Machine learning techniques to predict the risk of developing diabetic nephropathy: a literature review.

Purpose: Diabetes is a major public health challenge with widespread prevalence, often leading to complications such as Diabetic Nephropathy (DN)-a chronic condition that progressively impairs kidney function. In this context, it is important to evaluate if Machine learning models can exploit the inherent temporal factor in clinical data to predict the risk of developing DN faster and more accurately than current clinical models. Methods: Three different databases were used for this literature review: Scopus, Web of Science, and PubMed. Only articles written in English and published between January 2015 and December 2022 were included. Results: We included 11 studies, from which we discuss a number of algorithms capable of extracting knowledge from clinical data, incorporating dynamic aspects in patient assessment, and exploring their evolution over time. We also present a comparison of the different approaches, their performance, advantages, disadvantages, interpretation, and the value that the time factor can bring to a more successful prediction of diabetic nephropathy. Conclusion: Our analysis showed that some studies ignored the temporal factor, while others partially exploited it. Greater use of the temporal aspect inherent in Electronic Health Records (EHR) data, together with the integration of omics data, could lead to the development of more reliable and powerful predictive models.

Role of Brazilian bats in the epidemiological cycle of potentially zoonotic pathogens.

Bats (Chiroptera) are flying mammals of great biodiversity and habits. These characteristics contribute for them being natural reservoirs and part of the epidemiological cycle of several potentially zoonotic pathogens, such as viruses, protozoa, fungi and bacteria. Brazil hosts approximately 15% of the world's bat diversity, with 181 distinct species, 68 genera and 9 families. About 60% of infectious diseases in humans are of zoonotic origin and, in the last decades, the detection of zoonotic pathogens in bats and their environment has been reported, such as Rabies virus (RABV) and Histoplasma capsulatum. Thus, the aim of this work was to review the reports of zoonotic pathogens associated with bats in Brazil in the past ten years. We reviewed the main pathogenic microorganisms described and the species of bats most frequently involved in the epidemiological cycles of these zoonotic agents. The obtained data show an upward trend in the detection of zoonotic pathogens in Brazilian bats, such as RABV, Bartonella sp., Histoplasma capsulatum and Leishmania spp., with emphasis on the bat species Artibeus lituratus, Carollia perspicillata, Desmodus rotundus and Molossus molossus. These findings highlight the importance of monitoring bat-associated microrganisms to early identify pathogens that may threaten bat populations, including potentially zoonotic microrganisms, emphasizing the importance of the One Health approach to prevent and mitigate the risks of the emergence of zoonotic diseases.

A synchrotron computed tomography dataset for validation of longitudinal tensile failure models based on fibre break and cluster development.

We performed in-situ tensile tests on two carbon fibre/epoxy composites with continuous scanning using synchrotron computed tomography (CT). Both composites were cross-ply laminates, and two specimens were tested for each composite. The voxel size was sufficiently small to recognize individual fibres and fibre breaks. For each test, 16-19 volumes were reconstructed, cropped down to the 0° plies and analysed to track fibre break and cluster development. This dataset provides the last CT volume before failure for each of the four specimens as well as the individual fibre break locations in all reconstructed volumes. These data are then plotted against predictions from six state-of-the-art strength models. The target is that these data become a benchmark for the development of new models, inspiring researchers to set up refined experiments and develop improved models.

Double power-law viscoelastic relaxation of living cells encodes motility trends.

Living cells are constantly exchanging momentum with their surroundings. So far, there is no consensus regarding how cells respond to such external stimuli, although it reveals much about their internal structures, motility as well as the emergence of disorders. Here, we report that twelve cell lines, ranging from healthy fibroblasts to cancer cells, hold a ubiquitous double power-law viscoelastic relaxation compatible with the fractional Kelvin-Voigt viscoelastic model. Atomic Force Microscopy measurements in time domain were employed to determine the mechanical parameters, namely, the fast and slow relaxation exponents, the crossover timescale between power law regimes, and the cell stiffness. These cell-dependent quantities show strong correlation with their collective migration and invasiveness properties. Beyond that, the crossover timescale sets the fastest timescale for cells to perform their biological functions.

CD4+ T helper cells and regulatory T cells in active lupus nephritis: an imbalance towards a predominant Th1 response?

The objective of this study was to evaluate the frequency of CD4+ T cell subsets in peripheral blood mononuclear cells (PBMC), urine and renal tissue from patients with lupus nephritis (LN). PBMC and urinary cells were collected from 17 patients with active LN, 20 disease controls (DC) with primary glomerulonephritis and 10 healthy controls (HC) and were analysed by flow cytometry with markers for T helper type 1 (Th1), Th2, Th17 and regulatory T cells (Treg ) cells. T cell subsets were assessed by immunohistochemistry from LN biopsy specimens from 12 LN patients. T cell subtypes in PBMC were re-evaluated at 6 months of therapy. CD4+ T cells were decreased in PBMC in LN compared with DC and HC (P = 0·0001). No differences were observed in urinary CD4+ T cell subsets between LN and DC. The frequency of urinary Th17 cells was higher in patients with non-proliferative than in proliferative LN (P = 0·041). CD3+ and T-box 21 ( Tbet+) cells were found in glomeruli and interstitium of LN patients, while forkhead box protein 3 (FoxP3), retinoid-related orphan receptor gamma (ROR-γ) and GATA binding protein 3 (GATA-3) were present only in glomeruli. Th1 cells in PBMC were correlated negatively with urinary Th1 cells (Rho = -0·531; P = 0·028) and with Tbet in renal interstitium (Rho = -0·782; P = 0·004). At 6 months, LN patients showed an increase in Th17 cells in PBMC. In conclusion, the inverse association between Th1 cells from PBMC and urinary/renal tissue indicate a role for Th1 in LN pathophysiology. Urinary Th17 cells were associated with less severe LN, and Th17 increased in PBMC during therapy. Urinary CD4+ T cells were not different between LN and DC.

THESIS-ABSTRACT Supplementation levels of exogenous alpha-amylase in broilers diets.

This study aimed to evaluate the supplementation levels of an exogenous alpha-amylase in broilers diets and compare two indicators in determining the diets energy. The experiment was divided into two parallel evaluations, being one of performance and the other of metabolism. In performance assay, 1,700 one-day-old Cobb-500 male chicks were used. The animals were distributed in 50 experimental plots and evaluated five treatments with ten replicates in a completely randomized design (CRD). The treatments were: a positive control (PC), a negative control (NC) and three alpha-amylase supplementation levels 200, 400 and 600 g/t, and the NC was formulated with 50 and 90 kcal of energy reduction in relation to the PC to the phases from 1 to 21 days and from 22 to 42 days, respectively. In the metabolism assay were used 240 animals, 150 birds for stage from 14 to 21 days and 90 birds to stage from 35 to 42 days of age and the treatments were the same as the performance assay, with six replicates per treatment in CRD. All diets of metabolism test contained the digestibility indicators Lipe® (eucalyptus purified lignin) and chromic oxide (Cr2O3), in concentrations of 0.05 and 1.0%, respectively. In the period from 1 to 21 days old, no significant differences were observed in weight gain (WG) (P > 0.05), however, feed intake (FI) was found higher by using 200 ppm of enzyme (P < 0.05) and better feed conversion (FC) with the PC (P < 0.05). From 22 to 42 days, no significant differences were observed on the WG (P > 0.05), but were observed lower FI and better FC to PC treatment (P < 0.05). In the period from 1 to 42 days of age, significant differences were also not observed on the WG (P > 0.05), but there was lower FI and better FC for the PC treatment (P < 0.05). The AMEn (apparent metabolizable energy corrected for nitrogen balance), determined using the total collection, reaffirmed the values ​​calculated for the PC and NC with intermediate data obtained from the enzyme use (200, 400 and 600 ppm). Comparing the total collection using Lipe® and Cr2O3, a correlation was observed only for the PC results, that were always higher, and for the NC results, that were lower for the three methodologies. For IDE (ileal digestible energy), determined by Cr2O3, significant differences were observed (P < 0.05) and it presented higher values in the PC treatment and lower values in the NC. In IDE determining by Lipe®, significant was observed (P < 0.05), showing higher value in the PC treatment. It is concluded that in the metabolism assessments, exogenous alpha-amylase tested was effective in increasing the metabolizable energy (ME), however, not enough to be equivalent to the ME of PC treatment, a result that was better expressed in the total collection, AMEn by Lipe® and IDE by Lipe® methodologies. The performance results reflect the superiority of PC treatment, pointing enzyme limitation to supply the energy deficit practiced in this work.

Thesis Abstract Levels and forms of vitamin D in broilers diets.

This study aimed to evaluate the concentration effects of two vitamin D isoforms, cholecalciferol (D3) and 25-hydroxycholecalciferol (25-OHD3) in broilers diets on performance, bone and physiological features of these birds. Of a total of 1920 one-day-old male chicks Cobb-500 were used from commercial hatchery, reared under bed creation systems. The animals were distributed in six treatments and eight replicates with 40 birds per treatment in a completely randomized design. The following vitamin D supplementation levels were applied: 70 and 87.5 µg/kg feed in initial phase; 56 and 70 µg/kg feed during the growth phase, and 35 and 47.35 µg/kg of feed in final phase of creation, obtained from two forms (D3 and 25-OHD3). The treatments consisted of supplementation of two levels from each isolated source and their associations (60% D3 + 40% 25-OHD3) according to the study phases. In the metabolism assay, 480 birds (14 and 35 days of age) were separated to be used for evaluation of calcium (Ca) and phosphorus (P) retention and excretion during the periods of 19 to 21 days and 40 to 42 days of age. The diets were based on corn and soybean meal, with supplementation of phytase (500 FTU/kg). The performance, bone characteristics, plasma levels, bone radiographic density, carcass yield, and P and Ca retention were evaluated. In the initial creation phase, we observed an increased P excretion by broilers fed diets supplemented with vitamin D3 (P < 0.05). In addition, the association between the two vitamin D isoforms resulted in higher retention of Ca and P than the birds fed diets supplemented only with vitamin D3 (P < 0.05), and higher P retention when compared to birds fed diets supplemented with 25-OHD3 (P < 0.05). Dietary supplemental 25-OHD3 at 87.5 µg/kg resulted in higher plasma levels of Ca in relation to the same supplemented source with 70 µg/kg at 21 days of age (P < 0.05). In the final phase, the birds fed diets supplemented with vitamin D3 presented the lowest values of consumption and retention of Ca and P (P < 0.05). The association between the forms (D3 and 25-OHD3) reduced the excretion values of Ca and P (p < 0.05). The birds receiving a source of 25-OHD3 and the association had better feed conversion ratio and higher bone ash content (P < 0.05). At all stages studied D3 + 25-OHD3 combined supplementation increased tibial density of broilers in relation to supplementation of only vitamin D3 (P < 0.05). Results of this study indicate that the addition of 25-OHD3 in the feed supplemented with vitamin D3 improve the feed conversion, increase the Ca plasma levels, and also increases bone density, providing higher retention coefficients of Ca and P and lower P excretion, regardless of the development phase of these birds.

Thesis Abstract Bean (Phaseolus vulgaris L.) lines: chemical composition and protein digestibility.

The bean represents the main source of proteins for the low income populations, although the digestibility of those proteins is relatively low. Consequently, the programs of plant genetic breeding have been working on the search for new lines with higher protein levels. Thus, with the purpose of supplying information to the researchers, in this study, 21 bean (Phaseolus vulgaris L.) lines were analyzed for the centesimal and mineral composition, protein digestibility, phenolic compounds, and trypsin inhibitor. The entirely randomized experimental design was used with 21 treatments (lines) and three repetitions. All values were within the following ranges: 22.34 to 36.28 g crude protein/100 g dry matter (DM); 7.56 to 20.91 g neutral detergent fiber/100 g DM; 0.53 to 2.55 g fat/100 g DM and 2.97 to 4.87 g ashes/100 g DM. The levels of phosphorus, potassium, calcium, magnesium, and sulfur, in g/100 g DM, varied from 0.45 to 0.72; 1.51 to 2.48; 0.03 to 0.28; 0.18 to 0.34 and 0.28 to 0.45, respectively. Regarding copper, manganese, zinc and iron, the levels, in mg/kg DM, varied from 11.37 to 17.73; 14.93 to 28.90; 36.67 to 69.90 and 71.37 to 126.90, respectively. The in vitro protein digestibility varied from 18.03 to 48.32%. The levels of phenolic compounds varied from 0.28 to 1.08 mg acid tanic/100 g DM and the one of trypsin inhibitor from 59.93 to 151.07 trypsin inhibited units/mg DM. Among the lines with higher protein contents, "ESAL 569" (beige with brown stripe) presented the largest protein digestibility and considerable levels of minerals. "P-180" (beige with brown stripe) was one of the lines with higher crude protein contents and digestibilities, and also presented high levels for most of the minerals. No relation between protein digestibility and the contents of phenolic compounds or trypsin inhibitor was observed.

Follicular helper T cell in immunity and autoimmunity.

The traditional concept that effector T helper (Th) responses are mediated by Th1/Th2 cell subtypes has been broadened by the recent demonstration of two new effector T helper cells, the IL-17 producing cells (Th17) and the follicular helper T cells (Tfh). These new subsets have many features in common, such as the ability to produce IL-21 and to express the IL-23 receptor (IL23R), the inducible co-stimulatory molecule ICOS, and the transcription factor c-Maf, all of them essential for expansion and establishment of the final pool of both subsets. Tfh cells differ from Th17 by their ability to home to B cell areas in secondary lymphoid tissue through interactions mediated by the chemokine receptor CXCR5 and its ligand CXCL13. These CXCR5+ CD4+ T cells are considered an effector T cell type specialized in B cell help, with a transcriptional profile distinct from Th1 and Th2 cells. The role of Tfh cells and its primary product, IL-21, on B-cell activation and differentiation is essential for humoral immunity against infectious agents. However, when deregulated, Tfh cells could represent an important mechanism contributing to exacerbated humoral response and autoantibody production in autoimmune diseases. This review highlights the importance of Tfh cells by focusing on their biology and differentiation processes in the context of normal immune response to infectious microorganisms and their role in the pathogenesis of autoimmune diseases.

Endometrial transcriptional profiling of a bovine fertility model by Next-Generation Sequencing.

Studying the multitude of molecular networks and pathways that are potentially involved in a complex trait such as fertility requires an equally complex and broad strategy. Here, we used Next-Generation Sequencing for the characterization of the transcriptional signature of the bovine endometrial tissue. Periovulatory endocrine environments were manipulated to generate two distinctly different fertility phenotypes. Cycling, non-lactating, multiparous Nelore cows were manipulated to ovulate larger (> 13 mm; LF group; high fertility phenotype) or smaller (< 12 mm; SF group) follicles. As a result, greater proestrus estrogen concentrations, corpora lutea and early diestrus progesterone concentrations were also observed in LF group in comparison to SF group. Endometrial cell proliferation was estimated by the protein marker MKI67 on tissues collected 4 (D4) and 7 (D7) days after induction of ovulation. Total RNA extracts from D7 were sequenced and compared according to the transcriptional profile of each experimental group (LF versus SF). Functional enrichment analysis revealed that LF and SF endometria were asynchronous in regards to their phenotype manifestation. Major findings indicated an LF endometrium that was switching phenotypes earlier than the SF one. More specifically, a proliferating SF endometrium was observed on D7, whereas the LF tissue, which expressed a proliferative phenotype earlier at D4, seemed to have already shifted towards a biosynthetically and metabolically active endometrium on D7. Data on MKI67 support the transcriptomic results. RNA-Seq-derived transcriptional profile of the endometrial tissue indicated a temporal effect of the periovulatory endocrine environment, suggesting that the moment of the endometrial exposure to the ovarian steroids, E2 and P4, regulates the timing of phenotype manifestation. Gene expression profiling revealed molecules that may be targeted to elucidate ovarian steroid-dependent mechanisms that regulate endometrial tissue receptivity. Data was deposited in the SRA database from NCBI (SRA Experiment SRP051330) and are associated with the Bio-Project (PRJNA270391). An overview of the gene expression data has been deposited in NCBI's Gene Expression Omnibus (GEO) and is accessible through GEO Series accession number GSE65450. Further assessment of the data in combination with other data sets exploring the transcriptional profile of the endometrial tissue during early diestrus may potentially identify novel molecular mechanisms and/or markers of the uterine receptivity.

Follicular helper T cell in immunity and autoimmunity

The traditional concept that effector T helper (Th) responses are mediated by Th1/Th2 cell subtypes has been broadened by the recent demonstration of two new effector T helper cells, the IL-17 producing cells (Th17) and the follicular helper T cells (Tfh). These new subsets have many features in common, such as the ability to produce IL-21 and to express the IL-23 receptor (IL23R), the inducible co-stimulatory molecule ICOS, and the transcription factor c-Maf, all of them essential for expansion and establishment of the final pool of both subsets. Tfh cells differ from Th17 by their ability to home to B cell areas in secondary lymphoid tissue through interactions mediated by the chemokine receptor CXCR5 and its ligand CXCL13. These CXCR5+ CD4+ T cells are considered an effector T cell type specialized in B cell help, with a transcriptional profile distinct from Th1 and Th2 cells. The role of Tfh cells and its primary product, IL-21, on B-cell activation and differentiation is essential for humoral immunity against infectious agents. However, when deregulated, Tfh cells could represent an important mechanism contributing to exacerbated humoral response and autoantibody production in autoimmune diseases. This review highlights the importance of Tfh cells by focusing on their biology and differentiation processes in the context of normal immune response to infectious microorganisms and their role in the pathogenesis of autoimmune diseases.

The Receptive Endometrial Transcriptomic Signature Indicates an Earlier Shift from Proliferation to Metabolism at Early Diestrus in the Cow.

This study aimed to characterize the endometrial transcriptome and functional pathways overrepresented in the endometrium of cows treated to ovulate larger (≥13 mm) versus smaller (≤12 mm) follicles. Nelore cows were presynchronized prior to receiving cloprostenol (large follicle [LF] group) or not (small follicle [SF] group), along with a progesterone (P4) device on Day (D) -10. Devices were withdrawn and cloprostenol administered 42-60 h (LF) or 30-36 h (SF) before GnRH agonist treatment (D0). Tissues were collected on D4 (experiment [Exp.] 1; n = 24) or D7 (Exp. 2; n = 60). Endometrial transcriptome was obtained by RNA-Seq, whereas proliferation and apoptosis were assessed by immunohistochemistry. Overall, LF cows developed larger follicles and corpora lutea, and produced greater amounts of estradiol (D-1, Exp. 1, SF: 0.7 ± 0.2; LF: 2.4 ± 0.2 pg/ml; D-1, Exp. 2, SF: 0.5 ± 0.1; LF: 2.3 ± 0.6 pg/ml) and P4 (D4, Exp. 1, SF: 0.8 ± 0.1; LF: 1.4 ± 0.2 ng/ml; D7, Exp. 2, SF: 2.5 ± 0.4; LF: 3.7 ± 0.4 ng/ml). Functional enrichment indicated that biosynthetic and metabolic processes were enriched in LF endometrium, whereas SF endometrium transcriptome was biased toward cell proliferation. Data also suggested reorganization of the extracellular matrix toward a proliferation-permissive phenotype in SF endometrium. LF endometrium showed an earlier onset of proliferative activity, whereas SF endometrium expressed a delayed increase in glandular epithelium proliferation. In conclusion, the periovulatory endocrine milieu regulates bovine endometrial transcriptome and seems to determine the transition from a proliferation-permissive to a biosynthetic and metabolically active endometrial phenotype, which may be associated with the preparation of an optimally receptive uterine environment.

Água salina e biofertilizante bovino na produção de frutos e alocação de biomassa em noni (Morinda citrifolia L. ) / Saline water and bovine biofertilizer on yield and biomass allocation in noni plants (Morinda citrifolia L. )

RESUMO: Têm sido evidenciado que os compostos orgânicos exercem ação mitigadora dos efeitos tóxicos dos sais às plantas, promovendo maior crescimento e desenvolvimento em solos com excesso de sais. Nessa direção, um experimento foi desenvolvido para avaliar os efeitos do biofertilizante bovino na produção de frutos e alocação de biomassa pelos diversos órgãos de plantas de noni, irrigadas com águas salinas. Os tratamentos foram distribuídos em blocos ao acaso, com quatro repetições e duas plantas por parcela, adotando o arranjo fatorial 5 x 2, correspondente a salinidade das águas de irrigação de 0,5; 1,5; 3,0; 4,5; 6,0 dS m-1, em substratos sem e com biofertilizante bovino, aplicado uma única vez, em volume correspondente a 10% do volume do substrato, um dia antes da instalação do experimento. As variáveis avaliadas foram: salinidade do solo, expressa pela condutividade elétrica do extrato de saturação do solo; número de frutos; massa média de fruto; produção de frutos por planta; e alocação de biomassa pelas raízes, caules, folhas e frutos. O aumento da salinidade da água de irrigação elevou o caráter salino do solo desde "não salino" para "fortemente salino" e, inibiu a produção de frutos e a acumulação de biomassa seca das plantas de noni em geral, mas, com maior intensidade nos tratamentos sem o insumo orgânico. O biofertilizante estimula o crescimento e a produção de plantas de noni cultivadas sob irrigação com águas de alta salinidade.

ABSTRACT: Organic compounds have shown to exercise a mitigating action on salts in plants and they promote growth and development in salt stress environments. In this sense, an experiment was carried out in order to evaluate the effects of saline water irrigation and bovine biofertilizer on yield and biomass allocation by the various organs of noni plants. Treatments were arranged in a randomized block design with four replications and two plants per plot using the 5 x 2 factorial design, which correspond to the salinity levels of irrigation water of 0.5, 1.5, 3.0, 4.5 and 6.0 dS m-1 in substrates with and without bovine biofertilizer, applied to the soil once, in the volume corresponding to 10% of the volume of the substrate one day before the implementation of the experiment. The variables evaluated were soil salinity, expressed as electrical conductivity of the soil saturation extract, fruit number, fruit weight and fruit yield per plant, and biomass allocation by roots, stems, leaves and fruit. The increasing salinity of the water for irrigation increased the saline character of the soil from non-saline to saline soil and inhibited the production and accumulation of dry biomass in noni plants in general but with more intensity in the plants of the treatments without bovine biofertilizer applied to the soil in liquid form. The biofertilizer stimulates the growth and production of noni plants grown under irrigation with high salinity water.

Modulation of periovulatory endocrine profiles in beef cows: consequences for endometrial glucose transporters and uterine fluid glucose levels.

In beef cattle, proestrus estradiol and subsequent progesterone (P4) concentrations can regulate the endometrial characteristics and thereby determine maternal receptivity toward the embryo. However, the underlying mechanisms linking periovulatory endocrine profiles to receptivity, which is crucial to obtain pregnancy, need to be elucidated. We hypothesized that the size of the preovulatory follicle (POF) and subsequent circulating P4 concentrations, during early diestrus, modulate endometrial levels of glucose transporter transcripts and proteins, and subsequently affect the luminal glucose availability in the uterus. Therefore, follicle growth of Nelore cows was manipulated, and cows were assigned to 2 experimental groups: (1) large follicle and large corpus luteum (LF-LCL) group with a large POF and corpus luteum (CL); and (2) small follicle and small corpus luteum (SF-SCL) group with a small POF and CL. At day 7 post gonadotropin-releasing hormone induced ovulation (gonadotropin-releasing hormone treatment = day 0), animals were slaughtered (n = 18 per group), and uterine tissues and washings were collected for characterization of glucose transporters and glucose levels, respectively. The diameter of POF was larger (P < 0.05) in the LF-LCL cows compared with their SF-SCL counterparts (12.8 ± 0.4 vs 11.1 ± 0.4 mm). Furthermore, CL size (17.49 ± 0.88 vs 14.48 ± 0.52 mm) and circulating P4 concentrations at day 7 (4.5 ± 1.0 vs 3.3 ± 1.1 ng/mL, P < 0.05) were significantly higher in the LF-LCL cows compared with the SF-SCL cows. No differences (P > 0.05) were detected in gene expression patterns of SLC2A1, SLC2A3, SLC2A4, SLC2A5, SLC5A1, ATP1A2, ATP1B2, and SLC37A4. However, the protein abundance of endometrial SLC2A1was increased in the LF-LCL group compared with the SF-SCL group (P < 0.05). SLC2A1 and SLC2A4 protein products were mainly identified at the endometrial luminal and glandular epithelium membranes as well as in the endometrial stroma. Glucose concentrations in uterine washings were similar between groups. In conclusion, we provided information on the potential link between endocrine profiles and glucose transport pathways in the bovine endometrium. More specifically, our data reveal that the size of the POF, and subsequent P4 concentrations, do not functionally affect the main endometrial glucose transporter pathways or uterine fluid glucose concentrations during diestrus.

Impact of probing the reproductive tract during early pregnancy on fertility of beef cows.

This short communication reports the impact of endometrial biopsies, uterine flushings and follicular fluid aspiration procedures at day 6 post artificial insemination (AI) on pregnancy rates. In Experiment 1, cows were timed AI (TAI) and assigned to the following treatment groups: control (n = 37), uterine flushing (n = 35) and endometrial biopsy (n = 38). On day 30 post AI, pregnancy rates were 40.5%, 33% and 28.5%, respectively (p > 0.1). Pregnancy rate on day 60 was lower (p < 0.004) in flushed cows than in the controls. In Experiment 2, oestrus was detected and cows were assigned to flushing (n = 32) or biopsy (n = 33) treatments 6 days after AI, which resulted in pregnancy rates of 31% and 36%, respectively (p > 0.1). In Experiment 3, cows were, 6 days after TAI, randomly assigned to the following treatments: control (n = 84) or aspiration of the largest follicle (n = 73). Pregnancy rates on day 30 post AI were 63.5% for the control group and 53% for the aspirated group (p > 0.1). In conclusion, uterine flushing and endometrial biopsy negatively affect pregnancy rates, but neither procedure can be considered to be incompatible with pregnancy maintenance. Follicular aspiration during pregnancy does not interact with pregnancy success. The amount and quality of samples obtained are compatible with the use of cellular and molecular analysis of uterine variables from cows that failed or succeeded on maintaining pregnancy.

Manipulation of the periovulatory sex steroidal milieu affects endometrial but not luteal gene expression in early diestrus Nelore cows.

In beef cattle, the ability to conceive has been associated positively with size of the preovulatory follicle (POF). Proestrus estradiol and subsequent progesterone concentrations can regulate the endometrium to affect receptivity and fertility. The aim of the present study was to verify the effect of the size of the POF on luteal and endometrial gene expression during subsequent early diestrus in beef cattle. Eighty-three multiparous, nonlactating, presynchronized Nelore cows received a progesterone-releasing device and estradiol benzoate on Day-10 (D-10). Animals received cloprostenol (large follicle-large CL group; LF-LCL; N = 42) or not (small follicle-small CL group; SF-SCL; N = 41) on D-10. Progesterone devices were withdrawn and cloprostenol administered 42 to 60 hours (LF-LCL) or 30 to 36 hours (SF-SCL) before GnRH treatment (D0). Tissues were collected at slaughter on D7. The LF-LCL group had larger (P < 0.0001) POF (13.24 ± 0.33 mm vs. 10.76 ± 0.29 mm), greater (P < 0.0007) estradiol concentrations on D0 (2.94 ± 0.28 pg/mL vs. 1.27 ± 0.20 pg/mL), and greater (P < 0.01) progesterone concentrations on D7 (3.71 ± 0.25 ng/mL vs. 2.62 ± 0.26 ng/mL) compared with the SF-SCL group. Luteal gene expression of vascular endothelial growth factor A, kinase insert domain receptor, fms-related tyrosine kinase 1, steroidogenic acute regulatory protein, cytochrome P450, family 11, subfamily A, polypeptide 1, and hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 7 was similar between groups. Endometrial gene expression of oxytocin receptor and peptidase inhibitor 3, skin-derived was reduced, and estrogen receptor alpha 2, aldo-keto reductase family 1, member C4, and lipoprotein lipase expression was increased in LF-LCL versus SF-SCL. Results support the hypothesis that the size of the POF alters the periovulatory endocrine milieu (i.e., proestrus estradiol and diestrus progesterone concentrations) and acts on the uterus to alter endometrial gene expression. It is proposed that the uterine environment and receptivity might also be modulated. Additionally, it is suggested that increased progesterone secretion of cows ovulating larger follicles is likely due to increased CL size rather than increased luteal expression of steroidogenic genes.

Technical note: Evaluation of markers for estimating duodenal digesta flow and ruminal digestibility: Acid detergent fiber, sulfuric acid detergent lignin, and n-alkanes.

The amount of digesta flowing to the duodenum is a relevant measurement for the evaluation of nutrient supply to ruminants, which is usually estimated in animals fitted with a duodenal T-type cannula using internal or external markers. This study evaluated acid detergent fiber (ADF) compared with external (C32n-alkane) and internal [sulfuric acid lignin (ADL) and n-alkanes C31 and C33] markers for estimating duodenal flow and(or) ruminal digestibility of dry matter (DM) in cattle and sheep. In the first assay, 4 duodenally cannulated Holstein steers housed in metabolism cages, dosed with C32n-alkane, and fed Avena strigosa plus concentrate and increasing levels of tannin extract to reduce ruminal digestibility, were used in a Latin square design. The mobile-bag technique was used to measure the intestinal disappearance of ADL and ADF from forage (Avena strigosa, Pennisetum purpureum, Cynodon dactylon, and Medicago sativa) and concentrate (corn grain, soybean meal, and sunflower meal) samples that were previously incubated in the rumen of additional fistulated steer for 12, 24, 36, or 48 h. The ADF concentration in residues recovered in the feces was strongly related to the ADF concentration in residues at the duodenum (R(2)=0.93, standard deviation=30.0, n=901). This relationship showed a lower precision for ADL fraction (R(2)=0.88, standard deviation=12.6, n=590). In a second assay, duodenal flow and ruminal DM digestibility were calculated from the duodenal and fecal concentration of either marker. We observed a significant effect of marker type on ruminal DM digestibility values, and the effect of tannin treatments was observed only when ADF or ADL was used as the marker. The lowest residual error was obtained for ADF. Ruminal DM digestibility was, on average, higher for C31 and C(33)n-alkanes, and the use of dosed C(32)n-alkane resulted in a negative value. In the third assay, a data set of 235 individual observations was compiled from digestibility trials to compare ADF and ADL as markers for estimating duodenal digesta flow in wethers (n=204) and cattle (n=31). We observed a strong relationship between markers (R(2)=0.84 in sheep and R(2)=0.88 in cattle), but variance analysis within trials indicated that ADF was more precise than ADL. In conclusion, in digestibility trials in which fecal output was measured and spot samples of the duodenal digesta were obtained, duodenal flow and ruminal digestibility of the DM may be estimated from the relationship between the ADF concentration in feces and that in the duodenal digesta of ruminants.

Storage of bovine reproductive tissues and RNA extracts on ice for 24 h or repeated freeze-thaw cycles do not affect RNA integrity.

The aims of this study were to test (i) the effect of time of tissue and RNA extracts storage on ice and (ii) the effect of repeated freeze-thaw cycles on RNA integrity and gene expression of bovine reproductive tissues. Fragments of endometrium (ENDO), corpus luteum (CL) and ampulla (AMP) were subdivided and incubated for 0, 1, 3, 6, 12 or 24 h on ice. RNA extracts were incubated on ice for 0, 3, 12 or 24 h, or exposed to 1, 2, 4 or 6 freeze-thaw cycles. RNA integrity number (RIN) was estimated. Expression of progesterone receptor (PGR) and cyclophilin genes from RNA extracts stored on ice for 0 or 24 h, and 1 or 6 freeze-thaw cycles was measured by qPCR. Tissue and RNA extract incubation on ice, and repeated freeze-thaw cycles did not affect RIN values of RNA from ENDO, CL or AMP. Storage on ice or exposure to freeze-thaw cycles did not affect Cq values for PGR or cyclophilin genes. In conclusion, neither generalized RNA degradation nor specific RNA degradation was affected by storage of tissue or RNA extracts on ice for up to 24 h, or by up to 6 freeze-thaw cycles of RNA extracts obtained from bovine ENDO, CL and AMP.

Identifying individual common carp Cyprinus carpio using scale pattern.

Volunteers were able to match photographs of the same common carp Cyprinus carpio taken on two occasions. Images were identified correctly on 95·76% of occasions. Thus, scale patterns can be used for non-invasive identification of C. carpio over a period of time.

Effects of flunixin meglumine, recombinant bovine somatotropin and/or human chorionic gonadotropin on pregnancy rates in Nelore cows.

The objective was to compare pharmacological strategies aiming to inhibit prostaglandin F2 alpha (PGF(2α)) synthesis (flunixin meglumine; FM), stimulate growth of the conceptus (recombinant bovine somatotropin; bST) and progesterone (P(4)) synthesis (human chorionic gonadotropin; hCG), as well as their combinations, regarding their ability to improve pregnancy rates in beef cattle. Lactating Nelore cows (N = 975), 35 to 70 days postpartum, were synchronized and inseminated by timed artificial insemination (TAI) on Day 0. On Day 7, cattle were allocated into eight groups and received one of the following treatments: saline (S) on Days 7 and 16 (Group Control); S on Day 7 and FM on Day 16 (Group FM); bST on Day 7 and S on Day 16 (Group bST); bST on Day 7 and FM on Day 16 (Group bST + FM); hCG on Day 7 and S on Day 16 (Group hCG); hCG on Day 7 and FM on Day 16 (Group hCG + FM); bST and hCG on Day 7 and S on Day 16 (Group bST + hCG), or bST and hCG on Day 7 and FM on Day 16 (Group bST + hCG + FM). The aforementioned treatments were administered at the following doses: 2.2 mg/kg FM (Banamine®; Intervet Schering-Plough, Cotia, SP, Brazil), 500 mg bST (Boostin®; Intervet Schering-Plough), and 2500 IU hCG (Chorulon®; Intervet Schering-Plough). Pregnancy diagnosis was performed 40 days after TAI by transrectal ultrasonography. Pregnancy rates were not significantly different among treatments. However, there was a main effect of hCG treatment to increase pregnancy rates (63.0 vs. 55.4%; P = 0.001). Concentrations of P(4) did not differ significantly among groups on Day 7 or on Day 16. However, consistent with the higher pregnancy rates, hCG increased P(4) concentrations on Day 16 (10.6 vs. 9.6 ng/mL, respectively; P = 0.05). We concluded that hCG treatment 7 days after TAI improved pregnancy rates of lactating Nelore cows, possibly via a mechanism leading to induction of higher P(4) concentrations, or by reducing the luteolytic stimulus during maternal recognition of pregnancy.