Gene expression in the Longissimus dorsi muscle related to meat quality from tropical hair lambs.

The present study describes the expression of genes in the Longissimus dorsi muscle related to meat quality of hair lambs finished in an Integration Crop-Livestock system. Twenty-eight non-castrated lambs of two breeds, Somalis Brasileira and Santa Inês, at 120 ± 15 days of age, with an average initial live weight of 18 ± 3.1 kg, were kept in a pasture-based finishing system with supplementation. Upon reaching 28 kg body weight, animals were sent for slaughter. Samples of the Longissimus dorsi and Biceps femoris muscle were harvested for analyses of gene expression and physicochemical properties. Significant differences were detected between the breeds for tissue and chemical composition, whereas the physical aspects did not differ. We observed the expression of six genes related to lipid synthesis (acetyl-CoA carboxylase [ACACA], fatty acid synthase [FAS], stearoyl-CoA desaturase [SCD], lipoprotein lipase [LPL], cell death-inducing DFFA-like effector A [CIDEA], and thyroid hormone responsive [THRSP]) and six genes related to molecular synthesis (myostatin [MSTN], growth differentiation factor 8 [GDF8], insulin-like growth factor 1 [IGF1], insulin-like growth factor 2 [IGF2], delta-like 1 homolog [DLK1], and growth hormone receptor [GHr]) in both breeds. The Santa Inês breed and the Somalis Brasileira showed similar expression patterns of genes related to lipogenesis and myogenesis of the Longissimus dorsi muscle, with the exception of the THRSP gene, in which the Somalis Brasileira have more receptors for the action of thyroid hormones, which resulted in greater thickness of fat in the carcass (subcutaneous fat) and higher lipid content in the chemical composition of the meat.

Hindfoot Alignment in Flexible Cavovarus Deformity Under Orthostatic and Coleman Block Test Positions: A Weightbearing Computed Tomography Study.

BACKGROUND: Flexible cavovarus deformity is prevalent and the Coleman block test is frequently used to assess the first ray plantarflexion malpositioning in the overall deformity as well as the flexibility of the hindfoot. The objective was to assess and compare the weightbearing computed tomography (WBCT) 3-dimensional (3D) changes in clinical and bone alignment in flexible cavovarus deformity patients when performing the Coleman block test when compared to normal standing position and to controls. METHODS: Twenty patients (40 feet) with flexible cavovarus deformity and 20 volunteer controls (40 feet) with normal foot alignment underwent WBCT imaging of the foot and ankle. Cavovarus patients were assessed in normal orthostatic and Coleman block test positions. Foot and ankle offset (FAO), hindfoot alignment angle (HAA), talocalcaneal angle (TCA), subtalar vertical angle (SVA) and talonavicular coverage angle (TNCA) and a CT-simulated soft tissue envelope image, WBCT clinical hindfoot alignment angle (WBCT-CHAA), were evaluated by 2 readers. Measurements were compared between cavovarus nonstressed and stressed positions and to controls. P values of .05 or less were considered significant. RESULTS: The intra- and interobserver intraclass correlation coefficient were good or excellent for all WBCT measurements. Cavovarus patients demonstrated significant correction of WBCT-CHAA (9.7 ± 0.4 degrees), FAO (2.6 ± 0.4%), and TNCA (8.8 ± 1.8 degrees) when performing the Coleman block test (all P values <.0001). However, WBCT-CHAA and FAO measurements were still residually deformed and significantly different from controls (P values of .001 and <.0001, respectively). TNCA values corrected to values similar to healthy controls (P = .29). No differences were observed in cavovarus patients during Coleman block test for the coronal measures: HAA, TCA, and SVA measurements. CONCLUSION: In this study, we observed improvement in the overall 3D WBCT alignment (FAO), axial plane adduction deformity (TNCA), as well as CT simulated clinical hindfoot alignment (WBCT-CHAA) in flexible cavovarus deformity patients when performing a Coleman block test. However, we did not find improvement in measures of coronal alignment of the hindfoot, indicating continued varus positioning of the hindfoot in these patients. LEVEL OF EVIDENCE: Level III, retrospective comparative study.

Assessment of knee instability in ACL-injured knees using weight-bearing computed tomography (WBCT): a novel protocol and preliminary results.

OBJECTIVE: To propose a protocol for assessing knee instability in ACL-injured knees using weight-bearing computed tomography (WBCT). MATERIALS AND METHODS: We enrolled five patients with unilateral chronic ACL tears referred for WBCT. Bilateral images were obtained in four positions: bilateral knee extension, bilateral knee flexion, single-leg stance with knee flexion and external rotation, and single-leg stance with knee flexion and internal rotation. The radiation dose, time for protocol acquisition, and patients' tolerance of the procedure were recorded. A blinded senior radiologist assessed image quality and measured the anterior tibial translation (ATT) and femorotibial rotation (FTR) angle in the ACL-deficient and contralateral healthy knee. RESULTS: All five patients were male, aged 23-30 years old. The protocol resulted in a 16.2 mGy radiation dose and a 15-min acquisition time. The procedure was well-tolerated, and patient positioning was uneventful, providing good-quality images. In all positions, the mean ATT and FTR were greater in ACL-deficient knees versus the healthy knee, with more pronounced differences observed in the bilateral knee flexion position. Mean lateral ATT in the flexion position was 9.1±2.8 cm in the ACL-injured knees versus 4.0±1.8 cm in non-injured knees, and mean FTR angle in the bilateral flexion position was 13.5°±7.7 and 8.6°±4.6 in the injured and non-injured knees, respectively. CONCLUSION: Our protocol quantitatively assesses knee instability with WBCT, measuring ATT and FTR in diverse knee positions. It employs reasonable radiation, is fast, well-tolerated, and yields high-quality images. Preliminary findings suggest ACL-deficient knees show elevated ATT and FTR, particularly in the 30° flexion position.

Imaging of solitary and multiple osteochondromas: From head to toe - A review.

Osteochondromas account for 20%-50% of all benign bone lesions. These tumors may present as solitary non-hereditary lesions, which are the most common presentation, or as multiple tumors associated with hereditary conditions. Plain radiography is the imaging method of choice and demonstrates the typical cortical and medullary continuity of the tumor with the underlying bone. Magnetic resonance imaging is often performed to evaluate cartilage cap thickness, which correlates with malignant transformation. Other local complications include compression of adjacent neurovascular bundles, muscles, and tendons, bursitis, tendon tears, stalk fracture, and angular or rotational long bone deformities. Although the imaging features of osteochondromas are largely known, only a few papers in the literature have focused on their main complications and image-based follow-up. This paper aimed to illustrate the main complications of osteochondromas, suggest an image-based algorithm for management and follow-up and discuss differential diagnosis.

Miniaturized Carbon Fiber Paper Electrodes for In Situ High Resolution NMR Analyses.

Combining spectroscopic techniques with electrochemistry is a promising strategy, as it allows the detailed investigation of the species that are consumed and produced by the reaction in real time. However, as with any in situ coupling technique, the junction between NMR and electrochemistry presents some challenges, notably the distortion of NMR signals due to the placement of electrodes close to or within the detection region. In this work, miniaturized electrodes made of carbon fiber paper were developed and later modified with platinum. Platinum decoration by cathodic deposition was chosen, as platinum is a prominent element in electrocatalysis, able to catalyze a large variety of reactions. To evaluate the efficiency of this electrochemical system, the oxidation of ascorbic acid was used as a model reaction. It was observed that the electrodes caused substantial signal distortion when placed within the detection region (full width at half-maximum equal to 1.46 Hz), whereas no distortion was observed when the electrodes were placed 1 mm above the detection region (full width at half-maximum equal to 0.95 Hz). With this system, it was also possible to monitor the magnetoelectrolysis effect, caused by the interaction of the magnetic field with the flowing ions, leading to a doubling of the ascorbic acid oxidation rate, compared to the reaction performed without a magnetic field. In addition to its low cost and simplicity in preparation, the developed electrode system allows the electrode surface to be easily modified with other suitable catalysts.

Geometry and phenomenology of the living: Limits and possibilities of mathematization, complexity and individuation in biological sciences.

The central aim of this Special Issue, devoted to a multifocal study of the geometry and phenomenology of the living, is to show the need of working with models that integrate geometrical and topological objects and operations, dynamical variables and specific biological mechanisms and their relationships with one another. A multilevel and integrative approach has essentially to consider the fact that simply knowing the parts list of genes and proteins does not tell us much about how life's many biological processes works. The cellular organization is a complex dynamical system with hundreds of thousands of bio-molecules interacting with one another to execute organism's many related functions. It is argued that the production of complex living organisms owes much of its working to some topological mechanisms (i.e. four-dimensional transformations and deformations) which operate markedly on the three levels of the organization, regulation and evolution of biological systems. Thus, one can speak of a specific topology of the living acting very dynamically on the (non-fixed) substrate space of the physiological and metabolic activities of all complex living organisms. Philosophically, it is now clear that the genetic causality theory has several limitations, both intrinsic because of the multilevel complexity of biological processes, and extrinsic in that it disregards the influence of the phenotype on the genotype and in particular the possibility that certain acquired characteristics can be inherited. In a sense, we can say that the molecular biological conception of recent decades has limited or even misleadingly impacted our vision of the living world. New ideas are needed if we are to succeed in unravelling multifactorial genetic, epigenetic and environmental causation at higher levels of physiological function and so to explain fundamental living phenomena that genetics alone is unable to explain. Some of the most significant conclusions proposed in this issue are that (i) structural plasticity and biological functionality are deeply related, (ii) the biological information and signification is inherently spatial and temporal, it is not unidirectional, and it essentially evolve following a complex and changing network-like organization, (iii) finally, gene ontology is incomplete and it as to be supplied by integrating other fundamental levels of the organization and regulation of the living systems.

Composite Graphite-Epoxy Electrodes for In Situ Electrochemistry Coupling with High Resolution NMR.

The in situ coupling between electrochemistry and spectrometric techniques can help in the identification and quantification of the compounds produced and consumed during electrochemical reactions. The combination of electrochemistry with nuclear magnetic resonance is quite attractive in this respect, but it has some challenges to be addressed, namely, the reduction in the quality of the NMR signal when the metallic electrodes are placed close to or in the detection region. Since NMR is not a passive technique, the convective effect of the magnetic force (magnetoelectrolysis), which acts by mixing the solution and increasing the mass transport, has to be considered. In seeking to solve the aforementioned problems, we developed a system of miniaturized electrodes inside a 5 mm NMR tube (outer diameter); the working and counter electrodes were prepared with a mixture of graphite powder and epoxy resin. To investigate the performance of the electrodes, the benzoquinone reduction to hydroquinone and the isopropanol oxidation to acetone were monitored. To monitor the alcohol oxidation reaction, the composite graphite-epoxy electrode (CGEE) surface was modified through platinization. The electrode was efficient for in situ monitoring of the aforementioned reactions, when positioned 1 mm above the detection region of the NMR spectrometer. The magnetoelectrolysis effect acts by stirring the solution and increases the reaction rate of the reduction of benzoquinone, because this reaction is limited by mass transport, while no effect on the reaction rate is observed for the isopropanol oxidation reaction.

Imaging of progressive collapsing foot deformity with emphasis on the role of weightbearing cone beam CT.

The term progressive collapsing foot deformity (PCFD) is currently recommended as the replacement to adult-acquired flatfoot deformity and posterior tibial tendon dysfunction to better reflect its pathology, which consists of a complex three-dimensional deformity involving the foot and ankle. The new consensus has also provided a new classification that requires clinical and radiographic findings for patient stratification into each class. However, conventional radiographs are susceptible to errors resulting from the inadequate positioning of patients, incorrect angulation of the X-ray tube, and overlapping of bone structures. Weightbearing cone beam computed tomography (WBCBCT), which has greater diagnostic accuracy than conventional radiograph, is useful for evaluating progressive collapsing foot deformity to determine medial arch collapse, hindfoot alignment, peritalar subluxation, posterior subtalar joint valgus, intrinsic talus valgus, and lateral extra-articular bone impingement. The present review aimed to discuss the new recommendations for nomenclature, classification, and imaging evaluation of PCFD, with an illustrative and quantitative focus on the measurements used in conventional radiography and WBCBCT. The measurements presented here are important criteria for decision-making.

The limits of the mathematization of the living and the idea of formal morphology of the living world following Husserlian phenomenology.

Through a presentation and a commentary of Husserl's little-known analyses of mathematization in the life sciences and on morphology, this article proposes three goals. First, it aims at establishing the real meaning and results of the critical analyses of the mathematization in natural sciences and of exactness put forth as a standard of scientific knowledge that we read in the Krisis. As a result, it will appear that these analyses belong to the perspective of a project of a formal morphology, understood as an extension of mathesis. It is then to explain why this project only makes sense in the larger framework of the description of the "correlational a priori," i.e., the theory of constituting subjectivity, experiencing these morphologies, and engaging, theoretically, by induction, in the typification and categorial elaboration of possible explanatory models. After presenting the contours of this project and its achievements, we will conclude with some conjectural proposals concerning the profile of plausible mathematical structures likely to satisfy the minimal algebraic formal conditions for a model of stability and plasticity of the living and allowing to understand and express the dynamic stratification of morphological levels and the various forms of morphogenesis.

WEIGHT-BEARING CONE BEAM CT SCANS AND ITS USES IN ANKLE, FOOT, AND KNEE: AN UPDATE ARTICLE.

Imaging plays a key role in the preoperative diagnosis, surgical planning, and postsurgical assessment of the foot, ankle, and knee pathologies. Interpreting diagnostic imaging accurately is crucial for the clinical practice of orthopedic surgeons. Although among the most used imaging modalities, radiographic assessments are amenable to errors for various technical reasons and superposition of bones. Computed tomography (CT) is a conventional imaging procedure that provides high-resolution images, but fails in considering a truly weight-bearing (WB) condition. In an attempt to overcome this limitation, WB cone beam CT technology has being successfully employed in the clinical practice for the past decade. Besides economically viable and safe, the WB cone beam CT considers WB conditions and provides high-quality scans, thus allowing an equitable and correct interpretation. This review aims to address extensive description and discussion on WBCT, including imaging quality; costs; time consumption; and its applicability in common foot, ankle, and knee, conditions. With this technology increasing popularity, and considering the extensive literature on medical research, radiologists and orthopedic surgeons need to understand its potential applications and use it optimally. Level of Evidence III, Systematic review of level III studies.

Os exames de imagem são essenciais no diagnóstico, planejamento cirúrgico e avaliação pós-cirúrgica das patologias que envolvem pé, tornozelo e joelho. A interpretação acurada utilizando as tecnologias de diagnóstico por imagem disponíveis é crucial para os cirurgiões ortopédicos na sua prática clínica. Embora as radiografias convencionais estejam entre as modalidades de diagnóstico por imagem mais utilizadas, elas estão sujeitas a erros por várias razões técnicas e sobreposição de estruturas ósseas. Apesar de a tomografia computadorizada (TC) fornecer imagens de alta qualidade, ela falha em não considerar a carga corporal fisiológica. A TC de feixe cônico com carga vem sendo utilizada com sucesso desde a última década, superando a limitação da TC convencional. Além de ser econômica e segura, possibilita a aquisição de imagens de alta resolução, com carga, permitindo, assim, uma interpretação correta e equiparável. O objetivo principal dessa revisão é proporcionar uma discussão e descrição ampla de TC com carga, incluindo qualidade de imagem, custos financeiros, tempo consumido em exames, e suas aplicações em patologias comuns do pé, tornozelo e joelho. A TC com carga vem crescendo em popularidade, e é tema de um número extenso de pesquisas científicas, sendo necessário que radiologistas e cirurgiões ortopédicos entendam suas aplicações para melhor uso futuro. Nível de Evidência III, Revisão sistemática de Estudos de Nível III.

Arteriopathy in pediatric stroke: an underestimated clinical entity.

BACKGROUND: Pediatric arterial ischemic stroke (AIS), which was thought to be a rare disorder, is being increasingly recognized as an important cause of neurological morbidity, thanks to new advances in neuroimaging. OBJECTIVE: The aim of this study was to review the main etiologies of stroke due to arteriopathy in children. METHODS: Using a series of cases from our institution, we addressed its epidemiological aspects, physiopathology, imaging findings from CT, MR angiography, MR conventional sequences and MR DWI, and nuclear medicine findings. RESULTS: Through discussion of the most recent classification for childhood AIS (Childhood AIS Standardized Classification and Diagnostic Evaluation, CASCADE), we propose a modified classification based on the anatomical site of disease, which includes vasculitis, varicella, arterial dissection, moyamoya, fibromuscular dysplasia, Takayasu's arteritis and genetic causes (such as ACTA-2 mutation, PHACE syndrome and ADA-2 deficiency). We have detailed each of these separately. Conclusions: Prompt recognition of AIS and thorough investigation for potential risk factors are crucial for a better outcome. In this scenario, neurovascular imaging plays an important role in diagnosing AIS and identifying children at high risk of recurrent stroke.

Weight-bearing cone-beam computed tomography in the foot and ankle specialty: where we are and where we are going - an update.

Cone-beam computed tomography (CBCT) has been applied in dentistry and medicine for nearly two decades. Its application in the foot and ankle specialty has grown exponentially in recent years. Weight-bearing CBCT allows clinicians to obtain weight-bearing images that can be viewed in all three planes and to construct three-dimensional models, similar to those constructed from traditional CT scans, as well as exposing patients to less radiation than do traditional CT scans. This technology has revolutionized diagnoses, improving the understanding of various lesions and surgical planning in the foot and ankle specialty. Ongoing studies of the use of weight-bearing CBCT in foot and ankle surgery are focused on fully automated and semi-automated three-dimensional measurements, as well as bone segmentation, mapping of the distances/orientation of the joints, and the production of customized implants. The aims of this review article are to show the evolution of this emerging tool in the foot and ankle specialty, to update those in related specialties on its use in current clinical practice, and to indicate where the research community is heading.

A tomografia computadorizada (TC) de feixe cônico é aplicada na área da odontologia e medicina há cerca de duas décadas, e seu uso na especialidade do tornozelo e pé cresceu de forma exponencial nos últimos anos. A literatura demonstra como a TC de feixe cônico permite obter imagens com apoio do peso corporal total - weight-bearing cone-beam computed tomography - com cortes nos três planos e modelos tridimensionais semelhantes às TCs, associada a menor exposição à radiação, técnica que revolucionou o diagnóstico, o entendimento de diferentes lesões e o planejamento cirúrgico nessa área do conhecimento. As mensurações tridimensionais automáticas e semiautomáticas, a segmentação óssea, o mapeamento das distâncias e orientações articulares e a possibilidade de produção de implantes customizados são o interesse dos estudos em andamento na cirurgia do tornozelo e pé relacionados a essa ferramenta emergente. O objetivo deste artigo é mostrar a evolução do método, atualizar as especialidades de interface sobre uso na prática clínica atual e indicar para onde a comunidade científica está caminhando.

Arteriopathy in pediatric stroke: an underestimated clinical entity / Arteriopatia em crianças com acidente vascular cerebral: uma entidade clínica subestimada

ABSTRACT Background: Pediatric arterial ischemic stroke (AIS), which was thought to be a rare disorder, is being increasingly recognized as an important cause of neurological morbidity, thanks to new advances in neuroimaging. Objective: The aim of this study was to review the main etiologies of stroke due to arteriopathy in children. Methods: Using a series of cases from our institution, we addressed its epidemiological aspects, physiopathology, imaging findings from CT, MR angiography, MR conventional sequences and MR DWI, and nuclear medicine findings. Results: Through discussion of the most recent classification for childhood AIS (Childhood AIS Standardized Classification and Diagnostic Evaluation, CASCADE), we propose a modified classification based on the anatomical site of disease, which includes vasculitis, varicella, arterial dissection, moyamoya, fibromuscular dysplasia, Takayasu's arteritis and genetic causes (such as ACTA-2 mutation, PHACE syndrome and ADA-2 deficiency). We have detailed each of these separately. Conclusions: Prompt recognition of AIS and thorough investigation for potential risk factors are crucial for a better outcome. In this scenario, neurovascular imaging plays an important role in diagnosing AIS and identifying children at high risk of recurrent stroke.

RESUMO Introdução: O acidente vascular cerebral (AVC) pediátrico, considerado um distúrbio raro, está sendo cada vez mais reconhecido como importante causa de morbidade neurológica, graças aos novos avanços na neuroimagem. Objetivo: Revisar as principais etiologias do AVC por arteriopatia em crianças. Métodos: Utilizando-se de uma série de casos de nossa instituição, abordamos seus aspectos epidemiológicos, fisiopatológicos e de imagem na angiotomografia computadorizada e angiorressonância magnética, sequências convencionais e avançadas de ressonância magnética e medicina nuclear. Resultados: Com base na classificação mais recente de AVC na infância (Classificação Padronizada e Avaliação Diagnóstica do AVC na Infância - CASCADE) propusemos uma classificação modificada com base no local anatômico da doença, que inclui vasculite, varicela, dissecção arterial, Moyamoya, displasia fibromuscular, arterite de Takayasu e causas genéticas (como mutação ACTA-2, síndrome PHACE e deficiência de ADA-2), detalhando cada uma separadamente. Conclusões: O reconhecimento imediato do AVC na infância e a investigação minuciosa de possíveis fatores de risco são cruciais para um melhor resultado. Nesse cenário, a imagem neurovascular desempenha papel importante no diagnóstico de AVC e na identificação de crianças com alto risco de recorrência.

The role of MRI in evaluation of arthroscopic transtibial pullout repair for medial meniscus posterior root tears.

Medial meniscus posterior root tears (MMPRT) can lead to meniscal extrusion, loss of hoop tension, loss of load-sharing ability and increased contact pressure. Currently, the most commonly used technique for root repair is arthroscopic transtibial pullout repair (ATPR). This article aims to illustrate both normal and abnormal postoperative imaging findings of the MMPRT repair performed with ATPR, with emphasis on MRI. The radiologist must highlight the tunnel position, the reduction of the medial meniscus posterior root to its anatomical attachment, the signs of root healing (continuity and lower signal intensity), and eventual meniscal extrusion or signs of osteoarthritis.

Computed tomography (CT), X-ray, and MRI evaluation of two anterolateral knee reconstruction techniques: lateral extra-articular tenodesis (LET) and the anterolateral ligament (ALL) reconstruction.

The anterolateral ligament (ALL) and capsule of the knee are anatomical structures involved in rotational stability and pivot-shift control. As such, it has been demonstrated that the extra-articular anterolateral procedures improve clinical outcome when performed as an augmentation of the anterior cruciate ligament (ACL) reconstruction in specific groups of patients. This review describes the postoperative imaging findings of two techniques used to perform these procedures, using magnetic resonance imaging (MRI), computed tomography (CT), and radiography. The first technique described is the lateral extra-articular tenodesis (LET), which uses a strip of the iliotibial band that is harvested, passed underneath the lateral collateral ligament (LCL) and fixed posterior, and proximal to the lateral femoral epicondyle (LFE), preserving ITB insertion on Gerdy's tubercle. The second technique described is the ALL reconstruction, a procedure that attempts to recreate the anatomy of the ALL, using most often a gracilis autograft. In this procedure, femoral fixation is performed proximal and posterior to the LFE, and tibial fixation is slightly distal to the joint line, halfway from Gerdy's tubercle to the fibular head. The main objective of this review is to provide an overview of the postoperative imaging aspects of these two procedures with MRI, CT, and radiography and to describe possible complications. As they become more common, it is important for the radiologist and the orthopedic surgeon to understand their particularities in combination with the already well-known ACL reconstruction.

In vitro study of Withanolide D toxicity on goat preantral follicles and its effects on the cell cycle.

The Withanolide D is a chemotherapeutic potential against the human tumor cell. However, there is no report on the effect of this compound on ovarian function, especially on preantral folliculogenesis. The aim of this study was to evaluate the toxicity of a new candidate to anticancer drug, Withanolide D (WD) on morphologic integrity, development (activation and granulosa cell proliferation) and gene expression of ABCB1 protein of caprine preantral follicles. Ovarian fragments were cultured in vitro for 2 or 6 days in α-MEM or α-MEM added with paclitaxel (PTX -0.1 µg/mL; negative control) and different concentrations of WD (WD1.5, WD3.0 or WD6.0). The higher dose of WD showed a toxic effect similar to PTX and higher (P < 0.05) than other treatments after 2 and 6 days. In addition, WD6.0 reduced the cell proliferating compared to PTX or mild dose. The expression of ABCB1 remained unchanged in the presence of the chemotherapeutic agents (PTX and WD) throughout the culture period. In conclusion, WD exerted a toxic effect observed by decreasing follicular survival and cell proliferation, on the preantral caprine follicles similar to PTX, whose negative effect on folliculogenesis is already widely known.

Xenotransplantation of goat ovary as an alternative to analyse follicles after vitrification.

The aim of this study was to evaluate the caprine preantral follicles enclosed on vitrified/warmed ovarian cortex grafted to nude BALB/mice during 1 month. The ovarian cortex from goats was fragmented (3 × 3 × 0.5 mm) and divided into four groups: fresh control, vitrified control, fresh transplant and vitrified transplant. Follicular morphology, development and density, fibrosis as well as apoptosis, and tissue revascularization were evaluated. It was also observed a significant decrease in morphologically normal preantral (primordial, transition, primary and secondary) follicles in both vitrified control and vitrified transplant treatments when compared with both fresh control and fresh transplant. However, fresh control and fresh transplant exhibited a similar percentage of developing follicles. Additionally, Vitrified control showed a significant increase in developing follicles in comparison with both fresh control and fresh transplant. Follicular density significantly decreased in all treatments in comparison with fresh control. We observed high fibrosis in both fresh transplant and vitrified transplant. The mRNA expression of caspase 3 was lower in both fresh transplant and vitrified transplant in comparison with vitrified control. In conclusion, xenotransplantation is an excellent strategy to maintain normal preantral follicle morphology after vitrification/warming of goat ovarian tissue. Yet, in order to ensure the survival and development of these follicles, it is essential to improve the revascularization of the graft.

Gene and protein expression in the reproductive tract of Brazilian Somalis rams.

Brazilian Somalis is a locally-adapted breed of rams raised in tropical climate and native pastures. The present study was conducted to evaluate gene expression and proteome of the reproductive tract of such rams. Samples were collected from testes, epididymides, seminal vesicles and bulbourethral glands of four rams. Expression of clusterin (CLU), osteopontin (OPN) and prostaglandin D2 synthase (PGDS) genes were evaluated in all samples by real-time PCR. Shotgun proteomic analysis was performed using samples from the head, corpus and cauda epididymides and from all other structures as well. Gene ontology terms and protein interactions were obtained from UniProtKB databases and MetaCore v.6.8 platform. CLU trasncripts were detected in the testes, epididymides, seminal vesicles and bulbourethral glands of the Somalis rams. The initial region and body of the epididymis had the greatest CLU expression. OPN mRNA was localized in all tissues of the ram reproductive tract. PGDS mRNA was detected in the testes and epididymides. Lable-free mass spectrometry allowed the identification of 137 proteins in all samples. Proteins of the epididymis head mainly participate in cellular processes and response to stimulus, participating in catalityc activity and binding. Proteins of epididymis body acted as regulatory proteins and in cellular processes, with binding and catalytic activity. Cauda epididymis molecules were associated with cellular processes and regulation, with binding function and catalytic activity as well. Testis proteins were mainly linked to cell processes and response to stimuli, and had catalytic function. Seminal vesicle proteins were involved in regulation and mainly with binding functions. Most bulbourethral gland proteins participated in cellular processes. The present study is the first to evaluate the proteome and gene expressions in the reproductive tract of Brazilian Somalis rams. Such pieces of information bring significant cointribution for the understanding of the reproductive physiology of locally-adapted livestock.

Supplementation of in vitro culture medium with FSH to grow follicles and mature oocytes can be replaced by extracts of Justicia insularis.

The present study evaluated the effect of supplementing in vitro culture medium with J. insularis compared to FSH on isolated secondary follicles and in vitro maturation of oocytes from those follicles. Secondary follicles were isolated from sheep ovaries and individually cultured for 18 days in α-MEM+ (Control), α-MEM+ supplemented with 100 ng/mL recombinant bovine follicle stimulating hormone (FSH) or with 0.3, 1.25, or 2.5 mg/mL of J. insularis extract (JI0.3, JI1.25, and JI2.5, respectively). Culture medium collected every 2 days was used to measure ROS levels. At the end of the culture period, cumulus oocytes complex (COCs) were collected and matured in vitro. Follicular walls were used for mRNA quantitation. JI0.3 led to a higher (P < 0.05) percentages of intact follicles than other groups after 18 days of culture. While follicular diameter remained unchanged from Day 6 onwards with JI0.3 and FSH, percentages of antral cavity formation were higher (P < 0.05) with JI0.3 at Day 6 than in all other treatments. No differences were observed between controls and treatment groups regarding ROS levels and mRNA expression of genes. Viability of resulting oocytes was higher (P < 0.05) in JI0.3 compared to FSH. Interestingly, in control experiment, supplementation of maturation medium with JI0.3 led to higher (P < 0.05) percentages of metaphase II compared to controls. Although more validations will be needed, it seems that this natural extract could be used as a cheap and easily available alternative to commercial FSH.