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Mesenchymal Stem Cells, mesodermal origin and multipotent stem cells, have ability to differentiate into vascular endothelial cells. The cells are squamous in morphology, inlining, and protecting blood vessel tissue, as well as maintaining homeostatic conditions. ECs are essential in vascularization and blood vessels formation. The differentiation process, generally carried out in 2D culture systems, were relied on growth factors induction. Therefore, an artificial extracellular matrix with relevant mechanical properties is essential to build 3D culture models. Various 3D fabrication techniques, such as hydrogel-based and fibrous scaffolds, scaffold-free, and co-culture to endothelial cells were reviewed and summarized to gain insights. The obtained MSCs-derived ECs are shown by the expression of endothelial gene markers and tubule-like structure. In order to mimicking relevant vascular tissue, 3D-bioprinting facilitates to form more complex microstructures. In addition, a microfluidic chip with adequate flow rate allows medium perfusion, providing mechanical cues like shear stress to the artificial vascular vessels.
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Técnicas de Cultura de Células em Três Dimensões , Diferenciação Celular , Células Endoteliais , Células-Tronco Mesenquimais , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Técnicas de Cultura de Células em Três Dimensões/métodos , Alicerces Teciduais/química , Engenharia Tecidual/métodos , Animais , Hidrogéis/química , Técnicas de Cultura de Células/métodos , Técnicas de Cocultura/métodos , Matriz Extracelular/metabolismoRESUMO
OBJECTIVES: Several studies have attempted to prevent or improve oral mucositis (OM) but have not produced a qualified treatment yet. This study evaluates the effects of Carum carvi L. (caraway) hydroalcoholic extract (CHE) as one of the traditional medicinal plants in 5-fluorouracil (5-FU)-induced OM in golden hamsters. MATERIALS AND METHODS: OM was induced in 54 male golden hamsters by 5-FU and cheek pouch scratching. Starting from day 12, 500 and 1000 mg kg(-1) per day topical CHE were administered. Pouch histopathology score, malondialdehyde and reduced glutathione contents, and activity of myeloperoxidase plus microbial cultures of cheek pouch, antimicrobial properties of CHE, and essential oil constituents were evaluated. RESULTS: Lower histopathology score (0, 1, and 2) and malondialdehyde level, higher reduced glutathione level and activities of myeloperoxidase were detected in 1000 and 500 mg kg(-1) per day topical CHE and control groups, respectively (P < 0.001). The CHE was more potent against Staphylococcus epidermidis and Streptococcus intermedius. γ-Terpinene (37.2%) was identified as the main constituent of essential oil. CONCLUSION: The use of CHE in topical form may be associated with reduced intensity of OM. This may be due to appropriate antibacterial activity and terpinene contents.
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Carum/química , Extratos Vegetais/farmacologia , Estomatite/tratamento farmacológico , Animais , Anti-Infecciosos/farmacologia , Cricetinae , Método Duplo-Cego , Fluoruracila/administração & dosagem , Glutationa/metabolismo , Masculino , Malondialdeído/metabolismo , Mesocricetus , Mucosa Bucal/efeitos dos fármacos , Mucosa Bucal/patologia , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/química , Distribuição Aleatória , Staphylococcus epidermidis/efeitos dos fármacos , Estomatite/induzido quimicamente , Estomatite/metabolismo , Streptococcus intermedius/efeitos dos fármacosRESUMO
BACKGROUND: Breast cancer cells over-express the adenosine receptor A1 and in most of these cells, P53 gene is a wild type. Because of this finding and relationship between A1 receptor and cell apoptosis and proliferation, this study aimed to determine the effect of agonist and antagonist of A1 receptor on cell apoptosis and proliferation and recognize the relationship between this receptor and P53 expression. METHODS: We used a Real-Time PCR test for measuring expression of p53 gene also flow cytometry assay for apoptotic and survival cell rate after treatment of MCF-7 cells with A1 receptor agonist CPA (N6-Cyclopentyladenosine) and A1 receptor antagonist DPCPX (1,3-dipropyl-8-cyclopentylxanthine) in 24,48 and 72 hours. RESULTS: Our flow cytometry findings indicate that DPCPX significantly induces apoptosis in MCF-7. Also the expression of P53 becomes upregulated with time of DPCPX treatment. CPA treatment increased the survival cell rate and down-regulated this apoptosis-relevant gene P53 (p > 0.05). CONCLUSION: DPCPX can induce P53 expression which consequently promotes the cell apoptosis in MCF-7. Therefore, DPCPX could be used as an anti-cancer agent (Tab. 1, Fig. 3, Ref. 5).
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Adenosina/análogos & derivados , Neoplasias da Mama , Receptor A1 de Adenosina/metabolismo , Xantinas/farmacologia , Adenosina/farmacologia , Agonistas do Receptor A1 de Adenosina/farmacologia , Antagonistas do Receptor A1 de Adenosina/farmacologia , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Perfilação da Expressão Gênica , Genes p53/genética , Humanos , Células MCF-7RESUMO
Adipose derived stem cells (ADSCs) are multipotent and can transdifferentiate into neural stem cells. We investigated the transdifferentiation of ADSCs to neural phenotype (NP) cells using selegiline and two-dimensional electrophoresis (2-DE). The perinephric and inguinal fat of rats was collected and used to isolate ADSCs that were characterized by immunophenotyping using flow cytometry. The ADSCs were differentiated into osteogenic and lipogenic cells. The NP cells were generated using 10-9 mM selegiline and characterized by immunocytochemical staining of nestin and neurofilament 68 (NF-68), and by qRT-PCR of nestin, neurod1 and NF68. Total protein of ADSCs and NP cells was extracted and their proteome patterns were examined using 2-DE. ADSCs carried CD73, CD44 and CD90 cell markers, but not CD34. ADSCs were differentiated into osteocyte and adipocyte lineages. The differentiated NP cells expressed nestin, neuro d1 and NF-68. The proteome pattern of ADSCs was compared with that of NP cells and eight spots showed more than a two fold increase in protein expression. The molecular weights and isoelectric points of these highly expressed proteins were estimated using Melanie software. We compared these results with those of the mouse proteomic database using the protein isoelectric point database, and the functions of the eight proteins in differentiation of NP cells were predicted using the UniProt database. The probable identities of the proteins that showed higher expression in NP cells included cholinesterase, GFRa2, protein kinase C (PKC-eta) and RING finger protein 121. The sequences of the proteins identified from mouse database were aligned by comparing them with similar proteins in rat database using the Basic Local Alignment Search Tool (BLAST). The E values of all aligned proteins were zero, which indicates consistency of the matched protein. These proteins participate in differentiation of the neuron and their overexpression causes ADSCs transdifferentiation into NP cells.
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Diferenciação Celular/fisiologia , Eletroforese em Gel Bidimensional , Células-Tronco Neurais/citologia , Proteoma/metabolismo , Selegilina/metabolismo , Animais , Biomarcadores/metabolismo , Células Cultivadas , Eletroforese em Gel Bidimensional/métodos , Citometria de Fluxo/métodos , Neurônios/metabolismo , Osteogênese/fisiologia , Ratos Sprague-DawleyRESUMO
Coronavirus disease 2019 (COVID-19) infection can involve many organs, such as central nervous system, including in relapse. We describe the case of a 64-year-old woman with microbiologically confirmed COVID-19-induced respiratory distress whose treatment resulted in a negative nasopharyngeal swab reverse transcriptase PCR (RT-PCR) result for COVID-19. However, after a few weeks, relapse occurred, as indicated by symptoms of acute meningoencephalitis. Results of COVID-19 RT-PCR testing from her cerebrospinal fluid, nasopharyngeal and tracheal aspiration specimens became positive again, but COVID-19 serum antibodies were negative. We therefore note that symptoms with neurologic involvement can be one of COVID-19's first presentations, or they can appear at relapse. Regular evaluation of patients during convalescence is therefore necessary.
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BACKGROUND: Reducing the percentage of sperm anomalies in insemination samples remains a goal to be achieved in the intra-cytoplasmic sperm insemination (ICSI) procedure. The aim of this study was to evaluate the efficiency of density gradient centrifugation (DGC) and Zeta methods to recover sperm with intact chromatin, and to evaluate whether the combined DGC/Zeta procedure improved ICSI outcome. METHODS: In Experiment 1, DGC and Zeta methods were carried out on 60 unprocessed semen samples. The samples were then assessed by chromomycin A3 staining, acridine orange test, terminal deoxynucleotidyl transferase dUTP nick end labeling assay and the sperm chromatin dispersion test for protamine deficiency and DNA fragmentation. In Experiment 2, sibling oocytes from 30 ICSI candidates were divided into two groups; one group was inseminated with sperm processed by DGC and the second with sperm processed by DGC/Zeta. The outcomes of 30 ICSI cycles were compared between the two groups and also with 34 ICSI candidates whose oocytes were inseminated by DGC-processed sperm. RESULTS: Both procedures were efficient for the recovery of sperm with normal protamine content and low DNA fragmentation. However, the Zeta method yielded a greater number of sperm with less DNA fragmentation. Fertilization and pregnancy rates were improved following the combined DGC/Zeta procedure. Compared with DGC alone, the pregnancy rate appeared improved but this was not statistically significant (P = 0.091). CONCLUSIONS: Combining DGC and Zeta procedures improves the quality of semen samples which may increase fertilization rates and possibly pregnancy rates.
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Injeções de Esperma Intracitoplásmicas/métodos , Espermatozoides/citologia , Centrifugação com Gradiente de Concentração , Cromatina , Fragmentação do DNA , Feminino , Fertilização , Humanos , Masculino , Gravidez , Taxa de Gravidez , Protaminas/metabolismo , Espermatozoides/metabolismoRESUMO
We assessed the knowledge and attitude (K&A) toward Crimean-Congo haemorrhagic fever (CCHF) of occupationally at-risk healthcare workers (HCWs). A cross-sectional survey was performed in three referral hospitals in the Systan-Baluchestan and Isfahan provinces of Iran where CCHF is highly endemic. In all, 191/209 eligible HCWs were enrolled (response rate: 93%). All but 11 (5.8%) had heard of CCHF. The mean K&A scores of the respondents were 50.34% and 79.25%, respectively. The correlation between K&A was significant (correlation coefficient: 0.542; P<0.001). Being a physician, working in Isfahan (versus the relatively deprived Systan-Baluchestan) and relying on academic material rather than local media were independent factors significantly associated with more knowledge; higher education and laboratory staff with better attitude were also significant factors. Although HCWs showed the best K&A for preventive measures, only 44% wore gloves and masks for contact with CCHF patients and 22% failed to observe any safety measure. Those with a history of percutaneous contact (6.3%) had significantly lower knowledge scores (P=0.047). There is a need to establish professional education campaigns in highly endemic deprived areas in order to improve physicians' attitudes, encourage nurses' use of academic materials and increase the knowledge of less-educated HCWs.
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Atitude do Pessoal de Saúde , Conhecimentos, Atitudes e Prática em Saúde , Pessoal de Saúde , Febre Hemorrágica da Crimeia , Exposição Ocupacional , Adulto , Estudos Transversais , Feminino , Vírus da Febre Hemorrágica da Crimeia-Congo/patogenicidade , Febre Hemorrágica da Crimeia/prevenção & controle , Febre Hemorrágica da Crimeia/transmissão , Febre Hemorrágica da Crimeia/virologia , Hospitais Universitários , Humanos , Irã (Geográfico) , Masculino , Inquéritos e QuestionáriosRESUMO
BACKGROUND: Candida krusei is inherently resistant to fluconazole and is emerging as a frequent cause of fungemia in patients with hematologic malignant neoplasms. OBJECTIVE: To determine the risk and prognostic factors associated with C krusei fungemia in comparison with Candida albicans fungemia in patients with cancer. METHODS: Retrospective study of 57 cases of C krusei fungemia occurring at the M. D. Anderson Cancer Center, Houston, Tex, from 1989 to 1996. The C krusei cases were compared with 57 cases of C albicans fungemia with respect to demographics, underlying cancer, Acute Physiology and Chronic Health Evaluation II score, immunosuppression status, chemotherapy, and the use of central venous catheters, as well as fluconazole prophylaxis. RESULTS: At our institution, C krusei accounted for 5% of fungemias during 1989 through 1992 and for 10% during 1993 through 1996. Patients with C krusei fungemia more often had leukemia than patients with C albicans (77% vs 11%; P =.02), whereas catheter-related infections were more common among patients with C albicans fungemia (42% vs 0%; P<.001). Patients with C krusei fungemia had a lower response rate (51% vs 69%; P =.05), largely because they more frequently were neutropenic and had disseminated infection. Mortality related to fungemia was 49% in the cases with C krusei vs 28% in C albicans. Multiple logistic regression analysis showed that persistent neutropenia (P =.02) and septic shock (P =.002) were predictors of poor prognosis. CONCLUSION: In neutropenic patients, C krusei fungemia is associated with high mortality. It should be suspected in patients with leukemia who are receiving fluconazole prophylaxis and should be treated aggressively with an amphotericin B regimen.
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Antifúngicos/uso terapêutico , Fungemia/tratamento farmacológico , Fungemia/microbiologia , Neoplasias Hematológicas , Hospedeiro Imunocomprometido , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anfotericina B/uso terapêutico , Antineoplásicos/efeitos adversos , Candida albicans/efeitos dos fármacos , Estudos de Casos e Controles , Cateterismo Venoso Central/efeitos adversos , Cateteres de Demora/efeitos adversos , Criança , Pré-Escolar , Resistência Microbiana a Medicamentos , Feminino , Fluconazol/uso terapêutico , Fungemia/etiologia , Neoplasias Hematológicas/terapia , Humanos , Imunossupressores/efeitos adversos , Lactente , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Neutropenia/etiologia , Prognóstico , Estudos Retrospectivos , Fatores de Risco , Choque Séptico/etiologia , Resultado do TratamentoRESUMO
We compared the mortality rate among patients suspected of having Crimean-Congo hemorrhagic fever (CCHF) who received treatment with oral ribavirin and those who did not. Ninety-seven (69.8%) of 139 treated patients suspected of having CCHF survived, and 61 (88.9%) of 69 treated patients with confirmed CCHF survived. The efficacy of oral ribavirin was 80% among patients with confirmed CCHF and 34% among patients suspected of having CCHF. Considering the limitations of observational studies, we conclude that oral ribavirin is an effective treatment for the hemorrhagic form of CCHF.
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Antivirais/uso terapêutico , Febre Hemorrágica da Crimeia/tratamento farmacológico , Ribavirina/uso terapêutico , Administração Oral , Adulto , Surtos de Doenças , Feminino , Vírus da Febre Hemorrágica da Crimeia-Congo/efeitos dos fármacos , Febre Hemorrágica da Crimeia/epidemiologia , Febre Hemorrágica da Crimeia/mortalidade , Humanos , Irã (Geográfico)/epidemiologia , Masculino , Resultado do TratamentoRESUMO
From 1988 to 1998, we identified nine patients with Candida guilliermondii fungemia. Four of the five patients with nosocomial infection died, while all of the non-nosocomial cases survived, even though one half of them (2/4) did not receive any treatment Nosocomial C guilliermondii fungemia is often associated with poor outcome despite aggressive antifungal therapy.
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Candidíase/epidemiologia , Infecção Hospitalar/epidemiologia , Fungemia/epidemiologia , Leucemia/epidemiologia , Neoplasias/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Comorbidade , Febre/epidemiologia , Humanos , Hospedeiro Imunocomprometido , Pessoa de Meia-Idade , Neutropenia/epidemiologia , Infecções Oportunistas/epidemiologia , Pneumonia/epidemiologia , Estudos RetrospectivosRESUMO
Factors associated with failure of antifungal therapy were examined in 42 cancer patients with fusariosis (1987-1997). Thirty-six patients (86%) had leukemia and 39 (93%) were neutropenic. Disseminated infection was the most common presentation. The majority (83%) received amphotericin B-based therapy. Thirty patients (71%) failed therapy. No patient with persistent neutropenia responded.
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Fusarium , Neoplasias Hematológicas/complicações , Micoses/complicações , Micoses/diagnóstico , Neutrófilos/fisiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Neoplasias Hematológicas/microbiologia , Humanos , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Micoses/tratamento farmacológico , Micoses/imunologia , Neutropenia/complicações , Neutropenia/microbiologia , Neutrófilos/citologia , Resultado do TratamentoRESUMO
Sirtuin1 (SIRT1) is an enzyme that deacetylates histones and several nonhistone proteins including p53 during stress and plays an important role in the survival of tumor cells. Hereby, this study describes the potency of salermide as a SIRT1 inhibitor to induce apoptosis in the MCF-7 and MRC-5 cell lines. MCF7 and MRC-5 cell lines were cultured in RPMI-1640 and treated with or without salermide at concentration of 80.56 µmol/L, based on the half-maximal inhibitory concentration (IC50) index at different times (24, 48 and72 h). The IC50 value was established for the salermide in MCF-7. The percentage of apoptotic cells was measured by flow cytometry. Real-time quantitative RT-PCR was performed to estimate the mRNA expression of sirtuin1 in MCF-7 and MRC-5 with salermide at different times. ELISA and Bradford protein techniques were used to detect endogenous levels of total and acetylated p53 protein generated in MCF-7 and MRC-5 cells. Our findings indicated that salermide can induce apoptosis in MCF-7 significantly more effective than MRC-5 cells. We showed that the expression of SIRT1 was dramatically down-regulated by increasing the time of salermide treatment in MCF-7 but not MRC-5 and that the acetylated and total p53 protein levels were increased more in MCF-7 than MRC-5. Salermide, by decreasing the expression of sirtuin1 gene, can induce acetylation of P53 protein and consequently induce significant cell death in MCF-7 that was well tolerated in MRC-5.
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BACKGROUND: Oral Lichen planus (OLP) is a chronic lesion of the oral mucosa with unknown origin. Basement membrane changes are common in OLP and may be mediated by proteases such as matrix metalloproteinase (MMPs) and mast cell chymase. The aim of our study was to evaluate the level of serum MMP-3 in OLP com-pared to normal individuals and assess its clinical significance. METHODS: Thirty four serum samples from patients diagnosed with OLP (12 males, 22 females, age: 42.2±10.8 years) and 34 serum samples from healthy control subjects (11 males, 23 females, age: 42.5±13.3 years) were collected and MMP-3 concentration was measured by ELISA. RESULTS: The serum MMP-3 level in OLP patients was higher (21.64±24.31 ng/ml) compared with healthy con-trols (16.52±23.63 ng/ml), but showed no statistically significant difference. A statistically significant difference was demonstrated between the two types of OLP, being more pronounced in the erosive/atrophic form 6). CONCLUSION: The different clinical appearances of OLP are associated with significant differences in MMP-3 serum level.
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BACKGROUND AND OBJECTIVES: Aflatoxins are mycotoxins produced by Aspergillus flavus and Aspergillus parasiticus that can contaminate human and animal foods, including corn, wheat, rice, peanuts, and many other crops resulting in the illness or death of human and animal consumers. The aim of this study was to detect aflatoxin B1, B2, G1, G2 and total aflatoxin in Kashkineh, a traditional Iranian food. MATERIALS AND METHODS: This survey was conducted to detect aflatoxins on 41 samples of Kashkineh. The samples were randomly collected from traditional bazaars and supermarkets of Khorramabad city of Iran. The presence and quantity of aflatoxins was determined by high performance liquid chromatography (HPLC). RESULTS: The average concentrations of AFB1, AFB2, AFG1, and AFG2 in all samples and in a mixed sample of all samples were not detectable (ND). The only sample that showed aflatoxin contamination was sample number 29 of which the AFB1 concentration was 0.64 ng/g. CONCLUSION: Although some people believe Kashkineh is carcinogenic due to toxins, this study showed kashkineh is not contaminated with aflatoxins.
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Media used for tissue culture may have significant effects on the growth and morphology of the adipose tissue-derived stem cells (ADSCs). As fetal bovine serum (FBS) may induce an immunological reaction and health risks, this study was designed to evaluate and compare the effects of human placental serum (HPS) on the proliferation and morphology of hADSCs. We cultured hADSCs for at least three passages in four different culture media containing either FBS, HPS, autologous serum (AS) or human allogeneic serum (HAS). Morphological and immunophenotypic characteristics, as well as proliferation rates of the hADSCs were determined. The rates of proliferation of hADSCs seemed as follows: AS≥HPS>HAS>>FBS. Morphologically, hADSCs isolated and expanded in medium containing HPS were similar to those grown in medium containing AS, whereas the morphology of cells cultured in human sera was different in comparison with FBS-ADSCs cultures. The immunophenotypic markers of hADSCs grown up in medium containing placental serum such as CD44+, CD90+ and CD105+, were similar to hADSCs grown up in media containing other sera. These results indicate that medium enriched with HPS provided a better microenvironment for hADSCs in comparison with medium enriched with commercially available FBS, and other human sera.
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Tecido Adiposo/citologia , Proliferação de Células/efeitos dos fármacos , Meios de Cultura/farmacologia , Células-Tronco Mesenquimais/citologia , Placenta/irrigação sanguínea , Soro , Animais , Bovinos , Técnicas de Cultura de Células/métodos , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Feminino , Humanos , GravidezRESUMO
This study aims at evaluating on-admission serum level of d-dimer in patients with community-acquired pneumonia concerning the severity of the disease and in-hospital outcome of the patients. Sixty patients with community-acquired pneumonia were studied during a one-year period in Imam Khomeini and Sina Hospitals, Tabriz, Iran. On-admission serum d-dimer was measured by enzyme-linked immunoabsorbent assay and the severity of disease determined according to PORT grading system. In-hospital outcome was determined in regard to the level of serum d-dimer. Sixty patients with community-acquired pneumonia, 39 males and 21 females were enrolled. There were twelve patients with PORT one, eight patients with PORT two, eight patients with PORT three, twenty patients with PORT four and twelve patients with PORT five. The mean level of serum d-dimer was significantly higher in severe disease (p < 0.001), patients with hospital stay longer than one week (p = 0.003), patients with bronchopulmonary pattern (p = 0.012), cases in-need of mechanical ventilation (p < 0.001) and patients who expired during hospital stay (p = 0.022). On-admission level of serum d-dimer was significantly and independently higher in patients with severe disease (p < 0.001) and in cases with bronchopulmonary pattern on chest x-ray (p = 0.035). On-admission level of serum d-dimer may predict the severity of community-acquired pneumonia. Further studies are recommended for accurate cut-off points.
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Infecções Comunitárias Adquiridas/sangue , Produtos de Degradação da Fibrina e do Fibrinogênio/metabolismo , Pneumonia/sangue , Adulto , Infecções Comunitárias Adquiridas/mortalidade , Estudos Transversais , Feminino , Humanos , Irã (Geográfico) , Masculino , Pessoa de Meia-Idade , Pneumonia/mortalidade , Curva ROC , Índice de Gravidade de DoençaRESUMO
Fascioliasis is a zoonotic infection caused by Fasciola hepatica. Humans can become accidental hosts of this parasite by ingesting contaminated drinking water or plants in endemic area. The north of Iran is one of the regions. This disease is rarely seen with jaundice caused by obstruction of the biliary tree. We report a case of human fascioliasis with obstructive jaundice who was diagnosed using endoscopic retrograde cholangiopancreatography (ERCP). This report confirms the diagnostic role of ERCP in patients with obstructive jaundice caused by biliary fascioliasis.
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Sperm premature chromatin condensation (PCC) has been considered as the second cause of failed fertilization post-intracytoplasmic sperm injection (post-ICSI). Cytoplasmic factors, including oocyte cytoplasmic immaturity have been suggested to induce PCC sperm. However, recent studies suggest that sperm chromatin anomaly might also lead to PCC sperm. During this study, human sperm from infertile patients with protamine deficiency or with adequate amount of protamine assessed by chromomycin A3 were injected into metaphase II mouse oocyte, treated with colcemid. Chromatin analysis was carried out on the injected oocyte. The results of this study show that contrary to the percentage of intact sperm, percentage of PCC sperm was significantly higher in oocytes injected with protamine deficient sperm (36.43 +/- 4.46) compared to oocytes injected with sperm with an adequate amount of protamine (11.99 +/- 3.54, P < 0.001). A significant correlation was also observed between percentage of PCC sperm and protamine deficiency (r = 0.46, P = 0.004). Therefore, it can be suggested that oocytes injected with protamine deficient sperm have a higher chance of forming PCC sperm and may result in failed fertilization post-ICSI.
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Cromossomos de Mamíferos/fisiologia , Protaminas/metabolismo , Espermatozoides/metabolismo , Animais , Humanos , Masculino , Camundongos , Oócitos/fisiologia , Injeções de Esperma Intracitoplásmicas , Espermatozoides/fisiologiaRESUMO
PURPOSE: To consider the relationship between different sperm nuclear maturity tests and in vitro fertilization (IVF) rate, in order to select the most sensitive, specific, and independent factor(s) for prediction of in vitro fertilization. METHODS: Infertile couples (101) were randomly selected from IVF candidates referred to Isfahan Fertility and Infertility center. Semen samples were collected on the day of oocyte recovery. Following routine semen analysis, major portion of the semen was prepared for routine IVF insemination and the remaining was used for following sperm nuclear maturity tests: chromomycin A3 (CMA3), aniline blue, sodium dodecyl sulphate (SDS) test, and acridine orange test with or without heat shock (87 degrees C, 5 min). Sperms (200) were evaluated for each test. The results were recorded and analyzed for their correlation to fertilization rate, using correlation coefficient, logistic regression analysis, student t-test, and receiver operating characteristics (ROC) curve. RESULTS: Among these tests, aniline blue and CMA3, and semen parameters, sperm morphology, and sperm motility showed a significant correlation with fertilization rate. Using logistic regression analysis, sperm morphology and CMA3 were the only independent factors related to in vitro fertilization. ROC curves showed that among above tests, CMA3 is the most specific and sensitive for sperm nuclear maturity. CONCLUSION: Among CMA3, aniline blue, SDS test, and acridine orange, CMA3 was the most sensitive and specific test that can be used along with routine semen analysis for more precise prediction of fertilization rate.
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Compostos de Anilina , Núcleo Celular/fisiologia , Fertilização in vitro , Espermatozoides/citologia , Espermatozoides/fisiologia , Laranja de Acridina , Tamanho Celular , Cromomicinas/metabolismo , Feminino , Corantes Fluorescentes , Humanos , Masculino , Sêmen/citologia , Dodecilsulfato de Sódio , Contagem de Espermatozoides , Cabeça do Espermatozoide/fisiologia , Motilidade dos Espermatozoides , Espermatozoides/classificação , Coloração e Rotulagem , Estatística como AssuntoRESUMO
After aneuploidy, sperm premature chromatin condensation (PCC) is the next prevalent cause of fertilization failure. Therefore the aim of this study was to evaluate the effect of sperm protamine deficiency on sperm PCC formation post-ICSI. Chromatin analysis was carried out on failed fertilized oocytes post-ICSI and incidences of sperm PCC were evaluated and the results were compared with the extent of protamine deficiency assessed by chromomycin A3. The results show that incidence of sperm PCC was significantly different in failed fertilized oocytes injected from semen samples with greater or less than 30% CMA3 positivity (P = 0.04). However, except for fertilization rate (P < 0.001), the mean number of MII, MI and germinal vesicles oocytes and percentage normal sperm were not significantly different between the two groups (P > 0.05). A significant correlation was observed between sperm protamine deficiency with fertilization rate. Hence sperm protamine deficiency affects fertilization rate and possibly prones sperm to PCC post-ICSI.