozoospermic bulls' sperms from the freezing procedure and lead to higher quality of frozen-thawed bull sperm.
RéSUMé: CONTEXTE: L'asthénozoospermie est un facteur courant d'infertilité masculine, caractérisé par une diminution de la qualité du sperme. Il a été montré que les anti-oxydants amélioraient la fonction des spermatozoïdes, augmentaient les activités anti-oxydantes endogènes, et protégeaient les spermatozoïdes contre les dommages oxydatifs lors de la cryoconservation. Cette étude visait à évaluer les effets de la vitamine D sur la cinématique et l'apoptose des spermatozoïdes dans le sperme de taureaux qui présentaient une normozoospermie ou une asthénozoospermie après le processus de congélation-décongélation. À cette fin, 32 échantillons de sperme de quatre taureaux Holstein (normozoospermiques, mobilité progressive >70 %) et 32 échantillons de sperme de quatre taureaux (asthénozoospermiques ; mobilité progressive <40 %) ont été recueillis et regroupés séparément (normozoospermiques et asthénozoospermiques). Les échantillons ont ensuite été dilués en quatre aliquotes égales dans un milieu contenant différentes concentrations de vitamine D (0, 5, 10 et 50 ng/mL), puis aspirés dans une paille de 0.5 mL. RéSULTATS: Les pourcentages de mobilité progressive et de viabilité des spermatozoïdes étaient significativement plus élevés (p<0.05) avec 50 ng/mL de vitamine D dans le groupe normozoospermique. Dans les échantillons de sperme de taureaux normozoospermiques, les paramètres cinématiques des spermatozoïdes, incluant la vitesse curvilinéaire (VCL), la vitesse en ligne droite (VSL), et la vitesse moyenne du trajet (VAP), étaient significativement plus élevés dans le groupe traité par vitamine D à dose élevée (50 ng/mL) que dans le groupe traité par vitamine D à faible dose (5ng/mL). La supplémentation du milieu avec différentes concentrations de vitamine D n'a pas pu augmenter le taux d'intégrité de l'acrosome chez les taureaux normozoospermiques comparés au groupe témoin (p<0.05). Dans les échantillons de sperme de taureaux asthénozoospermiques, le groupe traité par vitamine D à la dose de 10 ng/mL a augmenté le taux d'intégrité de la membrane plasmique par comparaison au groupe traité par vitamine D à la dose de 5 ng/mL (p<0.05). Les pourcentages d'apoptose précoce (p=0.049) et d'apoptose tardive (p=0.005) étaient significativement plus élevés dans le groupe asthénozoospermique que le groupe normozoospermique. CONCLUSIONS: La présente étude a montré qu'une dose élevée (50 ng/mL) de vitamine D protégeait les spermatozoïdes des taureaux normozoospermiques lors de la procédure de congélation, et menait à une meilleure qualité des spermatozoïdes congelés-décongelés chez ces taureaux.
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INTRODUCTION: Approximately 1 percent of the tumors observed in the upper urinary tract are primary tumors of the ureter. The exact diagnosis is done by a histologic study. Most cases need surgery, which is done by complete resection of the tumoral segment of the ureter. PRESENTATION OF CASE: This study presents a 13-year-old boy who referred to Ali Asghar pediatric hospital with complaints of right flank pain and hematuria for four months. The ultrasonography revealed moderate-grade hydronephrosis. The retrograde urography confirmed a 1.5 cm lesion with filling defects in the proximal segment of the right ureter. The abdominal CT-scan confirmed a 1.5 cm filling defect lesion with a smooth margin in the right proximal ureter adjacent to the ureteropelvic orifice. On cystoscopy, a sessile mass developed in the proximal portion of right ureter and providing pronounced dilation of the ureter and ipsilateral hydronephrosis. Histopathology examination revealed a benign neoplasm composed of a delicate fibrovascular core covered by normal-appearing urothelium. The patient underwent a successful proximal segment resection of the ureter by surgery. DISCUSSION: Benign tumors of the ureter are much less frequent than malignant ones. Mostly, they involve the lower third of the ureter. Pain, hematuria, and hydronephrosis are the most clinical signs. The final diagnosis can be established with the histologic examination. The choice treatment in is segmental ureterectomy. CONCLUSION: Primary ureteral papilloma is extremely rare in the child. The histopathology study is essential for ruling out malignancy. The recurrence and progression of ureteral papilloma are controversial.