Triheptanoin, an odd-medium-chain triglyceride, impacts brain cognitive function in young and aged mice.

ABSTRACTThe brain aging process triggers cognitive function impairment, such as memory loss and compromised quality of life. Cognitive impairment is based on bioenergetic status, with reduced glucose uptake and metabolism in aged brains. Anaplerotic substrates are reported to promote mitochondrial ATP generation, having been tested in clinical trials for the treatment of neurological disorders and metabolic diseases.Objectives and Methods: To assess whether the improvement in oxidative capacity ameliorates cognitive function in adults (12 weeks), and aged (22-month-old) C57/6BJ mice, they received (1) a ketogenic diet, (2) a ketogenic diet supplemented with the anaplerotic substance, triheptanoin, or (3) a control diet for 12 weeks. Spontaneous alternation and time spent in a previously closed arm in the Y-maze test and time interacting with an unknown object in the novel object recognition test (NORT) were used to evaluate working memory. Acetylcholinesterase (AChE) activity in the prefrontal lobe, brain left hemisphere, and cerebellum was also evaluated. Glucose transporter 3 (GLUT3) expression in the prefrontal lobe was analyzed by western blotting.Results: The ketogenic diet (KD) reduced spontaneous alternation in aged mice, leading to lower AChE activity in the aged prefrontal lobe and cerebellum, and in the parieto-temporal-occipital lobe of adult mice. Furthermore, KD decreased GLUT3 protein expression in the frontal lobe of the adults.Discussion: Supplementation of KD with triheptanoin prevented memory impairment and showed similar values of AChE activity and GLUT3 expression compared to the controls. Our data suggest that triheptanoin has a potential role in the bioenergetic capacity of the brain, improving cognitive function.

Software testing in microbial bioinformatics: a call to action.

Computational algorithms have become an essential component of research, with great efforts by the scientific community to raise standards on development and distribution of code. Despite these efforts, sustainability and reproducibility are major issues since continued validation through software testing is still not a widely adopted practice. Here, we report seven recommendations that help researchers implement software testing in microbial bioinformatics. We have developed these recommendations based on our experience from a collaborative hackathon organised prior to the American Society for Microbiology Next Generation Sequencing (ASM NGS) 2020 conference. We also present a repository hosting examples and guidelines for testing, available from https://github.com/microbinfie-hackathon2020/CSIS.

Atrial matrix remodeling in atrial fibrillation patients with aortic stenosis.

BACKGROUND: This study aimed to evaluate atrium extracellular matrix remodeling in atrial fibrillation (AF) patients with severe aortic stenosis, through histological fibrosis quantification and extracellular matrix gene expression analysis, as well as serum quantification of selected protein targets. METHODS: A posthoc analysis of a prospective study was performed in a cohort of aortic stenosis patients. Between 2014 and 2019, 56 patients with severe aortic stenosis submitted to aortic valve replacement surgery in a tertiary hospital were selected. RESULTS: Fibrosis was significantly increased in the AF group when compared to sinus rhythm (SR) patients (p = 0.024). Moreover, cardiomyocyte area was significantly higher in AF patients versus SR patients (p = 0.008). Conversely, collagen III gene expression was increased in AF patients (p = 0.038). TIMP1 was less expressed in the atria of AF patients. MMP16/TIMP4 ratio was significantly decreased in AF patients (p = 0.006). TIMP1 (p = 0.004) and TIMP2 (p = 0.012) were significantly increased in the serum of AF patients. Aortic valve maximum (p = 0.0159) and mean (p = 0.031) gradients demonstrated a negative association with serum TIMP1. CONCLUSIONS: Atrial fibrillation patients with severe aortic stenosis present increased atrial fibrosis and collagen type III synthesis, with extracellular matrix remodelling demonstrated by a decrease in the MMP16/TIMP4 ratio, along with an increased serum TIMP1 and TIMP2 proteins.

Pontine Myelinolysis Caused by Hypovolemic Hypernatremia.

Introduction. Central pontine myelinolysis is characterized by the occurrence of acute demyelinating lesions of cells in the pons secondary to abrupt oscillations of serum osmolarity. Its exact incidence is not well defined, but studies show a prevalence of 0.25 to 0.5% in the general population, 2.5% in the intensive care unit, and up to 10% in patients with risk factors, such as chronic liver disease and hepatic transplantation, alcoholism, malnutrition, diuretic therapy, electrolyte imbalance, hypoglycemia, and hyperglycemia. Case Report. A 70-year-old white female with extranodal diffuse large B-cell non-Hodgkin's lymphoma (extensive mass on the left anterior chest wall), stage IVA, developed pontine myelinolysis secondary to hypovolemic acute hypernatremia, which occurred due to diarrhea caused by chemotherapy (rituximab, cyclophosphamide, doxorubicin, and vincristine). Discussion. Pontine myelinolysis occurs most often due to the rapid correction of chronic hyponatremia. But here, we describe a case of the disease secondary to the occurrence of hypovolemic acute hypernatremia in a patient with a hematological malignancy under treatment, who was on chronic treatment with thiazide diuretics and who presented with other electrolyte disturbances as risk factors for the development of pontine myelinolysis.

Diferentes momentos de aplicação de gonadotrofina coriônica equina em protocolo de inseminação artificial em tempo fixo para vacas de leite / Different times of application of equine chorionic gonadotropin in fixed-time artificial insemination protocols for dairy cows

Avaliaram-se diferentes momentos de aplicação da gonadotrofina coriônica equina (eCG) em protocolos de inseminação artificial em tempo fixo (IATF) para vacas de leite. Foram utilizadas 76 fêmeas, as quais receberam, no dia zero (D0) do protocolo, dispositivos intravaginais de progesterona, sendo esses retirados no D9, e os animais foram, então, distribuídos aleatoriamente em três tratamentos: T1 - aplicação de eCG no momento da retirada dos dispositivos; T2 e T3 - aplicação de eCG 48h e 24h antes da retirada dos dispositivos, respectivamente. No D10 os animais receberam 1mg de GnRH, e a IATF foi realizada 52 horas após a retirada do implante. Não houve diferença (P>0,05) para intervalo entre a retirada do implante à ovulação (72,56±3,92h), o diâmetro do maior folículo no D9 (10,88±1,49mm), o diâmetro do folículo ovulatório (15,15±1,16mm) e do segundo maior folículo (7,49±0,52mm), a taxa de crescimento folicular (1,38±0,04mm/dia), a taxa de ovulação (96,67%), o intervalo entre diâmetro final e inicial do folículo dominante (73,49±3,84h), a área de corpo lúteo (2,27±0,43cm²), a porcentagem de CL no ovário direito (53,00%) e no esquerdo (26,33%) e a taxa de gestação (33,33%). O momento da aplicação da eCG não influenciou na eficiência do protocolo. Recomenda-se a utilização da eCG no momento da retirada do implante por otimização do manejo.(AU)

Different times of application of equine chorionic gonadotropin (eCG) in Fixed-Time Artificial Insemination (FTAI) protocols for dairy cows were evaluated. A total of 76 females were used, which received intravaginal progesterone devices on day zero (D0) of the protocol, which were withdrawn on D9, and the animals were then randomly distributed in three treatments: T1- application of eCG at the time of device withdrawal; T2 and T3 - application of eCG 48h and 24h before withdrawal of the devices, respectively. On D10 the animals received 1mg of GnRH and the FTAI was carried out 52 hours after the removal of the implant. There was no difference (P>0.05) for interval from implant removal to ovulation (72.56±3.92h), diameter of the largest follicle of D9 (10.88±1.49mm), diameters of ovulatory follicle (15.15±1.16mm) and of the second largest follicle (7.49±0.52mm), follicle growth rate (1.38±0.04mm/day), ovulation rate (96.67%), interval between final diameter and initial diameter of dominant follicle (73.49±3.84h), corpus luteum area (2.27±0.43cm²), percentage of CL in right ovary (53.00%) and in left ovary (26.33%) and pregnancy rate (33.33%). The time of the application of eCG did not influence the efficiency of the protocol. We recommend the use of eCG at the time of the removal of the implant for management optimization.(AU)

Technical approaches to evaluate the surfactant-enhanced biodegradation of biodiesel and vegetable oils.

This research compared the effects of biosurfactant on the biodegradation of biodiesel and vegetable oils while validating two conceptually diverging methodologies. The two experimental setups were successfully modeled towards the effects of biosurfactants during biodegradation. We established the equivalence of both methodologies from the data output. As expected, the biosurfactants caused an increased oil uptake, thus increasing biodegradation performance. Cooking oils were favored by the microbial consortium as a carbon source when compared with biodiesel fuel, especially after use in food preparation. However, we found that biodiesel substrate standout with the highest biodegradation rates. Our results might indicate that a rapid metabolic change from the original compound initially favored biodiesels during the assimilation of organic carbon for a set specialized microbial inoculum. The data output was successfully combined with mathematical models and statistical tools to describe and predict the actual environmental behavior of biodiesel and vegetable oils. The models confirmed and predicted the biodegradation effectiveness with biosurfactants and estimated the required timeframe to achieve satisfactory contaminant removal.

Decay of the distance autocorrelation and Lyapunov exponents.

This work presents numerical evidence that for discrete dynamical systems with one positive Lyapunov exponent the decay of the distance autocorrelation is always related to the Lyapunov exponent. Distinct decay laws for the distance autocorrelation are observed for different systems, namely, exponential decays for the quadratic map, logarithmic for the Hénon map, and power-law for the conservative standard map. In all these cases the decay exponent is close to the positive Lyapunov exponent. For hyperchaotic conservative systems the power-law decay of the distance autocorrelation is not directly related to any Lyapunov exponent.

Qualidade seminal e perfil metabólico de caprinos alimentados com semente de linhaça na dieta / Semen quality and metabolic profile of goats fed flaxseed in the diet

O objetivo da presente pesquisa foi avaliar o efeito da inclusão de semente de linhaça na dieta sobre a qualidade seminal e o perfil metabólico de machos caprinos. Foram utilizados 16 machos da raça Saanen, distribuídos aleatoriamente em quatro grupos, recebendo níveis de inclusão de semente de linhaça (0, 4, 8 e 12%) na dieta, durante um período de 60 dias. As coletas e as avaliações seminais foram realizadas duas vezes por semana. Os animais foram submetidos a coletas quinzenais de sangue, durante todo o período experimental. Os dados foram avaliados por ANOVA e pela análise de regressão a 5% de significância. Houve comportamento cúbico para motilidade espermática progressiva, que apresentou o maior valor (82,30%) com a adição de 9,92% de semente de linhaça na dieta (P<0,05). Houve comportamento cúbico para concentração plasmática de triglicerídeos, e o nível de 8% de semente de linhaça na dieta apresentou média de 43,32mg dL-1, acima dos valores normais para a espécie caprina (P<0,05). A inclusão de semente de linhaça na dieta de machos caprinos proporcionou melhora na produção espermática e na qualidade seminal. No entanto, devem-se considerar os possíveis efeitos dos níveis superiores a 4% de semente de linhaça sobre o perfil metabólico dos animais.(AU)

This study aimed to evaluate the effect of flaxseed inclusion in the diet of male goats through the semen quality and metabolic profile. Randomly allocated 16 Saanen males were placed into four groups receiving flaxseed inclusion levels (0, 4, 8 and 12%) in the diet over a period of 60 days. The collections and seminal evaluations were performed twice a week. The animals were subjected to biweekly blood collections from the beginning to the end of the trial period. The data were evaluated by ANOVA and regression analysis at 5% significance. A cubic behavior for motility, with the highest value (82.30%) with the addition of 9.92% of flaxseed in the diet (P< 0.05) was detected. A cubic behavior for plasma triglyceride concentration and the level of 8% of flaxseed in the diet averaged 43.32mg dL -1 above normal values for goats (P< 0.05) was detected. The inclusion of flaxseed in the diet of male goats improved sperm production and quality. However, one should consider the possible effects of the levels above 4% of flaxseed on the metabolic profile of the animals.(AU)

Diagnostic accuracy of thromboelastometry and its correlation with the HPLC-MS/MS quantification test.

The aim of the study was to evaluate the diagnostic accuracy of thromboelastometry for assessing rivaroxaban concentrations. The accuracy of thromboelastometry was compared with the high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method, which is the gold standard for drug plasma monitoring (the reference standard). Forty-six clinically stable patients were treated with 10, 15, or 20 mg of rivaroxaban once daily (OD group) or 15 mg twice a day (BID group) (no particular indication for treatment). Patient samples were collected 2 h after the use of the medication (peak) and 2 h before the next dose (trough). The rivaroxaban plasma concentrations were determined via HPLC-MS/MS, and thromboelastometry was performed using a ROTEM® delta analyzer. There were significant prolongations in clotting time (CT) for the 10, 15, and 20 mg of rivaroxaban treatments in the OD groups. In the 15 mg BID group, the responses at the peak and trough times were similar. At the peak times, there was a positive correlation between the plasma concentration of rivaroxaban and CT (Spearman correlation rho=0.788, P<0.001) and clot formation time (rho=0.784, P<0.001), and a negative correlation for alpha angle (rho=-0.771, P<0.001), amplitude after 5 min (rho=-0.763, P<0.001), and amplitude after 10 min (rho=-0.680, P<0.001). The CT presented higher specificity and sensitivity using the cut-off determined by the receiver characteristics curve. ROTEM has potential as screening tool to measure possible bleeding risk associated with rivaroxaban plasma levels.

Localization-delocalization transition in discrete-time quantum walks with long-range correlated disorder.

We study the effects of spatially long-range correlated phase disorder on the Hadamard quantum walk on a line. The shift operator is built to exhibit an intrinsic disorder distribution featuring long-range correlations. To impose such, we resort to fractional Brownian motion with power-law spectrum 1/k^{2α} with α≥0 being the exponent that controls the degree of correlations. We discuss the scaling behavior of the walker's wave packet and report a localization-delocalization transition controlled by α. We unveil two intermediate dynamical regimes between exponential localization and full delocalization.

Detection of protamine 2 in bovine spermatozoa and testicles.

BACKGROUND: Sperm DNA integrity is crucial for transmission of genetic information to future generations and DNA damage can occur during chromatin packaging. Chromatin packaging involves the replacement of somatic nucleosomal histones by nuclear proteins called protamines. Protamine 1 (PRM1) is transcribed and translated in spermatids of all mammals; however, protamine 2 (PRM2) is transcribed in low levels in spermatids and it is not yet described in bull mature spermatozoa. OBJECTIVES: The aim of this study was to assess gene and protein expression of PRM2 and corroborate gene and protein expression of PRM1 in bull spermatozoa and testis. MATERIALS AND METHODS: For this purpose, absolute q-RT-PCR was performed to calculate the number of copies of PRM1 and PRM2 mRNAs in bovine epididymal spermatozoa and testicular tissue. Western blot and mass spectrometry were performed to identify PRM1 and PRM2 in samples of bovine epididymal spermatozoa. Samples of bovine testicular tissue were collected to identify PRM1 and PRM2 by immunohistochemistry. RESULTS: We evaluated that the number of PRM1 mRNA copies was about hundred times higher than PRM2 mRNA copies in sperm and testicular samples (p < 0.0001). In addition, we estimated the PRM1: PRM2 ratio based on mRNA number of copies. In spermatozoa, the ratio was 1: 0.014, and in testicle, the ratio was 1: 0.009. We also evaluated the immunolocalization for PRM1 and PRM2 in bovine testis, and both proteins were detected in spermatids. Western blot and mass spectrometry in bovine epididymal spermatozoa confirmed these results. CONCLUSION: Our work identifies, for the first time, PRM2 in bovine epididymal spermatozoa and in testis. Further studies are still needed to understand the role of PRM2 on the chromatin of the spermatozoa and to verify how possible changes in PRM2 levels may influence the bull fertility.

Combined open and laparoscopic approach for repair of flank hernias: technique description and medium-term outcomes of a single surgeon.

PURPOSE: A residual bulge in the lateral abdominal wall is a reason for patient dissatisfaction after flank hernia repair (FHR). We hypothesized that combining a laparoscopically-placed intraperitoneal mesh (IPOM) with onlay hernia repair performed through a small open incision would increase repair durability and decrease such residual bulges. We aim to report our medium-term outcomes with this technique. METHODS: Patients who have undergone FHR using the technique described above from March 2013 through June 2017 were identified in a prospectively maintained database. Outcomes of interest included surgical site infections (SSI), surgical site occurrences (SSO), surgical site occurrences requiring procedural intervention (SSOPI) and hernia recurrence. RESULTS: Sixteen patients were identified (62% females; mean age 59 ± 8 years, mean body mass index 29.5 kg/m2). Mean hernia width was. 6.4 ± 3 cm and 31% were recurrent hernias previously repaired through an onlay approach. Mean operative time was 159 ± 40 min, fascial closure was achieved in all cases, and there were no intraoperative complications. Median length of stay was 3 days (IQR 3-4), and there were no unplanned readmissions or reoperations. At a median 37-month follow-up (IQR 21-55), wound morbidity rate was 12.5% (2 seromas). There were no SSI/SSOPI and one hernia recurrence (6%) was detected at 12 months postoperatively. CONCLUSION: Combining laparoscopic IPOM with open onlay hernia repair resulted in low recurrence and acceptable wound morbidity rates, with no residual bulges noted at medium-term follow-up. Further studies with larger number of patients and other surgeon's experiences are necessary to determine the role of such technique in the surgical armamentarium for flank hernia repair.

Diagnostic accuracy of thromboelastometry and its correlation with the HPLC-MS/MS quantification test

The aim of the study was to evaluate the diagnostic accuracy of thromboelastometry for assessing rivaroxaban concentrations. The accuracy of thromboelastometry was compared with the high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method, which is the gold standard for drug plasma monitoring (the reference standard). Forty-six clinically stable patients were treated with 10, 15, or 20 mg of rivaroxaban once daily (OD group) or 15 mg twice a day (BID group) (no particular indication for treatment). Patient samples were collected 2 h after the use of the medication (peak) and 2 h before the next dose (trough). The rivaroxaban plasma concentrations were determined via HPLC-MS/MS, and thromboelastometry was performed using a ROTEM® delta analyzer. There were significant prolongations in clotting time (CT) for the 10, 15, and 20 mg of rivaroxaban treatments in the OD groups. In the 15 mg BID group, the responses at the peak and trough times were similar. At the peak times, there was a positive correlation between the plasma concentration of rivaroxaban and CT (Spearman correlation rho=0.788, P<0.001) and clot formation time (rho=0.784, P<0.001), and a negative correlation for alpha angle (rho=−0.771, P<0.001), amplitude after 5 min (rho=−0.763, P<0.001), and amplitude after 10 min (rho=−0.680, P<0.001). The CT presented higher specificity and sensitivity using the cut-off determined by the receiver characteristics curve. ROTEM has potential as screening tool to measure possible bleeding risk associated with rivaroxaban plasma levels.

Detection of protamine 2 in bovine spermatozoa and testicles

Background Sperm DNA integrity is crucial for transmission of genetic information to future generations and DNA damage can occur during chromatin packaging. Chromatin packaging involves the replacement of somatic nucleosomal histones by nuclear proteins called protamines. Protamine 1 (PRM1) is transcribed and translated in spermatids of all mammals; however, protamine 2 (PRM2) is transcribed in low levels in spermatids and it is not yet described in bull mature spermatozoa. Objectives The aim of this study was to assess gene and protein expression of PRM2 and corroborate gene and protein expression of PRM1 in bull spermatozoa and testis. Materials and methods For this purpose, absolute q-RT-PCR was performed to calculate the number of copies of PRM1 and PRM2 mRNAs in bovine epididymal spermatozoa and testicular tissue. Western blot and mass spectrometry were performed to identify PRM1 and PRM2 in samples of bovine epididymal spermatozoa. Samples of bovine testicular tissue were collected to identify PRM1 and PRM2 by immunohistochemistry. Results We evaluated that the number of PRM1 mRNA copies was about hundred times higher than PRM2 mRNA copies in sperm and testicular samples (p < 0.0001). In addition, we estimated the PRM1: PRM2 ratio based on mRNA number of copies. In spermatozoa, the ratio was 1: 0.014, and in testicle, the ratio was 1: 0.009. We also evaluated the immunolocalization for PRM1 and PRM2 in bovine testis, and both proteins were detected in spermatids. Western blot and mass spectrometry in bovine epididymal spermatozoa confirmed these results. Conclusion Our work identifies, for the first time, PRM2 in bovine epididymal spermatozoa and in testis. Further studies are still needed to understand the role of PRM2 on the chromatin of the spermatozoa and to verify how possible changes in PRM2 levels may influence the bull fertility.

Detection of protamine 2 in bovine spermatozoa and testicles

Abstract Background Sperm DNA integrity is crucial for transmission of genetic information to future generations and DNA damage can occur during chromatin packaging. Chromatin packaging involves the replacement of somatic nucleosomal histones by nuclear proteins called protamines. Protamine 1 (PRM1) is transcribed and translated in spermatids of all mammals; however, protamine 2 (PRM2) is transcribed in low levels in spermatids and it is not yet described in bull mature spermatozoa. Objectives The aim of this study was to assess gene and protein expression of PRM2 and corroborate gene and protein expression of PRM1 in bull spermatozoa and testis. Materials and methods For this purpose, absolute q-RT-PCR was performed to calculate the number of copies of PRM1 and PRM2 mRNAs in bovine epididymal spermatozoa and testicular tissue. Western blot and mass spectrometry were performed to identify PRM1 and PRM2 in samples of bovine epididymal spermatozoa. Samples of bovine testicular tissue were collected to identify PRM1 and PRM2 by immunohistochemistry. Results We evaluated that the number of PRM1 mRNA copies was about hundred times higher than PRM2 mRNA copies in sperm and testicular samples (p < 0.0001). In addition, we estimated the PRM1: PRM2 ratio based on mRNA number of copies. In spermatozoa, the ratio was 1: 0.014, and in testicle, the ratio was 1: 0.009. We also evaluated the immunolocalization for PRM1 and PRM2 in bovine testis, and both proteins were detected in spermatids. Western blot and mass spectrometry in bovine epididymal spermatozoa confirmed these results. Conclusion Our work identifies, for the first time, PRM2 in bovine epididymal spermatozoa and in testis. Further studies are still needed to understand the role of PRM2 on the chromatin of the spermatozoa and to verify how possible changes in PRM2 levels may influence the bull fertility.

Prevalence and predictive factors of moderate/severe liver steatosis in chronic hepatitis C (CHC) infected patients evaluated with controlled attenuation parameter (CAP).

A novel controlled attenuation parameter (CAP) using FibroScan® has been developed for assessment of liver steatosis. The aim was to evaluate the frequency and associated factors for moderate/severe steatosis evaluated by CAP in CHC patients submitted to transient elastography (TE) by FibroScan® . CHC patients underwent TE with CAP evaluation. The classification of steatosis was defined as: CAP < 222 dB/m  =  S0; CAP ≥ 222 dB/m and <233dB/m  =  S1; ≥233 dB/m < 290dB/m  =  S2 and >= 290 dB/m  =  S3. The prevalence of moderate/severe steatosis (CAP ≥ S2) and the related independent factors were identified by a logistic regression analysis. A significance level of 5% was adopted. 1104 CHC patients, 85% genotype-1 were included (mean age 55 ± 11 years; 46% male, mean BMI 25 ± 4 Kg/m2 ). Systemic arterial hypertension and type 2 diabetes mellitus prevalences were 39% and 17%, respectively. Liver stiffness measurement ≥ 9.5 kPa was observed in 39% of patients and steatosis was identified in 50% (S1 = 7%, S2 = 28% and S3 = 15%). The variables independently associated with moderate/severe steatosis were: male gender (OR=1.35; P = .037; 95% CI:1.01-1.81); systemic arterial hypertension (OR=1.57; P = .002; 95% CI:1.17-2.10) and BMI (OR=1.17; P < .01;95% CI:1.12-1.22). In conclusion, when CAP was adopted as a tool to detect steatosis, genotype 1 CHC patients presented a high prevalence of moderate/advanced steatosis. In these patients, liver steatosis was associated mostly to metabolic factors (arterial hypertension and high BMI).

Time collection and storage conditions of lipid profile.

The stability of samples is crucial for getting reliable concentrations of many analytes, including lipid profile. Thus, the goal of this study was to analyze lipid profile under different storage and temperature conditions. This was a prospective study with 809 patients of both genders. Total cholesterol, triglycerides, high-density lipoprotein cholesterol, low density lipoprotein cholesterol and non-high-density lipoprotein were measured within 1 h from collection at room temperature, after 2-3 h of refrigeration (8°C) and after 4-5 h at room temperature. The processing time and storage conditions did not affect the analytes measured. These findings are important for multicenter studies, because of the difficulties related to centrifugation and freezing of samples immediately after collection.

Counterclockwise or clockwise reciprocating motion for oval root canal preparation: a micro-CT analysis.

AIM: To evaluate oval root canal preparation using one or two instruments in counterclockwise or clockwise reciprocating motion. METHODOLOGY: The radiographic diameter of mandibular human incisors was evaluated, and oval canals were selected (2 ≤ Diameter Ratio ≤ 4). Fifty-seven teeth were assigned to root canal preparation (n = 19): Reciproc 40 (R40) in a counterclockwise reciprocating motion; Mtwo size 40, .06 taper (M 40.06) in a clockwise reciprocating motion or Mtwo size 20, .06 taper and size 40, .06 taper (M 20/40.06) in a clockwise reciprocating motion. Mtwo instruments were coupled to an ENDO DUAL motor, turning 150° clockwise and 30° counterclockwise. Scanning was performed before and after root canal preparation using a SkyScan 1176 micro-computed tomography. Volume, percentage of debris and percentage of uninstrumented surface were analysed in the entire root canal and in each third of the canal. Data were compared using anova and Tukey's tests or Kruskal-Wallis and Dunn tests. RESULTS: The Reciproc and Mtwo systems using different kinematics were associated with a similar increase in root canal volume. Additionally, both system had similar percentage of uninstrumented surface (P > 0.05). Mtwo size 20, .06 taper and size 40, .06 taper was associated with significantly lower debris (P < 0.05) in the middle third (0.56%) when compared to R40 (1.31%) and M size 40, .06 taper (1.54%). CONCLUSIONS: The conventional reciprocation motion for R40 and the clockwise reciprocation motion for Mtwo resulted in similar root canal preparations. Less remaining debris was present in the middle third when two instruments with different diameters were used.

Effect of bovine sperm chromatin integrity evaluated using three different methods on in vitro fertility.

In vitro fertility potential of individual bulls is still relatively uncharacterized. Classical sperm analysis does not include the evaluation of all sperm characteristics and thus, some cell compartments could be neglected. In humans, sperm DNA integrity has already proven to have major influence in embryo development and assisted reproduction techniques successfully. In bovine, some studies already correlated chromatin integrity with field fertility. However, none of those have attempted to relate DNA assessment approaches such as chromatin deficiency (CMA3), chromatin stability (SCSA; AO+) and DNA fragmentation (COMET assay) to predict in vitro bull fertility. To this purpose, we selected bulls with high and low in vitro fertility (n = 6/group), based on embryo development rate (blastocyst/cleavage rate). We then performed CMA3, SCSA test and COMET assay to verify if the difference of in vitro fertility may be related to DNA alterations evaluated by these assays. For the three tests performed, our results showed only differences in the percentage of cells with chromatin deficiency (CMA3+; high: 0.19 ± 0.03 vs low: 0.04 ± 0.04; p = 0.03). No difference for chromatin stability and any of COMET assay categories (grade I to grade IV) was observed between high and low in vitro fertility bulls. A positive correlation between AO + cells and grade IV cells was found. Despite the difference between groups in CMA3 analysis, our results suggest that protamine deficiency in bovine spermatozoa may not have a strong biological impact to explain the difference of in vitro fertility between the bulls used in this study.