Disrupting the wall accumulation of human sperm cells by artificial corrugation

Many self-propelled microorganisms are attracted to surfaces. This makes their dynamics in restricted geometries very different from that observed in the bulk. Swimming along walls is beneficial for directing and sorting cells, but may be detrimental if homogeneous populations are desired, such as in counting microchambers. In this work, we characterize the motion of human sperm cells $60 \mu m$ long, strongly confined to $25 \mu m$ shallow chambers. We investigate the nature of the cell trajectories between the confining surfaces and their accumulation near the borders. Observed cell trajectories are composed of a succession of quasi-circular and quasi-linear segments. This suggests that the cells follow a path of intermittent trappings near the top and bottom surfaces separated by stretches of quasi-free motion in between the two surfaces, as confirmed by depth resolved confocal microscopy studies. We show that the introduction of artificial petal-shaped corrugation in the lateral boundaries removes the tendency of cells to accumulate near the borders, an effect which we hypothesize may be valuable for microfluidic applications in biomedicine.Comment: 9 pages, latex. In accepted version on April 14, v2: abstract modified, information added to Sec. II.A and experiments added to Sec. III.A and Fig.3. Sec. III.C was deleted. Requested references adde

Continuous behavioural 'switching' in human spermatozoa and its regulation by Ca²⁺-mobilising stimuli

Human sperm show a variety of different behaviours (types of motility) that have different functional roles. Previous reports suggest that sperm may reversibly switch between these behaviours. We have recorded and analysed the behaviour of individual human sperm (180 cells in total), each cell monitored continuously for 3-3.5 min either under control conditions or in the presence of Ca2+-mobilising stimuli. Switching between different behaviours was assessed visually (1 s bins using four behaviour categories), and was verified by fractal dimension analysis of sperm head tracks. In the absence of stimuli, ~90% of cells showed at least one behavioural transition (mean rate under control conditions = 6.4 ± 0.8 transitions.min-1). Type 1 behaviour (progressive, activated-like motility) was most common, but the majority of cells (>70%) displayed at least three behaviour types. Treatment of sperm with Ca2+-mobilising agonists had negligible effects on the rate of switching but increased the time spent in type 2 and type 3 (hyperactivation-like) behaviours (P < 2∗10-8; chi-square). Treatment with 4-aminopyridine under alkaline conditions (pHo = 8.5), a highly-potent Ca2+-mobilising stimulus, was the most effective in increasing the proportion of type 3 behaviour, biasing switching away from type 1 (P < 0.005) and dramatically extending the duration of type 3 events (P < 10-16). Other stimuli, including 300 nM progesterone and 1% human follicular fluid, had qualitatively similar effects but were less potent. We conclude that human sperm observed in vitro constitutively display a range of behaviours and regulation of motility by [Ca2+]i, at the level of the single cell, is achieved not by causing cells to adopt a 'new' behaviour but by changing the relative contributions of those behaviours.Fil: Achikanu, Cosmas. University of Birmingham; Reino UnidoFil: Correia, Joao. University of Birmingham; Reino UnidoFil: Guidobaldi, Héctor Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Biología Celular y Molecular; ArgentinaFil: Giojalas, Laura Cecilia. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Biología Celular y Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Barratt, Christopher. University of Dundee; Reino UnidoFil: Da Silva, Sarah Martins. University of Dundee; Reino UnidoFil: Publicover, Stephen. University of Birmingham; Reino Unid

Geometrical guidance and trapping transition of human sperm cells

The guidance of human sperm cells under confinement in quasi-2D microchambers is investigated using a purely physical method to control their distribution. Transport property measurements and simulations are performed with diluted sperm populations, for which effects of geometrical guidance and concentration are studied in detail. In particular, a trapping transition at convex angular wall features is identified and analyzed. We also show that highly efficient microratchets can be fabricated by using curved asymmetric obstacles to take advantage of the spermatozoa specific swimming strategy.publishedVersionFil: Guidobaldi, Héctor Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina.Fil: Guidobaldi, Héctor Alejandro. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Biología Celular y Molecular; Argentina.Fil: Jeyaram, Y. Katholieke Universiteit Leuven. Institute for Nanoscale Physics and Chemistry; Bélgica.Fil: Berdakin, Ivan. Universidad Nacional de Córdoba. Facultad de Matemática, Astronomía y Física; Argentina.Fil: Berdakin, Ivan. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Física Enrique Gaviola; Argentina.Fil: Moshchalkov, V. V. Katholieke Universiteit Leuven. Institute for Nanoscale Physics and Chemistry; Bélgica.Fil: Condat, Carlos Alberto. Universidad Nacional de Córdoba. Facultad de Matemática, Astronomía y Física; Argentina.Fil: Condat, Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Física Enrique Gaviola; Argentina.Fil: Marconi, Verónica Iris. Universidad Nacional de Córdoba. Facultad de Matemática, Astronomía y Física; Argentina.Fil: Marconi, Verónica Iris. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Física Enrique Gaviola; Argentina.Fil: Giojalas, Laura Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina.Fil: Giojalas, Laura Cecilia. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Biología Celular y Molecular ; Argentina.Fil: Silhanek, Alejandro V. Université de Liège. Faculté des Sciences. Departement de Physique; Bélgica.Biofísic

Towards a dendrochronologically refined date of the Laacher See Eruption

The precise date of the Laacher See eruption (LSE), central Europe’s largest Late Pleistocene volcanic event that occurred around 12,900 years ago, is still unknown. Here, we outline the potential of combined high-resolution dendrochronological, wood anatomical and radiocarbon (14C) measurements, to refine the age of this major Plinian eruption. Based on excavated trees that were killed during the explosive LSE and buried under its pyroclastic deposits, we describe how a firm date of the eruption might be achieved, and how the resulting temporal precision would further advance our understanding of the environmental and societal impacts of this event. Moreover, we discuss the relevance of an accurate LSE date for improving the synchronization of European terrestrial and lacustrine Late Glacial to Holocene archives, and outline how the proposed, interdisciplinary dating approach can be applied to other large, yet undated, volcanic eruptions

Precise date for the Laacher See eruption synchronizes the Younger Dryas

The Laacher See eruption (LSE) in Germany ranks among Europe’s largest volcanic events of the Upper Pleistocene. Although tephra deposits of the LSE represent an important isochron for the synchronization of proxy archives at the Late Glacial to Early Holocene transition, uncertainty in the age of the eruption has prevailed. Here we present dendrochronological and radiocarbon measurements of subfossil trees that were buried by pyroclastic deposits that frmly date the LSE to 13,006 ± 9 calibrated years before present (bp; taken as ad 1950), which is more than a century earlier than previously accepted. The revised age of the LSE necessarily shifts the chronology of European varved lakes relative to the Greenland ice core record, thereby dating the onset of the Younger Dryas to 12,807 ± 12 calibrated years bp, which is around 130 years earlier than thought. Our results synchronize the onset of the Younger Dryas across the North Atlantic–European sector, preclude a direct link between the LSE and Greenland Stadial-1 cooling, and suggest a large-scale common mechanism of a weakened Atlantic Meridional Overturning Circulation under warming condition

Hypercholesterolemia Impaired Sperm Functionality in Rabbits

Hypercholesterolemia represents a high risk factor for frequent diseases and it has also been associated with poor semen quality that may lead to male infertility. The aim of this study was to analyze semen and sperm function in diet-induced hypercholesterolemic rabbits. Twelve adult White New Zealand male rabbits were fed ad libitum a control diet or a diet supplemented with 0.05% cholesterol. Rabbits under cholesterol-enriched diet significantly increased total cholesterol level in the serum. Semen examination revealed a significant reduction in semen volume and sperm motility in hypercholesterolemic rabbits (HCR). Sperm cell morphology was seriously affected, displaying primarily a “folded head”-head fold along the major axe-, and the presence of cytoplasmic droplet on sperm flagellum. Cholesterol was particularly increased in acrosomal region when detected by filipin probe. The rise in cholesterol concentration in sperm cells was determined quantitatively by Gas chromatographic-mass spectrometric analyses. We also found a reduction of protein tyrosine phosphorylation in sperm incubated under capacitating conditions from HCR. Interestingly, the addition of Protein Kinase A pathway activators -dibutyryl-cyclic AMP and iso-butylmethylxanthine- to the medium restored sperm capacitation. Finally, it was also reported a significant decrease in the percentage of reacted sperm in the presence of progesterone. In conclusion, our data showed that diet-induced hypercholesterolemia adversely affects semen quality and sperm motility, capacitation and acrosomal reaction in rabbits; probably due to an increase in cellular cholesterol content that alters membrane related events

Religious transformations in the Middle Ages: towards a new archaeological agenda

The study of religious change in Europe between the collapse of the Roman Empire and the Reformation forms one of the cornerstones of medieval archaeology but has been riven by period, denominational and geographical divisions. This paper lays the groundwork for a fundamental rethink of archaeological approaches to medieval religions, by adopting a holistic framework that places Christian, pagan, Islamic and Jewish case studies of religious transformation in a long-term, comparative perspective. Focused around the analytical themes of ‘hybridity and resilience’ and ‘tempo and trajectories’, our approach shifts attention away from the singularities of national narratives of religious conversion towards a deeper understanding of how religious beliefs, practices and identity were renegotiated by medieval people in their daily lives

Testing for BRAF (V600E) Mutation in Thyroid Nodules with Fine-Needle Aspiration (FNA) Read as Suspicious for Malignancy (Bethesda V, Thy4, TIR4): a Systematic Review and Meta-analysis.

In patients with thyroid fine-needle aspiration (FNA) report of suspicious for malignancy (SFM), both lobectomy and thyroidectomy might be considered. BRAF mutation analysis could guide towards accurate surgical therapy. The primary outcome was the reliability of BRAF (V600E) in detecting malignancy in nodules with FNA reading of SFM. The secondary outcome was to analyze its positive predictive value (PPV) and negative predictive value (NPV) considering the surgical histology as gold standard. A literature search of online databases was performed in June 2019. BRAF prevalence among thyroid nodules with FNA read as SFM according to the most popular classification systems (i.e., Bethesda V, Thy4, TIR4 category) was searched. The random-effects model was used. Three hundred sixty original articles were identified and 34 were finally included in the study. There were 1428 thyroid nodules with FNA read as SFM and 1287 (90.1%) lesions underwent surgery with a cancer rate 89.6%. The pooled prevalence of BRAF (V600E) mutation among all nodules with SFM cytology was 47% (95% CI = 40 to 54, I2 = 85.5%). Pooled PPV and NPV of BRAF testing were 99% (95% CI, 97-99) and 24% (95% CI, 16-32), respectively. BRAF (V600E) mutation was found in about one in two nodules with thyroid FNA read as SFM, its PPV to detect cancers was excellent, and its NPV was very poor. The routine BRAF testing in FNA read as SFM cannot be recommended. BRAF (V600E) test may be useful to extend surgical approach in selected cases with further suspicious clinical/ultrasound features