Effect of seminal plasma on hipoosmotic swelling test in fresh alpaca spermatozoa

A study was designed with the objective of evaluating the effect of seminal plasma on the response to the hypoosmotic swelling test (HOST) in alpaca spermatozoa, for which three experimental groups were organized as follows: Group 1(n=15) plasma free sperm seminal (obtained from the vas deferens, aspirated in PBS), Group 2(n=15) free seminal plasma sperm reconstituted with seminal plasma (obtained from the vas deferens, aspirated in PBS, mixed in 50/50% with seminal plasma) and Group 3(n=15) whole semen (obtained by artificial vagina), The samples were incubated in a hypoosmotic solution adjusted to 100mOsmol (sodium citrate+fructose+2H2Ocsp 100mL). 0.1mL of semen+0.9mL of hypoosmotic solution was mixed, incubated for 30minutes in a water bath at 37°C and the reaction was stopped with 0.1mL of 4% formaldehyde. A count of at least 200 spermatozoa was performed per sample, using an optical microscope with immersion objective (100X), the vitality was evaluated by supravital eosin staining (0.7%)-nigrosin(1%), the results indicate that it does not exist a detrimental effect of the seminal plasma on the endosmotic response, being, on the contrary, superior in the whole semen; the vitality of the spermatozoa with and without seminal plasma is similar, however it decreases when it is reconstituted with seminal plasma, possibly due to the seminal plasma of another animal; there is no positive correlation between endosmosis and vitality, indicating that the latter would not necessarily reflect the integrity of the membrane, which is why it is recommended to perform this test routinely in alpaca semen exams

Identification of an Imidazopyridine-based Compound as an Oral Selective Estrogen Receptor Degrader for Breast Cancer Therapy.

UNLABELLED: The pro-oncogenic activities of estrogen receptor alpha (ERα) drive breast cancer pathogenesis. Endocrine therapies that impair the production of estrogen or the action of the ERα are therefore used to prevent primary disease metastasis. Although recent successes with ERα degraders have been reported, there is still the need to develop further ERα antagonists with additional properties for breast cancer therapy. We have previously described a benzothiazole compound A4B17 that inhibits the proliferation of androgen receptor-positive prostate cancer cells by disrupting the interaction of the cochaperone BAG1 with the AR. A4B17 was also found to inhibit the proliferation of estrogen receptor-positive (ER+) breast cancer cells. Using a scaffold hopping approach, we report here a group of small molecules with imidazopyridine scaffolds that are more potent and efficacious than A4B17. The prototype molecule X15695 efficiently degraded ERα and attenuated estrogen-mediated target gene expression as well as transactivation by the AR. X15695 also disrupted key cellular protein-protein interactions such as BAG1-mortalin (GRP75) interaction as well as wild-type p53-mortalin or mutant p53-BAG2 interactions. These activities together reactivated p53 and resulted in cell-cycle block and the induction of apoptosis. When administered orally to in vivo tumor xenograft models, X15695 potently inhibited the growth of breast tumor cells but less efficiently the growth of prostate tumor cells. We therefore identify X15695 as an oral selective ER degrader and propose further development of this compound for therapy of ER+ breast cancers. SIGNIFICANCE: An imidazopyridine that selectively degrades ERα and is orally bioavailable has been identified for the development of ER+ breast cancer therapeutics. This compound also activates wild-type p53 and disrupts the gain-of-function tumorigenic activity of mutant p53, resulting in cell-cycle arrest and the induction of apoptosis

Historical trends in Hg, Pb and Zn sedimentation in the central shelf area of Portugal

Temporal records of excess 210Pb, and the determination of major (Al and Ca) and trace elements (Zn, Cr, Ni, Pb, Cu and Hg) in two sediment box-cores, collected in the central area of the Portuguese shelf of North of the Nazaré canyon (offshore from the Lis River), allow evaluation of the deposition of various chemical elements normally associated with anthropogenic activities. In order to compensate for the natural sediment variability, heavy metal contents were normalised to Al. Temporal variations of Hg, Pb and Zn (Al-normalised) show an increasing trend since the beginning of the 1920’s, recording the development of industrial activities. Enrichment factors (EF) were calculated to estimate the level of contamination in these sediments. Mercury is the element with the highest average EF values (EF = 3), followed by Pb (EF = 1.5) and Zn (EF = 1.2). The results indicate that since 1991 64% of total Hg, 44% of total Pb and 24% of total Zn are derived from anthropogenic sources. The average anthropogenic fluxes of Hg, Pb and Zn (0.008, 3, 6 μg cm-2yr-1, respectively) for the last 40 years in a ca. 400 km2 deposition area represent a total accumulation of approximately 30, 12000 and 24400 kg per year of Hg, Pb and Zn, respectively. These results indicate that despite the high-energy conditions and the generally sandy nature of the Portuguese shelf sediments, it is possible to identify significant anthropogenic enrichments in some areas of sediment accumulation. These contaminants are not necessarily related to immediate sources but may instead indicate atmospheric and or marine transport from more distant sources.Los registros temporales del 210Pb en exceso, elementos mayores (Al y Ca), y elementos traza (Zn, Cr, Ni, Pb, Cu y Hg) de 2 box-cores procedentes de la plataforma continental al norte del cañón de Nazaré han permitido caracterizar depósitos de elementos químicos asociados a actividades antropogénicas. Los contenidos de metales pesados han sido normalizados con Al para compensar la variabilidad natural. La variación temporal de Hg, Pb y Zn, normalizados con Al, muestran un incremento desde el inicio de la década de 1920 reflejando el desarrollo de la actividad humana. El nivel de contaminación de los sedimentos ha sido calculado en base a Factores de Enriquecimiento (EF). Los valores medios más altos de FE son de mercurio (EF = 3), seguidos de plomo (EF = 1.5) y zinc (EF = 1.2). Los resultados obtenidos indican que a partir de 1991 el 64% del total de Hg, 44% del total de Pb y 24% del total de Zn son de origen antropogénico. La media para los últimos 40 años de los flujos de Hg, Pb y Zn (0.008, 3, 6 μg cm-2 año-1, respectivamente), en un área de deposición de aproximadamente 400 km2, se traduce en la acumulación de 30, 12000 y 24400 kg por año de Hg, Pb y Zn. Sin embargo, a pesar de las condiciones de alta energía y el carácter arenoso de los sedimentos de la plataforma continental portuguesa, se pueden identificar importantes enriquecimientos antropogénicos en depocentros de sedimentos, no relacionados con fuentes antropogénicas próximas pero que pueden indicar transporte marino y atmosférico de fuentes de contaminación distantes

Transcriptomic data on the role of PEST-domain-enriched tyrosine phosphatase in the regulation of antigen-mediated activation and antiallergic action of glucocorticoids in mast cells

Protein tyrosine phosphatases and glucocorticoids are known to regulate allergic and antiallergic action in activated mast cells. Here we provide RNA sequencing and quantitative real-time PCR data from bone marrow derived mast cells, for wild-type and PEST-domain-enriched tyrosine phosphatase (PEP) null mice, activated by immunoglobulin E sensitization and dinitrophenol treatment, and additionally treated with the glucocorticoid dexamethasone. The transcriptomics experiment was performed in duplicate with a total of 16 samples (GSE108972). Keywords: RNA sequencing, Quantitative real-time PCR, Allergy, Gene expression, Phosphatas

Evening choruses in the Perth Canyon and their potential link with Myctophidae fishes

An evening chorus centered at near 2.2 kHz was detected across the years 2000 to 2014 from seabed receivers in 430-490 m depth overlooking the Perth Canyon, Western Australia. The chorus reached a maximum level typically 2.1 h post-sunset and normally ran for 2.1 h (between 3 dB down points). It was present at lower levels across most of the hours of darkness. Maximum chorus spectrum levels were 74-76 dB re 1 µPa2/Hz in the 2 kHz 1/3 octave band, averaging 6-12 dB and up to 30 dB greater than pre-sunset levels. The chorus displayed highest levels over April to August each year with up to 10 dB differences between seasons. The spatial extent of the chorus was not determined but exceeded the sampling range of 13-15 km offshore from the 300 m depth contour and 33 km along the 300 m depth contour. The chorus comprised short damped pulses. The most likely chorus source is considered to be fishes of the family Myctophidae foraging in the water column. The large chorus spatial extent and its apparent correlation with regions of high productivity suggest it may act as an acoustic beacon to marine fauna indicating regions of high biomass

The Impact of Biomechanics in Tissue Engineering and Regenerative Medicine

Biomechanical factors profoundly influence the processes of tissue growth, development, maintenance, degeneration, and repair. Regenerative strategies to restore damaged or diseased tissues in vivo and create living tissue replacements in vitro have recently begun to harness advances in understanding of how cells and tissues sense and adapt to their mechanical environment. It is clear that biomechanical considerations will be fundamental to the successful development of clinical therapies based on principles of tissue engineering and regenerative medicine for a broad range of musculoskeletal, cardiovascular, craniofacial, skin, urinary, and neural tissues. Biomechanical stimuli may in fact hold the key to producing regenerated tissues with high strength and endurance. However, many challenges remain, particularly for tissues that function within complex and demanding mechanical environments in vivo. This paper reviews the present role and potential impact of experimental and computational biomechanics in engineering functional tissues using several illustrative examples of past successes and future grand challenges.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/78125/1/ten.teb.2009.0340.pd

Bcl-2 associated athanogene 5 (Bag5) is overexpressed in prostate cancer and inhibits ER-stress induced apoptosis

Background The Bag (Bcl-2 associated athanogene) family of proteins consists of 6 members sharing a common, single-copied Bag domain through which they interact with the molecular chaperone Hsp70. Bag5 represents an exception in the Bag family since it consists of 5 Bag domains covering the whole protein. Bag proteins like Bag1 and Bag3 have been implicated in tumor growth and survival but it is not known whether Bag5 also exhibits this function. Methods Bag5 mRNA and protein expression levels were investigated in prostate cancer patient samples using real-time PCR and immunoblot analyses. In addition immunohistological studies were carried out to determine the expression of Bag5 in tissue arrays. Analysis of Bag5 gene expression was carried out using one-way ANOVA and Bonferroni’s Multiple Comparison test. The mean values of the Bag5 stained cells in the tissue array was analyzed by Mann-Whitney test. Functional studies of the role of Bag5 in prostate cancer cell lines was performed using overexpression and RNA interference analyses. Results Our results show that Bag5 is overexpressed in malignant prostate tissue compared to benign samples. In addition we could show that Bag5 levels are increased following endoplasmic reticulum (ER)-stress induction, and Bag5 relocates from the cytoplasm to the ER during this process. We also demonstrate that Bag5 interacts with the ER-resident chaperone GRP78/BiP and enhances its ATPase activity. Bag5 overexpression in 22Rv.1 prostate cancer cells inhibited ER-stress induced apoptosis in the unfolded protein response by suppressing PERK-eIF2-ATF4 activity while enhancing the IRE1-Xbp1 axis of this pathway. Cells expressing high levels of Bag5 showed reduced sensitivity to apoptosis induced by different agents while Bag5 downregulation resulted in increased stress-induced cell death. Conclusions We have therefore shown that Bag5 is overexpressed in prostate cancer and plays a role in ER-stress induced apoptosis. Furthermore we have identified GRP78/BiP as a novel interaction partner of Bag5.BioMed Central open acces

Is there a role for glucocorticoid receptor beta in asthma?

Glucocorticoids (GCs) are routinely used as anti-inflammatory drugs in the treatment of asthma. They act through binding to glucocorticoid receptor α (GRα), which represses numerous genes encoding pro-inflammatory mediators. A hormone binding deficient GR isoform named GRβ has been isolated in humans. When overexpressed by transfection, GRβ may function as a dominant negative modulator of GRα. However, to act as such, GRβ has to be more abundant than GRα, and conflicting data have been obtained concerning the relative levels of the two isoforms in cell lines and freshly isolated cells. Moreover, the dominant negative effect was not confirmed by independent laboratories. In GC-resistant asthmatics, GRβ was expressed by an increased number of peripheral blood mononuclear cells (PBMCs), airway T cells, and cells found in skin biopsies of tuberculin responses. However, the relative amounts of GRα and GRβ in these cells were not determined. In GC-dependent asthmatics, PBMCs expressed GRα predominantly. No cells containing higher levels of GRβ than GRα have yet been reported in asthmatics. Even if the existence of such cells is demonstrated, the role of GRβ in asthma will remain a matter of controversy because functional studies have given discrepant data

Oral anticoagulant underutilization among elderly patients with atrial fibrillation: insights from the United States Medicare database

Background Oral anticoagulants (OACs) mitigate stroke risk in patients with atrial fibrillation (AF). The study aim was to analyze prevalence and predictors of OAC underutilization. Methods Newly diagnosed AF patients with a CHA2DS2-VASc score ≥ 2 were identified from the US CMS Database (January 1, 2013–December 31, 2017). Patients were stratified based on having an OAC prescription versus not and the OAC prescription group was stratified by direct OAC (DOACs) versus warfarin. Multivariable logistic regression models were used to examine predictors of OAC underutilization. Results Among 1,204,507 identified AF patients, 617,611 patients (51.3%) were not prescribed an OAC during follow-up (mean: 2.4 years), and 586,896 patients (48.7%) were prescribed an OAC during this period (DOAC: 388,629 [66.2%]; warfarin: 198,267 [33.8%]). Age ≥ 85 years (odds ratio [OR] 0.55, 95% confidence interval [CI] 0.55–0.56), female sex (OR 0.96, 95% CI 0.95–0.96), Black race (OR 0.78, 95% CI 0.77–0.79) and comorbidities such as gastrointestinal (GI; OR 0.43, 95% CI 0.41–0.44) and intracranial bleeding (OR 0.29, 95% CI 0.28–0.31) were associated with lower utilization of OACs. Furthermore, age ≥ 85 years (OR 0.92, 95% CI 0.91–0.94), Black race (OR 0.78, 95% CI 0.76–0.80), ischemic stroke (OR 0.77, 95% CI 0.75–0.80), GI bleeding (OR 0.73, 95% CI 0.68–0.77), and intracranial bleeding (OR 0.72, 95% CI 0.65–0.80) predicted lower use of DOACs versus warfarin. Conclusions Although OAC therapy prescription is the standard of care for stroke prevention in AF patients, its overall utilization is still low among Medicare patients ≥ 65 years old, with specific patient characteristics that predict underutilization