RESUMO
Most tumours have an aberrantly activated lipid metabolism1,2 that enables them to synthesize, elongate and desaturate fatty acids to support proliferation. However, only particular subsets of cancer cells are sensitive to approaches that target fatty acid metabolism and, in particular, fatty acid desaturation3. This suggests that many cancer cells contain an unexplored plasticity in their fatty acid metabolism. Here we show that some cancer cells can exploit an alternative fatty acid desaturation pathway. We identify various cancer cell lines, mouse hepatocellular carcinomas, and primary human liver and lung carcinomas that desaturate palmitate to the unusual fatty acid sapienate to support membrane biosynthesis during proliferation. Accordingly, we found that sapienate biosynthesis enables cancer cells to bypass the known fatty acid desaturation pathway that is dependent on stearoyl-CoA desaturase. Thus, only by targeting both desaturation pathways is the in vitro and in vivo proliferation of cancer cells that synthesize sapienate impaired. Our discovery explains metabolic plasticity in fatty acid desaturation and constitutes an unexplored metabolic rewiring in cancers.
Assuntos
Ácidos Graxos/química , Ácidos Graxos/metabolismo , Redes e Vias Metabólicas , Neoplasias/metabolismo , Neoplasias/patologia , Animais , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Proliferação de Células , Ácidos Graxos Dessaturases/metabolismo , Feminino , Células HEK293 , Humanos , Masculino , Camundongos , Ácidos Oleicos/metabolismo , Palmitatos/metabolismo , Ácidos Palmíticos/metabolismo , Estearoil-CoA Dessaturase/metabolismoRESUMO
BACKGROUND: In recent decades, in vitro fertilization (IVF) has been widely used as a method of assisted reproductive technology (ART) to improve fertility in individuals. To be more successful in this laboratory method, we used the presence of two common types of antioxidants (melatonin and vitamin C) simultaneously and exclusively in IVF medium. METHODS: The cumulus-oocyte complexes (COCs) were obtained from Gonadotropin-releasing hormone (GnRH) and Human Chorionic Gonadotropin (HMG) -stimulated mice. Subsequently, metaphase II (MII) oocytes were fertilized in vitro. In the experiment, the IVF medium was randomly divided into two equal groups: The control group did not receive any antioxidants. In the treatment group, 100 µM melatonin and 5 mM vitamin C were added to the IVF medium. Finally, oocytes and putative embryos transferred into developmental medium and cultured 120 h after IVF to the blastocyst stage. After and before IVF, oocytes and putative embryos were stained with dichlorodihydrofluorescein diacetate (DCFDA) and the H2O2 level was measured with an inverted fluorescence microscope using ImageJ software. At the end of the fifth day after IVF, the expression of Bax and B cell lymphoma 2 (Bcl2) was evaluated using real-time PCR. RESULTS: The levels of reactive oxygen species (ROS) in oocytes and putative embryos observed in the treatment group demonstrated a significant reduce compared to the control group (p ≤ 0.01. (.Furthermore, the number of embryos in the blastocycte stage(P < 0.05), the expression level of the Bcl2 (P < 0.05) gene, the Bax unlike gene, significantly increased compared with the control group. CONCLUSION: We conclude that the presence of melatonin and vitamin C antioxidants simultaneously and exclusively in the IVF medium leads to a reduction in ROS and ,as a result, improves the growth of the embryo up to the blastocyst stage.
Assuntos
Melatonina , Humanos , Animais , Camundongos , Melatonina/farmacologia , Melatonina/uso terapêutico , Proteína X Associada a bcl-2/metabolismo , Ácido Ascórbico/farmacologia , Ácido Ascórbico/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Peróxido de Hidrogênio/metabolismo , Desenvolvimento Embrionário , Oócitos/metabolismo , Fertilização in vitroRESUMO
We quantified the life table parameters and predation capacity of a generalist predatory mite, Typhlodromus bagdasarjani Wainstein and Arutunjan on five monotypic diets, including Tetranychus urticae Koch (TSSM) eggs in the presence (SW) and absence (SN) of webs, Trialeurodes vaporariorum Westwood (GHWF) eggs (G), honeydew (H), and maize pollen (M) as well as three mixed diets, including SN + M, SN + G, and G + M. Our results showed that the individuals fed on the mixed diets had a considerably shorter developmental time and pre-oviposition period (APOP), higher oviposition days, higher fecundity and population growth rate than those raised on the monotypic diets. Furthermore, we found that the mixed diet of TSSM and GHWF eggs was the most favorable diet, resulted in the highest fecundity and population growth rate, shortest developmental time and APOP. While TSSM eggs alone in the presence of webs and honeydew were the worst diets resulted in the longest developmental time, lower oviposition day, higher fecundity and population growth rate. Our data determined that TSSM has more nutritional benefits than GHWF for T. bagdasarjani. We observed the positive effects of pollen addition to prey on the predatory mite's immature and adult life-history characters; however, it reduced the predation rate. Overall, maize pollen could enhance ecosystem services provided against spider mites and whiteflies by positively impacting the increase of T. bagdasarjani population. This predator may be more effective when two prey species are available than when only one species is present.
Assuntos
Hemípteros , Tetranychidae , Feminino , Animais , Comportamento Predatório , Crescimento Demográfico , Ecossistema , Dieta , Controle Biológico de Vetores/métodosRESUMO
Conventional sperm processing uses centrifugation has a negative effect on sperm parameters and DNA integrity. We designed and fabricated a novel microfluid device based on chemotaxis and thermotaxis, and compared it with the swim-up method. Twenty normal samples with high DNA fragmentation were included. Each sample was divided into four groups: Group 1, control, Group 2: sperm selection by thermotaxis, Group 3: sperm selection by chemotaxis, and Group 4: sperm selection with thermotaxis and chemotaxis. We used cumulus cells in a microfluid device to create chemotaxis, and, two warm stages to form a temperature gradient for thermotaxis. The spermatozoa were assessed based on the concentration, motility, and fine morphology using Motile Sperm Organelle Morphology Examination, mitochondrial membrane potential (MMP), acrosome reaction (AR), and sperm DNA fragmentation. Concentration (22.40 ± 5.39 vs. 66.50 ± 19.21; p < 0.001) and DNA fragmentation (12.30 ± 3.96% vs. 17.95 ± 2.89%; p < 0.001) after selection in the chemotaxis and thermotaxis microfluid device were significantly lower than control group. The progressive motility (93.75 ± 4.39% vs. 75.55 ± 5.86%, p < 0.001), normal morphology (15.45 ± 2.50% vs. 10.35 ± 3.36, p < 0.001), MMP (97.65 ± 1.81% vs. 94 ± 3.89%, p = 0.02), and AR status (79.20 ± 5.28% vs. 31.20 ± 5.24%, p < 0.001) in the chemotaxis and thermotaxis microfluid device were significantly increased compared to control group. According to these findings, spermatozoa that have penetrated the cumulus oophorus have better morphology and motility, as well as acrosome reactivity and DNA integrity.
Assuntos
Motilidade dos Espermatozoides , Resposta Táctica , Humanos , Masculino , Fragmentação do DNA , Dispositivos Lab-On-A-Chip , Sêmen , EspermatozoidesRESUMO
Sperm processing for assisted reproductive technologies (ART) aims to separate immotile and debris from the motile spermatozoa in the semen. The purpose of this study was to assess the effect of free centrifuge sorting (FCS) approach based on a combination of rheotaxis and swim-up on sperm biological characteristics and ICSI clinical outcomes. Each semen sample was splitted into two equal parts for 67 ICSI cycles with donation oocytes. Parts were processed with the Direct Swim Up (DSU) (control) and with the FCS method (experimental). Sperm quality was assessed in terms of motility, fine morphology, mitochondrial membrane potential, lipid peroxidation and sperm DNA fragmentation. Also Following ICSI, the clinical outcomes were compared between the groups. Sperm progressive motility (93.5 ± 4.1% vs. 78.6 ± 8.2%; p < 0.001), the fraction of Class I (good) morphology (30.2 ± 9.4% vs. 23.7 ± 8.5%; p < 0.0001) and the rate of mitochondrial membrane potential (77.4 ± 7.2% vs. 66.9 ± 5.7%; p < 0.0001) were significantly higher in the FCS compared to DSU groups. The level of lipid peroxidation (0.5 ± 0.05% vs. 0.6 ± 0.06%; p < 0.0001) and concentration of DNA fragmentation (DF) (7.4 ± 1.6% vs. 15.4 ± 2.6%; p < 0.0001) were lower in sperm from the FCS group compared to DSU group. There were higher rates of high-quality embryo formation (p < 0.001), implantation and clinical pregnancy rates (p = 0.03) in the FCS group compared to the control group. The processing of seminal samples using FCS collected spermatozoa with better biological quality and resulted in higher reproductive outcomes in ICSI cycles.
Assuntos
Sêmen , Espermatozoides , Fragmentação do DNA , Feminino , Humanos , Masculino , Oócitos , Gravidez , Taxa de Gravidez , Motilidade dos EspermatozoidesRESUMO
Semen parameters have been found to predict reproductive success poorly and are the most prevalent diagnostic tool for male infertility. There are few conflicting reports regarding the correlation of DNMT genes expression, mitochondrial DNA copy number (mtDNAcn) and deletion (mtDNAdel) with different sperm parameters. To investigate DNMT mRNA level, mtDNAcn and deletion in infertile men, with different sperm parameters, compared with fertile men, semen samples from 30 men with unknown male infertility and normal sperm parameters (experimental group I), 30 infertile patients with at least two abnormal sperm parameters (experimental group II) and 30 fertile normozoospermic men (control group) were collected. After semen analysis, total RNA and DNA were extracted. The isolated DNA was used for assessing the respective mtDNAcn and the presence of common 4977 bp deletion in mtDNA by applying real-time quantitative PCR and multiplex PCR, respectively. Synthesized cDNA from total RNAs was used to quantify DNMT1, DNMT3A and DNMT3B transcripts in study groups by using real-time quantitative reverse-transcription PCR. Significantly higher proportions of mtDNAcn were found in experimental group II. DNMT1 was significantly downregulated in both experimental groups and 4977 bp deletion was not detected. Progressive motility and normal morphology were significantly and negatively correlated with mtDNAcn. A significant positive correlation was detected between sperm parameters and DNMT1 mRNA levels. In conclusion, infertile men with different sperm parameter qualities showed elevated mtDNA content. Abnormal sperm parameters associated with DNMT1 gene expression indicate the possibility of changes in some epigenetic aspects of spermatogenesis in subfertile men with different sperm parameters.
Assuntos
Infertilidade Masculina , Sêmen , Variações do Número de Cópias de DNA , DNA Complementar , DNA Mitocondrial/genética , DNA Mitocondrial/metabolismo , Expressão Gênica , Humanos , Infertilidade Masculina/genética , Infertilidade Masculina/metabolismo , Masculino , Metiltransferases/genética , Metiltransferases/metabolismo , RNA/metabolismo , RNA Mensageiro/metabolismo , Sêmen/metabolismo , Motilidade dos Espermatozoides/genética , Espermatozoides/metabolismoRESUMO
This study aims to evaluate the expression of genes associated with the fertilisation potential and embryo development, sperm DNA fragmentation (SDF), and acrosome reaction in male partners of infertile couples with different sperm parameters compared to fertile men. First, male partners of infertile couples with abnormal (N = 25) and normal sperm parameters (N = 25), and fertile men (N = 10) were included in experimental groups I, II, and controls respectively. The mRNA levels of the Annexin A2 (ANXA2), Sperm protein 17 (SP17), Plasma serine protease inhibitor (SERPINA5), and Peroxiredoxin-2 (PRDX2) genes and SDF were evaluated. To evaluate the maturity of the sperm and oxidative stress, the acrosome reaction, the lipid peroxidation, and total antioxidant were measured. As result, SP17 showed a significantly lower expression in both experimental groups. SERPINA5 was significantly down-regulated in experimental group I that was aligned with the low rate of acrosome reaction. Significant overexpression of PRDX2 was found between experimental group II and controls. Significant higher rates of SDF were seen in both experimental groups compared to the controls. Finally, our data suggest that differentially gene expression of SP17 is a potential diagnostic biomarker in infertile men either with normal or abnormal sperm parameters. SDF is one of the causes of male infertility, independent of the sperm parameters.
Assuntos
Anexina A2 , Proteínas de Ligação a Calmodulina , Infertilidade Masculina , Proteínas de Membrana , Peroxirredoxinas , Inibidor da Proteína C , Anexina A2/genética , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Proteínas de Ligação a Calmodulina/genética , Fragmentação do DNA , Humanos , Infertilidade Masculina/etiologia , Masculino , Proteínas de Membrana/genética , Peroxirredoxinas/genética , Inibidor da Proteína C/genética , RNA Mensageiro/metabolismo , Sêmen/metabolismo , Espermatozoides/metabolismoRESUMO
Despite the beneficial effects of sperm cryopreservation, increased reactive oxygen species (ROS) production during this process can affect spermatozoon structure and function. Moreover, ROS production is associated with elevated DNA damage and alterations in DNA methylation. There is little information about the effects of cryopreservation on epigenetic modulation in sperm and the health of children born with frozen spermatozoa. Considering the potential consequences of cryopreservation in ART-conceived children, it is necessary to assure that cryopreservation does not modify sperm DNA methylation status. This review summarizes reports on epigenetic modifications of spermatozoa during cryopreservation and the probable effects of this process on offspring health. Contradictory results have reported the influence of sperm cryopreservation on DNA methylation in imprinted genes. Multiclinical studies with larger sample sizes under the same conditions of cryopreservation and DNA methylation analysis are needed to make any definitive conclusion about the effect of the cryopreservation process on sperm DNA methylation.
Assuntos
Preservação do Sêmen , Criança , Criopreservação/métodos , Metilação de DNA/genética , Humanos , Masculino , Espécies Reativas de Oxigênio/metabolismo , Sêmen , Preservação do Sêmen/métodos , Motilidade dos Espermatozoides , Espermatozoides/metabolismoRESUMO
Over the last few decades, using natural products has been increased to treat different diseases. Today, great attention has been pointed toward the usage of natural products such as flavonoids, especially Quercetin (QUR), in the treatment of diseases. QUR as a natural antioxidant has been traditionally used to prevent or treat a variety of diseases such as cancer, cardiovascular disease, polycystic ovary syndrome (PCOS), obesity, chronic inflammation, and reproductive system dysfunction. Several studies demonstrated that QUR acts as an anti-inflammatory, anti-apoptotic, antioxidant, and anticancer agent. With this in view, in this study, we intended to describe an overview of the biological effects of QUR on the ovary. QUR improves the quality of oocytes and embryos. It affects the proliferation and apoptosis and decreases the oxidative stress in granulosa cells (GCs). Furthermore, QUR can be used as a complementary and alternative therapy in ovarian cancer and it has beneficial effects in the treatment of PCOS patients. It seems that QUR as a supplementary factor has different activities for the treatment of different disorders and it also has bidirectional activities. However, further investigations are needed for understanding the efficacy of QUR in the treatment and improvement of gynecological patients.
Assuntos
Antioxidantes/uso terapêutico , Síndrome do Ovário Policístico/tratamento farmacológico , Quercetina/uso terapêutico , Animais , Antioxidantes/farmacologia , Feminino , Humanos , Quercetina/farmacologiaRESUMO
BACKGROUND: Antioxidants that used for the infertility treatment cannot have their complete effectiveness, because of their instability in the culture medium. SIGNIFICANCE: One of the most advances, in the drug delivery systems, is nanoliposomes-loaded, as biodegradable and bioavailable carriers. Hormonal and antioxidant agents encapsulating inside the nanoliposomes were used, to increase the effectiveness of antioxidants in the sperm culture medium. MATERIALS: Semen sample from 15 asthenospermia were divided into 10 equal parts. After preparation, the sperms were incubated with free form of drugs and nanocarriers contained resveratrol, catalase, resveratrol-catalase and testosterone for 45 min. All sperm parameters, sperm DNA and gene expressions were evaluated before and after freezing. RESULTS: Before freezing, all nanocarriers and free testosterone showed higher sperm motility compared to free drugs (p=.000). Free Testosterone and free resveratrol-catalase had higher DNA damage compared to nanocarriers (p=.000). Before freezing, the blank nanoliposome and testosterone nanoliposomes had the lowest HSP70 gene expression respectively (p = 0.005) (p = 0.001). After freezing, a significant reduction in sperm motility was observed in the free resveratrol-catalase group (p=.003). Also, a significant increase in sperm viability was observed in the free testosterone and nanoliposomes of blank and testosterone (p > 0.05). The least DNA fragmentation was related to catalase nanoliposomes (p=.000). All nanoliposomes, especially catalase, had the highest percentage of class I morphology compared to the control group (p=.000). CONCLUSIONS: Nanoliposomes could improve the sperm parameters and DNA integrity before and after freezing, by increasing the effectiveness of antioxidants. So, it can be recommended in the ART lab.
Assuntos
Antioxidantes , Preservação do Sêmen , Testosterona , Antioxidantes/farmacologia , Astenozoospermia , Catalase/farmacologia , Criopreservação , DNA , Expressão Gênica , Humanos , Lipossomos , Masculino , Nanopartículas , Resveratrol/farmacologia , Motilidade dos Espermatozoides , Espermatozoides , Testosterona/farmacologiaRESUMO
The box tree moth (BTM), Cydalima perspectalis and the fall webworm (FWW), Hyphantria cunea are two invasive pests of forest trees that have been recorded from Hyracinan forests in north Iran for the first time in 2016 and 2002, respectively. In a search for tentative native entomopathogenic nematode species (EPNs) with potential biocontrol ability against lepidopteran pests of forest trees in north Iran, Oscheius myriophilus was isolated by soil-baiting method from forests of Amlash in the east of Gilan province. The Iranian isolate of this species is characterized by 870-1,247 µm long hermaphrodites having 17-20 µm long stoma, vulva at 45.5-53.1% of body length, tail 90-126 µm long, common males with 38-49 µm long spicules and 583-791 µm long dauer larvae. Compared to the type and other populations, no remarkable differences were observed for this population. The phylogenetic affinities of this isolate with other rhabditid nematodes were studied using partial sequences of small, and the D2-D3 expansion segments of the large subunit ribosomal DNA (SSU and D2-D3 LSU rDNA). This is the first record of the species from Iran. The pathogenicity of Iranian isolate of O. myriophilus was evaluated on the larvae of two important aforementioned forest trees pests, BTM and FWW under laboratory conditions. The results indicated that the suspension of 500 infective juveniles per ml (IJs/ml) of the nematode was the most effective treatment on fifth instar larvae of BTM and FWW, causing 100 and 95% mortality after 48 h, respectively. The lethal concentration 50 (LC50) values of the nematode were 74.5, 152.7, 99.9, and 197.3 IJs/ml on fifth and fourth instar larvae of BTM, and fifth and fourth instar larvae of FWW, respectively, after 48 h at 25°C and 60% relative humidity. Together, present results corroborated the efficacy of the Iranian isolate of Oscheius myriophilus for biocontrolling of BTM and FWW in laboratory conditions.
RESUMO
The prevalence of COVID-19 disease continues to be a significant psychosocial status among health care workers (HCWs) and the general population worldwide. This cross sectional study aimed to compare the psychosocial status between healthcare workers and general population during the prevalence of COVID-19 disease in southeast Iran. Totally 415 health care workers of a medical service center for COVID-19 patients and 1023 people of general population participated in the study. An online socio-demographic characteristics questionnaire, General Health Questionnaire (GHQ -28), and Generalized Anxiety Disorder 7-item (GAD-7) were utilized to evaluate psychosocial status. According to GHQ, the psychological disorders of the HCWs were significantly higher than that of the general population (P < 0.001). According to GAD-7, no significant difference was found between general population and HCWs. Multivariate logistic analysis showed no difference between general population and HCWs in the psychological disorder. Although HCWs suffered from psychological disorders more than general population, nearly one third to half of the participants in both groups had psychosocial disorders.
RESUMO
The present study was carried out to unveil interactive relevance among consecutive and alternate members of a tritrophic system comprised of sugar beet genotypes, beet armyworm, Spodoptera exigua (Hübner), and its parasitoid, Habrobracon hebetor (Say) using demographic parameters. To do so, H. hebetor was reared on S. exigua fed on 10 sugar beet genotypes, including SB26; SB27; SB29; SB33; SB34; (7112*SB36)*Sh-1-HSF-5; FC220; FC301; SBSI006; and HM 1339RZ in a growth chamber at 25 ± 1 °C, 60 ± 5% RH, and 16:8 (L: D) h photoperiod. The data was analyzed based on the age-stage, two-sex life table theory. Our results revealed high variation in duration of different life stages of H. hebetor on S. exigua reared on different sugar beet genotypes examined. The shortest (10.605 days) and longest (13.721 days) pre-adult period of H. hebetor was on S. exigua reared on SB26 and SB34, respectively. The longest (17.2 days) and shortest adult longevity (7.5 days) was on S. exigua reared on SB26 and SB27, respectively. The highest values of the intrinsic rate of increase (r) (0.209 day-1) and finite rate of increase (λ) (1.233 day-1) were observed on S. exigua reared on SB34 and their lowest values (0.159 and 1.172 day-1, respectively) were recorded on SB27. Resistant and susceptible genotypes to S. exigua, FC301 and (7112*SB36)*Sh-1-HSF-5, respectively, were only genotypes on which H. hebetor had greater and approximately equal r compared with S. exigua. This finding indicates high capability of H. hebetor to be successfully employed against S. exigua on sugar beet genotypes which are extremely different in resistance to this pest.
Assuntos
Spodoptera/parasitologia , Vespas/crescimento & desenvolvimento , Vespas/fisiologia , Animais , Beta vulgaris/genética , Agentes de Controle Biológico , Feminino , Tábuas de Vida , Masculino , Spodoptera/fisiologiaRESUMO
PURPOSE: The aim was to assess the correlation of sperm apoptotic transcript levels with cleavage stage embryokinetic and pregnancy outcomes of intracytoplasmic morphologically selected sperm injection (IMSI) and ICSI methods in patients with male factor infertility. MATERIAL AND METHODS: Eighty male factor cases were divided into ICSI and IMSI groups. ICSI was done routinely, and for IMSI, sperm was selected at high magnification and injected. On day 3, time-lapse parameters were evaluated, and the best embryos were transferred and followed to delivery. In addition, sperm DNA fragmentation and apoptotic transcript levels were quantified using reverse transcription Q-PCR between the groups. RESULTS: IMSI selected spermatozoa had lower DNA fragmentation and apoptotic transcript levels compared with ICSI (p < 0.0001). Moreover, all cytokinetic variables and cleavage abnormalities were noticeably different between groups (p < 0.0001); the rates of clinical outcomes were higher in the IMSI group. The transcript levels of Caspase 3 showed a moderate negative correlation with s2 and s3 (rs = - 0.57, P = 0.008 and rs = - 0.51, p = 0.021, respectively) in the IMSI group. However, there was no relationship between sperm apoptotic transcript levels and clinical outcomes in two groups. CONCLUSIONS: Sperms selected at high magnification showed lower DNA fragmentation and apoptosis genes transcript. Also, better embryo kinetics and clinical outcomes were confirmed in IMSI than ICSI groups. Some time-lapse parameters may be associated with transcript levels of apoptosis genes. Therefore, these noninvasive techniques may be unique in assisting couples with male factor infertility. TRIAL REGISTRATION: This trial retrospectively registered on 4 July 2020 (IRCT20180130038561N1).
Assuntos
Apoptose/genética , Infertilidade Masculina/genética , Injeções de Esperma Intracitoplásmicas , Espermatozoides/metabolismo , Adulto , Fragmentação do DNA , Implantação do Embrião/genética , Feminino , Humanos , Infertilidade Masculina/epidemiologia , Infertilidade Masculina/patologia , Irã (Geográfico)/epidemiologia , Masculino , Gravidez , Resultado da Gravidez , Taxa de Gravidez , Análise do Sêmen , Espermatozoides/patologiaRESUMO
OBJECTIVE: Acute-on-chronic liver failure (ACLF) is associated with dysfunctional circulating monocytes whereby patients become highly susceptible to bacterial infections. Here, we identify the pathways underlying monocyte dysfunction in ACLF and we investigate whether metabolic rewiring reinstates their phagocytic and inflammatory capacity. DESIGN: Following phenotypic characterisation, we performed RNA sequencing on CD14+CD16- monocytes from patients with ACLF and decompensated alcoholic cirrhosis. Additionally, an in vitro model mimicking ACLF patient-derived features was implemented to investigate the efficacy of metabolic regulators on monocyte function. RESULTS: Monocytes from patients with ACLF featured elevated frequencies of interleukin (IL)-10-producing cells, reduced human leucocyte antigen DR isotype (HLA-DR) expression and impaired phagocytic and oxidative burst capacity. Transcriptional profiling of isolated CD14+CD16- monocytes in ACLF revealed upregulation of an array of immunosuppressive parameters and compromised antibacterial and antigen presentation machinery. In contrast, monocytes in decompensated cirrhosis showed intact capacity to respond to inflammatory triggers. Culturing healthy monocytes in ACLF plasma mimicked the immunosuppressive characteristics observed in patients, inducing a blunted phagocytic response and metabolic program associated with a tolerant state. Metabolic rewiring of the cells using a pharmacological inhibitor of glutamine synthetase, partially restored the phagocytic and inflammatory capacity of in vitro generated- as well as ACLF patient-derived monocytes. Highlighting its biological relevance, the glutamine synthetase/glutaminase ratio of ACLF patient-derived monocytes positively correlated with disease severity scores. CONCLUSION: In ACLF, monocytes feature a distinct transcriptional profile, polarised towards an immunotolerant state and altered metabolism. We demonstrated that metabolic rewiring of ACLF monocytes partially revives their function, opening up new options for therapeutic targeting in these patients.
Assuntos
Insuficiência Hepática Crônica Agudizada/tratamento farmacológico , Infecções Bacterianas/tratamento farmacológico , Glutamato-Amônia Ligase/antagonistas & inibidores , Imunossupressores/uso terapêutico , Monócitos/enzimologia , Insuficiência Hepática Crônica Agudizada/imunologia , Insuficiência Hepática Crônica Agudizada/metabolismo , Adulto , Infecções Bacterianas/metabolismo , Infecções Bacterianas/patologia , Células Cultivadas , Citocinas/metabolismo , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Monócitos/imunologia , Fagocitose , Estudos RetrospectivosRESUMO
Wolbachia are common intracellular bacteria that are generally found in arthropods, including a high proportion of insects and also some nematodes. This intracellular symbiont can affect sex ratio with a variety of reproductive anomalies in the host, including cytoplasmic incompatibility (CI) in haplodiploids. In this study, we questioned if the parasitoid wasp, Habrobracon hebetor (Hym.: Braconidae), an important biological control agent of many lepidopteran larvae, is infected with Wolbachia. To test this, DNA was extracted from adult insects and subjected to PCR using specific primers to Wolbachia target genes. The results showed a high rate of Wolbachia infection in this parasitoid wasp. To determine the biological function of Wolbachia in H. hebetor, we removed this bacterium from the wasps using antibiotic treatment (cured wasps). Results of crossing experiments revealed that Wolbachia induced CI in H. hebetor in which cured females crossed with infected males produced only males, while both male and female progeny were observed for other crosses. Also, we showed that the presence of Wolbachia in females increased fecundity and female offspring of this parasitoid wasp. The presence of Wolbachia in the males had no significant effect on fecundity and female production, but might have incurred costs. We also investigated the effect of Wolbachia on mate choice and found that Wolbachia affects mating behavior of H. hebetor. Together, we showed that Wolbachia induces CI in H. hebetor and affects host mating behavior in favor of its transmission. Wolbachia utilize these strategies to increase the frequency of infected females in the host population.
Assuntos
Interações entre Hospedeiro e Microrganismos , Transmissão Vertical de Doenças Infecciosas , Vespas/microbiologia , Wolbachia , Animais , Agentes de Controle Biológico , Feminino , Fertilidade , Masculino , Razão de Masculinidade , Comportamento Sexual Animal , Simbiose , Wolbachia/isolamento & purificaçãoRESUMO
Our objective was to evaluate the effect of IMSI on embryo kinetics and clinical outcomes in patients with different aetiologies of male infertility. A total of 150 couples with different aetiologies of male infertility were randomly divided into ICSI and IMSI treatment groups (n = 75). ICSI was done accordingly. For IMSI group, the sperm selection was done using MSOME criteria and then injected. The zygotes were cultured in time-lapse monitoring system (TLM) for 3 days. A total of 650 embryos were developed and assessed using TLM in two groups. Data showed the rate of fragmentation had significant correlation with different aetiologies (p = 0.01), and the timing of s1, t4, s2 and t5 occurred significantly later in oligoasthenoteratozoospermia (OAT) patients compared with others (p < 0.05). In IMSI group, there were no differences in the TLM parameters among different aetiologies (p > 0.05). The rates of chemical pregnancy and implantation (37.8% and 38.2% respectively) were insignificantly higher in OAT patients compare to others (p > 0.05). Also, the clinical pregnancy and live birth rates (32% and 32% respectively) were insignificantly higher in teratozoospermia (T) cases. Sperm selection with MSOME parameters and IMSI can improve the embryo morphokinetics and clinical outcomes in couples with male factor infertility, especially for OAT and T patients.
Assuntos
Embrião de Mamíferos/diagnóstico por imagem , Desenvolvimento Embrionário , Infertilidade Masculina/terapia , Injeções de Esperma Intracitoplásmicas/métodos , Adulto , Coeficiente de Natalidade , Técnicas de Cultura Embrionária , Embrião de Mamíferos/embriologia , Feminino , Humanos , Masculino , Microinjeções , Indução da Ovulação/métodos , Gravidez , Taxa de Gravidez , Análise do Sêmen/métodos , Imagem com Lapso de Tempo , Resultado do TratamentoRESUMO
The aim of this study was to comprise the effect of catalase on sperm parameters and chromatin in normospermic persons. Semen samples were obtained from fertile men. A certain amount of different concentrations of catalase (0.1, 1, 10, 50, 100, 150 and 200 IU.ml) was added to each vial containing semen. Control group had similar condition to treated groups without treatment. Treatment was done for one hour in incubator and 4 and 24 hr in room temperature. Sperm parameters (motility, viability and morphology) and chromatin were evaluated after incubation. The results show that percentage of motility was insignificantly increased at concentration of 100 IU.ml catalase. This increase was higher than other examined concentration in all incubation time. The increase in sperm motility had significant difference in concentrations of 100 IU.ml with other concentrations. Other parameters showed no significant difference in all concentrations. Regarding the health of sperm chromatin, low concentrations of catalase had significant effect on this variable. This effect was more in low concentrations than high concentrations. This study showed the use of lower concentrations of antioxidant can improve the sperm parameters and chromatin quality. The low concentrations of catalase led to protection of chromatin and optimisation of sperm parameters.
Assuntos
Antioxidantes/administração & dosagem , Catalase/administração & dosagem , Cromatina/metabolismo , Preservação do Sêmen/métodos , Espermatozoides/efeitos dos fármacos , Adulto , Células Cultivadas , Relação Dose-Resposta a Droga , Voluntários Saudáveis , Humanos , Masculino , Oligospermia/terapia , Técnicas de Reprodução Assistida , Contagem de Espermatozoides , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Adulto JovemRESUMO
Cumulus cells features and embryo developmental events can be considered as noninvasive indicators for embryo selection and clinical outcomes. A combination of time-lapse morphokinetic parameters and cumulus cell apoptosis in women with polycystic ovarian syndrome (PCOS) was evaluated for predicting pregnancy outcome. We assessed a total of 547 embryos from 100 intracytoplasmic sperm injection (ICSI) cycles. Time-lapse records were interpreted in time to pronuclear fading (tPNf), time to 2 to 8 cells (t2-t8), direct cleavage, reverse cleavage, and also for the presence of multinucleation. Percentages of apoptosis were identified in 100 associated cumulus cell samples using the TDT-mediated dUTP-biotin nick end-labeling assay. The significant decrease of apoptotic cumulus cells was detected in patients with chemical and clinical pregnancies as well as live birth among patients PCOS and in the tubal infertility group (p > 0.05). Furthermore, significantly higher implantation rate and also significantly lower cases of early pregnancy loss were observed in the group of oocytes with less apoptotic cumulus cells. Multivariate logistic regression analysis showed that tPNf together with cumulus cell apoptosis were independent prognostic factors of chemical pregnancy, clinical pregnancy rate, and live birth. Time-lapse embryo parameters may not reflect the cumulus cell apoptosis rate. However, the rate of apoptotic cumulus cells is significantly associated with ICSI outcome using Day 3 embryo transfer.