Animal models of male reproductive ageing to study testosterone production and spermatogenesis.

Ageing is the time-dependent gradual decline of the functional characteristics in an organism. It has been shown that it results in the loss of reproductive health and fertility. The age-dependent decline of fertility is a potential issue as the parenthood age is increasing in Western countries, mostly due to socioeconomic factors. In comparison to women, for whom the consequences of ageing are well documented and general awareness of the population is extensively raised, the effects of ageing for male fertility and the consequences of advanced paternal age for the offspring have not been widely studied. Studies with humans are welcome but it is hard to implement relevant experimental approaches to unveil the molecular mechanisms by which ageing affects male reproductive potential. Animal models have thus been extensively used. These models are advantageous due to their reduced costs, general easy maintenance in laboratory facilities, rigorous manipulation tools, short lifespan, known genetic backgrounds, and reduced ethical constraints. Herein, we discuss animal models for the study of male reproductive ageing. The most well-known and studied reproductive ageing models are rodents and non-human primates. The data collected from these models, particularly studies on testicular ageing, steroidogenesis, and genetic and epigenetic changes in spermatogenesis are detailed. Notably, some species challenge the currently accepted ageing theories and the concept of senescence itself, which renders them interesting animal models for the study of male reproductive ageing.

Lipopolysaccharide-induced epididymitis modifies the transcriptional profile of Wfdc genes in mice†.

Whey-acidic protein four-disulfide core domain (WFDC) genes display putative roles in innate immunity and fertility. In mice, a locus on chromosome 2 contains 5 and 11 Wfdc genes in its centromeric and telomeric subloci, respectively. Although Wfdc genes are highly expressed in the epididymis, their contributions to epididymal function remain elusive. Here, we investigated whether Wfdc genes are regulated in response to lipopolysaccharide (LPS)-induced epididymitis, an inflammatory condition that impairs male fertility. We induced epididymitis in mice via (i) interstitial LPS injection into epididymal initial segment and (ii) intravasal LPS injection into the vas deferens towards cauda epididymis. Interstitial and intravasal LPS induced a differential upregulation of inflammatory mediators (interleukin 1 beta, interleukin 6, tumor necrosis factor, interferon gamma, and interleukin 10) in the initial segment and cauda epididymis within 72 h post-treatment. These changes were accompanied by a time-dependent endotoxin clearance from the epididymis. In the initial segment, interstitial LPS upregulated all centromeric (Slpi, Wfdc5, Wfdc12, Wfdc15a, and Wfdc15b) and five telomeric (Wfdc2, Wfdc3, Wfdc6b, Wfdc10, and Wfdc13) Wfdc transcripts at 24 and 72 h. In the cauda epididymis, intravasal LPS upregulated Wfdc5 and Wfdc2 transcripts at 24 h, followed by a downregulation of Wfdc15b and three telomeric (Wfdc6a, Wfdc11, and Wfdc16) gene transcripts at 72 h. Pharmacological inhibition of nuclear factor kappa B activation prevented LPS-induced upregulation of centromeric and telomeric Wfdc genes depending on the epididymal region. We show that LPS-induced inflammation differentially regulated the Wfdc locus in the proximal and distal epididymis, indicating region-specific roles for the Wfdc family in innate immune responses during epididymitis.

Exercise training attenuates angiotensin II-induced vasoconstriction in the aorta of normotensive but not hypertensive rats.

NEW FINDINGS: What is the central question of this study? What are the effects of exercise on Ang II-induced vasoconstriction in aortas of normotensive rats and how do these effects occur in two-kidney-one-clip hypertensive animals? What is the main finding and its importance? In two-kidney rats, exercise training improves the Ang II-induced vasoconstriction by endothelium-derived NO released through AT2 R activation. This effect of exercise training on the Ang II-induced vasoconstriction is blunted in two-kidney-one-clip hypertensive animals, possibly as a consequence of oxidative stress. ABSTRACT: This study investigated the effects of both acute exercise and training on the Ang II-induced vasoconstriction in aorta of normotensive (two-kidney; 2K) and two-kidney-one-clip (2K1C) hypertensive rats, focusing on endothelial mechanisms related to nitric oxide (NO) and prostanoids. Aorta rings of 2K and 2K1C male Wistar rats, sedentary and trained, killed at rest and after acute exercise, were challenged with Ang II in either the absence or the presence of PD 123,319, a selective angiotensin receptor subtype 2 (AT2 R) antagonist; Nω -nitro-l-arginine methyl ester (l-NAME), a non-selective inhibitor of nitric oxide synthase; indomethacin, a non-selective inhibitor of cyclooxygenase; or Tiron, an analogue of superoxide dismutase. Aortas of sedentary and trained animals studied at rest were also submitted to histomorphometric analysis. Exercise training reduced the Ang II-induced vasoconstriction in aorta of 2K but not of 2K1C animals. This reduction of Ang II response in aortas of 2K animals was not found after endothelial removal or treatment with PD 123,319 or l-NAME. These results suggest that exercise training improves the modulation of Ang II-induced vasoconstriction in aorta of 2K animals, by endothelium-derived NO released due to the activation of AT2 R. No exercise-induced change of Ang II response occurred in 2K1C animals, except in the presence of Tiron, which was evidence for reduction of such responses only in resting trained 2K1C animals. In 2K1C animals, NO modulation of Ang II-induced vasoconstriction might be suppressed by local oxidative stress. Moreover, exercise training slightly reduced the media layer thickness in the aortas of the 2K1C, but not 2K animals, which may indicate cardiovascular protection of these animals.

The enigmatic role of cholinergic reflex in the pathogenesis of Chagas disease.

This study evaluated the inflammatory process in the colons of mice infected with Trypanosoma cruzi QM2 strain, through the analysis of muscle reactivity and the measurement of butyrylcholinesterase (BuChE) in plasma. "Swiss" mice were infected with T. cruzi QM2 strain and after 15 (G15), 30 (G30), 60 (G60), 90 (G90), and 210 (G210) days, each group had blood collected for the measurement of butyrylcholinesterase plasma concentrations ([BuChE]), a measure which functioned as an indicator of plasmatic Ach levels. All groups, except G15, had a segment of proximal colon removed to assess muscle reactivity to acetylcholine (Ach) and noradrenaline (NA) stimulation. After reactivity tests, the tissues were then fixed and stained with hematoxylin and eosin (HE) for histological evaluation of inflammatory response. The QM2 strain did induce inflammatory process in mice colon, and demonstrated differences in muscular contraction between the G60 and G210 groups, with p < 0.05. Plasma [BuChE] increased during the acute phase of infection (p < 0.05) with subsequent heterogeneous decline in the late chronic phase. These results show that the QM2 strain has tropism to the colon of mice and causes damage characteristic of megacolon; also, Ach has an enigmatic importance in the anti-inflammatory reflex over the course of T. cruzi infection.

Insights Into Antioxidant Strategies to Counteract Radiation-Induced Male Infertility.

Significance: Radiotherapy, which employs ionizing radiation to destroy or prevent the multiplication of tumor cells, has been increasingly used in the treatment of neoplastic diseases, especially cancers. However, radiation collaterally leads to prolonged periods of sperm count suppression, presumably due to impaired spermatogenesis by depleting the germ cell pool, which has long-term side effects for male reproduction. Recent Advances: Studies of antioxidant compounds as a potential strategy for male fertility preservation have been performed mainly from animal models, aiming to prevent and restore the male germinal tissue and its function, particularly against the oxidative stress effects of radiation. Evidence in preclinical and clinical trials has shown that inhibitors of the renin-angiotensin system and other drugs, such as statins and metformin, are candidates for ameliorating radiation-induced damage to several tissues, including the testis and prostate. Critical Issues: Research for developing an ideal radioprotective agent is challenging due to toxicity in the normal tissue, tumor radioresistance, cellular response to radiation, costs, regulation, and timeline development. Moreover, male radioprotection experiments in humans, mainly clinical trials, are scarce and use few individuals. This scenario is reflected in the slow progress of innovation in the radioprotection field. Future Directions: Expanding human studies to provide clues on the efficacy and safety of radioprotective compounds in the human reproductive system is necessary. Drug repurposing, frequently used in clinical practice, can be a way to shorten the development pipeline for innovative approaches for radioprotection or radiomitigation of the repercussions of radiotherapy in the male reproductive system.

Playful activity with robot for hand hygiene of elementary school students: quasi-experimental study.

OBJECTIVE: To evaluate the effectiveness of the hand hygiene process after elementary school students participated in an educational activity using an automated digital technology called Tutor Robot. METHOD: Quasi-experimental study developed in 2019 with elementary school students (n=203). Hand hygiene was performed with a fluorescent solution before and after participating in an educational activity with a tutor robot. The images were recorded in a dark chamber and the data related to area of residence, grade, gender, hand position and laterality were analyzed by Anova and Holm-Sidak Post-Hoc (p≤0,05). RESULTS: All conditions studied improved the hand hygiene process after activity with the tutor robot. There was no association between grade, gender, and dominant hand and the performance in the hand hygiene process, however, students from urban areas performed better than those from rural areas. CONCLUSION: The activity with the tutor robot represented an important resource for conducting health education actions on hand hygiene and can also be tested in other settings and populations.

Oral Infection and Survival of Trypanosoma cruzi in Sugarcane Juice Conditioned at Different Temperatures.

PURPOSE: The current most important form of transmission for Trypanosoma cruzi is the oral route, being responsible for high mortality during the acute phase in infected individuals. Thus, this study aimed to investigate the possibility of infection for this parasite using sugarcane juice in different temperatures employing metacyclic trypomastigotes obtained from xenodiagnosis performed in Swiss mice previously infected with T.cruzi Y strain, and then diluted in sugarcane juice. METHODS: For stomach histopathological analysis, 20 mice were infected with metacyclic trypomastigotes diluted in sugarcane juice and euthanized after the 2nd, 4th, 10th, and 15th days after infection. Concurrently, six batches of ten mice were fed using 1.5 mL of the mixture and kept for 12 h at the temperatures of - 80 ºC, - 20 ºC, + 2 ºC, + 28 ºC, + 60 ºC, and + 80 ºC, for later infection verification. RESULTS: Inflammatory infiltrate was found after the 2nd day of infection, and amastigotes nests were present after the 4th, 10th, and 15th day in the margo plicatus stomach region. Viable trypomastigotes were observed in the microtubes kept at - 80 ºC, - 20 ºC, and + 2 ºC, but the animal's infection was observed in the - 80 ºC and + 2 ºC groups. In vitro tests demonstrated the decrease of T. cruzi trypomastigote viability, which was negative after 120 h at -20 ºC and 144 h at + 2 ºC, in contrast to the maintenance of survival after 168 h at - 80 ºC. CONCLUSION: We observed the ability of survival and infection of T. cruzi packaged at - 80 ºC without the use of preservatives and, therefore, less suitable for storing food.

Losartan Attenuates Radiation-Induced Damage on Testes and Accelerates Tubular Regeneration.

Male germ cells are particularly susceptible to radiation; infertility being a common consequence after radiotherapy as it impairs spermatogenesis. This study aimed to test whether treatment with losartan (LOS), a selective antagonist of angiotensin II receptor subtype 1 (AT1R), can prevent or attenuate the acute and long-term radiation-induced damage to testes. Wistar rats were randomly distributed into six groups, three of which were studied on day 2 after irradiation: control (CTRL 2), irradiated non-treated (IR 2), and irradiated and treated with LOS (IRLOS 2); and three other groups that were studied on day 60 after irradiation: control (CTRL 60), irradiated non-treated (IR 60), and irradiated and treated with LOS (IRLOS 60). Seven consecutive days before and on the day of irradiation with 2.5 Gy directly administered in the scrotum, the animals were treated with LOS (34 mg/kg/two times/day). This treatment was continued 2 or 60 days after irradiation. The sperm quality was assessed from epididymis cauda. In addition, the testes were submitted to histopathological and morphometric-stereological analysis as well as the proliferating cell nuclear antigen (PCNA) quantification. Serum FSH and LH and plasma testosterone levels were also determined. The data obtained 2 days after the irradiation showed germ cell apoptosis, formation of vacuoles in the seminiferous epithelium, sloughing of germ cells into the lumen, and retention and phagocytosis of step-19 spermatids in Sertoli basal cytoplasm. The treatment with LOS in this period did not prevent or attenuate a radio-induced damage to the testes, illustrating that this drug does not protect against apoptosis derived from direct effects of radiation. On the other hand, 60 days after exposure, the data evidenced the deleterious effects of ionizing radiation on the testes as decreasing of testicular, epididymal, and seminal vesicle masses; tubular atrophy; reduction of cellular proliferation; and loss of germ cells. LOS was able to prevent some of those deleterious effects, promoting improvements in seminal vesicle mass, sperm vitality, plasma testosterone levels, vacuole number, and cell proliferation. In conclusion, inhibition of the AngII/AT1R axis by LOS is effective in protecting the indirect/delayed radiation damage resulting from oxidative stress established in the tissue.

Orchiectomy but not adjuvant-induced arthritis induces structural modifications in rat aortas.

PURPOSE: This study aimed to verify whether Adjuvant-Induced Arthritis (AIA) and/or Orchiectomy (ORX) modify the expression of the Nox1, Nox2 and Nox4 isoforms, the endothelial function or the structure of rat aortas. METHODS: Sixty-three Wistar rats were distributed into four groups: 1) Control; 2) ORX; 3) AIA; 4) Orchiectomy plus to Arthritis-induction (ORX/AIA). Thus, 21 days after the onset of AIA (by intradermal injection of Mycobacterium tuberculosis), the presence of Nox1, Nox2 and Nox4, the acetylcholine (ACh)-induced relaxation and the media layer thickness were assessed in the aorta taken from these animals. RESULTS: The Nox1, Nox2 and Nox4 were immunostained in intima, media and adventitia layers of aortas taken from all studied groups and AIA apparently increased this immunostaining. These modifications of Nox1, Nox2 or Nox4 expression, however, were not confirmed by Western blotting. In addition, neither AIA nor ORX changed the endothelial function, but ORX increased the media layer thickness in the studied aortas. CONCLUSION: The present study showed weak clues of increased expression of Nox1, Nox2 and Nox4 as a result of AIA, as well as of Nox1 reduction caused by ORX. In addition, the endothelial function was not modified in the aortas of these animals by both AIA and/or ORX. On the other hand, ORX increased significantly the aorta media layer thickness in the studied animals, which was apparently mitigated by AIA.

Reversal of diabetic-induced myopathy by swimming exercise in pregnant rats: a translational intervention study.

Gestational diabetes mellitus (GDM) plus rectus abdominis muscle (RAM) myopathy predicts long-term urinary incontinence (UI). Atrophic and stiff RAM are characteristics of diabetes-induced myopathy (DiM) in pregnant rats. This study aimed to determine whether swimming exercise (SE) has a therapeutic effect in mild hyperglycemic pregnant rats model. We hypothesized that SE training might help to reverse RAM DiM. Mild hyperglycemic pregnant rats model was obtained by a unique subcutaneous injection of 100 mg/kg streptozotocin (diabetic group) or citrate buffer (non-diabetic group) on the first day of life in Wistar female newborns. At 90 days of life, the rats are mated and randomly allocated to remain sedentary or subjected to a SE protocol. The SE protocol started at gestational day 0 and consisted of 60 min/day for 6 days/week in a period of 20 days in a swim tunnel. On day 21, rats were sacrificed, and RAM was collected and studied by picrosirius red, immunohistochemistry, and transmission electron microscopy. The SE protocol increased the fiber area and diameter, and the slow-twitch and fast-twitch fiber area and diameter in the diabetic exercised group, a finding was also seen in control sedentary animals. There was a decreased type I collagen but not type III collagen area and showed a similar type I/type III ratio compared with the control sedentary group. In conclusion, SE during pregnancy reversed the RAM DiM in pregnant rats. These findings may be a potential protocol to consider in patients with RAM damage caused by GDM.

Exercise-Induced Modulation of Angiotensin II Responses in Femoral Veins From 2-Kidney-1-Clip Hypertensive Rats.

The present study investigated the angiotensin II (Ang II) responses in rat femoral veins taken from 2-kidney-1clip (2K1C) hypertensive rats at 4 weeks after clipping, as well as the effects of exercise on these responses. In this manner, femoral veins taken from 2K1C rats kept at rest or exposed to acute exercise or to exercise training were challenged with Ang II or endothelin-1 (ET-1) in organ bath. Simultaneously, the presence of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) were determined in these preparations by western blotting. In these experiments, femoral veins exhibited subdued Ang II responses. However, after nitric oxide (NO) synthesis blockade, the responses were higher in the femoral veins taken from animals kept at rest [0.137(0.049-0.245); n = 10] than those obtained in trained animals kept at rest [0.008(0.001-0.041); n = 10] or studied after a single bout of exercise [0.001(0.001-0.054); n = 11]. In preparations in which, in addition to NO synthesis, both the local production of prostanoids and the action of ET-1 on type A (ETA) or B (ETB) receptors were inhibited, the differences induced by exercise were no longer observed. In addition, neither ET-1 responses nor the presence of COX-1 and COX-2 in these preparations were modified by the employed exercise protocols. In conclusion, NO maintains Ang II responses reduced in femoral veins of 2K1C animals at rest. However, vasodilator prostanoids as well as other relaxing mechanisms, activated by ETB stimulation, are mobilized by exercise to cooperate with NO in order to maintain controlled Ang II responses in femoral veins.

Influence of galantamine in the inflammatory process and tissular lesions caused by Trypanosoma cruzi QM2 strain.

INTRODUCTION: Trypanosoma cruzi infection triggers an inflammatory process with exacerbated production of cytokines that stimulate inflammatory and anti-inflammatory signals, including the efferent anti-inflammatory signal known as the anti-inflammatory cholinergic pathway. Thus, the use of anticholinesterase drugs, such as galantamine, could minimize the inflammatory process caused by this disease. METHODS: For the study at 30, 60, and 90 days, 120 Swiss mice were divided into three groups. Each group was subdivided into four subgroups: uninfected/untreated (CTRL), uninfected/treated (GAL), infected/untreated (INF), and infected/treated (GAL/INF). The infected groups were inoculated intraperitoneally with 0.1 ml of mouse blood containing 5 × 104 trypomastigote forms of the T. cruzi QM2 strain. The galantamine-treated groups received 5 mg/kg of galantamine orally, through pipetting. From each subgroup, the parameters of parasitemia, histopathological analysis, butyrylcholinesterase activity (BuChE), and functional study of the colon were evaluated. RESULTS: BuChE performance was observed when AChE was suppressed, with increased activity in the GAL/INF group similar to the INF group on the 30th day post infection, thus corroborating the absence of a significant difference in parasitic curves and histopathological analysis. CONCLUSIONS: The presence of an inflammatory process and nests of amastigotes, as well as evidence of reactivity to ACh and NOR, suggest that galantamine did not interfere with the colonic inflammatory response or even in colonic tissue parasitism at this stage of Chagas disease.

Análise comparativa da percepção de pessoas idosas e adultos hospitalizados sobre a qualidade da dieta hospitalar / Comparative analysis of older adults and hospitalized adults' perception of hospital diet quality

Resumo Objetivo analisar e comparar a percepção da qualidade da dieta hospitalar entre pessoas idosas e adultos hospitalizados. Método pesquisa quali-quantitativa, desenvolvida em um hospital público do interior paulista, com pessoas idosas (n=185) e adultos (n=185), hospitalizados há pelo menos três dias, recebendo dieta via oral geral/branda e distribuídos segundo o tipo de dieta prescrito: sem restrição de alimentos (GSR), para controle glicêmico (GDM) ou hipossódica (GHSS). Os dados foram coletados por entrevistas individuais, empregando-se questionário validado. A qualidade da dieta foi avaliada por meio da escala Likert, considerando-se as categorias sabor, temperatura, quantidade, aparência, horário das refeições, higiene e disponibilidade para substituição de alimentos. Os dados qualitativos foram submetidos à análise de conteúdo temática e, os quantitativos à análise descritiva e estatística pelos testes Kruskal-Wallis (variáveis quantitativas) e qui-quadrado (variáveis qualitativas). Resultados independentemente da dieta prescrita (p≥0,05), pessoas idosas e adultos hospitalizados consideraram a qualidade da dieta hospitalar satisfatória (Bom/Ótimo) entre as categorias de satisfação avaliadas, exceto o sabor, que para as pessoas idosas, apresentou associação significativa com a dieta prescrita (p=0,05). Três categorias temáticas emergiram dos relatos, mostrando que pessoas idosas e adultos compreendem a importância da dieta hospitalar para recuperação da saúde, mas ainda se vê a expectativa negativa quanto à refeição ofertada. Conclusão a dieta hospitalar com restrição influencia a percepção de sabor em pessoas idosas. Conhecer essa particularidade pode auxiliar na criação de estratégias de adequação e melhor aceitação da dieta hospitalar para esse grupo etário.

Abstract Objective To analyze and compare the perception of hospital diet quality between older adults and hospitalized adults. Method Quali-quantitative research conducted at a public hospital in the state of São Paulo, Brazil, involving older adults (n=185) and adults (n=185) who have been hospitalized for at least three days, receiving a general/soft oral diet, and classified according to the prescribed diet type: unrestricted diet (GSR), for glycemic control (GDM), or low-sodium (GHSS). The data were collected through individual interviews, utilizing a validated questionnaire. Diet quality was assessed using a Likert scale, considering the categories of taste, temperature, quantity, appearance, meal timing, hygiene, and availability for food substitution. The qualitative data were subjected to thematic content analysis, while the quantitative data underwent descriptive and statistical analysis using Kruskal-Wallis tests (for quantitative variables) and chi-square tests (for qualitative variables). Results Regardless of the prescribed diet (p≥0.05), both older adults and hospitalized adults considered the quality of hospital food satisfactory (Good/Excellent) across the assessed satisfaction categories, except for taste, which showed a significant association with the prescribed diet for older adults (p=0.05). Three thematic categories emerged from the reports, indicating that both older adults and adults understand the importance of hospital diet for health recovery, yet a negative expectation regarding the offered meal is still prevalent. Hospital diets with restrictions influence taste perception in older adults. Understanding this particularity can assist in the creation of strategies for adaptation and better acceptance of hospital diets for this age group.

Influence of the AT1 Receptor Antagonists Telmisartan and Losartan on Reproduction and Offspring After Paternal Exposure to Ionizing Radiation.

This study evaluated the repercussions of paternal exposure to radiation on reproduction and offspring in rats, as well as whether treatment with the angiotensin II type 1 (AT1) receptor antagonists telmisartan and losartan has a mitigating effect. Rats were randomly divided into 6 groups: control, radiation, telmisartan, losartan, radiation + telmisartan, and radiation + losartan. A single 5 Gy dose of radiation was administered directly into the scrotum, followed by treatment with telmisartan (12 mg/kg/d) or losartan (34 mg/kg/2 times per day) for 60 days in the groups receiving these medications. The reproductive ability of the test animals was assessed before and after exposure to radiation via fertility tests. The resulting offspring were analyzed for the presence of external and internal anomalies. Ionizing radiation significantly affected the rates of fertility, pre- and postimplantation losses, and implantation. Telmisartan and losartan did not significantly prevent this radiation-induced damage. The frequency of fetal anomalies was similar in offspring produced before and after paternal radiation exposure. Moreover, irradiated rats that received treatments and were able to generate offspring did not produce fetuses with morphological changes; this may represent a possible radioprotective effect AT1 antagonists have on offspring development, although few fetuses survived and were evaluated for malformations. Although the study findings indicate that these medications have a positive effect, further studies with longer treatment periods (extending beyond 1 rat spermatogenic cycle) are needed to determine whether these drugs significantly improve reproductive rates after paternal exposure to radiation, which may also reflect an increase in the number of viable fetuses.

CONTENT VALIDATION OF EDUCATIONAL MATERIAL ON HEALTHY EATING FOR CHILDREN UNDER TWO YEARS OF AGE. / VALIDAÇÃO DE CONTEÚDO DE MATERIAL EDUCATIVO SOBRE ALIMENTAÇÃO SAUDÁVEL PARA CRIANÇAS MENORES DE DOIS ANOS.

OBJECTIVE: To validate the content of an educational material aimed at mothers and caregivers on healthy feeding for children less than two years of age. METHODS: Quantitative study for content validation of an educational material containing three educational modules and respective folders, elaborated on the basis of official references for healthy feeding for children under two years old and adapted to the Brazilian context. Content validation was made through consensus conference in two stages by seven experts. RESULTS: In the first stage, an individual and anonymous evaluation was made and the items analyzed by the seven experts averaged seven or more, with standard deviation below three. However, some items in the educational modules (operational and adherence) and Leaflet I (motivation and culture) reached cut-off values. The second stage involved a face-to-face meeting with five of the seven experts, expressing their opinions and seeking for a new consensus. The items whose results were close to the cut-off had an expressive increase in importance and consensus level after the meeting. CONCLUSIONS: The quantitative data obtained after the consensus meetings were superior to the predefined cut-offs, and the content of the proposed educational material was consensually validated by all participating experts. The consensus conference was an efficient methodological technique to build and validate educational instruments.

OBJETIVO: Validar conteúdo de um material educativo sobre alimentação saudável para crianças menores de dois anos orientado às mães e aos cuidadores. MÉTODOS: Estudo quantitativo que envolveu a validação de conteúdo do material educativo, constituído por três módulos educacionais e seus respectivos folhetos, os quais foram elaborados com base em referenciais oficiais sobre alimentação saudável para crianças menores de dois anos adequados ao contexto brasileiro. Para validar o conteúdo concebido, empregou-se a conferência de consenso, em duas etapas, junto a sete especialistas. RESULTADOS: Na primeira etapa, de avaliação individual e anônima, os itens analisados pelos sete especialistas obtiveram média igual ou superior a sete, com desvio padrão inferior a três. Porém, itens dos módulos educacionais (operacional e adesão) e do Folheto I (motivação e cultura) alcançaram valores limítrofes ao critério de corte adotado. Na segunda etapa, em reunião presencial com cinco dos profissionais participantes, cada especialista pôde expressar suas considerações, visando à busca de novo consenso. Os itens com notas próximas do limite de corte obtiveram, após o debate, aumento expressivo de valor no grau de importância e nível de concordância dos especialistas. CONCLUSÕES: Os dados quantitativos obtidos após as etapas da conferência de consenso foram superiores ao limite de corte, sendo o conteúdo do material educativo proposto validado de forma consensual pelos especialistas participantes. A conferência consensual empregada revelou ser uma técnica metodológica eficiente para a construção e a validação de instrumentos educativos.

Effects of telmisartan and losartan on irradiated testes.

AIMS: To analyze the effects of radiation on the reproductive tissue of male Wistar rats and to evaluate whether treatment with the Ang II AT1 receptor antagonists telmisartan and losartan mitigate the dysfunctions resulting from this exposure. MAIN METHODS: Rats were randomly divided into groups: Control, Irradiated, Telmisartan, Losartan, Irradiated+Telmisartan, and Irradiated+Losartan. Single dose of 5Gy was administered directly into the scrotum, followed by treatment with telmisartan (12mg/kg/day) or losartan (34mg/kg/two times/day) for 60days. Testicular function parameters were evaluated from spermatozoa of the vas deferens. Testes were processed for histopathological and morphometric-stereological analysis. Proliferating cell nuclear antigen (PCNA) immunohistochemistry was evaluated. KEY FINDINGS: Radiation significantly reduced sperm motility, concentration, vitality, and increased the number of abnormal spermatozoa. Telmisartan and losartan did not significantly prevent these radiation-induced disorders. Seminiferous tubules were atrophied in both untreated and treated irradiated testes, and exhibited vacuoles, increased interstitial tissue and high number of blood vessels. However, several seminiferous tubules in recuperation were founded among damaged tubules in the testes of treated animals. The PCNA immunohistochemistry confirmed these outcomes. PCNA-positive cells were detected in dividing spermatogonia and spermatocytes from irradiated telmisartan and losartan treated rats whereas in the only-irradiated group, PCNA staining was observed in the nuclei of only the surviving spermatogonia. SIGNIFICANCE: Under these experimental conditions, the testicular function parameters showed that radiation produced marked damage that was not reversed by treatments. However, gonadal restructuring and recovery of spermatogenesis in treated animals may to reflect attenuation of radiation-induced damages and potential start of recovery.

Rivastigmine prevents injury induced by ischemia and reperfusion in rat liver.

PURPOSE: To evaluate whether pre-treatment with rivastigmine is able to attenuate the I/R induced lesions in rat liver. METHODS: SHAM animals or those submitted to I/R, non-treated or pre-treated with rivastigminine (2mg/kg) either 50 or 15 minutes before ischemia, were used. After I/R protocol, these animals were killed and their livers were harvested to measurement of the mitochondrial swelling as well as the malondialdehyde (MDA), nitrite and nitrate tissue concentration. Blood was also harvested for serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) determinations. RESULTS: I/R promoted a significant increase of mitochondrial swelling in the studied animals. This increase of mitochondrial swelling was partially prevented by rivastigmine, but only if administered 50 minutes before ischemia. No significant modification of MDA, nitrite or nitrate tissue concentrations was observed in consequence of I/R, followed or not by rivastigmine treatments. In addition, I/R elevated both AST and ALT. These elevations of serum enzymes were not reversed by the different rivastigmine treatments. CONCLUSIONS: Rivastigmine administered 50 minutes before ischemia attenuates I/R-induced mitochondrial swelling, that indicates liver injury. This protective effect may be related to a greater stimulation of α7nAChR present in the Kupffer cells by the non-methabolized ACh, leading to an attenuation of I/R-induced inflammation.

Playful activity with robot for hand hygiene of elementary school students: quasi-experimental study / Actividad lúdica con robot para la higiene de manos de estudiantes de educación primaria: estudio cuasi experimental / Atividade lúdica com robô para higienização das mãos de estudantes do ensino fundamental: estudo quase-experimental

ABSTRACT Objective: To evaluate the effectiveness of the hand hygiene process after elementary school students participated in an educational activity using an automated digital technology called Tutor Robot. Method: Quasi-experimental study developed in 2019 with elementary school students (n=203). Hand hygiene was performed with a fluorescent solution before and after participating in an educational activity with a tutor robot. The images were recorded in a dark chamber and the data related to area of residence, grade, gender, hand position and laterality were analyzed by Anova and Holm-Sidak Post-Hoc (p≤0,05). Results: All conditions studied improved the hand hygiene process after activity with the tutor robot. There was no association between grade, gender, and dominant hand and the performance in the hand hygiene process, however, students from urban areas performed better than those from rural areas. Conclusion: The activity with the tutor robot represented an important resource for conducting health education actions on hand hygiene and can also be tested in other settings and populations.

RESUMEN Objetivo: Evaluar la eficacia del proceso de higiene de manos tras la participación de estudiantes de educación primaria en una actividad educativa, utilizando una tecnología digital automatizada denominada Tutor Robot. Método: Estudio cuasi-experimental desarrollado en 2019 con estudiantes de primaria (n=203). Se realizó higiene de manos con una solución fluorescente antes y después de participar en una actividad educativa con un robot tutor. Las imágenes se grabaron en un cuarto oscuro y los datos relacionados con la zona de residencia, el grado, el sexo, la posición de la mano y la lateralidad se analizaron mediante Anova y Post-Hoc de Holm-Sidak (p≤0,05). Resultados: Todas las condiciones estudiadas mejoraron el proceso de higiene de manos después de la actividad con el robot tutor. No hubo asociación entre el grado, el sexo y la mano dominante y el desempeño en el proceso de higiene de manos, sin embargo, los estudiantes de la zona urbana se desempeñaron mejor que los de la zona rural. Conclusión: La actividad con el robot tutor representó un recurso importante para la realización de acciones de educación sanitaria sobre higiene de manos, puede también ser testeada en otros escenarios y poblaciones.

RESUMO Objetivo: Avaliar a efetividade do processo de higienização das mãos após a participação de estudantes do Ensino Fundamental em uma atividade educativa, com uso de uma tecnologia digital automatizada, denominada robô tutor. Método: Estudo quase-experimental desenvolvido em 2019 com estudantes do Ensino Fundamental I (n=203). Foi realizada higienização das mãos com solução fluorescente antes e após a participação em atividade educativa com um robô tutor. As imagens foram registradas em câmara escura e os dados relacionados à área de residência, série, sexo, posição das mãos e lateralidade foram analisados por Anova e Post-Hoc de Holm-Sidak (p≤0,05). Resultados: Todas as condições estudadas melhoraram o processo de higienização das mãos após atividade com o robô tutor. Não houve associação entre série, sexo e mão dominante e o desempenho no processo de higienização das mãos, porém, estudantes da área urbana apresentaram desempenho melhor do que os provenientes de área rural. Conclusão: A atividade com o robô tutor representou um recurso importante para a realização de ações de educação em saúde a respeito da higienização das mãos, e pode também ser testada em outros cenários e populações.

Influence of galantamine in the inflammatory process and tissular lesions caused by Trypanosoma cruzi QM2 strain

Abstract INTRODUCTION: Trypanosoma cruzi infection triggers an inflammatory process with exacerbated production of cytokines that stimulate inflammatory and anti-inflammatory signals, including the efferent anti-inflammatory signal known as the anti-inflammatory cholinergic pathway. Thus, the use of anticholinesterase drugs, such as galantamine, could minimize the inflammatory process caused by this disease. METHODS For the study at 30, 60, and 90 days, 120 Swiss mice were divided into three groups. Each group was subdivided into four subgroups: uninfected/untreated (CTRL), uninfected/treated (GAL), infected/untreated (INF), and infected/treated (GAL/INF). The infected groups were inoculated intraperitoneally with 0.1 ml of mouse blood containing 5 × 104 trypomastigote forms of the T. cruzi QM2 strain. The galantamine-treated groups received 5 mg/kg of galantamine orally, through pipetting. From each subgroup, the parameters of parasitemia, histopathological analysis, butyrylcholinesterase activity (BuChE), and functional study of the colon were evaluated. RESULTS: BuChE performance was observed when AChE was suppressed, with increased activity in the GAL/INF group similar to the INF group on the 30th day post infection, thus corroborating the absence of a significant difference in parasitic curves and histopathological analysis. CONCLUSIONS: The presence of an inflammatory process and nests of amastigotes, as well as evidence of reactivity to ACh and NOR, suggest that galantamine did not interfere with the colonic inflammatory response or even in colonic tissue parasitism at this stage of Chagas disease.

Follicular diameter range based on morphological features in Synbranchus marmoratus (Bloch, 1795) (Teleostei, Synbranchiformes, Synbranchidae) from the South-central region of Brazil.

The morphological characteristics of Synbranchus marmoratus female germ cells in various development stages were described in details; then measurements of ovarian follicle diameters were taken from primary and secondary growth as during these development stages the oocyte size varied considerably along the fish growth. The results were correlated to total fish length, using the individuals division in six size classes. It was possible to group oocytes by stages according to histological characteristics but not according to morphometric diameter, as there was a wide variation in diameter in each stage and overlap between different maturation stages. These data make available new information on the reproductive biology of Synbranchidae.