Effects of Hippotherapy on Postural Control in Children With Cerebral Palsy: A Systematic Review.

PURPOSE: To summarize and update the current knowledge on the effectiveness of hippotherapy on postural control in children with cerebral palsy. METHODS: Using a systematic review methodology, the electronic databases PubMed, Virtual Health Library, PEDro, Scielo, Embase, and Web of Science were searched for eligible articles from 2011 up to September 2021. Quality assessment of eligible studies was performed using the PEDro scale. RESULTS: There were 239 identified studies. Eight clinical trials were selected. The total sample consisted of 264 individuals, with 134 allocated to the experimental group (hippotherapy) and 130 individuals to the control group (conventional therapy). Most studies had moderate to high methodological quality. CONCLUSIONS: Hippotherapy can be an effective intervention to improve several aspects related to postural control, such as static balance (especially in the sitting posture), dynamic balance, and alignment (body posture) in children aged 3 to 16 years, particularly with spastic hemiplegia or diplegia. WHAT THIS ADDS TO THE EVIDENCE: This review summarizes studies that explore potential effects of hippotherapy on postural control in children with cerebral palsy.

Glandular odontogenic cysts: A collaborative investigation of 22 cases and proteins related to invasiveness.

BACKGROUND: A glandular odontogenic cyst (GOC) has an intriguing, aggressive behaviour whose mechanisms have not yet been clarified. OBJECTIVE: To conduct a collaborative cross-sectional study on the clinical, demographic, microscopic and immunohistochemical characteristics of GOCs, emphasizing the histopathological characteristics and expression of proteins related to invasiveness. METHODS: Twenty-two cases of GOC from three oral and maxillofacial pathology services in Brazil were selected from 1988 to 2018. Clinical and demographic data were collected. Histopathological features were evaluated in detail. Sixteen cases of GOC were also submitted to immunohistochemistry to detect MT1-MMP, TKS4, TKS5 and cortactin, the key regulators of invadopodia formation. RESULTS: Glandular odontogenic cysts were primarily seen in men over 40 years of age, in the posterior mandible and the anterior maxilla as a unilocular, radiolucent lesion. All cases presented hobnail cells, clear cells and variable thickness of the lining epithelium, 3 of the 10 key histopathological parameters to be evaluated in GOCs. Immunohistochemistry revealed a greater expression of the studied proteins in the GOCs than in the controls (p < 0.0001). CONCLUSION: Overexpression of proteins that regulate cell invasiveness was identified, and the present study's findings suggest that invadopodia activity is a possible mechanism used by GOCs to promote local invasion, which could partly explain its intriguing biological behaviour.

Impact of the COVID-19 pandemic on public University laboratories of oral and maxillofacial pathology: A Brazilian multicenter study.

OBJECTIVE: To evaluate the impact of the COVID-19 pandemic on the number of diagnoses of oral and maxillofacial lesions in public laboratories after one year of COVID-19 outbreak in Brazil. MATERIAL AND METHODS: This is a cross-sectional study. Biopsies submitted to histopathologic examination from March 2019 to February 2020 (pre-pandemic period) and from April 2020 to March 2021 (pandemic period) in nine Brazilian public oral pathology laboratories were retrieved and the number of diagnoses, types of lesion, and percentage changes during both periods were analyzed. RESULTS: There were 7389 diagnoses in the pre-pandemic period and 2728 in the pandemic era, indicating a reduction of 63.08%. The reduction was 64.23% for benign lesions and 49.48% for malignant lesions, with a 50.64% reduction in squamous cell carcinoma. The largest decreases were observed in April 2020 and January 2021. CONCLUSION: An important reduction in the diagnoses of benign and malignant lesions was noted in the Brazilian public oral pathology laboratories during the first year of the COVID-19 pandemic.

Effects of different amendments on the quality of burnt eucalypt forest soils - A strategy for ecosystem rehabilitation.

The magnitude of forest fires' impacts on the environment is directly related to the changes induced on soil physical, chemical and biological properties. Using available organic resources to rehabilitate burnt forest soils can help reduce post-fire soil fertility loss, accelerating ecosystem recovery. In the present study, the potential of four soil amendments: a mycotechnosol, a eucalypt residue mulch, dredged sediments from a freshwater lagoon and an organic-mineral biofertilizer, to improve the quality of burnt forest soils in terms of organic matter, carbon, nitrogen and phosphorus contents, was evaluated. Two experiments were set-up, one in a recently burnt eucalypt plantation and another in the laboratory using soils from the same area, to assess the effects of the amendments on soil quality, with both experiments lasting for 7 months. The effects of the amendments on nutrient leaching along the soil profile were also evaluated in the laboratory, to investigate possible negative impacts on groundwater and surface water quality. All amendments increased the organic matter and nutrient contents of burnt soils, confirming their potential for ecosystem rehabilitation. The biofertilizer, however, was found to promote nutrient losses by leaching, largely owing to its high solubility, increasing the risk of contamination of ground and surface waters. Using available organic resources to rehabilitate burnt forests as was done in the present work complies with the idea of a circular economy, being key for the sustainability of forest ecosystems.

Intravascular papillary endothelial hyperplasia in the oral mucosa and jawbones: A collaborative study of 20 cases and a systematic review.

BACKGROUND: Intravascular papillary endothelial hyperplasia is an unusual vascular lesion characterized by the proliferation of endothelial cells. The aim of this study was to determine the frequency and general features of this lesion. METHODS: Biopsy records of three oral pathology services were reviewed for intravascular papillary endothelial hyperplasia cases from 1959 to 2020. In addition, a systematic review of case reports and case series was carried out in eight electronic databases. RESULTS: Of the 65 205 retrieved cases, 20 (0.03%) were diagnosed as intravascular papillary endothelial hyperplasia. Mean patient age was 46.55 years, and females (12 cases/60%) were more affected. The lower lip (9 cases/47.36%) was the most commonly affected site, and the lesions were generally asymptomatic (7 cases/63.63%). Clinically, 90% of the lesions presented (18 cases) as a nodule, with a mean size of 1.13 cm. The clinical diagnostic hypotheses most frequently raised were mucocele (6 cases/37.50%) and hemangioma (5 cases/31.25%). An excisional biopsy was chosen in all cases for treatment. Forty-nine studies of the systematic review were included, yielding 105 cases. The literature showed similarity in all variables. CONCLUSION: Despite the uncommon frequency, clinicians and oral pathologists should familiarize themselves with the similarities between intravascular papillary endothelial hyperplasia and some other lesions in terms of clinical and histological features.

Metabolic exchanges between the oocyte and its environment: focus on lipids.

Finely regulated fatty acid (FA) metabolism within ovarian follicles is crucial to follicular development and influences the quality of the enclosed oocyte, which relies on the surrounding intra-follicular environment for its growth and maturation. A growing number of studies have examined the association between the lipid composition of follicular compartments and oocyte quality. In this review, we focus on lipids, their possible exchanges between compartments within the ovarian follicle and their involvement in different pathways during oocyte final growth and maturation. Lipidomics provides a detailed snapshot of the global lipid profiles and identified lipids, clearly discriminating the cells or fluid from follicles at distinct physiological stages. Follicular fluid appears as a main mediator of lipid exchanges between follicular somatic cells and the oocyte, through vesicle-mediated and non-vesicular transport of esterified and free FA. A variety of expression data allowed the identification of common and cell-type-specific actors of lipid metabolism in theca cells, granulosa cells, cumulus cells and oocytes, including key regulators of FA uptake, FA transport, lipid transformation, lipoprotein synthesis and protein palmitoylation. They act in harmony to accompany follicular development, and maintain intra-follicular homeostasis to allow the oocyte to accumulate energy and membrane lipids for subsequent meiotic divisions and first embryo cleavages.

Molecular phylogenetic study in Spirocercidae (Nematoda) with description of a new species Spirobakerus sagittalis sp. nov. in wild canid Cerdocyon thous from Brazil.

The nematode family Spirocercidae Chitwood and Wehr, 1932, comprises three subfamilies, Spirocercinae Chitwood and Wehr, 1932; Ascaropsinae Alicata and McIntosh, 1933; and Mastophorinae Quentin, 1970, which occur worldwide. Spirocercids infect canids and can cause severe illness. The crab-eating fox, Cerdocyon thous (Linnaeus, 1766), is a canid that inhabits most of South America, including Brazil and is a host for several parasitic worms, in particular, nematodes. However, few reports or genetic data are available on the spirocercids found in this host. In the present study, we describe a new species of Spirobakerus Chabaud and Bain 1981, from the intestine of two crab-eating foxes from two different biomes in Brazil. Spirobakerus sagittalis sp. nov. presents a) unequal spicules, with a long, thin left spicule with a lanceolated shape at the tip; b) a pair of sessile papillae and a median unpaired papillae located anteriorly of the cloaca, and c) a tuft without spines at the tip of the tail in females. The molecular phylogenetic analysis indicated that S. sagittalis sp. nov. is basal to the other species of the subfamily Ascaropsinae, which was not recovered as monophyletic. Our phylogenies also indicated that Spirocercidae is paraphyletic, given that Mastophorinae did not group with Ascaropsinae and Spirocercinae. We provide the first molecular data on the genus Spirobakerus and expand the molecular database of the spirocercids. However, further studies, including the sequences of other spirocercid taxa, are still needed to infer the relationships within this family more accurately.

The Defibrillation Conundrum: New Insights into the Mechanisms of Shock-Related Myocardial Injury Sustained from a Life-Saving Therapy.

Implantable cardiac defibrillators (ICDs) are recommended to prevent the risk of sudden cardiac death. However, shocks are associated with an increased mortality with a dose response effect, and a strategy of reducing electrical therapy burden improves the prognosis of implanted patients. We review the mechanisms of defibrillation and its consequences, including cell damage, metabolic remodeling, calcium metabolism anomalies, and inflammatory and pro-fibrotic remodeling. Electrical shocks do save lives, but also promote myocardial stunning, heart failure, and pro-arrhythmic effects as seen in electrical storms. Limiting unnecessary implantations and therapies and proposing new methods of defibrillation in the future are recommended.

Protein Palmitoylation in Bovine Ovarian Follicle.

Protein palmitoylation is a reversible post-translational modification by fatty acids (FA), mainly a palmitate (C16:0). Palmitoylation allows protein shuttling between the plasma membrane and cytosol to regulate protein stability, sorting and signaling activity and its deficiency leads to diseases. We aimed to characterize the palmitoyl-proteome of ovarian follicular cells and molecular machinery regulating protein palmitoylation within the follicle. For the first time, 84 palmitoylated proteins were identified from bovine granulosa cells (GC), cumulus cells (CC) and oocytes by acyl-biotin exchange proteomics. Of these, 32 were transmembrane proteins and 27 proteins were detected in bovine follicular fluid extracellular vesicles (ffEVs). Expression of palmitoylation and depalmitoylation enzymes as palmitoyltransferases (ZDHHCs), acylthioesterases (LYPLA1 and LYPLA2) and palmitoylthioesterases (PPT1 and PPT2) were analysed using transcriptome and proteome data in oocytes, CC and GC. By immunofluorescence, ZDHHC16, PPT1, PPT2 and LYPLA2 proteins were localized in GC, CC and oocyte. In oocyte and CC, abundance of palmitoylation-related enzymes significantly varied during oocyte maturation. These variations and the involvement of identified palmitoyl-proteins in oxidation-reduction processes, energy metabolism, protein localization, vesicle-mediated transport, response to stress, G-protein mediated and other signaling pathways suggests that protein palmitoylation may play important roles in oocyte maturation and ffEV-mediated communications within the follicle.

Intergenerational breastfeeding practices among parents and children: 1993 Pelotas (Brazil) birth cohort.

The objective of this study was to investigate intergenerational breastfeeding practices according to parental sex and age at delivery in the 1993 Pelotas (Brazil) birth cohort study. This is a prospective birth cohort study, and at the 22-year follow-up, a substudy with all children of the cohort members who had become parents was conducted (93Cohort-II). First generation breastfeeding data were collected at 3 months and 4-year-old follow-ups. In the 93Cohort-II, parents answered a questionnaire about their children's breastfeeding practices. Adjusted Tobit and Poisson regression models with robust variance were applied to estimate the association between predominant parental breastfeeding duration and exclusive breastfeeding duration of the children at 3 and 6 months. Out of 3,810 cohort participants, 955 (25%) had delivered at least one live-born infant, and 1,222 children were assessed. Fifty-four percent of parents were ≤19 years old. Direct effects of predominant parental breastfeeding duration on exclusive breastfeeding duration of their children were only observed when data were stratified by parental age: children born to parents aged ≥20 years old and who were predominantly breastfed for at least 3 months presented higher exclusive breastfeeding duration and higher prevalence of being exclusively breastfed for at least 3 months. When analyses were stratified by mothers and fathers, the result remained significant only among mothers. Longer predominant breastfeeding duration in the first generation was associated with longer exclusive breastfeeding duration in the second generation, but only among older mothers. Education and social support surrounding breastfeeding should be intensified among fathers and younger parents to create a positive environment supportive of breastfeeding.

Stability of extemporaneous sulfadiazine oral suspensions from commercially available tablets for treatment of congenital toxoplasmosis.

OBJECTIVES: To determine the physicochemical and microbiological stability of sulfadiazine suspensions (100 mg/mL) in simple syrup (A) and sorbitol (B) formulations prepared from commercially available tablets. METHODS: An ultra-performance liquid chromatographic assay was developed and validated to determine the chemical stability of sulfadiazine. Three samples were prepared and stored at 5 and 25 °C and assayed at 0, 7, 14 and 30 days. Physical parameters (appearance, pH, particle size and viscosity) were also monitored. Microbiological examination was performed through the suitable counting method. RESULTS: The formulations presented a sulfadiazine concentration of around 95% at the beginning at both temperatures. There was some variation in pH, viscosity and particle size distribution over time. The samples met the pharmacopoeia criteria of microbiological quality over 30 days, but only sulfadiazine formulated in syrup stored at 25 °C was suitable for use after one week. CONCLUSION: The sulfadiazine suspension in simple syrup was chosen as the most suitable formulation because it demonstrated stability for 14 days at room temperature, providing an alternative liquid dosage form of sulfadiazine for congenital toxoplasmosis treatment.

OBJECTIFS: Déterminer la stabilité physicochimique et microbiologique de suspensions de sulfadiazine (100 mg/mL) dans des formulations de sirop simple (A) et de sorbitol (B) préparées à partir de comprimés disponibles dans le commerce. MÉTHODES: Un test de chromatographie liquide ultra-performante a été développé et validé pour déterminer la stabilité chimique de la sulfadiazine. Trois échantillons ont été préparés et stockés à 5 ºC et à 25 ºC et analysés à 0, 7, 14 et 30 jours. Les paramètres physiques (apparence, pH, granulométrie et viscosité) ont également été contrôlés. Un examen microbiologique a été effectué par la méthode de comptage appropriée. RÉSULTATS: Les formulations présentaient une concentration en sulfadiazine d'environ 95% au début aux deux températures. Il y avait une certaine variation du pH, de la viscosité et de la distribution de la taille des particules au fil du temps. Les échantillons répondaient aux critères de pharmacopée pour la qualité microbiologique aprè 30 jours, mais seule la sulfadiazine formulée dans du sirop conservé à 25 ºC pouvait être utilisée après une semaine. CONCLUSION: La suspension de sulfadiazine dans un sirop simple a été choisie comme la formulation la plus appropriée car elle a démontré une stabilité à 14 jours à température ambiante, fournissant une forme galénique liquide alternative de sulfadiazine pour le traitement de la toxoplasmose congénitale.

Clinicopathological and immunohistochemical features of the oral lymphoepithelial cyst: A multicenter study.

BACKGROUND: Analyze the clinical, demographic, histopathological, and immunohistochemical features of oral lymphoepithelial cyst (OLEC). METHODS: Samples were retrospectively retrieved from five oral pathology services. Clinical and demographic data were collected from patient charts. Histopathological and immunohistochemical (CD3 and CD20) features were evaluated. Data analysis involved descriptive statistics and bivariate analyses (P ≤ .05). RESULTS: Seventy-seven cases were found among a total of 146 150 specimens (0.05%). OLEC was predominantly diagnosed in females (70.1%). Mean patient age was 46.51 years. The lesions arose mainly on the lateral border of the tongue (40.3%), measured up to 1 cm (61.0%), and were asymptomatic (64.9%). Twenty-four lesions (31.2%) were white. Forty-one cases (53.2%) presented lymphocytic inflammatory infiltrate with no specific arrangement. The cystic lining was composed of a non-keratinized stratified epithelium (59.7%) presenting hyperplasia (39.0%). Connection with the surface, epithelium was found in 23 cases (29.9%) and 31 (40.3%) cases had two or more cystic cavities. The lumen content was predominantly desquamated cells (48.1%). Subgemmal neurogenous plaque was found in 11/42 (26.2%) cases involving the tongue. CD20+ cells predominated in 36/63 cases (57.2%), and lymphocytic inflammatory infiltrate was not always continuous around the cystic cavity (52.4%). CONCLUSION: Lymphoepithelial cyst is an uncommon lesion of the oral cavity. The present study offers the largest sample of OLEC for which clinical, demographic, histopathological, and immunohistochemical features were evaluated. The clinical and demographic findings were similar to those described in previous reports, but the microscopic analyses revealed interesting aspects of the cystic epithelium and the lymphocytic inflammatory infiltrate in OLEC.

A Brazilian multicentre study of 2,497 isolated cases of odontogenic keratocysts.

We present the frequency of cases of isolated odontogenic keratocysts submitted to microscopic examination at 10 Brazilian referral centres in Oral and Maxillofacial Pathology. In a retrospective (1953-2017) analysis, data on clinicoradiographic features and treatment of these lesions were collected and analysed descriptively. Among the 258,867 cases retrieved, 2,497 (0.96%) were isolated odontogenic keratocysts. In summary, an overview of individuals affected with isolated odontogenic keratocysts is reported herein. This lesion showed predilection for the posterior mandible of young adult men.

Identification of 56 Proteins Involved in Embryo-Maternal Interactions in the Bovine Oviduct.

The bovine embryo develops in contact with the oviductal fluid (OF) during the first 4-5 days of pregnancy. The aim of this study was to decipher the protein interactions occurring between the developing embryo and surrounding OF. In-vitro produced 4-6 cell and morula embryos were incubated or not (controls) in post-ovulatory OF (OF-treated embryos) and proteins were then analyzed and quantified by high resolution mass spectrometry (MS) in both embryo groups and in OF. A comparative analysis of MS data allowed the identification and quantification of 56 embryo-interacting proteins originated from the OF, including oviductin (OVGP1) and several annexins (ANXA1, ANXA2, ANXA4) as the most abundant ones. Some embryo-interacting proteins were developmental stage-specific, showing a modulating role of the embryo in protein interactions. Three interacting proteins (OVGP1, ANXA1 and PYGL) were immunolocalized in the perivitelline space and in blastomeres, showing that OF proteins were able to cross the zona pellucida and be taken up by the embryo. Interacting proteins were involved in a wide range of functions, among which metabolism and cellular processes were predominant. This study identified for the first time a high number of oviductal embryo-interacting proteins, paving the way for further targeted studies of proteins potentially involved in the establishment of pregnancy in cattle.

MALDI-TOF Mass Spectrometry Revealed Significant Lipid Variations in Follicular Fluid and Somatic Follicular Cells but Not in Enclosed Oocytes between the Large Dominant and Small Subordinate Follicles in Bovine Ovary.

Lipid metabolism in ovarian follicular cells supports the preparation of an enclosed oocyte to ovulation. We aimed to compare lipid composition of a dominant large follicle (LF) and subordinated small follicles (SFs) within the same ovaries. Mass spectrometry imaging displayed the differences in the distribution of several lipid features between the different follicles. Comparison of lipid fingerprints between LF and SF by Matrix Assisted Laser Desorption/Ionisation Time-Of-Flight (MALDI-TOF) mass spectrometry revealed that in the oocytes, only 8 out of 468 detected lipids (1.7%) significantly changed their abundance (p < 0.05, fold change > 2). In contrast, follicular fluid (FF), granulosa, theca and cumulus cells demonstrated 55.5%, 14.9%, 5.3% and 9.8% of significantly varied features between LF and SF, respectively. In total, 25.2% of differential lipids were identified and indicated potential changes in membrane and signaling lipids. Tremendous changes in FF lipid composition were likely due to the stage specific secretions from somatic follicular cells that was in line with the differences observed from FF extracellular vesicles and gene expression of candidate genes in granulosa and theca cells between LF and SF. In addition, lipid storage in granulosa and theca cells varied in relation to follicular size and atresia. Differences in follicular cells lipid profiles between LF and SF may probably reflect follicle atresia degree and/or accumulation of appropriate lipids for post-ovulation processes as formation of corpus luteum. In contrast, the enclosed oocyte seems to be protected during final follicular growth, likely due in part to significant lipid transformations in surrounding cumulus cells. Therefore, the enclosed oocyte could likely keep lipid building blocks and energy resources to support further maturation and early embryo development.

Osteonecrosis of the jaw as a late complication of herpes zoster infection: a case report.

Occurrences of rare oral complications following herpes zoster (HZ) infection have been reported. In the present case, a 57-year-old man was referred for periodontal evaluation due to gingival bleeding. His medical history included diagnoses of chronic lymphocytic leukemia and HZ infection. Intraoral examination revealed necrosis and alveolar bone exposure around the mandibular left lateral incisor, and the patient reported spontaneous exfoliation of the maxillary and mandibular left central incisors. Conservative surgical and antibiotic therapies were provided to the patient, and a diagnosis of osteonecrosis of the jaw following HZ infection was established. There were no signs of recurrence in 17 months of follow-up. Clinicians should be aware of unusual complications related to a previous HZ infection.

Binder of Sperm Proteins 1 and 5 have contrasting effects on the capacitation of ram spermatozoa.

Binder of Sperm Proteins (BSPs) are the most abundant seminal plasma protein family in the ram and bull. They have been extensively studied in the bull but less is known about their function in ovine seminal plasma and current knowledge suggests that BSPs may have different effects in these two species. In the bull, they facilitate capacitation and destabilize the sperm membrane during in vitro handling, whereas in the ram, they appear to stabilize the sperm membrane and prevent cryopreservation-induced capacitation-like changes. Further investigation into the effects of BSPs on ram spermatozoa under capacitating conditions is required to further clarify their physiological roles in the ram. We investigated the effects of Binder of Sperm Proteins 1 and 5 on epididymal ram spermatozoa in conditions of low, moderate, and high cAMP. BSPs had minimal effects on sperm function in low-cAMP conditions, but caused significant changes under cAMP upregulation. BSP1 stabilized the membrane and qualitatively reduced protein tyrosine phosphorylation, but significantly increased cholesterol efflux and induced spontaneous acrosome reactions. BSP5 slightly increased spontaneous acrosome reactions and caused sperm necrosis. However, BSP5 had minimal effects on membrane lipid order and cholesterol efflux and did not inhibit protein tyrosine phosphorylation. These findings demonstrate that under maximal cAMP upregulation, BSP1 affected ram spermatozoa in a manner comparable to bull spermatozoa, while BSP5 did not.

Odontogenic myxoma: A 63-year retrospective multicenter study of 85 cases in a Brazil population and a review of 999 cases from literature.

BACKGROUND: Odontogenic myxoma (OM) is an uncommon neoplasm of the jaws. Considering the importance of defining the relative incidence and demographic profile of these lesions in South America, the aim of this study was to analyze the clinical and imagiological features of OM from three South American oral pathology services and to discuss these findings in light of the literature. METHODS: Data regarding age, gender, anatomic site, and imagiological features from 85 cases of OM were collected. Additionally, we did a review of OM studies published in three electronic databases. RESULTS: Among 63 450 oral biopsies, 1178 (1.85%) were odontogenic tumors (World Health Organization - 2017), of which 85 (7.21%) met the criteria of OM. The mean age was 30.7 years (range: 10-61 years; SD: 12.22). Forty-five (52.9%) cases occurred in females and 40 (47.1%) in males (ratio: 1:1.12). Maxilla was affected in 44 cases (53%) and mandible in 39 (47%). Of the 41 informed cases (48.2%), all of them were radiolucent lesions. The literature review indicated a majority of mean ages in third decade and a predilection for females, mandible, and multilocular radiolucent lesions. CONCLUSIONS: The features of OM samples have strong similarity to that reported in studies from other continents. It is possible to infer that geographic variation does not help to explain some differences observed in the clinical features of OM.

Intact Cell MALDI-TOF MS on Sperm: A Molecular Test For Male Fertility Diagnosis.

Currently, evaluation of sperm quality is primarily based on in vitro measures of sperm function such as motility, viability and/or acrosome reaction. However, results are often poorly correlated with fertility, and alternative diagnostic tools are therefore needed both in veterinary and human medicine. In a recent pilot study, we demonstrated that MS profiles from intact chicken sperm using MALDI-TOF profiles could detect significant differences between fertile/subfertile spermatozoa showing that such profiles could be useful for in vitro male fertility testing. In the present study, we performed larger standardized experimental procedures designed for the development of fertility- predictive mathematical models based on sperm cell MALDI-TOF MS profiles acquired through a fast, automated method. This intact cell MALDI-TOF MS-based method showed high diagnostic accuracy in identifying fertile/subfertile males in a large male population of known fertility from two distinct genetic lineages (meat and egg laying lines). We additionally identified 40% of the m/z peaks observed in sperm MS profiles through a top-down high-resolution protein identification analysis. This revealed that the MALDI-TOF MS spectra obtained from intact sperm cells contained a large proportion of protein degradation products, many implicated in important functional pathways in sperm such as energy metabolism, structure and movement. Proteins identified by our predictive model included diverse and important functional classes providing new insights into sperm function as it relates to fertility differences in this experimental system. Thus, in addition to the chicken model system developed here, with the use of appropriate models these methods should effectively translate to other animal taxa where similar tests for fertility are warranted.

Binder of Sperm Proteins protect ram spermatozoa from freeze-thaw damage.

Cryopreservation causes sub-lethal damage which limits the fertility of frozen thawed spermatozoa. Seminal plasma has been investigated as a cryoprotectant, but has yielded inconsistent results due to considerable variation in its constituents. Individual seminal plasma proteins offer an ideal alternative to whole seminal plasma, and several have been correlated with freezing success. Binder of Sperm Proteins (BSPs) are abundant ram seminal plasma proteins which have been suggested to have significant protective effects on ram spermatozoa during cold shock. This is in direct opposition to bull spermatozoa, where BSPs cause sperm deterioration during in vitro handling. We investigated the potential of BSP1 and BSP5 to prevent freezing associated damage to important functional parameters of ram spermatozoa. BSPs purified by size exclusion chromatography improved post thaw motility and penetration through artificial mucus. Highly purified BSP1 and BSP5, isolated by gelatin affinity and RP-HPLC, improved motility and membrane integrity, and reduced post thaw protein tyrosine phosphorylation. Exposure to BSP5 before freezing increased the amount of phosphatidylethanolamine on the sperm surface after thawing. Neither BSP1 nor BSP5 prevented freezing associated changes in membrane lipid disorder. These results suggest that BSPs may significantly improve freezing outcomes of ram spermatozoa.