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1.
Theriogenology ; 225: 55-66, 2024 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-38795511

RESUMEN

This study aims to evaluate the effects of adding alpha lipoic acid (ALA) to the in vitro ovarian tissue culture medium, either fresh or after vitrification/warming. For this purpose, 10 ovaries from five adult sheep were used. Each pair of ovaries gave rise to 16 fragments and were randomly distributed into two groups: fresh (n = 8) and vitrified (n = 8). Two fresh fragments were fixed immediately and considered the control, while another six were cultured in vitro for 14 days in the absence; presence of a constant (100 µM/0-14 day) or dynamic (50 µM/day 0-7 and 100 µM/day 8-14) concentration of ALA. As for the vitrified fragments, two were fixed and the other six were cultured in vitro under the same conditions described for the fresh group. All the fragments were subjected to morphological evaluation, follicular development and stromal density (classical histology), DNA fragmentation (TUNEL), senescence (Sudan Black), fibrosis (Masson's Trichome), and endoplasmic reticulum stress (immunofluorescence). Measurements of the antioxidant capacity against the free radicals 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and estradiol (E2) levels in the culture medium was performed. The results showed that in the absence of ALA, in vitro culture of vitrified ovarian fragments showed a significant reduction (P < 0.05) in follicular morphology and increased the presence of senescence and tissue fibrosis (P < 0.05). Dynamic ALA maintained E2 levels unchanged (P > 0.05) until the end of vitrified ovarian tissue culture and controlled the levels of ABTS and DPPH radicals in fresh or vitrified cultures. Therefore, it is concluded that ALA should be added to the vitrified ovarian tissue in vitro culture medium to reduce the damage that leads to loss of ovarian function. To ensure steroidogenesis during in vitro culture, ALA should be added dynamically (different concentrations throughout culture).


Asunto(s)
Ácido Tióctico , Técnicas de Cultivo de Tejidos , Animales , Femenino , Ácido Tióctico/farmacología , Ovinos , Técnicas de Cultivo de Tejidos/veterinaria , Ovario/efectos de los fármacos , Folículo Ovárico/efectos de los fármacos , Antioxidantes/farmacología , Vitrificación , Criopreservación/veterinaria
2.
Theriogenology ; 215: 78-85, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38016304

RESUMEN

During in vitro maturation (IVM) cumulus-oocyte complexes (COCs) are exposed to conditions that can trigger oxidative stress, thus, reducing oocyte maturation and viability. Aiming to mitigate these detrimental conditions, the effects of IVM medium supplementation with anethole have been tested. Anethole, also known as trans-anethole (1-methoxy-4 [1-propenyl]-benzene), is a naturally occurring phenylpropanoid with various pharmacological properties, including antioxidant effects. However, no study has examined anethole effect on goat COCs during IVM. Thus, the aim of this study was to evaluate the effects of different anethole concentrations on oocyte maturation, oxidative stress, and in vitro development of caprine embryos after parthenogenetic activation. Goat COCs were selected and randomly distributed into the following treatments: TCM-199+ medium (control), or TCM-199+ medium supplemented with 30 µg/mL (AN30); 300 µg/mL (AN300) or 2000 µg/mL (AN2000) of anethole. After IVM, part of the COCs was chosen for oocyte viability and chromatin configuration, intracellular reactive oxygen species levels, and mitochondrial membrane potential assessment. Another part of COCs was parthenogenetically activated, and presumptive zygotes were cultured for 7 days. Results demonstrated that anethole at 30 µg/mL increased oocyte maturation and cleavage rates when compared to the other treatments (P < 0.05), as well as oocyte viability and in vitro embryo production when compared to the control treatment (P < 0.05). Additionally, treatment with anethole at 2000 µg/mL decreased oocyte nuclear maturation and cleavage rates when compared to other treatments (P < 0.05) and embryo production if compared to control and AN30 treatments (P < 0.05). Moreover, anethole at 2000 µg/mL increased mitochondrial membrane potential when compared to the other treatments (P < 0.05). In conclusion, anethole exerts a concentration-dependent effect during goat COCs IVM. For a more desirable outcome of oocyte viability and maturation, and in vitro embryo production, the use of anethole at 30 µg/mL is recommended.


Asunto(s)
Cabras , Técnicas de Maduración In Vitro de los Oocitos , Animales , Femenino , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Técnicas de Maduración In Vitro de los Oocitos/métodos , Cabras/fisiología , Oocitos/fisiología , Suplementos Dietéticos , Células del Cúmulo
3.
Theriogenology ; 211: 151-160, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37639997

RESUMEN

This study aims to define the best method (slow freezing or vitrification) and fragment size (1, 5, or 9 mm³) for prepubertal goat testis cryopreservation, as well as to evaluate testicular morphological integrity after cryopreservation and in vitro culture (IVC). Initially (experiment I), 1, 5, or 9 mm³ testis fragments were cryopreserved by slow freezing using a Mr. Frosty container with 20% Dimethylsulfoxide (DMSO) or vitrified using the Ovarian Tissue Cryosystem (OTC) device, (Equilibration solution - ES: 10% DMSO and 10% ethylene glycol - EG; Vitrification solution - VS: 20% DMSO and 20% EG) and then subjected to morphological analysis, type I and III collagen quantification and gene expression (Oct4, C-kit, Bax, and Bcl-2). Subsequently, (experiment II), fresh or cryopreserved by slow freezing testis fragments were cultured in vitro and submitted to morphological analysis by scanning electron microscopy. The data from the experiment I revealed fewer morphological alterations in 1 and 5 mm³ fragments after vitrification and slow freezing, respectively. The percentage of type I collagen fibers in 5 and 9 mm³ frozen was higher than in fresh or vitrified fragments. For type III collagen, fresh or frozen fragments of 1 and 5 mm3 showed a higher percentage than fragments of 9 mm3. Gene expression for Oct4 and C-kit after slow freezing or vitrification in the 5 mm3 fragments was lower than that observed in the fresh fragments. The Bax:Bcl-2 ratio in the 1 and 9 mm³ fragments was lower than in the 5 mm³ fragments for fresh fragments or after freezing. In experiment II, fragments cultured in vitro, previously frozen or not, showed more morphological alterations than fresh or frozen fragments. We concluded that slow freezing of 5 mm³ fragments was the best protocol for cryopreserving prepubertal goat testis and although the results of IVC are encouraging, it still needs improvement to restore testicular function after cryopreservation.


Asunto(s)
Dimetilsulfóxido , Cabras , Animales , Masculino , Proteína X Asociada a bcl-2 , Criopreservación/veterinaria , Proteínas Proto-Oncogénicas c-bcl-2 , Proteínas Proto-Oncogénicas c-kit
4.
Lett Appl Microbiol ; 75(6): 1639-1644, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36073022

RESUMEN

This study aimed to evaluate the performance of accelerated hydrogen peroxide® wipes (HPW) for decontamination of the chimpanzee adenovirus AZD1222 vaccine strain used in the production of recombinant COVID-19 vaccine in a pharmaceutical industry. Two matrices were tested on stainless-steel (SS) and low-density-polyethylene (LDP) surfaces: formulated recombinant COVID-19 vaccine (FCV) and active pharmaceutical ingredient (API). The samples were spiked, dried and the initial inoculum, possible residue effect (RE) and titre reduction after disinfection with HPW were determined. No RE was observed. The disinfection procedure with HPW resulted in complete decontamination the of AZD1222 adenovirus strain in FCV (≥7·46 and ≥7·49 log10 infectious unit [IFU] ml-1 for SS and LDP carriers respectively) and API (≥8·79 and ≥8·78 log10 IFU ml-1 for SS and LDP carriers respectively). In conclusion, virucidal activity of HPW was satisfactory against the AZD1222 adenovirus strain and can be a good option for disinfection processes of SS and LPD surfaces in pharmaceutical industry facilities during recombinant COVID-19 vaccine production. This procedure is simple and can be also applied on safety unit cabins and sampling bags made of LDP as well.


Asunto(s)
COVID-19 , Desinfectantes , Humanos , Peróxido de Hidrógeno/farmacología , Desinfectantes/farmacología , ChAdOx1 nCoV-19 , Vacunas contra la COVID-19 , Adenoviridae/genética , Descontaminación/métodos , COVID-19/prevención & control , Desinfección/métodos , Acero Inoxidable , Industria Farmacéutica
5.
Theriogenology ; 188: 135-144, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35689943

RESUMEN

Recent in vitro follicle culture (IVFC) studies in caprine have yielded lower maturation rates using late preantral follicles compared to early antral follicles. Thus, research focusing on developing stage-specific customized culture systems able to improve the efficiency of IVFC for late preantral follicles are warranted. This study aimed to compare the morphometric features, estradiol production, and gene expression between early antral caprine follicles produced in vitro and in vivo. In vitro-derived early antral follicles were produced after a 6-day in vitro culture of late preantral follicles, while in vivo-derived early antral follicles were yielded immediately after isolation from the ovaries; antral follicles were, thereafter, cultured for 18 days. In vitro-derived antral follicles were cultured either in a medium developed for preantral follicles (PF medium) or in a medium developed for antral follicles (AF medium). In vivo-derived early antral follicles, on the other hand, were cultured in AF medium (Control treatment). Results demonstrated that in vitro-derived antral follicles cultured in PF medium produced higher estradiol concentration, and m-RNA expression for matrix metalloproteinase-9 (MMP-9), and insulin receptor when compared to both in vitro- and in vivo-derived antral follicles cultured in AF medium. Remarkably, in vitro-derived antral follicles cultured in PF medium had similar MII and oocytes ≥110 µm rates compared with in vivo-derived antral follicles (Control treatment). In conclusion, when cultured in a single and appropriate medium (i.e., PF medium), in vitro-derived early antral follicles had comparable oocyte maturation rates to the in vivo-derived early antral follicles.


Asunto(s)
Cabras , Folículo Ovárico , Animales , Estradiol/metabolismo , Estradiol/farmacología , Femenino , Hormona Folículo Estimulante , Cabras/metabolismo , Técnicas de Maduración In Vitro de los Oocitos/métodos , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Oocitos/metabolismo
6.
Zygote ; 30(1): 144-147, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33988116

RESUMEN

Ovine ovarian fragments (3 × 3 × 1 mm) were fixed in neutral buffered formalin (NBF), Carnoy's solution (CAR), Davidson's solution (DAV), or paraformaldehyde (PFA) for 12 h or 24 h. After this fixation time, each fragment was prepared for histological analysis. Although fixative and fixation period did not affect follicular and stromal cells density, the percentages of morphologically normal primordial and primary follicles was affected by the fixative type and period of fixation. Paraformaldehyde was not indicated as a fixative for ovarian fragments. Formalin was a suitable fixative only when the period of fixation was 12 h, while Carnoy was efficient after a fixation period of 24 h. In conclusion, the most indicated fixative for the morphological evaluation of ovarian preantral follicles was DAV, regardless of the fixation period, that is 12 or 24 h.


Asunto(s)
Folículo Ovárico , Ovario , Animales , Femenino , Fijadores/farmacología , Ovinos , Fijación del Tejido
7.
Zygote ; 30(3): 419-422, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-34689852

RESUMEN

The aim of this study was to evaluate the effect of 1 µmol/l zearalenone (ZEN) and 1 µmol/l matairesinol (MAT), alone or in combination, on the morphology of in vitro-cultured ovarian preantral follicles. Ovaries from four adult sheep were collected at a local slaughterhouse and fragmented, and the ovarian pieces were submitted to in vitro culture for 3 days in the presence or absence of the test compounds. The morphology of primordial and primary follicles was impaired by ZEN. The plant lignan MAT alone did not maintain the morphology of the ovarian follicles; its combination with ZEN counteracted the negative effects observed when follicles were cultured in the presence of the mycotoxin alone. However, MAT was not able to promote the in vitro development of the ovarian follicles.


Asunto(s)
Lignanos , Zearalenona , Animales , Femenino , Furanos , Lignanos/farmacología , Folículo Ovárico , Ovario , Ovinos , Zearalenona/toxicidad
8.
Theriogenology ; 177: 127-132, 2022 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-34700069

RESUMEN

ATP-binding cassette (ABC) transporters perform multiple functions in reproductive tissues. During ovarian tissue vitrification, the plasma membrane has important functions in the influx or efflux of water, and substances such as cryoprotectants and channel proteins that are required in this process. Thus, the present study aimed to verify the relative abundance of mRNA transcript of ABC transporters ABCB1, ABCG2, and MRP2 after vitrification and in vitro culture (IVC) of ovine ovarian tissue. For this study, the ovarian cortex fragments were proportioned into four groups as fresh control, vitrified control, fresh culture, and vitrified culture groups. After vitrification and in vitro culture, the ovarian tissue was evaluated using morphological procedures. Further, relative abundance of ABCB1, ABCG2, and MRP2 transporter mRNA transcripts in the ovarian cortex subjected to aforementioned treatment conditions were evaluated using qPCR. Our results showed a negative association between degenerated follicles and mRNA transcript abundances of ABCB1 and ABCG2. In addition, the percentage of growing follicles in the ovine ovarian cortex after vitrification was similar to that of the fresh control tissue without in vitro culture. The in vitro culture of fresh and vitrified tissue however, showed a significant decrease in the percentage of growing follicles. To the best of our knowledge, we believe that our data for the first time has studied the relative abundances of ABCB1 and ABCG2 mRNA transcripts in the ovine ovarian cortex. In addition, alterations of these protein channels may be indicative of a deleterious effect of osmotic stress on follicular survival during vitrification. Furthermore, these effects were detectable only after the IVC of the ovarian tissues. Nonetheless, further studies are required to investigate the functions of ABC transporters in ovine folliculogenesis, especially after in vitro culture of ovarian tissue.


Asunto(s)
Transportadoras de Casetes de Unión a ATP , Vitrificación , Transportadoras de Casetes de Unión a ATP/genética , Animales , Criopreservación/veterinaria , Crioprotectores/farmacología , Regulación hacia Abajo , Femenino , Ovinos
9.
Lett Appl Microbiol ; 74(4): 536-542, 2022 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-34921425

RESUMEN

This study aimed to evaluate the performance of hydrogen peroxide vapour (HPV) to inactivate the chimpanzee adenovirus AZD1222 vaccine strain used in the production of recombinant COVID-19 vaccine for application in cleaning validation in pharmaceutical industries production areas. Two matrixes were tested: formulated recombinant COVID-19 vaccine (FCV) and active pharmaceutical ingredient (API). The samples were dried on stainless steel and exposed to HPV in an isolator. One biological indicator with population >106 Geobacillus stearothermophilus spores was used to validate the HPV decontamination cycle as standard. HPV exposure resulted in complete virus inactivation in FVC (≥5·03 log10 ) and API (≥6·40 log10 ), showing HPV efficacy for reducing chimpanzee adenovirus AZD1222 vaccine strain. However, the optimum concentration and contact time will vary depending on the type of application. Future decontamination studies scaling up the process to the recombinant COVID-19 vaccine manufacturing areas are necessary to evaluate if the HPV will have the same or better virucidal effectivity in each specific production area. In conclusion, HPV showed efficacy for reducing AZD1222 chimpanzee adenovirus strain and can be a good choice for pharmaceutical industries facilities disinfection during recombinant COVID-19 vaccine production.


Asunto(s)
COVID-19 , Desinfectantes , Adenoviridae , Animales , Vacunas contra la COVID-19 , ChAdOx1 nCoV-19 , Industria Farmacéutica , Humanos , Peróxido de Hidrógeno/farmacología , Industria Manufacturera , Pan troglodytes , Preparaciones Farmacéuticas
10.
Theriogenology ; 174: 124-130, 2021 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-34428678

RESUMEN

The aim of this study was to evaluate the effect of 1 µmol/L zearalenone (ZEN) and 1 µmol/L enterolactone (ENL), alone or in combination, on the survival and morphology of in vitro cultured ovarian preantral follicles. Ovaries from 10 sheep were collected at a local abattoir and fragmented, and the ovarian pieces were submitted to in vitro culture for 3 days in the presence or absence of the test compounds. The morphology of primordial and primary follicles was impaired by ZEN, whereas that of cultured secondary follicles was improved by ENL. However, the combination of ENL with ZEN impaired the quality of primary and secondary follicles. Both ZEN and ENL induced apoptosis, but only ZEN was responsible for oocyte autophagy. None of these xenoestrogens affected endoplasmic reticulum stress as observed by the unaltered expression of ERP29. Differently from ZEN, ENL increased the expression of the efflux transporter ABCG2. In conclusion, although ENL can counteract the negative effects of ZEN on primordial and primary follicles, this positive effect is not similar to that observed in ovarian tissue cultures in the presence of ENL alone.


Asunto(s)
Zearalenona , 4-Butirolactona/análogos & derivados , Animales , Femenino , Lignanos , Oocitos , Folículo Ovárico , Ovario , Ovinos , Zearalenona/toxicidad
11.
Respir Med Res ; 80: 100812, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34224956
12.
Dermatol Online J ; 26(3)2020 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-32609451

RESUMEN

Eosinophilic dermatosis of hematologic malignancy (EDHM) is a dermatosis characterized by tissue eosinophilia that has been previously reported as insect bite-like reaction. It is a rare condition with a wide variety of clinical presentations ranging from papules, nodules, or blisters that simulate arthropod bites, to the formation of plaques of differing sizes. We report a case of eosinophilic dermatosis of hematologic malignancy in a patient with a hematoproliferative disorder.


Asunto(s)
Eosinofilia/patología , Síndromes Mielodisplásicos/complicaciones , Síndromes Paraneoplásicos/patología , Enfermedades de la Piel/patología , Anciano , Eosinofilia/etiología , Humanos , Masculino , Enfermedades de la Piel/etiología
13.
Braz J Med Biol Res ; 53(4): e8604, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32294697

RESUMEN

Maraba virus is a member of the genus Vesiculovirus of the Rhabdoviridae family that was isolated in 1983 from sandflies captured in the municipality of Maraba, state of Pará, Amazônia, Brazil. Despite 30 years having passed since its isolation, little is known about the neuropathology induced by the Maraba virus. Accordingly, in this study the histopathological features, inflammatory glial changes, cytokine concentrations, and nitric oxide activity in the encephalon of adult mice subjected to Maraba virus nostril infection were evaluated. The results showed that 6 days after intranasal inoculation, severe neuropathological-associated disease signs appeared, including edema, necrosis and pyknosis of neurons, generalized congestion of encephalic vessels, and intra- and perivascular meningeal lymphocytic infiltrates in several brain regions. Immunolabeling of viral antigens was observed in almost all central nervous system (CNS) areas and this was associated with intense microglial activation and astrogliosis. Compared to control animals, infected mice showed significant increases in interleukin (IL)-6, tumor necrosis factor (TNF)-α, interferon (INF)-γ, MCP-1, nitric oxide, and encephalic cytokine levels. We suggest that an exacerbated inflammatory response in several regions of the CNS of adult BALB/c mice might be responsible for their deaths.


Asunto(s)
Meningoencefalitis/complicaciones , Estomatitis Vesicular/complicaciones , Animales , Astrocitos/metabolismo , Brasil , Citocinas/análisis , Modelos Animales de Enfermedad , Citometría de Flujo , Masculino , Meningoencefalitis/patología , Ratones , Ratones Endogámicos BALB C , Microglía/metabolismo , Óxido Nítrico/análisis , Estomatitis Vesicular/patología , Vesiculovirus
14.
J Public Health (Oxf) ; 42(2): 395-402, 2020 05 26.
Artículo en Inglés | MEDLINE | ID: mdl-32166309

RESUMEN

BACKGROUND: Heatwaves can lead to increased mortality. In the Portuguese heat-health warning system (HHWS), ÍCARO, a daily report with heat-related mortality prediction is sent to heat-health action plan (HHAP) practitioners. HHAP practitioners assess risk and implement measures to prevent heatwave-related impact, but ÍCARO's use and understanding are unknown. We assessed ÍCARO's use and understanding by key HHAP practitioners. METHODS: We conducted semi-structured interviews with national/regional HHAP practitioners. Interviews were recorded, transcribed and analysed using thematic content analysis. To maximize credibility a validation process was implemented through researcher triangulation; a sample of 30 segments was recorded by independent researchers. RESULTS: We conducted six interviews with nine professionals (mean time 52 min) from five regions. We identified four categories: report's content and presentation, report's reception and communication, ÍCARO and risk assessment and other issues. Practitioners use ÍCARO and perceived it as relevant; they raised issues on its interpretation and felt these were not fully addressed, given researchers' use of statistical/epidemiological terms. We identified the need for improved communication and report's clarity. CONCLUSIONS: Our study stresses the need for collaboration between experts within HHWS/HHAP. Despite ÍCARO's understanding being challenging, practitioners consider it a relevant tool. Researchers should use less statistical language and clarify ÍCARO's interpretation. Practitioners' needs should be considered when developing/revising tools.


Asunto(s)
Calor , Lenguaje , Comunicación , Planificación en Salud , Humanos , Portugal
15.
Theriogenology ; 147: 10-17, 2020 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-32074494

RESUMEN

Numerous studies have reported the importance of thyroid hormones on the development of later preantral and antral follicles, but their interactions with other hormones and effects in regulating early preantral follicle growth remain unclear. Here we investigated the in vitro effects of thyroxine combined with insulin on caprine preantral follicle survival and development. Sliced ovarian tissues were cultured for 1 or 7 days using 10 ng/mL (low) or 10 µg/mL (high) insulin in the presence of thyroxine at 0, 0.5, 1 or 2 µg/mL. Post-culture, we evaluated the follicular survival and development, assessed the expression of apoptotic-related genes (Bcl2/Bax) and receptors of insulin and thyroid hormones, and quantified the estradiol and reactive oxygen species (ROS) production levels. Follicular survival in low-insulin culture conditions was enhanced by the presence of 0.5 µg/mL thyroxine (P < 0.05) as compared to the thyroxine-free medium but remained similar to non-cultured control in the presence of 2 µg/mL (P > 0.05). Significantly higher ROS production was measured from Day 1 to Day 7 in low-insulin culture media containing 0.5 or 2 µg/mL thyroxine (P < 0.05). When compared to high insulin level, the presence of thyroxine in low insulin culture conditions yielded higher stromal cell density (P < 0.05), increased estradiol production on Day 1, and higher Bcl2/Bax ratio on Day 7. Cultures with high levels of both insulin and thyroxine led to follicles and oocytes with larger diameters (P < 0.05). The RNA transcript levels of insulin and thyroid receptors were reduced in the presence of high insulin cultures when compared to controls (non-cultured). In conclusion, the combination of low concentrations of insulin and thyroxine better maintained follicle survival, while high levels ensured better follicular development.


Asunto(s)
Cabras , Insulina/farmacología , Folículo Ovárico/fisiología , Tiroxina/farmacología , Animales , Relación Dosis-Respuesta a Droga , Estradiol/biosíntesis , Femenino , Regulación de la Expresión Génica , Insulina/administración & dosificación , Especies Reactivas de Oxígeno , Tiroxina/administración & dosificación , Técnicas de Cultivo de Tejidos
16.
Reprod Fertil Dev ; 32(5): 531-537, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-32087765

RESUMEN

Ovary fragments from six sexually mature cats were vitrified in the presence or absence of betaine or ascorbic acid, loaded (7.4 or 74µM betaine; 20 or 200µM ascorbic acid) or not (1mM betaine or 0.3mM ascorbic acid) into CaCO3 microparticles, and assessed for follicular morphology, oxidative stress and mitochondrial activity Feline ovarian tissue was successfully preserved after vitrification in the presence of 74µM betaine loaded in CaCO3 microparticles, as confirmed by morphological analysis and the density of preantral follicles and stromal cells, as well as by the increased mitochondrial activity and decreased production of reactive oxygen species.


Asunto(s)
Betaína/farmacología , Carbonato de Calcio/farmacología , Criopreservación , Mitocondrias/efectos de los fármacos , Folículo Ovárico/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Animales , Ácido Ascórbico/farmacología , Gatos , Supervivencia Celular/efectos de los fármacos , Femenino , Mitocondrias/metabolismo , Mitocondrias/patología , Folículo Ovárico/metabolismo , Folículo Ovárico/patología , Vitrificación
17.
Sci Rep ; 10(1): 903, 2020 Jan 16.
Artículo en Inglés | MEDLINE | ID: mdl-31942035

RESUMEN

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

18.
J Ethnopharmacol ; 251: 112571, 2020 Apr 06.
Artículo en Inglés | MEDLINE | ID: mdl-31935494

RESUMEN

BACKGROUND: Senecio biafrae is a medicinal plant widely used in traditional medicine to cure female infertility. Some effects have been pharmacologically demonstrated on immature female rats but in vivo and in vitro investigations are still necessary for determining its mechanism of action. The aim of the present study was to evaluate the estrogenic and FSH-like effects of the plant extracts and fractions on some fertility parameters in immature female rats and on in vitro survival and growth of swine preantral follicles. METHODS: 21-23 days old female Wistar rats orally received extracts and fractions of S. biafrae at 0, 8 and 64 mg/kg doses over 20 days. The LH, FSH, estradiol and progesterone serum levels were evaluated as well as the ovarian cholesterol, uterus and ovaries masses and proteins. The numbers of follicles at different developmental stages were recorded in ovarian cortexes after histology. Slices of swine ovarian cortexes were cultured along 1 or 7 days in alpha-minimum essential medium (α-MEM) and fixed for morphological analysis of preantral follicles. The fresh control, cultured control (CIV control) and different Senecio biafrae-treated ovarian fragments were analyzed for preantral follicles development. Treatments that showed the best follicle growth in culture were submitted to AgNOR test. The aqueous and MeOH/CH2Cl2 extracts as well as the ethyl acetate and hexane fractions of S. biafrae were submitted to the HPLC for analysis of polyphenolic secondary metabolites. RESULTS: Ovarian and uterine proteins were significantly high (p < 0.01) in animals treated with the two dosages of ethyl acetate and n-butanol fractions. The same result was recorded with uterine proteins in animals treated with the hexane fraction. The FSH level significantly dropped with all ethanolic extract doses and with the 64 mg/kg dosage of the methanol/methylene chloride (MeOH/CH2Cl2) extract while LH was reduced (p < 0.01) in almost all the treated groups. Estradiol level was significantly increased (p < 0.001) in the three groups receiving the extracts, but reduced (p < 0.001) in the three groups receiving the fractions of the plant. The progesterone level increased with almost all the treated groups. Primary and secondary follicles augmented (p < 0.01) in MeOH/CH2Cl2 extract and n-butanol fraction while tertiary follicles increased with the same extract and the ethyl acetate fraction (p < 0.05). Treatments with aqueous and ethanolic extracts as well as ethyl acetate fraction led to a significant increase (p < 0.05) in the number of morphologically normal follicles after 7 days of culture as compared to the CIV control. The number of AgNOR dots per follicle was significantly low (p < 0.05) in all cultured groups as compared to the fresh control, except the ethyl acetate 2.8 ng/ml dosage. The same observation was done with AgNOR dots per cell in the 2.8 ng/ml dosage aqueous extract-treated fragments. The phenolic compounds mainly encountered in the plant, independently of the extract or fraction are apigenin, eugenol and rutin. CONCLUSION: Extracts and fractions of S. biafrae have an important FSH-like effect which induces follicular survival and growth.


Asunto(s)
Ovario/efectos de los fármacos , Extractos Vegetales/farmacología , Senecio , Animales , Colesterol/metabolismo , Estradiol/sangre , Femenino , Hormona Folículo Estimulante/sangre , Hormona Luteinizante/sangre , Ovario/metabolismo , Progesterona/sangre , Ratas Wistar , Porcinos
19.
Braz. j. med. biol. res ; 53(4): e8604, 2020. graf
Artículo en Inglés | LILACS | ID: biblio-1100926

RESUMEN

Maraba virus is a member of the genus Vesiculovirus of the Rhabdoviridae family that was isolated in 1983 from sandflies captured in the municipality of Maraba, state of Pará, Amazônia, Brazil. Despite 30 years having passed since its isolation, little is known about the neuropathology induced by the Maraba virus. Accordingly, in this study the histopathological features, inflammatory glial changes, cytokine concentrations, and nitric oxide activity in the encephalon of adult mice subjected to Maraba virus nostril infection were evaluated. The results showed that 6 days after intranasal inoculation, severe neuropathological-associated disease signs appeared, including edema, necrosis and pyknosis of neurons, generalized congestion of encephalic vessels, and intra- and perivascular meningeal lymphocytic infiltrates in several brain regions. Immunolabeling of viral antigens was observed in almost all central nervous system (CNS) areas and this was associated with intense microglial activation and astrogliosis. Compared to control animals, infected mice showed significant increases in interleukin (IL)-6, tumor necrosis factor (TNF)-α, interferon (INF)-γ, MCP-1, nitric oxide, and encephalic cytokine levels. We suggest that an exacerbated inflammatory response in several regions of the CNS of adult BALB/c mice might be responsible for their deaths.


Asunto(s)
Animales , Masculino , Conejos , Estomatitis Vesicular/complicaciones , Meningoencefalitis/complicaciones , Brasil , Astrocitos/metabolismo , Citocinas/análisis , Vesiculovirus , Microglía/metabolismo , Modelos Animales de Enfermedad , Estomatitis Vesicular/patología , Citometría de Flujo , Meningoencefalitis/patología , Ratones Endogámicos BALB C , Óxido Nítrico/análisis
20.
Biotechnol Rep (Amst) ; 24: e00365, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31463206

RESUMEN

Buriti oil nanoemulsions were prepared using non-interesterified buriti oil or buriti oil interesterified for 6 or 24 h (NBO, NBO6h, and NBO24 h), respectively. The aim was to investigate the effects of interesterified oils on the physicochemical and biological properties of nanoemulsions. Samples were stored at 4 and 25 °C for 30 days, and their physicochemical properties and biological activities were evaluated. The mean droplet diameter of nanoemulsions ranged from 196 to 270 nm. NBO24 h had the smallest droplet size and was the most stable during the storage period. Furthermore, NBO24 h demonstrating the good oxidative stability, had a high antioxidant capacity, and was less susceptible to droplet aggregation. NBO and NBO24 h had similar biological activity against Gram-negative bacteria (Escherichia coli O157: H7); bacterial growth was inhibited by at least 60% at 3.12 mg mL-1. The nanoemulsions have interesting properties for the production of pharmaceutical, cosmetic, and food formulations with antimicrobial activity.

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