Use of alternative diluent ACP-Lact® and coconut water powder (ACP) in cryopreservation of goat semen and fixed-time artificial insemination (FTAI) in half-breed goats.

The aim of the present study was to evaluate semen cryopreservation with ACP-Lact® diluent, which consists of coconut water powder (ACP) added to goat milk powder. After thawing, the samples were evaluated for sperm kinetics, membrane evaluation and in vivo insemination. For cryopreservation, a pool was made with the ejaculate of six goats, diluted in four equal aliquots for the respective treatments: T1 (ACP-Lact®); T2 (ACP-Lact® 50%); T3 (ACP + 2.5% egg yolk) and T4 (Tris + 2.5% egg yolk). After dilution of the treatments, the samples were placed in 0.5 ml straws and chilled at a rate of -1.07°C/min. After reaching 4°C and stabilizing for one hour, the straws were placed in nitrogen vapour at -60°C for 15 minutes and then immersed in liquid nitrogen (-196ºC). The straws were thawed in a 37°C water bath and kinetic assessments were performed immediately using a computerized semen analysis program (CSA), viability (EN), membrane functionality (HOST), mitochondrial activity (DAB) and DNA integrity assessment of spermatozoa. For the in vivo experiment, ten goats were inseminated, divided into two groups of five goats each, G1 inseminated with ACP-Lact® and G2 with ACP, by fixed-time artificial insemination (FTAI). Regarding the kinetic parameters, the ACP-Lact® treatment showed higher progressive motility (PM) and sperm velocity than the other treatments (36.77%). In the VSL parameter the ACP-Lact diluent was superior to ACP and Tris. In viability the treatment with ACP-Lact® was superior to the treatment with Tris, 95% and 83% respectively. In FTAI two goats were born out of the 5 goats inseminated with ACP-Lact®. It was concluded that the use of ACP-Lact® for cryopreservation of caprine semen is efficient in maintaining seminal parameters during thawing in vitro and in vivo and proved to be a good alternative extender for the caprine species.

Freeze-dried Platelet-rich Plasma and Stem Cell-conditioned Medium for Therapeutic Use in Horses.

This study investigated platelet-rich plasma (PRP) and adipose stem cell-conditioned medium (ASC-CM) use as a strategy to accelerate tissue healing. Platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) were quantified in fresh and freeze-dried PRP and ASC-CM, and a stability test was performed in the freeze-dried samples (90 and 180 days of storage). A cell proliferation test was performed using equine mesenchymal stem cell culture in reconstituted PRP gel mesh after freeze-drying. In vivo PRP, ASC-CM applications, or their association were performed in induced wounds at 15 and 9-day intervals, according to the treatments: saline solution (control), PRP, ASC-CM, or ASC-CM + PRP. Horses were monitored through photographs and wound area measurements on days 5, 7, 15, and 24 after lesion induction. Skin biopsies were obtained on days 15 and 24 of the experiment. PDGF and VEGF quantification did not differ between fresh or freeze-dried treatments, was similar after freeze-drying or 90 days of storage, but showed a significant reduction after 180 days of storage. Comparing all treatments, no differences were observed in the histopathological analyses. For inflammation, fibroplasia, and collagen formation, only the time effect between the first and second biopsies was significant. The cell proliferation test revealed intense multiplication in the PRP gel mesh. Healing time was similar among all treatments. In conclusion, our results showed the possibility to produce and maintain freeze-dried PRP and ASC-CM for 90 days. Further studies are needed to better explore the in vivo therapeutic PRP and ASC-CM effects.

Assessment of coconut water added to Numida meleagris egg yolk as cryoprotectant for goat semen.

Effects were assessed of the dilutants TRIS and ACP - 101c® with the addition of different guinea fowl (Numida meleagris) egg yolk concentrations. Fifteen ejaculates were collected from five goats of the Anglo Nubian breed. The ejaculates were pooled and then divided into 12 groups, two control groups (GC1 TRIS, with 2.5% Gallus gallus domesticus hen egg yolk GOGD), (GC2 Control Group ACP - 101c®, with the addition of 2.5% Gallus gallus domesticus hen egg yolk GOGD) and ten experimental groups (EG), containing TRIS and ACP added with different concentrations of egg yolk from guinea hen (Numida meleagris) (TRIS 2,5% GONM; TRIS 5% GONM; TRIS 10% GONM; TRIS 15% GONM; TRIS 20% GONM; ACP® 2,5% GONM; ACP® 5% GONM; ACP® 10% GONM; ACP® 15% GONM; ACP® 20% GONM). Then cryopreservation was carried out and the samples stored in liquid nitrogen (-196 °C). After seven days, the samples were thawed and assessed for spermatic kinetics, immunofluorescence and sperm morphology. Analysis of GOMN by the CASA system showed that the various parameters were similar to those of GOGD (P>0.05). The membrane integrity, mitochondrial potential and the acrosome were not influenced by the treatment (P>0.05) nor by the dilutant used for cryopreservation (P>0.05). The spermatic morphology was also preserved by the different GOGD and GONM concentrations in the ACP® and TRIS dilutants, with no statistically significant differences (P<0.05). It was concluded that Numida meleagris egg yolk, as external membrane cryoproctant added to the dilutants ACP-101c® and TRIS, improved goat semen quality.

Extra Virgin Coconut Oil as a Cryoprotector for Cryopreservation of Caprine Semen.

The study aimed to evaluate the effect of powdered coconut water-based diluent (ACP-101c) associated with extra virgin coconut oil (CO) as an external cryoprotectant in the conservation of cryopreserved buck sperm. For cryopreservation, the ejaculates from four bucks were pooled and divided into three aliquots and diluted at 37°C for treatments T1 (ACP-101c + 2.5% egg yolk +7% glycerol), T2 (ACP-101c + 2.5% CO +7% glycerol), and T3 (ACP-101c + 5% CO +7% glycerol). Then, the samples were packaged and cooled at a rate of 1.07°C/min decrease. Upon reaching 4°C, the samples were stored in a refrigerator at 4°C for 30 minutes for stabilization. After this period, the straws were frozen in nitrogen vapor for 15 minutes and then immersed and stored in liquid nitrogen at -196°C. After thawing, the samples were evaluated for sperm kinetics, plasma membrane integrity, acrosomal integrity, membrane functionality, mitochondrial activity (MA), and sperm morphology. In this study, no statistically significant differences were observed between the three treatments regarding the kinetic parameters (p > 0.05; Table 1). However, in relation to the velocities, a reduction was observed beyond the expected. There were no statistically significant differences between the diluents T1, T2, and T3 for the three velocities (curvilinear velocity [VCL], linear velocity [VSL], average path velocity [VAP]). Furthermore, no statistically significant differences were observed (p > 0.05) among treatments regarding the evaluation of membrane integrity, the functional membrane, MA, and sperm morphology after thawing. In conclusion, the use of CO in concentrations of 2.5% and 5.0% is effective in maintaining goat sperm quality, presenting itself as an alternative diluent for international programs of artificial insemination and embryo transfers. [Table: see text].

Sêmen ovino e caprino imunossexado e diluído em meio de conservação à base de água de coco em pó (ACP101/102c) / Ram and goat semen immunosexed and diluted in powdered coconut water-based preservation medium (ACP101/102c)

Sperm sexing aims to separate sperm populations in carriers of the "X" or "Y" chromosome. Currently, flow cytometry is a technique that allows greater accuracy; however, it causes structural changes in sperm, reduces viability, and has a high cost. As a result, other methods have been researched, including immunosexing, which uses monoclonal antibodies to detect sex-specific surface antigens. Thus, the objective of this study was to evaluate the immunosexing technique using a monoclonal antibody against sex-specific protein (HY) in the conservation of ram and goat semen in ACP101/102c. Ejaculates from five rams and five goats were collected with the aid of an artificial vagina; they were evaluated and submitted to the immunosexing protocol, according to the manufacturer's recommendations, using the Monoclonal Antibody Kit specific for mammalian sperm with "Y" chromosomes (HY; HY Biotechnology, Rio de Janeiro, RJ, Brazil). After sexing, the supernatant was resuspended in the cryopreservation diluent: ACP ram (ACP101/102c + 20% egg yolk + 7% glycerol) and ACP goat (ACP101/102c + 2.5% egg yolk + 7% glycerol), packaged in 0.25 mL straws, refrigerated at 4°C, stabilized for 30 min, frozen in liquid nitrogen vapor (-60°C) for 15 min, immersed in liquid nitrogen, and stored in cryogenic cylinders. The samples were evaluated in natura (T1), after immunosexing (T2) and after thawing (T3) for sperm motility subjectively using conventional microscopy (40x). Plasma membrane integrity (IMP) and sperm cell morphology were evaluated by the smear staining technique using eosin-nigrosine dye, and the percentages of healthy and morphologically defect spermatozoa were determined. In the evaluation of ram semen regarding sperm motility and IMP, no statistically significant differences were observed between treatments after sexing in the evaluation of absolute data (P > 0.05), with the difference being observed only between T1 and T2, and T3 (P < 0.05). Regarding the relative percentage and sperm morphology, no statistically significant differences were observed (P > 0.05). Regarding the evaluation of goat semen samples, the motility parameters were consistent with the technique submitted; however, the IMP data did not appear as expected, requiring further evaluation for a better assessment of the technique for this species. The data obtained from ram semen submitted to the immunosexing protocol, regarding the absolute evaluation of motility and IMP, demonstrated that the non-sexed semen (T1) was superior to the sexed treatments (T2 and T3); however, it is noteworthy that freezing started with approximately 50% of the cells, since the immunosexing technique results in a loss of viability of approximately 50% of the sperm, which corresponds to the ratio of sperm carrying the X chromosome. In addition, when the data in this study were transformed into relative values, no statistical differences were observed, indicating that the immunosexing protocol, as well as the freezing protocol, did not significantly affect the quality of ram sperm cells. In relation to the immunosexing of goat semen, future studies should be conducted in vitro to define a more appropriate protocol for the species and, in addition, in vivo studies should be performed to prove the quality of the technique. It was concluded that the immunosexing process using a monoclonal antibody against sex-specific protein (HY) associated with the use of powdered coconut water diluent (ACP101/102c) in the cryopreservation of semen proved to be efficient in the in vitro evaluation of ovine species.(AU)

Alterações posturais: a caracterização de discentes de Fisioterapia em uma instituição do ensino superior em Fortaleza/CE / Postural changes: the characterization of Physiotherapy students in a higher education institution in Fortaleza/CE

Na postura padrão, a coluna apresenta curvaturas normais e os ossos dos membros inferiores ficam em alinhamento. Quando o componente estrutural é alterado, o corpo humano modifica-se para desempenhar tal situação da melhor forma possível, o que pode ocasionar em um desvio postural. O objetivo desta pesquisa é avaliar a postura e seus comprometimentos em discentes de um curso de Fisioterapia, traçando o perfil clínico socioeconômico, conhecendo os comprometimentos posturais dos participantes da pesquisa. Tratou-se de um estudo transversal, descritivo e quantitativo realizado entre os meses de agosto de 2015 a junho de 2016, no Centro Universitário Estácio do Ceará com 32 discentes do curso de Fisioterapia. Os dados foram coletados através de uma ficha de avaliação postural desenvolvida para o inquérito e analisada no programa Excel® 2013 e confrontados com a literatura vigente sobre o assunto. O gênero feminino foi predominante em 75% da amostra. Em relação ao estado civil dos participantes, 87,5% eram solteiros. A média de idade foi de 24,4 anos. Em relação ao Índice de Massa Corpórea, foi evidenciado que 28 participantes (87,5%) apresentaram um peso normal, de acordo com sua altura e peso. Para verificar a presença de escoliose, foi realizado o Teste de Adams. Evidenciou-se que 72% dos participantes apresentaram gibosidade, 12,50% apresentaram gibosidade lombar, 43,75% apresentaram gibosidade torácica e 15,63% apresentaram gibosidade toracolombar. A maioria dos estudantes apresentou escoliose. No entanto, estudos com uma amostra maior devem ser realizados, a fim de que resultados mais abrangentes possam ser obtidos. (AU)

In the standard posture, the spine has normal curvatures and the bones of the lower limbs are in alignment. When the structural component is changed, the human body changes to perform this situation in the best possible way, which can cause a postural deviation. The objective of this study was to evaluate the posture and its commitments in students of a Physical therapy course, tracing the socioeconomic clinical profile, knowing the postural impairments of the participants. This was a cross-sectional, descriptive and quantitative study carried out between August 2015 and June 2016, at the Estácio do Ceará University Center with 32 students. The data were collected through a postural assessment form developed for the survey and analyzed using the Excel® program and compared with the current literature. The female gender was predominant in 75% of the sample. Regarding the participants' marital status, 87.5% were single. The average age was 24.4 years. Regarding the Body Mass Index, it was evidenced that 28 participants (87.5%) had a normal weight, according to their height. To test for the presence of scoliosis, the Adams Test was performed. 72% of the participants presented gibosity, 12.50% lumbar gibosity, 43.75% thoracic gibosity and 15.63% thoraco-lumbar gibosity. Most of the students had scoliosis. However, studies with a larger sample should be carried out to obtain more accurate results. (AU)

Supplemented powdered coconut water (ACP-406®) promotes growth of goat secondary follicles and oocyte meiotic resumption.

The objective of this study was to test the efficiency of powdered coconut water (ACP-406®) base-medium without or with the addition of supplements on in vitro culture of isolated goat secondary follicles. Follicles were cultured for 18 days in α-MEM or in ACP-406®, both without supplements (referred to as α-MEM and ACP, respectively), or both supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred to as α-MEM+ and ACP+). Follicular morphology, antrum formation, follicular and oocyte diameter, levels of glutathione (GSH), and chromatin configuration after in vitro maturation were evaluated. At the end of culture, ACP-406® base-medium (without or with supplements) showed a higher (P < 0.05) percentage of normal follicles than α-MEM (without or with supplements). Antrum formation was similar among α-MEM+, ACP and ACP+, and significantly higher than α-MEM without supplements. The follicular diameter was greater in ACP+ than α-MEM, and similar to other treatments. Moreover, fully and daily grown rates were higher (P < 0.05) in ACP-406® base-medium (without or with supplements) than α-MEM (without or with supplements). Levels of GSH were similar between ACP+ and α-MEM+ treatments. Both ACP+ and α-MEM+ allowed meiotic resumption without a significant difference between the two groups. In conclusion, supplemented ACP-406® base-medium maintained follicular survival and promoted the development as well as meiotic resumption of isolated goat secondary follicles cultured in vitro for 18 days.

Estudo comparativo da prevalência de alterações posturais na coluna vertebral em escolares do ensino fundamental do município de Quixadá/CE / Comparative study of the impact of postural changes on spine in elementary school of Quixadá/CE

Introdução: A má postura caracteriza-se pela relação defeituosa entre as várias partes do corpo que produz uma maior tensão nas estruturas de apoio. Objetivo: Comparar a prevalência de alterações posturais da coluna vertebral em alunos do ensino fundamental de duas escolas, uma da rede privada e outra da rede pública, do município de Quixadá/CE. Métodos: Avaliaram-se 40 alunos voluntários de ambas as escolas, sendo 48% do sexo masculino e 52% do sexo feminino, com idades entre 11 e 15 anos. Analisaram-se com ajuda de bolas de isopor sobrepostas nos principais relevos anatômicos e em seguida fotografias, nas quatro vistas, anterior, posterior, lateral esquerda e lateral direita, de forma estática, posicionados atrás do simetrógrafo, analisando através do software Coreldraw, marcando as alterações mais evidentes. Resultados: Demonstraram a saúde deficiente da coluna vertebral em escolares, com a identificação de várias alterações posturais, sendo mais prevalente na instituição da rede municipal de Quixadá/CE. Sendo elas: hipercifose, desequilíbrio pélvico, desequilíbrio escapular, inclinação lateral cervical, anteriorização cervical e elevação unilateral do ombro. Entretanto, os alunos da instituição da rede privada apresentaram as mesmas alterações, sendo em um menor índice. Conclusão: Observou-se que o fator socioeconômico das instituições e dos alunos que as compõe pode provocar futuras alterações posturais, devido às mobílias e a forma de transporte de materiais serem inadequadas. O estudo sugere a presença de um fisioterapeuta no ambiente escolar onde seria possível intervir na evolução dessas alterações e a fixação dos desvios, visando o trabalho preventivo e educacional. (AU)

Introduction: Bad posture is characterized by the defective relationship between the various parts of the body that produces a greater tension in the supporting structures. Objective: To compare the prevalence of postural alterations of the spine in elementary school students from two schools, one private and another public, from the municipality of Quixadá/CE. Methods: A total of 40 students from both schools were evaluated, 48% male and 52% female, aged 11 to 15 years. We analyzed with the aid of styrofoam balls overlapped in the main anatomical reliefs and then photographied, in the four views, anterior, posterior, left lateral and right lateral, in a static way, positioned behind the symetrograph, analyzing through Coreldraw software, marking the most evident changes. Results: The results showed poor health of the spine in schoolchildren, with the identification of several postural alterations, being more prevalent in the public institution. These include: hyperkyphosis, pelvic imbalance, scapular imbalance, cervical lateral inclination, anterior cervical and unilateral shoulder elevation. However, the students from the private institution showed the same changes, but in a lower level. Conclusion: We observed that the socioeconomic factor of institutions and students can cause future postural alterations, due to the furniture and the inadequate way of transportation of school materials. The study suggests the presence of a physiotherapist in the school environment where it would be possible to modify the evolution of these alterations and the establishment of deviations, aiming at preventive and educational work. (AU)

Replanted teeth stored in a newly developed powdered coconut water formula.

BACKGROUND/AIM: A newly developed powdered coconut water formulae (ACP-404) has been proposed to preserve periodontal ligament cell viability. The aim of this study was to investigate the periodontal healing of dog teeth replanted after storage in ACP-404 with 2 different osmolalities. MATERIALS AND METHODS: Sixty teeth of 10 adult beagle dogs were extracted and randomly subjected to the following protocols: immediate replantation after extraction (positive control); dry conditions (negative control); and storage in ACP-404 I (G1), ACP-404 II (G2), in coconut water long shelf life (G3) or in pH-neutralized coconut water long shelf life (G4) for 50 minutes before replantation. The animals were euthanized 28 days post-operatively and processed for histomorphometric analysis using an optical microscope. The groups were subjected to histomorphometric analysis compared with immediate replantation and dry conditions. The data were statistically analysed using the Kruskal-Wallis test with post-analysis using Dunnett's test. RESULTS: The normal periodontal area was significantly higher for the G1 than for the G2 and G3 groups (P < .001). The positive control group presented similar normal periodontal areas compared to the experimental groups, except for the G3 and negative control groups (P < .001). G1 presented the lowest level of external root replacement resorption compared to the other groups (P < .05). All storage solutions had similar periodontal conditions for repaired resorption areas, inflammatory root resorption, ankylosis and replacement resorption (P > .05). CONCLUSIONS: ACP-404 I (G1) was better overall and was able to preserve the normal periodontium and had less external root replacement resorption.