Making the most of sperm activation responses: experiments with boar spermatozoa and bicarbonate

2018 ◽  
Vol 30 (6) ◽  
pp. 842 ◽  
Author(s):  
William V. Holt ◽  
Nana Satake
Keyword(s):  
Semen Quality ◽  
Current Knowledge ◽  
Sperm Quality ◽  
Environmental Chemicals ◽  
Information Value ◽  
In Vitro Tests ◽  
Sperm Activation ◽  
Activation Mechanisms

Attempting to extract useful and reliable information about semen quality and its fertility potential remains a difficult exercise, partly because the sperm heterogeneity within samples often renders simple statistical analyses rather meaningless. In fact, a mean and standard deviation may reflect neither the very fast swimming activities of the most active cells nor the slow and sluggish activities of others. Herein we propose that the information value within semen samples can be maximised if current knowledge about sperm activation mechanisms is exploited before undertaking the measurements. We explain, using boar semen as an example, that estimating and defining relative sperm subpopulation sizes, after activation by bicarbonate, provides a means of quantifying sperm quality. Although such estimates may indeed be related to in vivo fertility, the general approach also suggests potential new avenues that could be exploited for the elaboration of novel in vitro tests for the characterisation of toxic environmental chemicals and, indeed, to reduce the number of animals used in such testing programs.

scholarly journals Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo

Animals ◽  
2019 ◽  
Vol 9 (6) ◽  
pp. 304 ◽  
Author(s):  
Giusy Rusco ◽  
Michele Di Iorio ◽  
Pier Paolo Gibertoni ◽  
Stefano Esposito ◽  
Maurizio Penserini ◽  
...  
Keyword(s):  
Brown Trout ◽  
Semen Quality ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Dna Integrity ◽  
Experimental Conditions ◽  
C 30 ◽  
Thawing Rate

The aim of our study was to test the effects of different non-permeating cryoprotectants (NP-CPAs), namely low-density lipoproteins (LDLs), sucrose, and egg yolk, and thawing rates on the post-thaw semen quality and fertilizing ability of the native Mediterranean brown trout. Pooled semen samples were diluted 1:3 (v:v) with 2.5%, 5%, 10%, or 15% LDL; 0.05, 0.1, or 0.3 M sucrose; or 10% egg yolk. At the moment of analysis, semen was thawed at 30 °C/10 s or 10 °C/30 s. The post-thaw semen quality was evaluated, considering motility, the duration of motility, viability, and DNA integrity. Significantly higher values of motility and viability were obtained using egg yolk/10 °C for 30 s, across all treatments. However, LDL and sucrose concentrations affected sperm cryosurvival, showing the highest post-thaw sperm quality at 5% LDL and 0.1 M sucrose. Based on the in vitro data, egg yolk, 5% LDL, and 0.1 M sucrose thawed at 10 °C or 30 °C were tested for the in vivo trial. The highest fertilization and hatching rates were recorded using egg yolk/10 °C (p < 0.05). According to these in vitro and in vivo results, egg yolk emerged as the most suitable NP-CPA and 10 °C/30 s as the best thawing rate for the cryopreservation of this trout sperm, under our experimental conditions.

scholarly journals 15 EFFECT OF SEMEN THAW METHOD ON CONCEPTION RATE IN DAIRY HEIFER HERDS

2005 ◽  
Vol 17 (2) ◽  
pp. 157 ◽  
Author(s):  
M. Kaproth ◽  
H. Rycroft ◽  
G. Gilbert ◽  
G. Abdel-Azim ◽  
B. Putnam ◽  
...  
Keyword(s):  
Mixed Model ◽  
Linear Mixed Model ◽  
Semen Quality ◽  
Sperm Quality ◽  
Field Trials ◽  
Conception Rate ◽  
Warm Water ◽  
Dairy Heifer

Semen processed with procedures permitting a flexible thaw method is used to breed millions of cows yearly. “Pocket thawing” is widely used as an alternative to warm-water thawing with such semen. To pocket thaw, a straw is retrieved from cryostorage, immediately wrapped in a folded paper towel, and moved to a thermally protected pocket for 2 to 3 min of thawing within the pocket before AI gun loading. Published field data are lacking for comparisons of such a thaw method with those for semen prepared to permit flexible-thawing. We measured the effect of warm-water or pocket thaw on conception rate in four dairy heifer herds using semen prepared with methods previously optimized for flexible-thawing success. Semen processing (Anderson S et al. 1994 J. Dairy Sci. 77, 2302–2307) includes two-step whole-milk extension, static vapor tank freezing (0.5-mL straws), and IMV Digitcool mechanical freezing (0.25-mL straws). It is unclear which specific processing steps permit flexible thawing. These procedures have been developed using breeding results from decades of field trials by professional inseminators using both pocket and warm-water thaw. Semen prepared from each of 12 sires produced equal straw units at 10 and 15 million total sperm per straw, in both 0.5- and 0.25-mL straw packages. Professional inseminators used each combination evenly over 16 months. Additional commercial semen (55% of total) from the same source was used. The thaw methods alternated weekly. Thaw effect on conception status, from 70 day non-return data for 11,215 services (67.6% conception rate), was estimated by a generalized linear mixed model. Neither thaw method nor total sperm per straw significantly affected conception rate (P = 0.658, 0.769, respectively). Bull, herd, inseminator within herd, year, season, and straw size did significantly affect conception rate (P < 0.05). No thaw method interactions with herd, sperm number, season, and straw package size were significant (P = 0.297, 0.526, 0.365, 0.723, respectively). This suggests that if semen has been prepared with procedures specific to flexible-thawing, it can be either pocket thawed or warm-water thawed within a range of herdsman or inseminator practices, season, or straw packaging choices. Even at 10 million, the lowest total sperm per straw, pocket thaw was equally as successful as warm-water thaw. We generally observe that in vitro sperm quality, as expected, is maximal for rapidly thawed straws, with slower thawing resulting in lower values. However, while it appears that conventional measures of in vitro semen quality are improved with fast thaw rates, these measures do not appear to correspond to higher in vivo fertility for semen prepared intentionally to be flexibly thawed. We conclude that, for semen prepared with procedures that permit flexible thawing, the thaw method, whether pocket or warm-water thaw, does not affect conception under commercial conditions and with routine semen handling methods. We thank the herd owners and their staff, the inseminators, and Hap Allen, Ron Hunt, Gordon Nickerson, and Bryan Krick of Genex for their help and cooperation.

Endocrine Disruptors: A Critical Review of In Vitro and In Vivo Testing Strategies for Assessing Their Toxic Hazard to Humans

2000 ◽  
Vol 28 (1) ◽  
pp. 81-118 ◽  
Author(s):  
Robert D. Combes
Keyword(s):  
Endocrine Disruptors ◽  
Human Exposure ◽  
Mechanisms Of Action ◽  
Environmental Chemicals ◽  
Toxicity Tests ◽  
In Vitro Tests ◽  
Wide Range ◽  
The Us

Currently, there is much concern that a wide range of both synthetic and naturally occurring environmental chemicals can act as endocrine disruptors (EDs), and can adversely affect humans and wildlife. Many in vivo and in vitro tests have been proposed for screening EDs, and several regulatory agencies, including the US Environmental Protection Agency (EPA), have recommended tier-testing schemes. Unfortunately, most of the proposed toxicity tests have substantial problems, including non-specificity and lack of reproducibility. There is also uncertainty concerning their relevance for generating useful hazard data for risk assessment purposes, in view of the diversity of the possible ED mechanisms of action (for example, receptor binding, steroidogenesis and modulation of the homeostatic processes which regulate endogenous responses to hormones). Moreover, most of the suggested test methods have yet to be validated according to internationally accepted criteria, although the OECD and the US EPA have defined tests for validation, and an interlaboratory “prevalidation” exercise has been initiated by the OECD. All this is compounded by the lack of information regarding human exposure levels to EDs, and a lack of direct evidence for a causal link between exposure and the development of adverse human health effects. In addition, the regulatory testing of EDs has important negative implications for animal welfare, as some of the proposed in vivo tests require large group sizes of animals and stressful procedures. From a detailed analysis of the available published literature, it is concluded that it is impossible to assess the relative values of currently available in vitro and in vivo toxicity tests for EDs, or to recommend any test or test battery. Any plans for the widespread testing of EDs are therefore premature and might be unnecessary, at least for detecting possible human effects. Several recommendations are made for rectifying this unsatisfactory situation, including the postponement of screening programmes pending: a) more information on human exposure; b) further details of the mechanisms of action of EDs; and c) the development of improved tests, followed by their proper scientific validation.

Limitations on the Reliability of In Vitro Predictive Toxicity Models to Predict Pulmonary Toxicity in Rodents

2016 ◽  
Author(s):  
Jeremy M. Gernand
Keyword(s):  
Meta Analysis ◽  
Information Value ◽  
Screening Tests ◽  
In Vitro Tests ◽  
Screening Methods ◽  
Toxicity Screening ◽  
Animal Testing ◽  
Risk Models

Given the rapidly proliferating varieties of nanomaterials and ongoing concerns that these novel materials may pose emerging occupational and environmental risks, combined with the possibility that each variety might pose a different unique risk due to the unique combination of material properties, researchers and regulators have been searching for methods to identify hazards and prioritize materials for further testing. While several screening tests and toxic risk models have been proposed, most have relied on cellular-level in vitro data. This foundation enables answers to be developed quickly for any material, but it is yet unclear how this information may translate to more realistic exposure scenarios in people or other more complex animals. A quantitative evaluation of these models or at least the inputs variables to these models in the context of rodent or human health outcomes is necessary before their classifications may be believed for the purposes of risk prioritization. This paper presents the results of a machine learning enabled meta-analysis of animal studies attempting to use significant descriptors from in vitro nanomaterial risk models to predict the relative toxicity of nanomaterials following pulmonary exposures in rodents. A series of highly non-linear random forest models (each made up of an ensemble of 1,000 regression tree models) were created to assess the maximum possible information value of the in vitro risk models and related methods of describing nanomaterial variants and their toxicity in rat and mouse experiments. The variety of chemical descriptors or quantitative chemical property measurements such as bond strength, surface charge, and dissolution potential, while important in describing observed differences with in vitro experiments, proved to provide little indication of the relative magnitude of inflammation in rodents (explained variance amounted to less than 32%). Important factors in predicting rodent pulmonary inflammation such as primary particle size and chemical type demonstrate that there are critical differences between these two toxicity assays that cannot be captured by a series of in vitro tests alone. Predictive models relying primarily on these descriptors alone explained more than 62% of the variance of the short term in vivo toxicity results. This means that existing proposed nanomaterial toxicity screening methods are inadequate as they currently stand, and either the community must be content with the slower and more expensive animal testing to evaluate nanomaterial risks, or further conceptual development of improved alternative in vitro screening methodologies is necessary before manufacturers and regulators can rely on them to promote safer use of nanotechnology.

scholarly journals Environmental chemicals impact dog semen quality in vitro and may be associated with a temporal decline in sperm motility and increased cryptorchidism

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Richard G. Lea ◽  
Andrew S. Byers ◽  
Rebecca N. Sumner ◽  
Stewart M. Rhind ◽  
Zulin Zhang ◽  
...  
Keyword(s):  
Polychlorinated Biphenyl ◽  
Semen Quality ◽  
Sperm Quality ◽  
Temporal Trends ◽  
Reproductive Function ◽  
Environmental Chemicals ◽  
Dna Integrity ◽  
Diethylhexyl Phthalate ◽  
Number Of Males

Abstract Adverse temporal trends in human semen quality and cryptorchidism in infants have been associated with exposure to environmental chemicals (ECs) during development. Here we report that a population of breeding dogs exhibit a 26 year (1988–2014) decline in sperm quality and a concurrent increased incidence of cryptorchidism in male offspring (1995–2014). A decline in the number of males born relative to the number of females was also observed. ECs, including diethylhexyl phthalate (DEHP) and polychlorinated biphenyl 153 (PCB153), were detected in adult dog testes and commercial dog foods at concentrations reported to perturb reproductive function in other species. Testicular concentrations of DEHP and PCB153 perturbed sperm viability, motility and DNA integrity in vitro but did not affect LH stimulated testosterone secretion from adult testis explants. The direct effects of chemicals on sperm may therefore contribute to the decline in canine semen quality that parallels that reported in the human.

scholarly journals THE CLINICAL EFFICACY OF SUBLINGUAL ALLERGEN-SPECIFIC IMMUNOTHERAPY IN CHILDREN AGED 3-5 YEARS

2016 ◽  
Vol 6 ◽  
pp. 3-9
Author(s):  
Elena Sharikadze ◽  
Elena Okhotnikova ◽  
Serhii Yuriev
Keyword(s):  
House Dust ◽  
Specific Immunotherapy ◽  
Dermatophagoides Pteronyssinus ◽  
Information Value ◽  
House Dust Mites ◽  
In Vitro Tests ◽  
Dust Mites ◽  
Allergen Specific Immunotherapy

The aim of this study was to determine the efficacy of sublingual allergen–specific immunotherapy (SLIT) in Ukrainian children younger than 5 years old with allergic rhinitis and bronchial asthma sensitized to house dust mite allergens. Material and methods: Four hundred and fifty children aged 28 months up to 5 years with rhinitis or asthma were examined. One hundred and twenty five children sensitized to house dust mites Dermatophagoides pteronyssinus and/or Dermatophagoides farina were included. In vivo and in vitro tests were made with a standard inhalant allergens panel. Results: The high information value of molecular diagnostics methods applied prior to prescription of the given therapy in children is analyzed. It has been found that in children under 5 sensitized to allergens of house dust mites Dermatophagoides pteronyssinus and/or Dermatophagoides farinae the application of sublingual allergen–specific immunotherapy therapy allows gaining control over the symptoms of the disease during the first 6 months. Conclusion: The high safety of SLIT in children has been proven. Comparative analysis in the group of patients not receiving SLIT shows a high frequency of symptoms of the disease after “free-of-symptoms interval” against full or partial baseline therapy denial.

Comparison of High Purity Factor IX Concentrates and a Prothrombin Complex Concentrate in a Canine Model of Thrombogenicity

1991 ◽  
Vol 66 (05) ◽  
pp. 609-613 ◽  
Author(s):  
I R MacGregor ◽  
J M Ferguson ◽  
L F McLaughlin ◽  
T Burnouf ◽  
C V Prowse
Keyword(s):  
High Purity ◽  
Time Course ◽  
Prothrombin Complex Concentrate ◽  
Degradation Products ◽  
Canine Model ◽  
Factor Ix ◽  
In Vitro Tests ◽  
Albumin Infusion

SummaryA non-stasis canine model of thrombogenicity has been used to evaluate batches of high purity factor IX concentrates from 4 manufacturers and a conventional prothrombin complex concentrate (PCC). Platelets, activated partial thromboplastin time (APTT), fibrinogen, fibrin(ogen) degradation products and fibrinopeptide A (FPA) were monitored before and after infusion of concentrate. Changes in FPA were found to be the most sensitive and reproducible indicator of thrombogenicity after infusion of batches of the PCC at doses of between 60 and 180 IU/kg, with a dose related delayed increase in FPA occurring. Total FPA generated after 100-120 IU/kg of 3 batches of PCC over the 3 h time course was 9-12 times that generated after albumin infusion. In contrast the amounts of FPA generated after 200 IU/kg of the 4 high purity factor IX products were in all cases similar to albumin infusion. It was noted that some batches of high purity concentrates had short NAPTTs indicating that current in vitro tests for potential thrombogenicity may be misleading in predicting the effects of these concentrates in vivo.

A Comparison of the in Vitro and in Vivo Thrombogenic Activity of Factor IX Concentrates Using Stasis (Wessler) and Non-Stasis Rabbit Models

1980 ◽  
Vol 44 (02) ◽  
pp. 081-086 ◽  
Author(s):  
C V Prowse ◽  
A E Williams
Keyword(s):  
Platelet Rich Plasma ◽  
Thrombus Formation ◽  
Factor Ix ◽  
Coagulation System ◽  
In Vitro Tests ◽  
Clinical Use ◽  
Low Activity

SummaryThe thrombogenic effects of selected factor IX concentrates were evaluated in two rabbit models; the Wessler stasis model and a novel non-stasis model. Concentrates active in either the NAPTT or TGt50 in vitro tests of potential thrombogenicity, or both, caused thrombus formation in the Wessler technique and activation of the coagulation system in the non-stasis model. A concentrate with low activity in both in vitro tests did not have thrombogenic effects in vivo, at the chosen dose. Results in the non-stasis model suggested that the thrombogenic effects of factor IX concentrates may occur by at least two mechanisms. A concentrate prepared from platelet-rich plasma and a pyrogenic concentrate were also tested and found to have no thrombogenic effect in vivo.These studies justify the use of the NAPTT and TGt50 in vitro tests for the screening of factor IX concentrates prior to clinical use.

Inhibition of Fibrinolysis and Fibrinogenolysis in Man: Comparison of ε-Aminocaproic Acid and Kallikrein Inhibitor

1963 ◽  
Vol 10 (01) ◽  
pp. 106-119 ◽  
Author(s):  
E Beck ◽  
R Schmutzler ◽  
F Duckert ◽  
Keyword(s):  
Aminocaproic Acid ◽  
Side Reactions ◽  
In Vitro Tests ◽  
Factor V ◽  
Clinical Use ◽  
Strong Inhibitor ◽  
Kallikrein Inhibitor ◽  
Therapeutic Possibilities

SummaryInhibitor of kallikrein and trypsin (KI) extracted from bovine parotis was compared with ε-aminocaproic acid (EACA): both substances inhibit fibrinolysis induced with streptokinase. EACA is a strong inhibitor of fibrinolysis in concentrations higher than 0, 1 mg per ml plasma. The same amount and higher concentrations are not able to inhibit completely the proteolytic-side reactions of fibrinolysis (fibrinogenolysis, diminution of factor V, rise of fibrin-polymerization-inhibitors). KI inhibits well proteolysis of plasma components in concentrations higher than 2,5 units per ml plasma. Much higher amounts of KI are needed to inhibit fibrinolysis as demonstrated by our in vivo and in vitro tests.Combination of the two substances for clinical use is suggested. Therapeutic possibilities are discussed.

Role of in vivo and in vitro Tests in the Diagnosis of Severe Cutaneous Adverse Reactions (SCAR) to Drug

2019 ◽  
Vol 25 (36) ◽  
pp. 3872-3880 ◽  
Author(s):  
Marcel M. Bergmann ◽  
Jean-Christoph Caubet
Keyword(s):  
Adverse Reactions ◽  
Lymphocyte Transformation ◽  
Stevens Johnson Syndrome ◽  
In Vitro Tests ◽  
High Morbidity ◽  
Patch Tests ◽  
Severe Cutaneous Adverse Reactions ◽  
Cutaneous Adverse Reactions

Severe cutaneous adverse reactions (SCAR) are life-threatening conditions including acute generalized exanthematous pustulosis (AGEP), Stevens-Johnson Syndrome (SJS), toxic epidermal necrolysis (TEN) and drug reaction with eosinophilia and systemic symptoms (DRESS). Diagnosis of causative underlying drug hypersensitivity (DH) is mandatory due to the high morbidity and mortality upon re-exposure with the incriminated drug. If an underlying DH is suspected, in vivo test, including patch tests (PTs), delayed-reading intradermal tests (IDTs) and in vitro tests can be performed in selected patients for which the suspected culprit drug is mandatory, or in order to find a safe alternative treatment. Positivity of in vivo and in vitro tests in SCAR to drug varies depending on the type of reaction and the incriminated drugs. Due to the severe nature of these reactions, drug provocation test (DPT) is highly contraindicated in patients who experienced SCAR. Thus, sensitivity is based on positive test results in patients with a suggestive clinical history. Patch tests still remain the first-line diagnostic tests in the majority of patients with SCAR, followed, in case of negative results, by delayed-reading IDTs, with the exception of patients with bullous diseases where IDTs are still contra-indicated. In vitro tests have shown promising results in the diagnosis of SCAR to drug. Positivity is particularly high when the lymphocyte transformation test (LTT) is combined with cytokines and cytotoxic markers measurement (cyto-LTT), but this still has to be confirmed with larger studies. Due to the rarity of SCAR, large multi-center collaborative studies are needed to better study the sensitivity and specificity of in vivo and in vitro tests.

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