Characterization of purinergic signaling in tumor-infiltrating lymphocytes from lower- and high-grade gliomas.

Malignant gliomas are highly heterogeneous glia-derived tumors that present an aggressive and invasive nature, with a dismal prognosis. The multi-dimensional interactions between glioma cells and other tumor microenvironment (TME) non-tumoral components constitute a challenge to finding successful treatment strategies. Several molecules, such as extracellular purines, participate in signaling events and support the immunosuppressive TME of glioma patients. The purinergic signaling and the ectoenzymes network involved in the metabolism of these extracellular nucleotides are still unexplored in the glioma TME, especially in lower-grade gliomas (LGG). Also, differences between IDH-mutant (IDH-Mut) versus wild-type (IDH-WT) gliomas are still unknown in this context. For the first time, to our knowledge, this study characterizes the TME of LGG, high-grade gliomas (HGG) IDH-Mut, and HGG IDH-WT patients regarding purinergic ectoenzymes and P1 receptors, focusing on tumor-infiltrating lymphocytes. Here, we show that ectoenzymes from both canonical and non-canonical pathways are increased in the TME when compared to the peripheral blood. We hypothesize this enhancement supports extracellular adenosine generation, hence increasing TME immunosuppression.

Secretome of stem cells from human exfoliated deciduous teeth (SHED) and its extracellular vesicles improves keratinocytes migration, viability, and attenuation of H2 O2 -induced cytotoxicity.

Therapies for wound healing using the secretome and extracellular vesicles (EVs) of mesenchymal stem/stromal cells have been shown to be successful in preclinical studies. This study aimed to characterise the protein content of the secretome from stem cells from human exfoliated deciduous teeth (SHED) and analyse the in vitro effects of SHED-conditioned medium (SHED-CM) and SHED extracellular vesicles (SHED-EVs) on keratinocytes. EVs were isolated and characterised. The keratinocyte viability and migration of cells treated with SHED-EVs and conditioned medium (CM) were evaluated. An HaCaT apoptosis model induced by H2 O2 in vitro was performed with H2 O2 followed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and live/dead assays. Finally, the expression of vascular endothelial growth factor (VEGF) in keratinocytes treated with secretome and EVs was evaluated by immunofluorescence staining and confirmed with RT-qPCR. SHED-EVs revealed a cup-shaped morphology with expression of the classical markers for exosomes CD9 and CD63, and a diameter of 181 ± 87 nm. The internalisation of EVs by HaCaT cells was confirmed by fluorescence microscopy. Proteomic analysis identified that SHED-CM is enriched with proteins related to stress response and development, including cytokines (CXCL8, IL-6, CSF1, CCL2) and growth factors (IGF2, MYDGF, PDGF). The results also indicated that 50% CM and 0.4-0.6 µg/mL EVs were similarly efficient for improving keratinocyte viability, migration, and attenuation of H2 O2 -induced cytotoxicity. Additionally, expression of VEGF on keratinocytes increased when treated with SHED secretome and EVs. Furthermore, VEGF gene expression in keratinocytes increased significantly when treated with SHED secretome and EVs. Both SHED-CM and SHED-EVs may therefore be promising therapeutic tools for accelerating re-epithelialization in wound healing.

Preclinical evaluation of bozepinib in bladder cancer cell lines: modulation of the NPP1 enzyme.

Bladder cancer (BC) is the most common cancer of the urinary tract. Bozepinib (BZP), a purine-derived molecule, is a potential compound for the treatment of cancer. Purinergic signaling consists of the activity of nucleosides and nucleotides present in the extracellular environment, modulating a variety of biological actions. In cancer, this signaling is mainly controlled by the enzymatic cascade involving the NTPDase/E-NPP family and ecto-5'-nucleotidase/CD73, which hydrolyze extracellular adenosine triphosphate (ATP) to adenosine (ADO). The aim of this work is to evaluate the activity of BZP in the purinergic system in BC cell lines and to compare its in vitro antitumor activity with cisplatin, a chemotherapeutic drug widely used in the treatment of BC. In this study, two different BC cell lines, grade 1 RT4 and the more aggressive grade 3 T24, were used along with a human fibroblast cell line MRC-5, a cell used to predict the selectivity index (SI). BZP shows strong antitumor activity, with notable IC50 values (8.7 ± 0.9 µM for RT4; 6.7 ± 0.7 µM for T24), far from the SI for cisplatin (SI for BZP: 19.7 and 25.7 for RT4 and T24, respectively; SI for cisplatin: 1.7 for T24). BZP arrests T24 cells in the G2/M phase of the cell cycle, inducing early apoptosis. Moreover, BZP increases ATP and ADP hydrolysis and gene/protein expression of the NPP1 enzyme in the T24 cell line. In conclusion, BZP shows superior activity compared to cisplatin against BC cell lines in vitro.

Chicken embryo model for in vivo acute toxicological and antitumor efficacy evaluation of lipid nanocarrier containing doxorubicin.

Nanoencapsulation of chemotherapeutics, including doxorubicin, can endow the formulations with unique properties, such as a decrease in adverse effects and toxicity. The chicken embryo model is an alternative and well-accepted strategy for evaluating the toxicity and efficacy of drugs and nanoformulations. Therefore, this study proposes the development of a new lipid nanocarrier for doxorubicin delivery (NanoLip-Dox) and posterior evaluation of toxicological profile and antitumoral efficacy against a breast tumor in chicken embryos. NanoLip-Dox showed a unimodal particle size (< 150 nm), negative zeta potential (-19.5 mV), absence of drug crystals, drug content of 0.099 mg·mL-1, and high entrapment efficiency (95%). NanoLip-Dox did not cause toxicity in the chicken embryos; in contrast, doxorubicin hydrochloride induced moderate irritation in the chorioallantoic membrane (at 862.1 µmol·L-1), a survival rate of 50% (at 1.7 µmol·L-1), and an increase in aspartate aminotransferase (at 862.1, 344.8, and 172.4 µmol·L-1). In addition, NanoLip-Dox (at 1.7 µmol·L-1) showed potent antitumor efficacy with a high tumor remission percentage (40.9 ± 9.7%) compared to the control group (8.6 ± 14.8%). These findings together with the absence of toxicity concerning morphological characteristics, weights of embryos and organs, hematologic parameters, and enzymatic activity (alanine aminotransferase, aspartate aminotransferase, and creatinine) suggest the safety and efficacy of NanoLip-Dox.

Caracterização genética, clínica e epidemiológica de pacientes com Covid-19 em uma região do Sul do Brasil / Genetic, clinical and epidemiological characterization of Covid-19 patients in South Brazil

Caracterizar o perfil epidemiológico, clínico e genético de pacientes com Covid-19. Realizou-se um estudo observacional e transversal com voluntários que tiveram diagnóstico de Covid-19 no período de abril de 2020 a maio de 2021 no município de Santa Cruz do Sul (RS, Brasil), no qual foram coletados dados clínicos e epidemiológicos, além de amostras de sangue para a identificação de polimorfismos no gene ACE2. Foram recrutados 87 indivíduos e destes, 16,7% necessitaram de internação hospitalar, sendo a maioria do sexo masculino. A obesidade foi a comorbidade mais frequente, no entanto, doenças cardiovasculares, hipertensão e diabetes apresentaram maior significância quando associadas às internações. Em relação à características genéticas, entre os voluntários não foram encontrados polimorfismos no gene ACE2. A pesquisa sugere que o sexo masculino e presença de comorbidades são importantes fatores de risco para a severidade da Covid-19.

Current paper characterizes the epidemiological, clinical and genetic profile of patients with Covid-19. An observational and cross-sectional study was carried out with volunteers diagnosed with Covid-19 between April 2021 and May 2021 in the municipality of Santa Cruz do Sul, Brazil; a blood sample also identified polymorphism in the ACE2 gene. 87 patients were recruited; 6.7% required hospitalization, the majority being male. Although obesity was a more frequent co-morbidity, cardiovascular diseases, hypertension and diabetes were more significant when associated with hospitalizations. In the case of genetic characteristics, polymorphisms were found in the ACE2 gene among volunteers. Important research suggests male gender and co-morbidities are risk factors for the severity of Covid-19.

Potential Ameliorative Effects of Chromium Supplementation on Glucose Metabolism, Obesity, and Genomic Stability in Prediabetic Rat Model.

Chromium (III) (Cr(III)) effect on improving glucose, body mass loss, and genomic stability has been extensively studied in models of type 2 diabetes. However, there is a lack of studies evaluating its effect on prediabetes. Thus, this study evaluates the effects of Cr(III) as dietetic supplementation on glucose metabolism, obesity, and genomic stability on prediabetic rat model using high-invert sugar. Male Wistar rats were divided randomly into four treatment groups: (1) control, receiving standard diet (control); (2) prediabetic (PD), receiving a 32% of invert sugar; (3) Cr(III), receiving chromium (III) chloride (CrCl3•6H2O) (58.4 mg/L); and (4) Cr(III) + PD, receiving CrCl3•6H2O in combination with high-invert sugar. Cr(III) supplementation significantly reduced blood glucose (123.00 ± 8.29 mg/dL vs. 115.30 ± 9.31 mg/dL, p = 0.015) and partially reduced area under the 120-min blood glucose response curve (AUC) in PD rats (p = 0.227). Moreover, Cr(III) attenuated weight gain (187.29 ± 38.56 g vs. 167.22 ± 29.30 g, p = 0.004), significantly reducing body mass index (0.68 ± 0.04 g/cm2 vs. 0.63 ± 0.04 g/cm2, p < 0.001), Lee index (0.30 ± 0.01 vs. 0.28 ± 0.01, p < 0.001), and peritoneal fat (p < 0.001). Regarding genomic stability, high-invert sugar, Cr(III), or the combination of both did not produce changes in oxidative stress, DNA damage in pancreas, or cytotoxicity markers. These data suggest that Cr(III) supplementation improved partially glucose metabolism and reduced obesity in rat model PD due to high-invert sugar without influence in genomic stability.

Evaluation of the Efficacy of Musical Vibroacupuncture in Pain Relief: A Randomized Controlled Pilot Study.

OBJECTIVES: To investigate if skin vibration employing consonant frequencies emitted by skin transducers attached to a combination of acupuncture points and according to musical harmony (musical chord) produces more significant pain relief compared to just a single frequency. MATERIALS AND METHODS: Skin vibrostimulation produced by five electromagnet transducers was applied at five acupoints traditionally used to pain relief and anxiety in 13 pain-free healthy volunteers using the cold pressor test (CPT). The study consisted of three randomized sessions conducted on alternate days, with participants receiving either simultaneous frequencies of 32, 48, and 64 Hz that equate those used in a musical chord, hereby defined as musical vibroacupuncture (MVA), a single frequency of 32 Hz, set as vibroacupuncture (VA) and sham procedure (SP). CPT scores for pain thresholds and pain tolerance were assessed using repeated-measures ANOVAs. Pain intensity was evaluated using a numerical rating scale (NRS), while sensory and affective aspects of pain were rated using the short-form McGill Pain Questionnaire (SF-MPQ) and State-Trait Anxiety Inventory (STAI) Y-Form. RESULTS: Pain thresholds did not vary significantly between trials. Pain tolerance scores were markedly higher in MVA compared to baseline (p = 0.0043) or SP (p = 0.006) but not for VA. Pain intensity for MVA also differed significantly from the baseline (p = 0.007) or SP (p = 0.027), but not for VA. No significant differences were found in SF-MPQ and STAI questionnaires. CONCLUSIONS: These results suggest that MVA effectively increased pain tolerance and reduced pain intensity when compared with all groups, although not significant to the VA group.

Ready for the journey: a comparative proteome profiling of porcine cauda epididymal fluid and spermatozoa.

In the present study, we describe the proteome of porcine cauda epididymis fluid and spermatozoa by means of Multidimensional Protein Identification Technology (MudPIT). Ten sexually mature healthy boars were surgically castrated and epididymides were dissected to obtain the cauda epididymal content. Polled protein extracts of cauda epididymal fluid (CEF) and spermatozoa (CESperm) were loaded in an Agilent 1100 quaternary HPLC and peptides eluted from the microcapillary column were electro-sprayed directly into a LTQ Orbitrap XL mass spectrometer. Using bioinformatics, identified proteins were classified by their molecular functions, involvement in biological processes and participation in relevant metabolic pathways associated with spermatozoa physiology, fertility potential and protection. A total of 645 proteins were identified in the CEF, with epididymal-specific lipocalin-5, beta-hexosaminidase subunit beta precursor and phosphatidylethanolamine-binding protein 4 being the most abundant proteins found. A total of 2886 proteins were identified in the CESperm proteome with 81 proteins being considered more abundant (spectral counts > 100). CEF and CESperm data were compared and 345 proteins were present in both proteomes. Phosphatidylethanolamine-binding protein 4 precursor was the only protein found most abundant in both CEF and CESperm proteomes. Based on Gene Ontology analysis, we identified CEF and CESperm proteins associated with sperm protection against ROS and immune mediated response, glycosaminoglycan degradation, ubiquitin-proteasome system, metabolic process and maturation, modulation of acrosome reaction and ZP binding and oocyte penetration. These results provide a better comprehension about the molecular process and biological pathways involved in sperm epididymis maturation and establishment of the cauda epididymis sperm reservoir.

Deoxyribonucleic acid damage and repair response in the chemotherapy of lung cancer: cross-sectional study.

This study evaluated 24 patients with lung cancer (CA) and 23 individuals with no smoking history or cancer in the family and without respiratory disease in childhood (CO). Peripheral blood lymphocytes was used to perform alkaline comet assay and to assess DNA damage as well as to evaluate methyl methane sulfonate (MMS) DNA repair after one hour and three hours at 37 ºC. The percentage of residual damage (RD) after three hours of MMS treatment, for each patient was assessed. The majority of patients were in the CA group, male patients, former smokers, with a history of smoking for 15 years and without associated comorbidities. Alkaline and residual damages were higher in the CA group when compared to controls (alkaline damage P = 0.015 and RD P = 0.05). After one hour of MMS treatment the DNA damage of the CA increased indicating failure to repair it, compared to the controls, and after three hours DNA repair was observed in both groups. Patients with lung cancer are mostly men, former smokers and with more than 15 years of tobacco consumption, undergoing chemotherapy, have high rates of DNA damage and deficiency in their ability to repair against induced damage when compared to controls.

Structural modeling and mRNA expression of epididymal ß-defensins in GnRH immunized boars: A model for secondary hypogonadism in man.

Human secondary hypogonadism is associated with impaired testicular function, however, little is known about its impact on sperm epididymal maturation. Endocrine disruption in the epididymis could impair the secretion of key proteins, such as ß-defensins, responsible for spermatozoa maturation during epididymal transit. This study evaluated the sequence and structural similarities between porcine epididymal ß-defensins porcine ß-defensins (pBD3), pBD4, pBD125, and pEP2C and their human homologs using bioinformatics integrated with information derived from protein databanks. We then verified whether the expression of pBD3, pBD4, pBD125, and pEP2C genes in the testis and epididymis are influenced by disruption of the hypothalamic-pituitary-testicular (HPT) axis in a pig model for male human secondary hypogonadism. Upon modeling porcine ß-defensins, structural and functional analysis confirmed the presence of motifs associated with ß-defensin function, validating the models generated in silico. pBD3 and pBD4 showed acceptable structural alignments with human ß-defensins BDEF103 and BDEF110, respectively. In addition, evaluation of hormonal regulation of ß-defensins was assessed by analyzing the expression of these four ß-defensins in adult boars immunized against gonadotropin-releasing hormone (GnRH). Our results indicate that HPT axis disruption modifies the expression of pBD3, pBD4, pBD125, and pEP2C in boar testis and epididymis, suggesting an endocrine-dependent regulation of ß-defensins in swine epididymis. In conclusion, sequence and structural homology between pBD3 and pBD4 and their human homologs provide a basis for using the pig as a model for the study of human secondary hypogonadism. Further investigation of the human homologs in hypogonadal men could elucidate the connections between fertility and epididymal expression of ß-defensins.

Prevalência de anticorpos e fatores de risco associados à infecção por Toxoplasma gondii em galinhas domésticas da zona rural de Santa Maria, Rio Grande do Sul / Prevalence of antibodies and risk factors associated with infection by Toxoplasma gondii in free range chickens of rural area of Santa Maria, Rio Grande do Sul, Brazil

RESUMO: Toxoplasma gondii é um protozoário apicomplexa que infecta animais de sangue quente, podendo ser considerado um dos principais parasitas capazes de infectar os seres humanos. Galinhas domésticas podem ser facilmente infectadas por protozoários, uma vez que estas podem ingerir oocistos encontrados no solo, sendo consideradas boas indicadoras de contaminação ambiental por T. gondii. O objetivo deste estudo foi determinar a presença de anticorpos anti-T. gondii em galinhas domésticas criadas extensivamente e avaliar os fatores de risco associados ao protozoário, na zona rural de Santa Maria, RS, Brasil. No período de março de 2013 a fevereiro 2014 foram coletadas 597 amostras de sangue de galinhas domésticas em 74 propriedades, oriundas de nove estratos que representam cada distrito da zona rural. Para avaliar os fatores de risco, nessas propriedades foi aplicado um questionário epidemiológico aos moradores. As amostras de soro foram testadas por imunofluorescência indireta, e 49,2% (294/597) foram positivas para anticorpos anti-T. gondii, com títulos variando de 16 a 4096. Das 74 propriedades analisadas, em 63 (85,1%) houve relatos que os gatos têm acesso ao deposito de alimentos, com associação significativa quando associado à presença de galinhas positivas (p=0,04) e o OR de 4,07. A variável "abate de animais" (aves e bovinos), em 51 (68,9%) das propriedades foi relatado o abate de bovinos e aves na propriedade, com valor de p significativo (p=0,05). A maioria das propriedades 59 (79,7%) foi relatada a presença de gatos domésticos, o que poderia estar associada com a alta soroprevalência encontrada em galinhas e a taxa de contaminação ambiental. A elevada prevalência de anticorpos encontrada neste estudo, além da alta frequência de propriedades com casos positivos, sugere uma grande contaminação ambiental nos distritos pesquisados, sendo assim um risco potencial para a saúde humana e animal.

ABSTRACT: Toxoplasma gondii is an apicomplex protozoan that infects warm-blooded animals and can be considered a major parasite capable of infecting humans. Domestic chickens can be easily infected by protozoa, since they can ingest oocysts found in the soil and are considered good indicators of environmental contamination by T. gondii. The aim of this study was to determine the presence of anti-T. gondii antibodies in free range chickens and to evaluate the risks factors associated with the protozoan in rural area of Santa Maria, RS, Brazil. From March 2013 to February 2014, 597 blood samples from domestic chickens were collected from 74 farms, from nine layers representing each district in the rural area. To evaluate the risk factors, in these farms an epidemiological questionnaire was applied to the residents. Serum samples were tested by indirect immunofluorescence and 49.2% (294/597) were positive for anti-T.gondii antibodies, with titres varying from 16 to 4096. Of the 74 analyzed farms, 63 (85.1%) reported that cats had access to food deposits, with a significant association when positive chickens were present (p = 0.04) and the OR of 4.07. The variable "slaughter of animals" (poultry and cattle) in 51 (68.9%) of the farms was reported the slaughter of cattle and birds in the farm, with significant p value (p = 0.05). Most farms 59 (79.7%) reported the presence of domestic cats, which could be associated with the high seroprevalence found in chickens and the rate of environmental contamination. The high prevalence of antibodies found in this study, in addition to the high frequency of farms with positive cases, suggests a great environmental contamination in the studied districts, thus being a potential risk to human and animal health.

DNA damage and repair capacity in lymphocyte of chronic obstructive pulmonary diseases patients during physical exercise with oxygen supplementation.

BACKGROUND: We hypothesized that the use of oxygen supplementation during aerobic exercise induces less DNA damage than exercise alone. The aim of this study is to assess the level of DNA damage induced by physical exercise with and without oxygen supplementation in chronic obstructive pulmonary diseases (COPD) patients. METHODS: Peripheral blood was collected before and after aerobic exercise in two conditions: (I) aerobic exercise without oxygen supplementation (AE group) and (II) with oxygen supplementation (AE-O2 group). Lymphocytes were collected to perform the alkaline version of the Comet Assay. To assess the susceptibility to exogenous DNA damage, the lymphocytes were treated with methyl methanesulphonate (MMS) for 1-h or 3-h. After 3-h treatment, the percentage of residual damage was calculated assuming the value of 1-h MMS treatment as 100%. RESULTS: AE group showed lower induced damage (1 h of MMS treatment) and consequently less DNA repair compared to AE-O2 group. AE-O2 group showed an increase in the induced DNA damage (1 h of MMS treatment) and an increased DNA repair capacity. Within the AE-O2 group, in the post-exercise situation the induced DNA damage after 1 h of MMS treatment was higher (p = 0.01) than in the pre-exercise. CONCLUSION: COPD patients who performed physical exercise associated with oxygen supplementation had a better response to DNA damage induced by MMS and a better DNA repair when compared to the condition of physical exercise without oxygen supplementation. TRIAL REGISTRATION: UNISC N374.298. Registered 04 JUN 2013 (retrospectively registered).

Oral infection of neonate gerbils by Neospora caninum tachyzoites / Infecção oral de gerbils neonatos com taquizoítos de Neospora caninum

ABSTRACT: Neosporosis is a parasitic disease caused by the protozoan Neospora caninum which results in major economic losses for cattle breeding due to abortion and other reproductive disorders. Gerbils (Meriones unguiculatus) are commonly used as experimental models for neosporosis due to their high susceptibility to N. caninum infection, both by oocysts ingestion as by tachyzoites/bradyzoites parenteral inoculation. However, the risk of transmission by tachyzoites ingestion is not fully elucidated. In this study, infection of neonate gerbils by N. caninum (NC-1 strain) tachyzoites inoculated by the oral route and the parasite distribution in gerbils' tissues were evaluated by protozoan DNA detection. Seventeen neonate gerbils, aged 4-5 days, were inoculated with 4x105 tachyzoites by the oral route and one gerbil was kept as uninfected control. N. caninum DNA was detected in 100% of the inoculated gerbils, showing that the oral route is effective as a potential route of infection of neonates by N. caninum tachyzoites. N. caninum DNA was reported in all organs evaluated (heart, lungs, kidneys, liver, spleen and brain), with different frequencies. These results showed systemically distributed infection of neonate gerbils after oral inoculation of tachyzoites.

RESUMO: Neosporose é uma doença parasitária causada pelo protozoário Neospora caninum e resulta em grandes perdas econômicas para a bovinocultura, o que se deve a abortos e outras desordens reprodutivas. Gerbils (Meriones unguiculatus) são comumente utilizados como modelos experimentais para neosporose, devido a sua alta susceptibilidade à infecção por N. caninum, seja pela ingestão de oocistos ou pela inoculação parenteral de taquizoítos/bradizoítos. Porém, o risco de transmissão pela ingestão de taquizoítos não está totalmente elucidado. Neste estudo, foram avaliadas a infecção de gerbils neonatos por taquizoítos de N. caninum (cepa NC-1) inoculados por via oral e a distribuição do parasita nos tecidos desses gerbils, através da detecção de DNA do protozoário. Dezessete gerbils neonatos, com idade de 4-5 dias, foram inoculados com 4x105 taquizoítos por via oral e um gerbil foi mantido como controle não-infectado. DNA de N. caninum foi detectado em 100% dos gerbils inoculados, demonstrando a eficácia da via oral como potencial rota de infecção de neonatos por taquizoítos de N. caninum. DNA de N. caninum foi encontrado, em diferentes frequências, em todos os órgãos avaliados (coração, pulmões, rins, fígado, baço e cérebro). Esses resultados demonstraram infecção sistêmica dos gerbils neonatos após a inoculação oral de taquizoítos.

DNA damage and cellular abnormalities in tuberculosis, lung cancer and chronic obstructive pulmonary disease.

BACKGROUND: Tuberculosis (TB), Lung Cancer (LC) and Chronic Obstructive Pulmonary Diseases (COPD) affect millions of individuals worldwide. Monitoring of DNA damage in pathological situations has been investigated because it can add a new dimension to clinical expression and may represent a potential target for therapeutic intervention. The aim of this study was to evaluate DNA damage and the frequency of cellular abnormalities in TB, LC and COPD patients by comparing them to healthy subjects. METHODS: The detection of DNA damage by a buccal micronucleus cytome assay was investigated in patients with COPD (n = 28), LC (n = 18) and TB (n = 22) and compared to control individuals (n = 17). RESULTS: The COPD group had a higher frequency of apoptotic cells compared to TB and LC group. The TB group showed a higher frequency of DNA damage, defect in cytokinesis, apoptotic and necrotic cells. Patients with LC had low frequency of chromosomal aberrations than TB and COPD patients. CONCLUSION: COPD patients showed cellular abnormalities that corresponded to cell death by apoptosis and necrosis, while patients with TB presented defects in cytokinesis and dysfunctions in DNA repair that resulted in the formation of micronucleus (MN) besides apoptotic and necrotic cells. Patients with COPD, TB and LC had a low frequency of permanent DNA damage.

Effects of music in combination with vibration in acupuncture points on the treatment of fibromyalgia.

OBJECTIVE: Fibromyalgia (FM) is a disease that causes widespread pain and increased sensitivity to pain because of a dysfunction in the central nervous system. This study investigated the effect of music combined with vibration on acupuncture points for the treatment of FM. METHODS: A total of 120 patients with FM were allocated randomly to four groups (30 patients each). One group listened to a sequence of Bach's compositions. The second group was subjected to vibratory stimuli on a combination of acupuncture points on the skin. The third group (complete) underwent both procedures in a simultaneous and synchronized manner, with inclusion of binaural beats. The fourth group (control) received no stimulation. The participants underwent the experimental procedure during five sessions performed on alternate days. They were assessed by the Fibromyalgia Impact Questionnaire (FIQ) and the Health Assessment Questionnaire (HAQ) before the first session and after the last session (20 days). RESULTS: All groups showed a significant improvement in FIQ and HAQ scores at the evaluation after the intervention. The complete group exhibited the best result on both the FIQ and HAQ (p<0.001), and the improvement in HAQ score was significant (p<0.004). CONCLUSIONS: The results suggest that the placebo effect in FM may be substantial. However, comparison between groups revealed that the complete group had the greatest reduction in both FIQ and HAQ, with a significant improvement in HAQ, suggesting that the combined use of music and vibration exerts a greater effect on FM symptoms.

Seroprevalence and risk factors for Neospora caninum in goats in Santa Catarina state, Brazil.

Neosporosis is an infectious disease caused by the parasite Neospora caninum. Knowledge regarding neosporosis in goats is still quite limited, especially in the state of Santa Catarina (SC), southern Brazil. Therefore, this study aimed to assess the seroprevalence and risk factors for N. caninum in goats in the western and mountain regions of SC. Blood samples were collected from 654 goats in 57 municipalities. The indirect immunofluorescence test was used for antibody detection against N. caninum. Thirty samples (4.58%) were seropositive, with titers ranging from 1:50 to 1:6400. An epidemiological survey was also conducted in order to identify risk factors for neosporosis in goats. It was found that reproductive problems on the farms, as well as the diet and direct contact with dogs were casual risks for neosporosis. These results indicate that N. caninum infects goats in these regions, which may lead to reproductive problems.

Seroprevalence and risk factors for Neospora caninum in goats in Santa Catarina state, Brazil / Soroprevalência e fatores de risco para Neospora caninum em caprinos no estado Santa Catarina, Brasil

Neosporosis is an infectious disease caused by the parasite Neospora caninum. Knowledge regarding neosporosis in goats is still quite limited, especially in the state of Santa Catarina (SC), southern Brazil. Therefore, this study aimed to assess the seroprevalence and risk factors for N. caninum in goats in the western and mountain regions of SC. Blood samples were collected from 654 goats in 57 municipalities. The indirect immunofluorescence test was used for antibody detection against N. caninum. Thirty samples (4.58%) were seropositive, with titers ranging from 1:50 to 1:6400. An epidemiological survey was also conducted in order to identify risk factors for neosporosis in goats. It was found that reproductive problems on the farms, as well as the diet and direct contact with dogs were casual risks for neosporosis. These results indicate that N. caninum infects goats in these regions, which may lead to reproductive problems.

Neosporose é uma doença infecciosa causada pelo parasito Neospora caninum. Nos caprinos, as informações sobre neosporose ainda são bastante limitadas, especialmente no Estado de Santa Catarina (SC), Sul do Brasil. Em virtude disso, este estudo teve como objetivo avaliar a soroprevalência e os fatores de risco envolvidos na infecção por N. caninum em caprinos das regiões Oeste e Planalto Serrano do Estado de SC. Amostras de sangue foram coletadas de 654 caprinos em 57 cidades. A técnica de imunofluorescência indireta (RIFI) foi usada para a detecção sorológica de anticorpos contra N. caninum. Trinta amostras (4,58%) foram soropositivas para o agente infeccioso com títulos variando entre 1:50 a 1:6400. Durante a coleta de sangue, um levantamento epidemiológico foi realizado para identificar fatores de risco para neosporose em caprinos. Problemas reprodutivos presentes nas fazendas, bem como a dieta e o contato direto com cães foram detectados como fatores de risco para a doença. Os resultados indicam que N. caninum infecta cabras nas regiões avaliadas, o que pode estar relacionado a problemas reprodutivos nesta espécie.

Improving liveweight gain of lambs infected by multidrug-resistant nematodes using a FECRT-based schedule of treatments.

The aim of this study was to compare the liveweight gain of lambs, infected by multidrug-resistant nematodes, treated by conventional schemes of helminth control or using a schedule based on fecal egg count reduction test (FECRT). The flock was selected after a FECRT (experiment 1) which revealed a parasite population resistant to benzimidazoles, imidazothiazoles, macrocyclic lactones (ivermectin), salicylanilides, nitrophenols, and organophosphates. Despite the parasite resistance to ivermectin (an avermectin), the moxidectin (a milbemycin) was effective against the gastrointestinal nematodes (PR > 90%). In experiment 2, 48 suckling lambs were distributed in four randomized blocks (G1, G2, G3, and G4) by previous body weighings. G1 was kept as untreated control; G2 was treated following a FECRT-based schedule with drugs chosen based on fecal analysis (first drench with moxidectin, second drench with a combination of moxidectin and levamisole, and third drench with praziquantel, an anti-cestode drug); G3 and G4 received three drenches with ivermectin or disophenol, respectively. Body weighings and fecal analysis of these lambs were performed every 2 weeks over a 98-day period. An effective control of gastrointestinal nematodes was obtained with two nematicidal drenches following the FECRT-based schedule of treatments. On the other hand, eggs per gram of feces (EPG) counts were no different among untreated control, G3, and G4. Lambs treated using the FECRT-based schedule had the greatest liveweight gain among the groups tested. Additionally, liveweight gain was no different among the groups G3, G4, and G1. The FECRT-based schedule of anthelmintic treatments was beneficial regarding productivity and sustainability of helminth control in lambs infected by multidrug-resistant nematodes.

Detecção de ácidos nucleicos em tecidos de gerbils submetidos à infecção aguda por Neospora caninum / Detection of nucleic acids in tissues of gerbils submitted to acute infection with Neospora caninum

Neospora caninum é um protozoário de grande importância na pecuária, por determinar problemas reprodutivos principalmente em bovinos. Os gerbils (Meriones unguiculatus) podem atuar como modelos experimentais para reproduzir a neosporose aguda de bovinos. Neste trabalho, foram formados dois grupos de gerbils (n=10), inoculados com taquizoítos de N. caninum (cepa NC-1) nas doses de 5x10(6) taquizoítos ml-1 (G1) ou de 5x10(5) taquizoítos ml-1 (G2), doses capazes de induzir infecção aguda. Cérebro, medula espinhal, coração, pulmão, fígado, rins e baço foram coletados e a técnica de PCR foi realizada a partir das amostras de tecidos e órgãos. Na maioria dos animais, o DNA do N. caninum foi detectado pelo menos em cinco tecidos, considerando ambos os grupos (12/20; 60%). No grupo 1, a frequência de detecção de DNA, na totalidade das amostras, foi maior (52/70; 74,28%) quando comparada ao grupo 2 (38/70; 54,28%). A partir desses resultados, pode-se afirmar que o protozoário replicou eficientemente após inoculação e se disseminou pelos tecidos. Além disso, demonstrou-se que gerbils podem ser utilizados como modelo de infecção aguda pelo N. caninum, apresentando sinais clínicos da neosporose.

Neospora caninum is a protozoan of great importance for livestock, mainly by causing reproductive diseases in cattle. Gerbils (Meriones unguiculatus) may be a model to reproduce experimental acute neosporosis of cattle. In this study two groups of gerbils were formed (n=10) and gerbils were inoculated with N. caninum tachyzoites (NC-1 strain) at two different doses appropriate to cause acute neosporosis: 5x10(6) tachyzoites ml-1 (G1) or 5x10(5) tachyzoites ml-1 (G2). Brain, spinal cord, heart, lung, liver, kidneys and spleen were collected and PCR was performed using samples of these tissues and organs. DNA of N. caninum was detected in at least five tissues for most animals (12/20; 60%) considering both groups. In group 1, the frequency of DNA detection, evaluating all samples, was higher (52/70; 74.28%) compared to group 2 (38/70; 54.28%). These results showed that the protozoan replicated efficiently after inoculation spreading by several tissues. Also, was demonstrated that gerbils can be used as model for acute infection with N. caninum showing clinical signs of neosporosis.