ABSTRACT
In brief: Elevated temperatures disturbed sperm physiology. Bovine sperm cells exposed to heat shock led to diminished mitochondrial activity, fertilizing ability, increased oxidative stress and caspase activity concomitant with a delay in embryonic developmental kinetics and modulation of sperm-borne microRNAsmiRNAs. Abstract: Sperm function is susceptible to adverse environmental conditions. It has been demonstrated that in vivo and in vitro exposure of bovine sperm to elevated temperature reduces sperm motility and fertilizing potential. However, the cascade of functional, cellular, and molecular events triggered by elevated temperature in the mature sperm cell remains not fully understood. Therefore, the aim of this study was to determine the effect of heat shock on mature sperm cells. Frozen-thawed Holstein sperm were evaluated immediately after Percoll purification (0 h non-incubation control) or after incubation at 35, 38.5, and 41°C for 4 h. Heat shock reduced sperm motility after 3-4 h at 41°C while mitochondrial activity was reduced by 38.5 and 41°C when compared to the control. Heat shock also increased sperm reactive oxygen species production and caspase activity. Heat-shocked sperm had lower fertilizing ability, which led to diminished cleavage and blastocyst rates. Preimplantation embryo developmental kinetics was also slowed and reduced by sperm heat shock. The microRNA (miR) profiling identified >300 miRs in bovine sperm. Among these, three and seven miRs were exclusively identified in sperm cells exposed to 35 and 41°C, respectively. Moreover, miR-181d was enriched in sperm cells exposed to higher temperatures. Hence, elevated temperature altered the physiology of mature sperm cells by perturbing cellular processes and the miR profile, which collectively led to lower fertilizing ability and preimplantation development.
Subject(s)
MicroRNAs , Semen Preservation , Animals , Caspases , Cattle , Heat-Shock Response , Male , MicroRNAs/genetics , Reactive Oxygen Species , Semen , Sperm Motility , Spermatozoa/physiologyABSTRACT
Alzheimer's disease (AD) is the most common form of dementia worldwide, being characterized by the deposition of senile plaques, neurofibrillary tangles (enriched in the amyloid beta (Aß) peptide and hyperphosphorylated tau (p-tau), respectively) and memory loss. Aging, type 2 diabetes (T2D) and female sex (especially after menopause) are risk factors for AD, but their crosslinking mechanisms remain unclear. Most clinical trials targeting AD neuropathology failed and it remains incurable. However, evidence suggests that effective anti-T2D drugs, such as the GLP-1 mimetic and neuroprotector liraglutide, can be also efficient against AD. Thus, we aimed to study the benefits of a peripheral liraglutide treatment in AD female mice. We used blood and brain cortical lysates from 10-month-old 3xTg-AD female mice, treated for 28 days with liraglutide (0.2 mg/kg, once/day) to evaluate parameters affected in AD (e.g., Aß and p-tau, motor and cognitive function, glucose metabolism, inflammation and oxidative/nitrosative stress). Despite the limited signs of cognitive changes in mature female mice, liraglutide only reduced their cortical Aß1-42 levels. Liraglutide partially attenuated brain estradiol and GLP-1 and activated PKA levels, oxidative/nitrosative stress and inflammation in these AD female mice. Our results support the earlier use of liraglutide as a potential preventive/therapeutic agent against the accumulation of the first neuropathological features of AD in females.
Subject(s)
Alzheimer Disease/metabolism , Amyloid beta-Peptides/metabolism , Brain/metabolism , Hypoglycemic Agents/pharmacology , Inflammation/metabolism , Liraglutide/pharmacology , Peptide Fragments/metabolism , Animals , Behavior, Animal , Cyclic AMP-Dependent Protein Kinases/metabolism , Diabetes Mellitus, Type 2/metabolism , Estradiol/metabolism , Female , Glucagon-Like Peptide 1/metabolism , Glycolysis , Maze Learning , Memory Disorders , Mice , Neurofibrillary Tangles/metabolism , Nitrosative Stress , Oxidative Stress , Phenotype , Plaque, Amyloid/metabolismABSTRACT
Macrophages play a very important role in the conduction of several regenerative processes mainly due to their plasticity and multiple functions. In the muscle repair process, while M1 macrophages regulate the inflammatory and proliferative phases, M2 (anti-inflammatory) macrophages direct the differentiation and remodelling phases, leading to tissue regeneration. The aim of this study was to evaluate the effect of red and near infrared (NIR) photobiomodulation (PBM) on macrophage phenotypes and correlate these findings with the repair process following acute muscle injury. Wistar rats were divided into 4 groups: control; muscle injury; muscle injury + red PBM; and muscle injury + NIR PBM. After 2, 4 and 7 days, the tibialis anterior muscle was processed for analysis. Macrophages phenotypic profile was evaluated by immunohistochemistry and correlated with the different stages of the skeletal muscle repair by the qualitative and quantitative morphological analysis as well as by the evaluation of IL-6, TNF-α and TGF-ß mRNA expression. Photobiomodulation at both wavelengths was able to decrease the number of CD68+ (M1) macrophages 2 days after muscle injury and increase the number of CD163+ (M2) macrophages 7 days after injury. However, only NIR treatment was able to increase the number of CD206+ M2 macrophages (Day 2) and TGF-ß mRNA expression (Day 2, 4 and 7), favouring the repair process more expressivelly. Treatment with PBM was able to modulate the inflammation phase, optimize the transition from the inflammatory to the regeneration phase (mainly with NIR light) and improve the final step of regeneration, enhancing tissue repair.
Subject(s)
Low-Level Light Therapy , Muscle Development/radiation effects , Muscles/radiation effects , Regeneration/radiation effects , Animals , Antigens, CD/genetics , Antigens, Differentiation, Myelomonocytic/genetics , Cell Differentiation/radiation effects , Humans , Macrophages/pathology , Macrophages/radiation effects , Muscle, Skeletal/growth & development , Muscle, Skeletal/injuries , Muscle, Skeletal/radiation effects , Muscles/injuries , Muscles/pathology , Rats , Receptors, Cell Surface/genetics , Wound Healing/physiology , Wound Healing/radiation effectsABSTRACT
[Purpose] The aim of the present study was to evaluate the relationship between upper limb impairment and oral health impact in individuals with hemiparesis stemming from a stroke. [Subjects and Methods] The study subjects were conducted with a sample of 27 stroke survivors with complete or partial hemiparesis with brachial or crural predominance. The 14-item short version of the Oral Health Impact Profile was used to evaluate perceptions of oral health. The Brazilian version of the Stroke Specific Quality of Life Scale was used to evaluate perceptions regarding quality of life. [Results] A statistically significant association was found between the upper extremity function subscale of the SSQOL-Brazil and the impact of oral health evaluated using the OHIP-14, with a strong correlation found for the physical pain subscale, moderate correlations with the functional limitation, psychological discomfort, physical disability, social disability and social handicap subscales as well as a weak correlation with the psychological disability subscale. Analyzing the OHIP-14 scores with regard to the impact of oral health on quality of life, the most frequent classification was weak impact, with small rates of moderate and strong impact. [Conclusion] Compromised upper limb function and self-perceived poor oral health, whether due to cultural resignation or functional disability, exert a negative impact on the quality of life of individuals with hemiparesis stemming from a stroke.
ABSTRACT
In recent years, Candida auris has emerged as a hazardous hospital-acquired pathogen. Its resistance to antifungal treatments makes it challenging, requiring new approaches to manage it effectively. Herein, we aimed to assess the impact of photodynamic inactivation mediated by methylene blue (MB-PDI) or 1,9-dimethyl MB (DMMB-PDI) combined with a red LED against C. auris. To evaluate the photoinactivation of yeasts, we quantified colony-forming units and monitored ROS production. To gain some insights into the differences between MB and DMMB, we assessed lipid peroxidation (LPO) and mitochondrial membrane potential (ΔΨm). After, we verified the effectiveness of DMMB against biofilms by measuring metabolic activity and biomass, and the structures were analyzed through scanning electron microscopy and optical coherence tomography. We also evaluated the cytotoxicity in mammalian cells. DMMB-PDI successfully eradicated C. auris yeasts at 3 µM regardless of the light dose. In contrast, MB (100 µM) killed cells only when exposed to the highest dose of light. DMMB-PDI promoted higher ROS, LPO and ΔΨm levels than those of MB. Furthermore, DMMB-PDI was able to inhibit biofilm formation and destroy mature biofilms, with no observed toxicity in fibroblasts. We conclude that DMMB-PDI holds great potential to combat the global threat posed by C. auris.
ABSTRACT
Radiotherapy is a well-established cancer treatment; it is estimated that approximately 52% of oncology patients will require this treatment modality at least once. However, some tumors, such as triple-negative breast cancer (TNBC), may present as radioresistant and thus require high doses of ionizing radiation and a prolonged period of treatment, which may result in more severe side effects. Moreover, such tumors show a high incidence of metastases and decreased survival expectancy of the patient. Thus, new strategies for radiosensitizing TNBC are urgently needed. Red light therapy, photobiomodulation, has been used in clinical practice to mitigate the adverse side effects usually associated with radiotherapy. However, no studies have explored its use as a radiosensitizer of TNBC. Here, we used TNBC-bearing mice as a radioresistant cancer model. Red light treatment was applied in three different protocols before a high dose of radiation (60 Gy split in 4 fractions) was administered. We evaluated tumor growth, mouse clinical signs, total blood cell counts, lung metastasis, survival, and levels of glutathione in the blood. Our data showed that the highest laser dose in combination with radiation arrested tumor progression, likely due to inhibition of GSH synthesis. In addition, red light treatment before each fraction of radiation, regardless of the light dose, improved the health status of the animals, prevented anemia, reduced metastases, and improved survival. Collectively, these results indicate that red light treatment in combination with radiation could prove useful in the treatment of TNBC.
Subject(s)
Radiation-Sensitizing Agents , Triple Negative Breast Neoplasms , Humans , Animals , Mice , Triple Negative Breast Neoplasms/radiotherapy , Triple Negative Breast Neoplasms/drug therapy , Triple Negative Breast Neoplasms/pathology , Disease Models, Animal , Cell Line, Tumor , Radiation-Sensitizing Agents/pharmacology , LightABSTRACT
Radiotherapy is a well-established cancer treatment; it is estimated that approximately 52% of oncology patients will require this treatment modality at least once. However, some tumors, such as triple-negative breast cancer (TNBC), may present as radioresistant and thus require high doses of ionizing radiation and a prolonged period of treatment, which may result in more severe side effects. Moreover, such tumors show a high incidence of metastases and decreased survival expectancy of the patient. Thus, new strategies for radiosensitizing TNBC are urgently needed. Red light therapy, photobiomodulation, has been used in clinical practice to mitigate the adverse side effects usually associated with radiotherapy. However, no studies have explored its use as a radiosensitizer of TNBC. Here, we used TNBC-bearing mice as a radioresistant cancer model. Red light treatment was applied in three different protocols before a high dose of radiation (60 Gy split in 4 fractions) was administered. We evaluated tumor growth, mouse clinical signs, total blood cell counts, lung metastasis, survival, and levels of glutathione in the blood. Our data showed that the highest laser dose in combination with radiation arrested tumor progression, likely due to inhibition of GSH synthesis. In addition, red light treatment before each fraction of radiation, regardless of the light dose, improved the health status of the animals, prevented anemia, reduced metastases, and improved survival. Collectively, these results indicate that red light treatment in combination with radiation could prove useful in the treatment of TNBC.
ABSTRACT
Lamellar alpha-zirconium phosphate was synthesized by direct precipitation and also directly expanded with octadecylamine, through alcoholic solution. To produce a nanocomposite, it was incorporated in linear low-density polyethylene in the molten state, using a counterrotating twin-screw extruder set at 170-190 degrees C and 100 rpm. The differential scanning calorimetry analysis revealed a decrease in the polyolefin melting temperature and crystallinity degree. The higher onset temperature of the zirconium phosphate modified with octadecylamine and linear low density polyethylene composite indicated an increasing of thermal stability and it suggests that some polyethylene chains entered into the filler's spacing. Dynamic-mechanical analysis evidenced an increase in both moduli (storage and loss). Wide-angle X-ray diffraction showed additional peaks--diffraction angles appeared in the region beneath 12 degrees--which were attributed to partial intercalation of polyethylene chains between filler interlamellar spacing. By hydrogen low-field nuclear magnetic resonance, the two low intensity relaxation time peaks shifted to higher values, strongly suggesting interaction between the octadecylamine and polymer matrix into the filler galleries. From these results, it may be postulated that a partially intercalated and/or exfoliated nanostructure in the zirconium phosphate modified with octadecylamine and linear low density polyethylene composite was achieved.
ABSTRACT
Nitric oxide (NO) is a reactive nitrogen species (RSN) that, over the years, has been shown to be integrated with biological and physiological events, including reproductive processes. NO can affect the functionality of spermatozoa through free radical scavenging, deactivating and inhibiting the production of superoxide anions (O2.-). However, the role of NO in mammalian spermatozoa physiology seems paradoxical. The aim of this study was to investigate the effects of NO on motility, hyperactivation, membrane integrity, peroxidation, and capacitation in cryopreserved equine sperm. Ejaculates were collected and cryopreserved. After thawing, samples were centrifuged, suspended in an in vitro fertilization (IVF) medium and incubated with the following treatments: 1) Câ¯=â¯control (IVF); 2) Aâ¯=â¯l-arginine (10â¯mM - In); 3) Lâ¯=â¯L-NAME (1â¯mM - Ih); 4) Mâ¯=â¯methylene blue (100â¯mM - Re); 5) ALâ¯=â¯L-arginineâ¯+â¯L-NAME (In + Ih); 6) AMâ¯=â¯L-arginineâ¯+â¯methylene blue (In + Re). The samples were evaluated for spermatic kinetics by CASA and other analyses [plasma and acrosomal membranes used the propidium iodide (PI) and Pisum sativum agglutinin (PSA), detection of tyrosine residues phosphorylation in the membrane (F0426), nitric oxide (DAF-2/DA), lipid peroxidation (C11-BODIPY581/591)] by flow cytometry. The l-arginine treatments reduced MOT, PROG, RAP and LIN only at time 0â¯min compared to the control and L-NAME. These treatments (MT and MP, VAP, VSL, LIN, RAP) also reduced the sperm movement characteristics but only at the beginning of the incubation period. After this period of incubation, motility recovered. NO removal by methylene blue almost completely inhibited sperm motility, but these treatments had the highest percentages of intact membranes. l-arginine treatments improved acrosome reactions and differed from M and AM. NO production, tyrosine phosphorylation and lipid peroxidation did not differ among treatments, except for M and AM, where a reduction in these variables was detected. Therefore, equine sperm capacitation and the acrosome reaction are part of an oxidative process that involves the participation of ROS, and NO plays an important role in the maintenance and regulation of motility, hyperactivation, induction of acrosome reaction and possibly in capacitation, which are indispensable processes for the fertility of equine sperm.
Subject(s)
Cell Membrane/drug effects , Horses , Nitric Oxide/pharmacology , Sperm Capacitation/drug effects , Sperm Motility/drug effects , Animals , Cryopreservation/veterinary , Image Processing, Computer-Assisted , Male , Semen Preservation/veterinaryABSTRACT
Visible and near-infrared radiation is now widely employed in health science and technology. Pre-clinical trials are still essential to allow appropriate translation of optical methods into clinical practice. Our results stress the importance of considering the mouse strain and gender when planning pre-clinical experiments that depend on light-skin interactions. Here, we evaluated the optical properties of depilated albino and pigmented mouse skin using reproducible methods to determine parameters that have wide applicability in biomedical optics. Light penetration depth (δ), absorption (µa), reduced scattering (µ's) and reduced attenuation (µ't) coefficients were calculated using the Kubelka-Munk model of photon transport and spectrophotometric measurements. Within a broad wavelength coverage (400-1400nm), the main optical tissue interactions of visible and near infrared radiation could be inferred. Histological analysis was performed to correlate the findings with tissue composition and structure. Disperse melanin granules present in depilated pigmented mouse skin were shown to be irrelevant for light absorption. Gender mostly affected optical properties in the visible range due to variations in blood and abundance of dense connective tissue. On the other hand, mouse strains could produce more variations in the hydration level of skin, leading to changes in absorption in the infrared spectral region. A spectral region of minimal light attenuation, commonly referred as the "optical window", was observed between 600 and 1350nm.
Subject(s)
Skin/chemistry , Spectroscopy, Near-Infrared/methods , Animals , Female , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BLABSTRACT
BACKGROUND: Handgrip strength is currently considered a predictor of overall muscle strength and functional capacity. Therefore, it is important to find reliable and affordable instruments for this analysis, such as the modified sphygmomanometer test (MST). OBJECTIVES: To assess the concurrent criterion validity of the MST, to compare the MST with the Jamar dynamometer, and to analyze the reproducibility (i.e. reliability and agreement) of the MST in individuals with Parkinson's disease (PD). METHOD: The authors recruited 50 subjects, 24 with PD (65.5 ± 6.2 years of age) and 26 healthy elderly subjects (63.4 ± 7.2 years of age). The handgrip strength was measured using the Jamar dynamometer and modified sphygmomanometer. The concurrent criterion validity was analyzed using Pearson's correlation coefficient and a simple linear regression test. The reproducibility of the MST was evaluated with the coefficient of intra-class correlation (ICC(2,1)), the standard error of measurement (SEM), the minimal detectable change (MDC), and the Bland-Altman plot. For all of the analyses, α ≤ 0.05 was considered a risk. RESULTS: There was a significant correlation of moderate magnitude (r ≥ 0.45) between the MST and the Jamar dynamometer. The MST had excellent reliability (ICC(2,1) ≥ 0.7). The SEM and the MDC were adequate; however, the Bland-Altman plot indicated an unsatisfactory interrater agreement. CONCLUSIONS: The MST exhibited adequate validity and excellent reliability and is, therefore, suitable for monitoring the handgrip strength in PD. However, if the goal is to compare the measurements between examiners, the authors recommend that the data be interpreted with caution.
Subject(s)
Hand Strength , Parkinson Disease/physiopathology , Sphygmomanometers , Aged , Equipment Design , Female , Humans , Male , Middle Aged , Reproducibility of ResultsABSTRACT
BACKGROUND: Macrophages play a major role among the inflammatory cells that invade muscle tissue following an injury. Low-level laser therapy (LLLT) has long been used in clinical practice to accelerate the muscle repair process. However, little is known regarding its effect on macrophages. OBJECTIVE: This study evaluated the effect of LLLT on the mitochondrial activity (MA) of macrophages. METHOD: J774 macrophages were treated with lipopolysaccharide (LPS) and interferon - gamma (IFN-γ) (activation) for 24 h to simulate an inflammatory process, then irradiated with LLLT using two sets of parameters (780 nm; 70 mW; 3 J/cm2 and 660 nm; 15 mW; 7.5 J/cm2). Non-activated/non-irradiated cells composed the control group. MA was evaluated by the cell mitochondrial activity (MTT) assay (after 1, 3 and 5 days) in three independent experiments. The data were analyzed statistically. RESULTS: After 1 day of culture, activated and 780 nm irradiated macrophages showed lower MA than activated macrophages, but activated and 660 nm irradiated macrophages showed MA similar to activated cells. After 3 days, activated and irradiated (660 nm and 780 nm) macrophages showed greater MA than activated macrophages, and after 5 days, the activated and irradiated (660 nm and 780 nm) macrophages showed similar MA to the activated macrophages. CONCLUSIONS: These results show that 660 nm and 780 nm LLLT can modulate the cellular activation status of macrophages in inflammation, highlighting the importance of this resource and of the correct determination of its parameters in the repair process of skeletal muscle.
Subject(s)
Low-Level Light Therapy , Macrophages/metabolism , Macrophages/radiation effects , Mitochondria/radiation effects , Cells, CulturedABSTRACT
BACKGROUND: Handgrip strength is currently considered a predictor of overall muscle strength and functional capacity. Therefore, it is important to find reliable and affordable instruments for this analysis, such as the modified sphygmomanometer test (MST). OBJECTIVES: To assess the concurrent criterion validity of the MST, to compare the MST with the Jamar dynamometer, and to analyze the reproducibility (i.e. reliability and agreement) of the MST in individuals with Parkinson's disease (PD). METHOD: The authors recruited 50 subjects, 24 with PD (65.5±6.2 years of age) and 26 healthy elderly subjects (63.4±7.2 years of age). The handgrip strength was measured using the Jamar dynamometer and modified sphygmomanometer. The concurrent criterion validity was analyzed using Pearson's correlation coefficient and a simple linear regression test. The reproducibility of the MST was evaluated with the coefficient of intra-class correlation (ICC2,1), the standard error of measurement (SEM), the minimal detectable change (MDC), and the Bland-Altman plot. For all of the analyses, α≤0.05 was considered a risk. RESULTS: There was a significant correlation of moderate magnitude (r≥0.45) between the MST and the Jamar dynamometer. The MST had excellent reliability (ICC2,1≥0.7). The SEM and the MDC were adequate; however, the Bland-Altman plot indicated an unsatisfactory interrater agreement. CONCLUSIONS: The MST exhibited adequate validity and excellent reliability and is, therefore, suitable for monitoring the handgrip strength in PD. However, if the goal is to compare the measurements between examiners, the authors recommend that the data be interpreted with caution. .
Subject(s)
Humans , Male , Female , Middle Aged , Aged , Parkinson Disease/physiopathology , Hand Strength , Sphygmomanometers , Reproducibility of Results , Equipment DesignABSTRACT
BACKGROUND: Macrophages play a major role among the inflammatory cells that invade muscle tissue following an injury. Low-level laser therapy (LLLT) has long been used in clinical practice to accelerate the muscle repair process. However, little is known regarding its effect on macrophages. OBJECTIVE: This study evaluated the effect of LLLT on the mitochondrial activity (MA) of macrophages. METHOD: J774 macrophages were treated with lipopolysaccharide (LPS) and interferon - gamma (IFN-γ) (activation) for 24 h to simulate an inflammatory process, then irradiated with LLLT using two sets of parameters (780 nm; 70 mW; 3 J/cm2 and 660 nm; 15 mW; 7.5 J/cm2). Non-activated/non-irradiated cells composed the control group. MA was evaluated by the cell mitochondrial activity (MTT) assay (after 1, 3 and 5 days) in three independent experiments. The data were analyzed statistically. RESULTS: After 1 day of culture, activated and 780 nm irradiated macrophages showed lower MA than activated macrophages, but activated and 660 nm irradiated macrophages showed MA similar to activated cells. After 3 days, activated and irradiated (660 nm and 780 nm) macrophages showed greater MA than activated macrophages, and after 5 days, the activated and irradiated (660 nm and 780 nm) macrophages showed similar MA to the activated macrophages. CONCLUSIONS: These results show that 660 nm and 780 nm LLLT can modulate the cellular activation status of macrophages in inflammation, highlighting the importance of this resource and of the correct determination of its parameters in the repair process of skeletal muscle. .
CONTEXTUALIZAÇÃO: O macrófago tem papel de destaque dentre as células inflamatórias que invadem o músculo após as lesões. Por outro lado, o laser em baixa intensidade (LBI) tem sido muito utilizado na clínica para acelerar o reparo muscular, e pouco se conhece sobre seu efeito nos macrófagos. OBJETIVO: Avaliar o efeito do LBI sobre a atividade mitocondrial (AM) de macrófagos ativados para simular um processo inflamatório. MÉTODO: Macrófagos J774 foram tratados com lipopolissacarídeo (LPS) e IFN-gamma (ativação) por 24 horas para simular um processo inflamatório e então foram irradiados com LBI (780 nm; 70 mW; 3 J/cm(2) e 660 nm; 15mW; 7,5 J/cm(2)). A AM foi avaliada pela técnica MTT após um, três e cinco dias das irradiações. Foram realizados três experimentos independentes, e os dados, submetidos à análise estatística. RESULTADOS: Após um dia de cultivo, os macrófagos ativados e irradiados com o laser de 780 nm mostraram AM menor que os somente ativados, já os macrófagos ativados e irradiados com o laser de 660 mostraram AM semelhante aos somente ativados. Após três dias, os macrófagos ativados e irradiados (660 e 780 nm) mostraram AM maior que os macrófagos ativados; já após cinco dias, os grupos ativados e irradiados (660 e 780 nm) mostraram AM semelhante aos macrófagos somente ativados. CONCLUSÕES: Esses resultados mostram que tanto o LBI de 660 nm como o de 780 nm são capazes de modular a ativação celular de macrófagos em situação de inflamação, ressaltando a importância desse recurso e da determinação de seus parâmetros dosimétricos no processo de reparo do músculo esquelético. .